WorldWideScience

Sample records for abl-related gene arg

  1. [Investigation of Antibiotic Resistance Genes (ARGs) in Landfill].

    Li, Lei; Xu, Jing; Zhao, You-cai; Song, Li-yan

    2015-05-01

    Antibiotic resistant genes (ARGs), an emerging contaminant, have been detected worldwide in various environments such as sediments and river. However, little is known about ARGs distribution in landfill. In this study, we investigated five ARGs [sulfonamides resistant genes (sulI and sulII), chloramphenicols resistant gene (cat), β-lactams resistant gene (bla-SHV), and tetracyclines resistant gene (tetW)] in refuse samples collected from jiangeungou landfill (Xi'an, China) by real-time PCR. We then correlated the ARGs and physiochemical properties of refuse to examine the link between them. Results showed that all tested ARGs have been detected in all samples, suggesting that landfill served as ARGs reservoir. The highest copies numbers of sulII, sulI, tetW, bla-SHV, and cat were (3.70 ± 0.06) x 10(8) copies · g(-1) ( dry refuse), (9.33 · 0.06) x 10(6) copies · g(-1) (dry refuse), (2.27 0.08) x 10(5) copies · g(-1) (dry refuse), (3.68 ± 0.09) x 10(4) copies · g(-1) (dry refuse), and (1.39 ± 0.10) x 10(4) copies · g(-1) (dry refuse), respectively. Further, sulI, sulII, and cat positively correlated to moisture and sulI and cat negatively correlated to pH. PMID:26314129

  2. [Effects of Thermophilic Composting on Antibiotic Resistance Genes (ARGs) of Swine Manure Source].

    Zheng, Ning-guo; Huang, Nan; Wang, Wei-wei; Yu, Man; Chen, Xiao-yang; Yao, Yan-lai; Wang, Wei-ping; Hong, Chun-lai

    2016-05-15

    To investigate the effects of thermophilic composting process on antibiotic resistance genes (ARGs) of swine manure source at a field scale, the abundance of four erythromycin resistance genes (ermA, ermB, ermC and ermF), three β-lactam resistance genes (blaTEM, blaCTX and blaSHV) and two quinolone resistance genes (qnrA and qnrS) were quantified by quantitative PCR ( qPCR) during the composting process. The results suggested that the erm genes' copy numbers were significantly higher than those of the bla and qnr genes in the early stage of composting (P < 0.01). The maximum abundance of erm genes was ermB (9.88 x 10⁸ copies · g⁻¹), following by ermF (9.4 x 10⁸ copies · g⁻¹). At the end of the composting process, bla and qnr genes were at low levels, while erm genes were still at high levels. Even through ermF was proliferated comparing with the initial copies. These results indicated that thermophilic composting process could not effectively remove all ARGs. For some ARGs, compost may be a good bioreactor resulting in their proliferation. Application of composting products on farmland may cause transference of ARGs. PMID:27506057

  3. Mutation of the Erwinia amylovora argD gene causes arginine auxotrophy, nonpathogenicity in apples, and reduced virulence in pears.

    Ramos, Laura S; Lehman, Brian L; Peter, Kari A; McNellis, Timothy W

    2014-11-01

    Fire blight is caused by Erwinia amylovora and is the most destructive bacterial disease of apples and pears worldwide. In this study, we found that E. amylovora argD(1000)::Tn5, an argD Tn5 transposon mutant that has the Tn5 transposon inserted after nucleotide 999 in the argD gene-coding region, was an arginine auxotroph that did not cause fire blight in apple and had reduced virulence in immature pear fruits. The E. amylovora argD gene encodes a predicted N-acetylornithine aminotransferase enzyme, which is involved in the production of the amino acid arginine. A plasmid-borne copy of the wild-type argD gene complemented both the nonpathogenic and the arginine auxotrophic phenotypes of the argD(1000)::Tn5 mutant. However, even when mixed with virulent E. amylovora cells and inoculated onto immature apple fruit, the argD(1000)::Tn5 mutant still failed to grow, while the virulent strain grew and caused disease. Furthermore, the pCR2.1-argD complementation plasmid was stably maintained in the argD(1000)::Tn5 mutant growing in host tissues without any antibiotic selection. Therefore, the pCR2.1-argD complementation plasmid could be useful for the expression of genes, markers, and reporters in E. amylovora growing in planta, without concern about losing the plasmid over time. The ArgD protein cannot be considered an E. amylovora virulence factor because the argD(1000)::Tn5 mutant was auxotrophic and had a primary metabolism defect. Nevertheless, these results are informative about the parasitic nature of the fire blight disease interaction, since they indicate that E. amylovora cannot obtain sufficient arginine from apple and pear fruit tissues or from apple vegetative tissues, either at the beginning of the infection process or after the infection has progressed to an advanced state. PMID:25172854

  4. An association between TRP64ARG polymorphism of the B3 adrenoreceptor gene and some metabolic disturbances

    Abilova Samai S

    2011-10-01

    Full Text Available Abstract Backgrounds B3 adrenoreceptors (ADRB3 are abundant in adipose tissue and play the role in its metabolism and lipolysis. Some variants of the ADRB3 gene may predispose subjects for the development obesity and metabolic abnormalities in the setting of modern sedentary lifestyle. ADRB3 gene polymorphism association with metabolic disturbances has never been studied before in the ethnic Kyrgyz population. Aim To study an association between Trp64Arg polymorphism of the ADRB3 and metabolic syndrome (MS components in an ethnic Kyrgyz group. Materials and methods 213 Ethnic Kyrgyz volunteers over the age of 30 were enrolled in the study. The assessment plan for each individual comprised of general physical and anthropometric exams as well as laboratory tests (glucose, lipid panel, insulin and genotyping by Trp64Arg polymorphism of the ADRB3. MS diagnosis was consistent with modified ATP III criteria (2005. Logistic regression analysis was performed to test the potential independent association between Arg64 allele with obesity, abdominal obesity (AO and arterial hypertension (AH. Results Trp64Arg polymorphism of the ADRB3 was assessed in 213 individuals (145 men, 68 women aged 30-73 (mean age 50.7 ± 7.6. Arg64 allele frequency was 0.239; ADRB3 genotype distribution among participants was: Trp64 homozygotes 54.5%, Trp64Arg 43.2% and Arg64 homozygotes 2.3%. There was an association between Trp64Arg и Arg64Arg genotypes and higher BMI, WC and obesity frequency (p Conclusion Arg64 allele of the ADRB3 gene in the studied group has an association with MS components such as obesity, AO and decreased HDL-C level.

  5. Sequence analysis and complementation studies of the argJ gene encoding ornithine acetyltransferase from Neisseria gonorrhoeae.

    Martin, P R; Mulks, M H

    1992-01-01

    Clinical isolates of Neisseria gonorrhoeae frequently are deficient in arginine biosynthesis. These auxotrophs often have defects in the fifth step of the arginine biosynthetic pathway, the conversion of acetylornithine to ornithine. This reaction is catalyzed by the enzyme ornithine acetyltransferase, which is a product of the argJ gene. We have cloned and sequenced the gonococcal argJ gene and found that it contains an open reading frame of 1,218 nucleotides and encodes a peptide with a ded...

  6. Characterization of the Saccharomyces cerevisiae ARG7 gene encoding ornithine acetyltransferase, an enzyme also endowed with acetylglutamate synthase activity.

    Crabeel, M; Abadjieva, A; Hilven, P; Desimpelaere, J; Soetens, O

    1997-12-01

    We have cloned by functional complementation and characterized the yeast ARG7 gene encoding mitochondrial ornithine acetyltransferase, the enzyme catalyzing the fifth step in arginine biosynthesis. While forming ornithine, this enzyme regenerates acetylglutamate, also produced in the first step by the ARG2-encoded acetylglutamate synthase. Interestingly, total deletion of the genomic ARG7 ORF resulted in an arginine-leaky phenotype, indicating that yeast cells possess an alternative route for generating ornithine from acetylornithine. Yeast ornithine acetyltransferase has been purified and characterized previously as a heterodimer of two subunits proposed to derive from a single precursor protein [Liu, Y-S., Van Heeswijck R., Hoj, P. & Hoogenraad, N. (1995) Eur. J. Biochem. 228, 291-296]; those authors further suggested that the internal processing of Arg7p, which is a mitochondrial enzyme, might occur in the matrix, while the leader peptide would be of the non-cleavable-type. The characterization of the gene (a) establishes that Arg7p is indeed encoded by a single gene, (b) demonstrates the existence of a cleaved mitochondrial prepeptide of eight residues, and (c) shows that the predicted internal processing site is unlike the mitochondrial proteolytic peptidase target sequence. Yeast Arg7p shares between 32-43% identity in pairwise comparisons with the ten analogous bacterial ArgJ enzymes characterized. Among these evolutionarily related enzymes, some but not all appear bifunctional, being able to produce acetylglutamate not only from acetylornithine but also from acetyl-CoA, thus catalyzing the same reaction as the apparently unrelated acetylglutamate synthase. We have addressed the question of the bifunctionality of the eucaryotic enzyme, showing that overexpressed ARG7 can complement yeast arg2 and Escherichia coli argA mutations (affecting acetylglutamate synthase). Furthermore, Arg7p-linked acetylglutamate synthase activity was measurable in an assay. The

  7. Distribution of antibiotic resistance genes (ARGs) in anaerobic digestion and land application of swine wastewater.

    Sui, Qianwen; Zhang, Junya; Chen, Meixue; Tong, Juan; Wang, Rui; Wei, Yuansong

    2016-06-01

    Swine farm and the adjacent farmland are hot spots of ARGs. However, few studies have investigated the on-site occurrence of ARGs distributed in the process of anaerobic digestion (AD) followed by land application of swine wastewater. Two typical swine farms, in southern and northern China respectively, with AD along with land application were explored on ARG distributions. ARGs were highly abundant in raw swine wastewater, AD effectively reduced the copy number of all detected ARGs (0.21-1.34 logs removal), but the relative abundance with different resistance mechanisms showed distinctive variation trends. The reduction efficiency of ARGs was improved by stable operational temperature and longer solid retention time (SRT) of AD. ARGs in soil characterized the contamination from the irrigation of the digested liquor. The total ARGs quantity in soil fell down by 1.66 logs in idle period of winter compared to application period of summer in the northern region, whereas the total amount was steady with whole-year application in south. Some persistent (sul1 and sul2) and elevated ARGs (tetG and ereA) in AD and land application need more attention. PMID:27038206

  8. TRP64ARG polymorphism of the beta 3-adrenergic receptor gene and obesity risk: effect modification by a sedentary lifestyle.

    Marti, A; Corbalan, M. (M.S.); Martinez-Gonzalez, M.A. (Miguel Angel); Martinez, J. A.

    2002-01-01

    Aim: We performed a case–control study to assess the association between obesity risk and the Trp64Arg polymorphism of the β3-adrenergic receptor gene. Methods: Obese subjects [n = 159; body mass index (BMI) > 30 kg/m2] and controls (n = 154; BMI < 25 kg/m2) were compared using multivariable logistic regression to control for potential confounders. Results: A higher obesity risk (adjusted OR: 2.98; 95% CI: 1.00–8.56; p = 0.05) was associated with the Trp64Arg polymorphism among sedentar...

  9. The Polymorphism of DNA Repair Gene ERCC2/XPD Arg156Arg and Susceptibility to Breast Cancer in a Chinese Population

    Yin, J. Y.; Liang, D. H.; Vogel, Ulla Birgitte;

    2009-01-01

    association between ERCC2/XPD Arg156Arg and susceptibility to breast cancer in a Chinese population, we conducted a hospital-based case-control study consisting of 129 patients with breast cancer and 205 controls matched by age, gender, and ethnicity. PCR-RFLP was used for genotyping. No associations were...... (mean age 45 years) than cases with the wild CC genotype (mean age 50 years) (P = 0.05). There were no differences in risk estimates in relation to menopause and occurrence of breast cancer. Our findings do not suggest that ERCC2/XPD Arg156Arg contributes to breast cancer susceptibility in a Chinese...

  10. [Regional features of obesity-associated gene polymorphism (rs9939609 FTO gene and gene Trp64Arg ADRB3) in Russian population].

    Baturin, A K; Sorokina, E Iu; Pogozheva, A V; Peskova, E V; Makurina, O N; Tutel'ian, V A

    2014-01-01

    Recent studies have shown a significant association with obesity polymorphisms: rs9939609 gene due to fat mass and obesity FTO in European and some Asian and African American populations Trp64Arg ADRB3 gene in several European populations. Association of variants rs9939609 and Trp64Arg obesity was studied in 1244 the inhabitants of Moscow and Sverdlovsk regions. Genotyping was performed using allele-specific amplification, detection results in real time using TaqMan-probes complementary DNA polymorphic sites. The frequency of the mutant allele of the FTO gene in the population of Moscow and Sverdlovsk region was 45.1%, with the TT genotype was detected in 30.2% of cases, AT--49.5%, AA--20.3%. Women had the presence of the mutant allele more likely than men (48.4 vs. 42.5%). People with obesity were more genotypes AA (26.3%) and AT (52.8%) compared to the surveyed with a BMI of less than 30 kg/m2 (respectively 18.1 and 50.7%). A significantly higher incidence of risk allele A was found in individuals with obesity (52.6 and 43.4%). The presence of the mutant allele of the gene ADRB3 among the population of Moscow and Sverdlovsk regions was noted in 7.4% of cases. While 15.5% of patients had a heterozygous genotype Trp64Arg ADRB3, that is consistent with international research. The frequency of the risk allele and genotype Arg64 Trp64Arg in women (9.3 and 18.5%) was significantly higher than men (6.2 and 12.2%). The presence of the mutant allele and genotype Trp64Arg ADRB3 (respectively, 9.1 and 18.1%) were significantly more marked in the examined obese compared with those with a body mass index less than 30 kg/m2 (7.4 and 14.9%), but these differences were not statistically significant. The results of these studies suggest that genetic variants of the FTO gene rs9939609 genotype and Trp64Arg ADRB3 contribute to the development of obesity among residents of Moscow and Sverdlovsk Region of Russia. The risk of obesity increases in the case of combined polymorphisms in

  11. An association between TRP64ARG polymorphism of the B3 adrenoreceptor gene and some metabolic disturbances

    Abilova Samai S; Zalesskaya Yulia V; Moldokeeva Cholpon B; Lunegova Olga S; Kerimkulova Alina S; Mirrakhimov Aibek E; Sovhozova Nurmira A; Aldashev Almaz A; Mirrakhimov Erkin M

    2011-01-01

    Abstract Backgrounds B3 adrenoreceptors (ADRB3) are abundant in adipose tissue and play the role in its metabolism and lipolysis. Some variants of the ADRB3 gene may predispose subjects for the development obesity and metabolic abnormalities in the setting of modern sedentary lifestyle. ADRB3 gene polymorphism association with metabolic disturbances has never been studied before in the ethnic Kyrgyz population. Aim To study an association between Trp64Arg polymorphism of the ADRB3 and metabol...

  12. Trp64Arg polymorphism in ADRB3 gene is associated with elite endurance performance

    Santiago Dorrego, Catalina; Ruiz, Jonatan R.; Buxens, Amaya; Artieda, Marta; Arteta, David; González-Freire, Marta; Rodríguez Romo, Gabriel; Altmäe, Signe; Lao, José I.; Gómez Gallego, Félix; Lucía Mulas, Alejandro

    2011-01-01

    In this study, allele and genotype frequencies of the ADRB1 Arg389Gly (rs1801253), ADRB2 Gly16Arg (rs1042713) and Gln27Glu (rs1042714), and ADRB3 Trp64Arg (rs4994) variations were compared in the following three groups of Spanish (Caucasian) men: (1) world-class endurance athletes (E; runners and cyclists, n=100), (2) elite power athletes (P; sprinters, jumpers and throwers, n=53) and (3) non-athletic controls (C; n=100). No significant differences were observed in genotype and allele distrib...

  13. [The Trp64Arg polymorphism of beta3-adrenoreceptor gene study in persons with overweight and obesity].

    Baturin, A K; Pogozheva, A V; Sorokina, E Iu; Makurina, O N; Tutel'ian, V A

    2012-01-01

    The development of obesity is determined by lifestyle and genetic mechanisms. In particular, the polymorphisms in the adrenergic receptor genes (ADRB) have been extensively studied for association with obesity-related phenotypes. ADRB3 is an obvious candidate gene given its involvement in the regulation of lipolysis and thermogenesis. ADRB3 Trp64Arg polymorphism, a missense mutation in the first transmembrane domain of the R3-adrenergic receptor is associated with visceral obesity and insulin resistance in the Pima Indian, French, and Finnish populations. The recent meta-analysis that combined data of 6582 individuals from Japanese populations showed significant association the Arg64 allele with increased BMI. There are tested the polymorphisms in the beta3-Adrenoreceptor (ADRB3) gene in associated with body mass index (BMI), fat mass and biochemical parameters.We have been examined 91 persons from Moscow region with BMI >25 kg/m2. The Trp64Arg polymorphism of ADRB3 genes were genotyped with the use of an allelic discrimination assay. The TaqMan-based real-time PCR method was applied. There have been estimated of anthropometric and biochemicalparameters. The frequencies of the Trp64Trp and Trp64Arggenotypes of ADRB3 gene were 82% and 12%, respectively, the frequencies of mutant allele was 6%. Trp64Arg genotypes of ADRB3 compared to Trp64Trp genotypes had significantly higher body fat percentage (respectively 48,6 +/- 0,96% and 43,8 +/- 1,72%, pADRB3 gene polymorphisms can be used for the personalization of diet in persons with obesity. PMID:22774474

  14. Expression of arg genes of Escherichia coli during arginine limitation dependent upon stringent control of translation.

    Williams, M.G.; Rogers, P

    1987-01-01

    The transcription and translation of operons for arginine biosynthetic enzymes after arginine removal (arginine down shift) were studied in relA and relA+ strains of Escherichia coli. After arginine down shift, derepression of synthesis of the arginine biosynthetic enzymes ornithine carbamoyltransferase (argF) and argininosuccinate lyase (argH) began at about 15 min in relA+ cells but was delayed in relA cells for more than 2 h. However, both relA+ and relA cells accumulated high levels of ar...

  15. Association of beta3-adrenergic receptor (ADRB3) Trp64Arg gene polymorphism with obesity and metabolic syndrome in the Balinese: a pilot study

    Trimarsanto Hidayat; Oktavianthi Sukma; Suastika Ketut; Saraswati Made R; Malik Safarina G; Sudoyo Herawati

    2011-01-01

    Abstract Background Prevalence of obesity is increasing all over the world. ADRB3 Trp64Arg gene polymorphism was proposed to be associated with obesity, although inconsistent findings and differences of the Arg64 allele frequency among various ethnics were reported. Westernization was reported to increase the prevalence of obesity in developing world. In this study we determined the prevalence of obesity and metabolic syndrome among urban and rural Balinese, and studied the association of ADR...

  16. Congenital erythropoietic porphyria with two mutations of the uroporphyrinogen III synthase gene (Cys73Arg, Thr228Met

    Zoran Gucev

    2011-01-01

    Full Text Available Congenital erythropoietic porphyria (CEP is an autosomal recessive inborn error of metabolism that results from the markedly deficient activity of uroporphyrinogen III synthase (UROS. We describe a 14-year-old girl with red urine since infancy, progressive blistering and scarring of the skin, and moderate hemolytic anemia. After years of skin damage, her face is mutilated; she has a bald patch on the scalp, hypertrichosis of the neck, areas of skin darkening, and limited joint movements of the hands. Total urine excretion and fecal total porphyrin were both markedly raised above normal levels. Sequencing of the UROS gene identified two mutations causing CEP (Cys73Arg, Thr228Met. The patient lesions are progressing. Bone marrow transplantation and/or gene therapy are proposed as the next steps in her treatment. In brief, we describe a CEP with confirmed two pathogenic mutations, severe phenotype and discuss the various treatment options available.

  17. Gene encoding γ-carbonic anhydrase is cotranscribed with argC and induced in response to stationary phase and high CO2 in Azospirillum brasilense Sp7

    Mishra Mukti N

    2010-07-01

    Full Text Available Abstract Background Carbonic anhydrase (CA is a ubiquitous enzyme catalyzing the reversible hydration of CO2 to bicarbonate, a reaction underlying diverse biochemical and physiological processes. Gamma class carbonic anhydrases (γ-CAs are widespread in prokaryotes but their physiological roles remain elusive. At present, only γ-CA of Methanosarcina thermophila (Cam has been shown to have CA activity. Genome analysis of a rhizobacterium Azospirillum brasilense, revealed occurrence of ORFs encoding one β-CA and two γ-CAs. Results One of the putative γ-CA encoding genes of A. brasilense was cloned and overexpressed in E. coli. Electrometric assays for CA activity of the whole cell extracts overexpressing recombinant GCA1 did not show CO2 hydration activity. Reverse transcription-PCR analysis indicated that gca1 in A. brasilense is co-transcribed with its upstream gene annotated as argC, which encodes a putative N-acetyl-γ-glutamate-phosphate reductase. 5'-RACE also demonstrated that there was no transcription start site between argC and gca1, and the transcription start site located upstream of argC transcribed both the genes (argC-gca1. Using transcriptional fusions of argC-gca1 upstream region with promoterless lacZ, we further demonstrated that gca1 upstream region did not have any promoter and its transcription occurred from a promoter located in the argC upstream region. The transcription of argC-gca1 operon was upregulated in stationary phase and at elevated CO2 atmosphere. Conclusions This study shows lack of CO2 hydration activity in a recombinant protein expressed from a gene predicted to encode a γ-carbonic anhydrase in A. brasilense although it cross reacts with anti-Cam antibody raised against a well characterized γ-CA. The organization and regulation of this gene along with the putative argC gene suggests its involvement in arginine biosynthetic pathway instead of the predicted CO2 hydration.

  18. Energy expenditure, body composition and insulin response to glucose in male twins discordant for the Trp64Arg polymorphism of the beta3-adrenergic receptor gene

    Højlund, K; Christiansen, C; Bjørnsbo, K S;

    2006-01-01

    secretion could play a role. METHODS: In 10 male twin pairs discordant for the Trp64Arg polymorphism, we examined insulin response to glucose by an oral glucose tolerance test (OGTT), a frequently sampled intravenous glucose tolerance test (FSIGT), body composition by the bioimpedance method, dual-energy X......AIM: The tryptophan to arginine change in position 64 (Trp64Arg) polymorphism of the beta3-adrenergic receptor (beta3AR) gene has been associated with an increased prevalence of obesity, insulin resistance and type 2 diabetes. In this, decreased rates of energy expenditure and impaired insulin......-ray absorptiometry scanning and energy expenditure by indirect and direct calorimetry. RESULTS: Twins heterozygous for the Trp64Arg polymorphism showed significantly lower fat mass independent of the method used, and significantly lower fasting insulin and glucose concentrations compared with their homozygous wild...

  19. Antithrombin gene Arg197Stop mutation-associated venous sinus thrombosis in a Chinese family

    Ang Li; Dexin Wang; Qiming Xue; Baoen Wang; Tianhui Liu; Zhandong Liu; Jimei Li; Chunling Zhang; Jun Chen; Jinmei Sun; YanfeiHan; Lili Wang

    2011-01-01

    This study sought to elucidate the genetic correlation of cerebral venous sinus thrombosis caused by a hereditary antithrombin deficiency in a Chinese family, at the genetic and protein levels. A nonsense mutation from C to T on locus 6431 in exon 3B of the antithrombin gene was observed,leading to an arginine (CGA) to stop codon (TGA) change in the protein. This is the first report of this mutation in China. Ineffective heparin therapy in the propositus patient is associated with a lack of heparin binding sites after antithrombin gene mutation. Characteristic low intracranial pressure in the acute phase might be specific to this patient with cerebral venous sinus thrombosis.

  20. The ARG1-LIKE2 gene of Arabidopsis functions in a gravity signal transduction pathway that is genetically distinct from the PGM pathway

    Guan, Changhui; Rosen, Elizabeth S.; Boonsirichai, Kanokporn; Poff, Kenneth L.; Masson, Patrick H.

    2003-01-01

    The arl2 mutants of Arabidopsis display altered root and hypocotyl gravitropism, whereas their inflorescence stems are fully gravitropic. Interestingly, mutant roots respond like the wild type to phytohormones and an inhibitor of polar auxin transport. Also, their cap columella cells accumulate starch similarly to wild-type cells, and mutant hypocotyls display strong phototropic responses to lateral light stimulation. The ARL2 gene encodes a DnaJ-like protein similar to ARG1, another protein previously implicated in gravity signal transduction in Arabidopsis seedlings. ARL2 is expressed at low levels in all organs of seedlings and plants. arl2-1 arg1-2 double mutant roots display kinetics of gravitropism similar to those of single mutants. However, double mutants carrying both arl2-1 and pgm-1 (a mutation in the starch-biosynthetic gene PHOSPHOGLUCOMUTASE) at the homozygous state display a more pronounced root gravitropic defect than the single mutants. On the other hand, seedlings with a null mutation in ARL1, a paralog of ARG1 and ARL2, behave similarly to the wild type in gravitropism and other related assays. Taken together, the results suggest that ARG1 and ARL2 function in the same gravity signal transduction pathway in the hypocotyl and root of Arabidopsis seedlings, distinct from the pathway involving PGM.

  1. 钝齿棒杆菌argR基因克隆、表达及其重组菌发酵产精氨酸的探讨%Study on Gene cloning and expression of argR and its recombinant strain producing arg through fermentation in Corynebacterium crenatum

    石磊

    2013-01-01

    目的 探讨N-乙酰鸟氨酸的转氨酶于钝齿棒杆菌当中,其精氨酸合成环节所起到的作用,并验证其酶学性质,从而为优化提高培养基的成分,以及发酵过程的工艺条件,和精氨酸的产量提供理论及实验依据.方法 于精氨酸的高产菌株中,使其钝齿棒杆菌的染色体(SYPA5-5)扩增,从而获取ACOAT的编码基因-argD,其全长为1176bp,编码氨基酸为390个,均于C.crenatum SYPA与Escherichia coli BL21(DE3)上成功表达.选择Ni柱进行亲和层析并纯化后,能够得到重组的蛋白比酶活可达到108.2U/g,并研究其酶学性质.再构建重组的钝齿棒杆菌,以加强其精氨酸的合成途径中ACOAT的蛋白表达量,同时对重组的菌产精氨酸予以发酵分析.结果 重组的钝齿棒杆菌同出发菌株进行对比,其胞内的ACOAT的酶活得到增强;并且重组菌中CCD1其精氨酸的平均产量达到了39.7g/L,其产酸最终提高了14.7%.结论 通过研究,不仅对高产精氨酸其重组基因菌工程的构建提供以理论依据,而且还具有非常重要的指导意义.%Objective N - acety - L - omithine transaminase which reacted at the step of arg biosynthesis was not only discussed, but also its enzyme properties were proved. It even could provide theoretical and experimental basis for improving the culture, optimizing technological conditions and increasing output of Arg. Methods Through amplifying chromosome SYPA5 -5 of Corynebacterium crenatum in high - producing Arg strain,the ArgD - code gene of ACOAT was achived. The gene was 1176 bp in full length,encoding 390 amino acid residues. At last,all of them were expressed in C. Crenatum SYPA and Escherichia coli BL21( DE3). By selecting NI column to do the research, we had got recombinant protein whose enzyme activity was as high as 108. 2U/g after affinity purification. And then we had study its enzymatic properties. At last, recombinant Corynebacterium crenatum had been construct in order to

  2. Trp64Arg Polymorphism in Beta3-Adrenergic Receptor Gene Is Associated with Decreased Fat Oxidation Both in Resting and Aerobic Exercise in the Japanese Male

    Emiko Morita

    2009-01-01

    Full Text Available The purpose of our study was to investigate whether the Trp64Arg polymorphism in β3-AR gene and the −3826A/G polymorphism in the UCP1 gene were associated with the reduction in energy expenditure and fat oxidation both in resting and aerobic exercise in Japanese. Eighty-six nonobese young healthy Japanese were recruited. Energy expenditure was measured using indirect calorimetry. The subjects performed an aerobic exercise program at 60% of their maximal heart rate for 30 minutes. The level of fat oxidation at rest and aerobic exercise of the male subjects with Trp/Arg of the β3-AR gene was significantly lower than that of the Trp/Trp genotype. No difference in FO0-30 was observed in the female subjects. There was no association between UCP-1 polymorphism and energy expenditure during aerobic exercise. It was revealed that the Trp64Arg polymorphism in β3-AR gene is associated with reduction of fat oxidation both in resting and aerobic exercise in healthy, young Japanese males.

  3. Genome-wide identification of rubber tree (Hevea brasiliensis Muell. Arg.) aquaporin genes and their response to ethephon stimulation in the laticifer, a rubber-producing tissue

    Zou, Zhi; Gong, Jun; An, Feng; Xie, Guishui; Wang, Jikun; Mo, Yeyong; Yang, Lifu

    2015-01-01

    Background Natural rubber, an important industrial raw material, is specifically synthesized in laticifers located inside the rubber tree (Hevea brasiliensis Muell. Arg.) trunk. Due to the absence of plasmodesmata, the laticifer water balance is mediated by aquaporins (AQPs). However, to date, the characterization of H. brasiliensis AQPs (HbAQPs) is still in its infancy. Results In this study, 51 full-length AQP genes were identified from the rubber tree genome. The phylogenetic analysis assi...

  4. 钝齿棒杆菌argR基因缺失株构建及其缺失对精氨酸生物合成途径相关基因转录水平的影响%Construction of Corynebacterium crenatum AS 1.542△argR and analysis of transcriptional levels of the related genes of arginine biosynthetic pathway

    陈雪岚; 汤立; 焦海涛; 徐峰; 熊勇华

    2013-01-01

    [Objective] ArgR, coded by the argR gene from Corynebacterium crenatum AS 1. 542, acts as a negative regulator in arginine biosynthetic pathway. However, the effect of argR on transcriptional levels of the related biosynthetic genes has not been reported. Here, we constructed a deletion mutant of argR gene; C. Crenatum AS 1.542 △argR using marker-less knockout technology, and compared the changes of transcriptional levels of the arginine biosynthetic genes between the mutant strain and the wild-type strain. [Methods] We used marker-less knockout technology to construct C. Crenatum AS 1. 542△argR and analyzed the changes of the relate genes at the transcriptional level using real-time fluorescence quantitative PCR. [Results] C. Crenatum AS 1. 542△argR was successfully obtained and the transcriptional level of arginine biosynthetic genes in this mutant increased significantly with an average of about 162. 1 folds. [Conclusion] The arginine biosynthetic genes in C. Crenatum are clearly controlled by the negative regulator ArgR. However, the deletion of this regulator does not result in a clear change in arginine production in the bacteria.%[目的]钝齿棒杆菌AS 1.542中argR基因编码的蛋白ArgR在精氨酸生物合成途径中扮演负调控的角色,但其对相关基因在转录水平的影响还未见报道.因此,本课题组构建了钝齿棒杆菌argR基因缺失株,并在转录水平上比较野生株与缺失株精氨酸生物合成途径相关基因的变化.[方法]采用无痕敲除的方法构建了钝齿棒杆菌argR基因缺失株,并采用荧光定量PCR方法分析缺失株和野生株精氨酸生物合成途径相关基因在转录水平的变化.[结果]利用pK18mobsacB质粒中蔗糖致死基因sacB反向筛选标记及PCR方法成功筛选到钝齿棒杆菌argR基因缺失株;荧光定量PCR结果表明,argR基因缺失株精氨酸生物合成途径中相关基因在转录水平获得大量提高,平均约上调162.13倍.[结论]

  5. Trp64Arg Polymorphism in Beta3-Adrenergic Receptor Gene Is Associated with Decreased Fat Oxidation Both in Resting and Aerobic Exercise in the Japanese Male

    Emiko Morita; Hiroshi Taniguchi; Motoyoshi Sakaue

    2009-01-01

    The purpose of our study was to investigate whether the Trp64Arg polymorphism in β3-AR gene and the −3826A/G polymorphism in the UCP1 gene were associated with the reduction in energy expenditure and fat oxidation both in resting and aerobic exercise in Japanese. Eighty-six nonobese young healthy Japanese were recruited. Energy expenditure was measured using indirect calorimetry. The subjects performed an aerobic exercise program at 60% of their maximal heart rate for 30 min...

  6. Effects of Arc/Arg3.1 gene deletion on rhythmic synchronization of hippocampal CA1 neurons during locomotor activity and sleep.

    Malkki, Hemi A I; Mertens, Paul E C; Lankelma, Jan V; Vinck, Martin; van Schalkwijk, Frank J; van Mourik-Donga, Laura B; Battaglia, Francesco P; Mahlke, Claudia; Kuhl, Dietmar; Pennartz, Cyriel M A

    2016-05-01

    The activity-regulated cytoskeletal-associated protein/activity regulated gene (Arc/Arg3.1) is crucial for long-term synaptic plasticity and memory formation. However, the neurophysiological substrates of memory deficits occurring in the absence of Arc/Arg3.1 are unknown. We compared hippocampal CA1 single-unit and local field potential (LFP) activity in Arc/Arg3.1 knockout and wild-type mice during track running and flanking sleep periods. Locomotor activity, basic firing and spatial coding properties of CA1 cells in knockout mice were not different from wild-type mice. During active behavior, however, knockout animals showed a significantly shifted balance in LFP power, with a relative loss in high-frequency (beta-2 and gamma) bands compared to low-frequency bands. Moreover, during track-running, knockout mice showed a decrease in phase locking of spiking activity to LFP oscillations in theta, beta and gamma bands. Sleep architecture in knockout mice was not grossly abnormal. Sharp-wave ripples, which have been associated with memory consolidation and replay, showed only minor differences in dynamics and amplitude. Altogether, these findings suggest that Arc/Arg3.1 effects on memory formation are not only manifested at the level of molecular pathways regulating synaptic plasticity, but also at the systems level. The disrupted power balance in theta, beta and gamma rhythmicity and concomitant loss of spike-field phase locking may affect memory encoding during initial storage and memory consolidation stages. PMID:27038743

  7. Association of beta3-adrenergic receptor (ADRB3 Trp64Arg gene polymorphism with obesity and metabolic syndrome in the Balinese: a pilot study

    Trimarsanto Hidayat

    2011-05-01

    Full Text Available Abstract Background Prevalence of obesity is increasing all over the world. ADRB3 Trp64Arg gene polymorphism was proposed to be associated with obesity, although inconsistent findings and differences of the Arg64 allele frequency among various ethnics were reported. Westernization was reported to increase the prevalence of obesity in developing world. In this study we determined the prevalence of obesity and metabolic syndrome among urban and rural Balinese, and studied the association of ADRB3 Trp64Arg polymorphism with obesity and MetS. Findings A total of 528 Balinese (urban 282, rural 246 were recruited. Body mass index (BMI and waist circumference (WC were determined; high-density lipoprotein cholesterol (HDL-C, triglyceride (TG, systolic and diastolic blood pressure (SBP and DBP, and fasting plasma glucose (FPG were measured using standard procedures. BMI and WC classifications were based on WHO classifications for Asian. Metabolic syndrome (MetS was defined as described in the Joint Interim Statement. Chi-square test was employed to test the association between the ADRB3 Trp64Arg genotype and disease traits. Urban have higher BMI (p = 2.8 × 10-13, WC ( p -16, TG (p = 0.0028, DBP (p = 1.8 × 10-5, and lower HDL-C (p = 0.0376 when compared to rural. Abdominal obesity and MetS prevalence were significantly higher in urban as compared to rural (both p p = 0.041 for BMI and p = 0.012 for WC, and consequently higher abdominal obesity prevalence (p = 0.007. Comparison between male and female, as well as urban and rural, showed different prevalence of MetS co-morbidities. Abdominal obesity and hypertriglyceridaemia were consistently appeared in all groups, suggesting to play a role as determinant of MetS in both urban and rural. Conclusions Prevalence of obesity and MetS in urban were two times higher when compared to rural. Abdominal obesity and hypertriglyceridaemia appears to be the key determinant of MetS in both urban and rural Balinese

  8. Beta2-adrenoceptor gene variant Arg16Gly is associated with idiopathic ventricular outflow-tract tachycardia

    RAN Yu-qin; LI Ning; YANG Ying; CHEN Jing-zhou; FENG Li; ZHANG Shu; PU Jie-lin

    2010-01-01

    Background Imbalance of the sympathetic nervous system was involved in the pathogenesis of idiopathic ventricular outflow-tract tachycardia (IVOT). We aimed to investigate whether the major genetic variants in β1-and β2-adrenoceptors and GNB3 C825T were associated with IVOT and verapamil sensitive idiopathic left ventricular tachycardia (ILVT).Methods Patients with IVOT and ILVT from December 2005 to December 2007 were consecutively enrolled into this study. Controls were randomly selected from the community-based inhabitants. Five genetic variants, Ser49Gly and Gly389Arg in the β1-adrenoceptor, Arg16Gly and Gln27Glu in the β2-adrenoceptor and GNB3 C825T, were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis.Results A total of 227 patients with IVOT and 110 patients with ILVT were included. Genotyping revealed that the 16Gly allele of Arg16Gly variant of β2-adrenoceptor was associated with a higher risk of IVOT (OR:1.40, 95% CI: 1.12-1.75,P=0.003 in the addictive model and OR:. 1.62, 95% CI: 1.14-2.31, P=0.007 in the dominant model). Patients with Gly16Gln27 haplotype also had a higher risk of IVOT (OR: 1.38, 95% CI: 1.11-1.73, P=0.012). Other four variants,including Ser49Gly and Arg389Gly in β1-adrenoceptor, GIn27Glu in β2-adrenoceptor and GNB3 C825T, did not differ between patients with IVOT and controls. In patients with ILVT, no significant difference was found in these five variants compared with controls.Conclusions Arg16Gly in β2-adrenoceptor is significantly associated with IVOT in Chinese Han population. Major genetic variants in β1- and β2-adrenoceptor and GNB3 C825T may not be associated with ILVT. These data suggest a different arrhythmogenic mechanism in IVOT and ILVT.

  9. Association of Gln27Glu and Arg16Gly polymorphisms in Beta2-adrenergic receptor gene with obesity susceptibility: a meta-analysis.

    Hongxiu Zhang

    Full Text Available BACKGROUND: The beta2-adrenergic receptor (ADRB2 gene polymorphism has been implicated in susceptibility to obesity, but study results are still controversial. OBJECTIVE: The present meta-analysis is performed to determine whether there are any associations between the Gln27Glu (rs1042714 or the Arg16Gly (rs1042713 polymorphisms in ADRB2 and obesity susceptibility. METHODS: The PubMed (1950-2014, Embase (1974-2014, and China National Knowledge Infrastructure (CNKI, 1994-2014 databases were searched using the search terms ("Beta2-adrenergic receptor", "β2-adrenergic receptor" or "ADRB2", "polymorphism," and "obesity". Fixed- or random-effects pooled measures were determined on the bias of heterogeneity tests across studies. Publication bias was examined by Egger's test and the modified Begg's test. RESULTS: Eighteen published articles were selected for meta-analysis. Overall analyses showed that rs1042714 (Gln27Glu was associated with significantly increased obesity risk in the heterozygote model (Gln/Glu vs. Gln/Gln: OR: 1.16, 95% CI: 1.04-1.30, I2 = 49%, P = 0.009 and the dominant model (Gln/Glu + Glu/Glu vs. Gln/Gln: OR: 1.2, 95% CI: 1.00-1.44, I2 = 55%, P = 0.04, whereas no significant association was found in the other models for rs1042714. Also, no significant association was found between the rs1042713 (Arg16Gly gene polymorphism and the risk of obesity in all genetic models. In addition, neither rs1042713 (Arg16Gly nor rs1042714 (Gln27Glu showed any significant association with obesity susceptibility when the population were stratified based on gender. CONCLUSION: Our meta-analysis revealed that the rs1042714 (Gln27Glu polymorphism is associated with obesity susceptibility. However, our results do not support an association between rs1042713 (Arg16Gly polymorphisms and obesity in the populations investigated. This conclusion warrants confirmation by more case-control and cohort studies.

  10. Association between essential hypertension and polymorphisms of beta 1 adrenergic receptor gene G1165C (Gly389Arg) in Chinese Mongolian population

    Rile Hu; Shigang Zhao; Guangming Niu; Chunyu Zhang; Zhiguang Wang; Mingfang Jiang

    2006-01-01

    BACKGROUND: The prevalences of hypertension, cerebrovascular diseases, etc. are higher in Mongolian population because of the influence of various factors including genetics, geography, diet, etc. Therefore, it is helpful to develop researches on the genetics of various diseases including hypertension in Mongolian population.OBJECTIVE: To analyze the association between the polymorphism of beta1 adrenergic receptor (β1-AR)gene G1165C (Arg389Gly), an important candidate gene for various diseases of cardiovascular system, and essential hypertension in Mongolian population.DESIGN: A cross-sectional study.SETTINGS: Department of Neurology, the First Affiliated Hospital of Inner Mongolia Medical College; Wulate Houqi Red Cross Society.PARTICIPANTS: The survey was carried out from February 2003 to March 2005. Totally 239 Mongolian residents, whose blood relations of 3 generations were all Mongolians, were selected from Wulate Houqi, Inner Mongolia, and they were all informed with the survey and detected items. Based on the diagnostic standard of hypertension set by WHO in 1999, the subjects were divided into two groups according to the level blood pressure: ① Normal blood pressure group (n=117): systolic blood pressure (SBP) < 140 mm Hg (1 mm Hg =0.133 kPa), diastolic blood pressure (DBP) < 90 mm Hg, and those having histories of cerebrovascular disease, heart disease, diseases of liver, kidney and tiroides, and diabetes mellitus were excluded. ② Essential hypertension group (n=122): including 51 patients with simple high SBP. All the enrolled subjects had no blood relationship with each other, and had no history of miscegenation.METHODS: The body height, body mass, waist circumference and blood lipids were measured routinely, and their habits of smoking and drinking were also investigated. Peripheral venous blood (5 mL) was drawn, the genome DNA was extracted, and the polymorphisms of the β1-AR G1165C (Gly389Arg) genotype were detected with the Sequenom system

  11. A non-synonymous coding change in the CYP19A1 gene Arg264Cys (rs700519 does not affect circulating estradiol, bone structure or fracture

    Wang Jenny Z

    2011-12-01

    Full Text Available Abstract Background The biosynthesis of estrogens from androgens is catalyzed by aromatase P450 enzyme, coded by the CYP19A1 gene on chromosome 15q21.2. Genetic variation within the CYP19A1 gene sequence has been shown to alter the function of the enzyme. The aim of this study is to investigate whether a non-synonymous Arg264Cys (rs700519 single nucleotide polymorphism (SNP is associated with altered levels of circulating estradiol, areal bone mineral density or fracture. Methods This population- based study of 1,022 elderly Caucasian women (mean age 74.95 ± 2.60 years was genotyped for the rs700519 SNP were analyzed to detect any association with endocrine and bone phenotypes. Results The genotype frequencies were 997 wildtype (97.6%, 24 heterozygous (2.3% and 1 homozygous (0.1%. When individuals were grouped by genotype, there was no association between the polymorphism and serum estradiol (wildtype 27.5 ± 16.0; variants 31.2 ± 18.4, P = 0.27. There was also no association seen on hip bone mineral density (wildtype 0.81 ± 0.12; 0.84 ± 0.14 for variants, P = 0.48 or femoral neck bone mineral density (0.69 ± 0.10 for wildtype; 0.70 ± 0.12 for variants, P = 0.54 before or after correction of the data with age, height, weight and calcium therapy. There were also no associations with quantitative ultrasound measures of bone structure (broadband ultrasound attenuation, speed of sound and average stiffness. Conclusions In a cohort of 1,022 elderly Western Australian women, the presence of Arg264Cys (rs700519 polymorphism was not found to be associated with serum estradiol, bone structure or phenotypes.

  12. Polymorphism Trp64Arg of beta 3 adrenoreceptor gene: allelic frequencies and influence on insulin resistance in a multicenter study of Castilla-León Polimorfismo TRP64ARG del gen receptor beta 3: frecuencia alélica e influencia en la resistencia a la insulina en un estudio multicéntrico de Castilla y León

    D. A. de Luis

    2010-04-01

    Full Text Available Background and objective: The genetic variant (Trp64Arg is a missense mutation located within the beta3 adrenoreceptor (Beta3AR. The aim of our study was to investigate the influence of Trp64Arg polymorphism in the Beta3AR gene on insulin resistance in obese patients and the allelic distribution of this polymorphismin a geographic area of Spain. Design: A population of 264 obese patients was analyzed. A bioimpedance, blood pressure, an assessment of nutritional intake, and biochemical parameters were measured. The beta 3 adrenoreceptor gene polymorphism(Trp64Arg was genotyped. Results: Two hundred and twenty six patients (77 males/149 females (85.6% had the genotype Trp64/Trp64 (wild type group with and average age of 41.12 ± 13.1 years and 38 patients (16 males/22 females Trp64/Arg64 (14.4% (mutant type group with an average age of 40.5 ± 12.7 years. High frequencies of Arg64 allele were observed in Salamanca and Valladolid. In the mutant type group, HOMA (3.75 ± 2.77 vs 5.27 ± 5.4; p Introducción y objetivos: La variante genética (Trp64Arg es una mutación localizada en el adrenoreceptor Beta 3 (Beta3AR. El objetivo de nuestro trabajo es evaluar la influencia de el polimorfismo Trp64Arg del gen de Beta3AR sobre la resistencia a la insulina en pacientes obesos, así como la distribución alélica de este polimorfismo en un área geográfica de España. Diseño: Una muestra de 264 pacientes obesos fue analizada. Se realizó una bioimpedancia, evaluación nutricional y análisis bioquímico. Se genotiparon a los pacientes en función delpolimorfismos Tr64Arg del gen adrenoreceptor-beta 3. Resultados: Un total de 227 pacientes (77 varones/149 mujeres (85,6% presentaron el genotipo Trp64/Trp64 (grupo genotipo salvaje, con una media de edad de 41,12 ± 13,1 años y un total de 38 pacientes (16 varones/22 mujeres Trp64/Arg64 (14,4% (grupo genotipo mutante con una edad media de 40,5 ± 12,7 años. Se detectó una alta frecuencia alélica (Arg64

  13. X-ray repair cross-complementing gene 1 Arg399GIn polymorphism and glioma risk among Asians A meta-analysis based on 2326 cases and 3610 controls

    Liang Zhang; Zhiqun Qiu; Jiaohua Luo; Weiqun Shu

    2012-01-01

    OBJECTIVE:Previous reports have demonstrated that X-ray repair cross-complementing gene 1 (XRCC1) Arg399GIn polymorphism is a possible risk factor for several cancers.Published data on the association of XRCC1Arg399GIn polymorphism with glioma susceptibility have generated conflicting results.This study is designed to precisely estimate the relationship.DATA RETRIEVAL:A computer-based online retrieval of Medline,EMBASE,OVID,Sciencedirect,and Chinese National Knowledge Infrastructure was performed to search papers regarding association of XRCC1 Arg399GIn polymorphisms with glioma published up to April 2012.SELECTION CRITERIA:Two investigators selected data independently.Meta analysis was then performed for the selected studies using STATA 11.0 software after strict selection.Heterogeneity test,sensitivity analysis and publication bias assessments were then conducted.MAIN OUTCOME MEASURES:Association of XRCC1 Arg399GIn polymorphism with glioma risk.RESULTS:A total of nine case-controlled studies comprising 2 326 cases and 3 610 controls were selected for final analysis.The overall data failed to indicate a significant association of XRCC1 Arg399GIn polymorphism with glioma risk (GIn/GIn vs.Arg/Arg:odds ratio (OR) =1.11; 95% confidence interval (Cl) =0.94-1.31; dominant model:OR =1.06; 95%Cl=0.95-1.18; recessive model:OR =1.04; 95%Cl=0.81-1.34).However,subgroup analysis regarding ethnicity showed an increased risk among Asians (GIn/GIn vs.Arg/Arg:OR =1.70; 95%Cl=1.17-2.46; dominant model:OR =1.40; 95%Cl=1.10-1.78; recessive model:OR =1.46; 95%Cl =1.04-2.05) but not Caucasians or mixed ethnicities.CONCLUSION:XRCC1 Arg399GIn polymorphism might modify the susceptibility to glioma among Asians but not Caucasians.Further large and well-designed studies are needed to confirm this conclusion.

  14. Identification of a differentially expressed thymidine kinase gene related to tapping panel dryness syndrome in the rubber tree (Hevea brasiliensis Muell. Arg. by random amplified polymorphic DNA screening

    Arjunan Thulaseedharan

    2010-01-01

    Full Text Available Tapping panel dryness (TPD syndrome is one of the latex yield affecting factors in the rubber tree (Hevea brasiliensis Mull. Arg.. Therefore, identification of a DNA marker will be highly useful for screening progenies in breeding programs. The major goal of this study was to detect genetic variations and/or identification of gene fragments among 37 Hevea clones by the random amplified polymorphic DNA “fingerprinting” technique. Different levels of DNA polymorphism were detected with various primers and a distinct polymorphic band (2.0 kb was obtained with OPA-17 primer. It was cloned into a plasmid vector for further sequence characterization and the nucleotide sequence shows homology with a novel putative plant thymidine kinase (TK gene, designated as HbTK (Hevea brasiliensis thymidine kinase; GenBank accession number AY130829. The protein HbTK has 67%, 65%, 64%, and 63% similarity to TK genes of Medicago, Oryza, Arabidopsis, and Lyco- persicon, respectively, and it was highly conserved in all species analyzed. The predicted amino acid sequence contained conserved domains of TK proteins in the C-terminal half. Southern blot analysis indicated that HbTK is one of the members of a small gene family. Northern blot results revealed that the expression of the HbTK gene was up-regulated in mature bark tissues of the healthy tree while it was down-regulated in the TPD-affected one. These results suggest that this gene may play important roles in maintaining active nucleotide metabolism during cell division at the tapped site of bark tissues in the healthy tree under stress (tapping conditions for normal latex biosynthesis.

  15. Studies of associations between the Arg389Gly polymorphism of the beta1-adrenergic receptor gene (ADRB1) and hypertension and obesity in 7677 Danish white subjects

    Gjesing, A P; Andersen, G; Albrechtsen, A; Glümer, C; Borch-Johnsen, K; Jørgensen, T; Hansen, T; Pedersen, O

    2007-01-01

    Activation of the beta(1)-adrenergic receptor (ADRB1) causes increased lipolysis in adipose tissue and enhances cardiac output. Analysis of the association of the functional ADRB1 Arg389Gly variant with obesity and hypertension has given ambiguous results. To clarify the potential impact of this...... variant on obesity and hypertension in the general population, we examined the Arg389Gly variant in a relatively large-scale population-based study....

  16. Transcriptional Cross-Regulation between Gram-Negative and Gram-Positive Bacteria, Demonstrated Using ArgP-argO of Escherichia coli and LysG-lysE of Corynebacterium glutamicum

    Marbaniang, Carmelita N.; J.Gowrishankar

    2012-01-01

    The protein-gene pairs ArgP-argO of Escherichia coli and LysG-lysE of Corynebacterium glutamicum are orthologous, with the first member of each pair being a LysR-type transcriptional regulator and the second its target gene encoding a basic amino acid exporter. Whereas LysE is an exporter of arginine (Arg) and lysine (Lys) whose expression is induced by Arg, Lys, or histidine (His), ArgO exports Arg alone, and its expression is activated by Arg but not Lys or His. We have now reconstituted in...

  17. An Investigation and Clinical Analysis on Trp64Arg Mutation Frequency of ADR β3 Gene%β3肾上腺素能受体基因Trp64Arg突变率调查分析

    徐文玲; 王明琴; 崔立芳; 曲云东

    2001-01-01

    Objective To investigate the influence of Trp64Arg mutation of β3-adrenergic receptor on obesity, plasma lipid, blood pressure and heart rate.Methods Genotype of Trp64Arg mutation of ADR β3 gene and several clinical variables including body mass index BMI, plasma glucose, plasma lipid, blood pressure, heart rate and so on were determined in 198 Chinese who took their physical examinations.Results The frequency of Trp64Arg mutation of ADR β3 gene in above 198 Chinese was 20.20%. The mutation of ADR β3 gene was found to have some associations with the increase of BMI. Individuals with Trp64Arg mutation of ADR β3 gene were also found to have lower plasma lipid levels.Conclusion The mutation of ADR β3 seems to be related with over-weight. It also seems to have association with plasma lipid regulation. But it has no relation with blood pressure or heart rate.%目的研究中国人β3肾上腺素能受体(ADR β3)基因Trp64Arg突变对肥胖、血脂、血压、心率等的影响。方法对受检者198人进行ADR β3基因Trp64Arg突变基因型的检测,并测体重指数(BMI)、血糖、血脂、血压,心率等。结果本调查组β3肾上腺素能受体基因Trp64Arg突变率为20.20%,BMI增高与其基因突变有关。基因突变者血脂偏低。结论 ADR β3基因突变参与中国人体重的增加和血脂水平的调节,与血压、心率无明显相关性。

  18. The Trp64Arg mutation of the beta3 adrenergic receptor gene has no effect on obesity phenotypes in the Québec Family Study and Swedish Obese Subjects cohorts.

    Gagnon, J; Mauriège, P; S Roy; Sjöström, D; Chagnon, Y. C.; Dionne, F.T.; Oppert, J.M.; Pérusse, L.; Sjöström, L.; Bouchard, C

    1996-01-01

    The beta adrenergic system plays a key role in regulating energy balance through the stimulation of both thermogenesis and lipid mobilization in brown and white adipose tissues in human and various animal models. Recent studies have suggested that a missense Trp64Arg mutation in the beta3 adrenergic receptor (ADRB3) gene was involved in obesity and insulin resistance. We have investigated the effect of this mutation on obesity-related phenotypes in two cohorts: the Québec Family Study (QFS) a...

  19. Interaction between Arg16Gly Polymorphism of ADRA2B Gene and Shift Work on Essential Hypertension among Coal Workers%煤矿工人ADRA2B基因Arg16Gly多态性和倒班的交互作用与原发性高血压的关系

    姚向晖; 范红敏; 王东; 宋士勋; 秦天榜; 郭素娟; 袁聚祥

    2013-01-01

    [目的]探讨β2肾上腺素能受体(β2-AR)ADRA2B基因Arg16Gly多态性及与倒班的交互作用对原发性高血压发病的影响. [方法]以某煤矿进行职业病健康检查的井下作业工人为研究对象,采用成组病例对照研究方法.选择病例组511例,对照组456例,调查内容包括工人年龄、工龄、倒班、吸烟、饮酒情况等危险因素.采集血液标本,分离淋巴细胞,提取DNA,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测ADRA2B基因Arg16Gly多态性.采用多因素logistic回归模型分析基因与倒班的交互作用对原发性高血压发病的影响. [结果]病例组和对照组的ADRA2B基因Arg16Gly位点AA、AG、GG基因型频率分别为27.59%(141/511)、48.73%(249/511)、23.68%(121/511);30.01%(137/456)、53.95%(246/456)、16.04%(73/456).病例组中携带A和G等位基因的频率分别为51.96%、48.04%;对照组为57.02%、42.98%.病例组、对照组基因型和等位基因分布差别皆有统计学意义(P<0.05).两组倒班情况分布差异具有统计学意义(P<0.05).在调整年龄、工龄、高血压家族史、食盐、食用水果、食用蔬菜、体质指数、体育锻炼等影响因素后,携带GG基因型工人患原发性高血压OR是AA型的1.702倍(95%CI:1.130~2.565);携带G等位基因型患原发性高血压OR是A等位基因的1.307倍(95%CI:1.104~1.972);而倒班者患原发性高血压OR是不倒班者的1.388倍(95%CI:1.049~1.835).两者同时存在时对原发性高血压影响的交互作用无统计学意义(P>0.05). [结论] ADRA2B基因Arg16Gly多态性可能是煤矿工人原发性高血压发生的易感因素,但尚未发现倒班和ADRA2B基因Arg16Gly多态性的交互作用与原发性高血压发生有关.%To explore the role of the interaction between Arg16Gly polymorphism of beta 2 adrenergic receptor (β2-AR) gene (ADRA2B) and shift work on essential hypertension (EH) occurrence

  20. Enhanced Expression of (His) 6-Arg-Arg-human Proinsulin in Escherichia coli%(His)6-Arg-Arg-人胰岛素原在大肠杆菌中的高效表达

    李晓红; 余蓉; 曾蓉; 彭先凤; 杨继虞

    2003-01-01

    为了提高胰岛素原的表达量,用pQE-40质粒构建了(His)6-Arg-Arg-人胰岛素原[(His)6-Arg-Arg-human Proinsulin,RRhPI]的大肠杆菌表达系统.通过培养条件的优化,在摇瓶培养的条件下,目标产物RRhPI以包涵体形式获得了高效表达,每升培养基收获湿菌体约27g,包涵体6g(干重1.8g),RRhPI约540mg.该水平已超过现有文献报道的摇瓶培养的最高水平.%To enhance the expression level of human proinsulin, Escherichia coli ( E. coli ) M15 harbouring a plasmid(His)6-Arg -Arg -human proinsulin (RRhPI)/pQE-40 for human proinsulin production was cultured using flask. As a result,high expression level of the gene product, which accumulated as inclusion bodies, was obtained. 27g wet E. coli cells wereharvested from 1L culture medium, and 6g inclusion body also. The harvest of our designed gene product RRhPI was about540mg/L culture medium.

  1. Evaluation of the frequency of polymorphisms in XRCC1 (Arg399Gln) and XPD (Lys751Gln) genes related to the genome stability maintenance in individuals of the resident population from Monte Alegre, PA/Brazil municipality

    The human exposure to ionizing radiation coming from natural sources is an inherent feature of human life on Earth. Ionizing radiation is a known genotoxic agent, which can affect biological molecules, causing DNA damage and genomic instability. The cellular system of DNA repair plays an important role in maintaining genomic stability by repairing DNA damage caused by genotoxic agents. However, genes related to DNA repair may have their role committed when presenting a certain polymorphism. This study intended to analyze the frequency of single nucleotide polymorphisms (SNPs) in genes of DNA repair XRCC1 (Arg39-9Gln) and XPD (Lys751Gln) in a: population of the city of Monte Alegre, that resides in an area of high exposure to natural radioactivity. Samples of saliva were collected from individuals of the population of Monte Alegre, in which 40 samples were of male and 46 female. Through the use of RFLP (length polymorphism restriction fragment) the frequency of homozygous genotypes and / or heterozygous was determined for polymorphic genes. The XRCC1 gene had 65.4% of the presence of the allele 399Gln and XPD gene had 32.9% of the 751Gln allele. These values are similar to those found in previous studies for the XPD gene, whereas XRCC1 showed a frequency much higher than described in the literature. The. influence of these polymorphisms, which are involved in DNA repair and consequent genotoxicity induced by radiation depends on dose and exposure factors such as smoking, statistically a factor in public health surveillance in the region. This study gathered information and molecular epidemiology for risk assessment of cancer in the population of Monte Alegre. (author)

  2. Asymmetric allosteric activation of the symmetric ArgR hexamer.

    Jin, Lihua; Xue, Wei-Feng; Fukayama, June Wong; Yetter, Jaclyn; Pickering, Michael; Carey, Jannette

    2005-02-11

    Hexameric arginine repressor, ArgR, bound to L-arginine serves both as the master transcriptional repressor/activator at diverse regulons in a wide range of bacteria and as a required cofactor for resolution of ColE1 plasmid multimers. Multifunctional ArgR is thus unusual in possessing features of specific gene regulators, global regulators, and non-specific gene organizers; its closest functional analog is probably CAP, the cyclic AMP receptor/activator protein. Isothermal titration calorimetry, surface plasmon resonance, and proteolysis indicate that binding of a single L-argine [corrected] per ArgR hexamer triggers a global conformation [corrected] change and resets the affinities of the remaining five sites, making them 100-fold weaker. The analysis suggests a novel thermodynamic signature for this mechanism of activation. PMID:15663926

  3. Evaluation of the frequency of polymorphisms in XRCC1 (Arg399Gln) and XPD (Lys751Gln) genes related to the genome stability maintenance in individuals of the resident population from Monte Alegre, PA/Brazil municipality; Avaliacao da frequencia de polimorfismos nos genes XRCC1 (Arg399Gln) e XPD (Lys751Gln) relacionados a manutencao da estabilidade do genoma em individuos da populacao residente no municipio de Monte Alegre, PA

    Duarte, Isabelle Magliano

    2010-07-01

    The human exposure to ionizing radiation coming from natural sources is an inherent feature of human life on Earth. Ionizing radiation is a known genotoxic agent, which can affect biological molecules, causing DNA damage and genomic instability. The cellular system of DNA repair plays an important role in maintaining genomic stability by repairing DNA damage caused by genotoxic agents. However, genes related to DNA repair may have their role committed when presenting a certain polymorphism. This study intended to analyze the frequency of single nucleotide polymorphisms (SNPs) in genes of DNA repair XRCC1 (Arg39-9Gln) and XPD (Lys751Gln) in a: population of the city of Monte Alegre, that resides in an area of high exposure to natural radioactivity. Samples of saliva were collected from individuals of the population of Monte Alegre, in which 40 samples were of male and 46 female. Through the use of RFLP (length polymorphism restriction fragment) the frequency of homozygous genotypes and / or heterozygous was determined for polymorphic genes. The XRCC1 gene had 65.4% of the presence of the allele 399Gln and XPD gene had 32.9% of the 751Gln allele. These values are similar to those found in previous studies for the XPD gene, whereas XRCC1 showed a frequency much higher than described in the literature. The. influence of these polymorphisms, which are involved in DNA repair and consequent genotoxicity induced by radiation depends on dose and exposure factors such as smoking, statistically a factor in public health surveillance in the region. This study gathered information and molecular epidemiology for risk assessment of cancer in the population of Monte Alegre. (author)

  4. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum

    CHEN Xue Lan; ZHANG Bin; TANG Li; JIAO Hai Tao; XU Heng Yi; XU Feng; XU Hong; WEI Hua; XIONG Yong Hua

    2014-01-01

    Objective Corynebacterium crenatum MT, a mutant from C. crenatum AS 1.542 with a lethal argR gene, exhibits high arginine production. To confirm the effect of ArgR on arginine biosynthesis in C. crenatum, an intact argR gene from wild-type AS 1.542 was introduced into C. crenatum MT, resulting in C. crenatum MT. sp, and the changes of transcriptional levels of the arginine biosynthetic genes and arginine production were compared between the mutant strain and the recombinant strain. Methods Quantitative real-time polymerase chain reaction was employed to analyze the changes of the related genes at the transcriptional level, electrophoretic mobility shift assays were used to determine ArgR binding with the argCJBDF, argGH, and carAB promoter regions, and arginine production was determined with an automated amino acid analyzer. Results Arginine production assays showed a 69.9%reduction in arginine from 9.01±0.22 mg/mL in C. crenatum MT to 2.71±0.13 mg/mL (P Conclusion The arginine biosynthetic genes in C. crenatum are clearly controlled by the negative regulator ArgR, and intact ArgR in C. crenatum MT results in a significant descrease in arginine production.

  5. Human migration activities drive the fluctuation of ARGs: Case study of landfills in Nanjing, eastern China.

    Sun, Mingming; Ye, Mao; Schwab, Arthur P; Li, Xu; Wan, Jinzhong; Wei, Zhong; Wu, Jun; Friman, Ville-Petri; Liu, Kuan; Tian, Da; Liu, Manqiang; Li, Huixin; Hu, Feng; Jiang, Xin

    2016-09-01

    Landfills are perfect sites to study the effect of human migration on fluctuation of antibiotic resistance genes (ARGs) as they are the final destination of municipal waste. For example, large-scale human migration during the holidays is often accompanied by changes in waste dumping having potential effects on ARG abundance. Three landfills were selected to examine fluctuation in the abundance of fifteen ARGs and Intl1 genes for 14 months in Nanjing, eastern China. Mass human migration, the amount of dumped waste and temperature exerted the most significant effects on bimonthly fluctuations of ARG levels in landfill sites. As a middle-sized cosmopolitan city in China, millions of college students and workers migrate during holidays, contributing to the dramatic increases in waste production and fluctuation in ARG abundances. In line with this, mass migration explained most of the variation in waste dumping. The waste dumping also affected the bioaccessibility of mixed-compound pollutants that further positively impacted the level of ARGs. The influence of various bioaccessible compounds on ARG abundance followed the order: antibiotics>nutrients>metals>organic pollutants. Concentrations of bioaccessible compounds were more strongly correlated with ARG levels compared to total compound concentrations. Improved waste classification and management strategies could thus help to decrease the amount of bioaccessible pollutants leading to more effective control for urban ARG dissemination. PMID:27179703

  6. A Trp33Arg Mutation at Exon 1 of the MYH9 Gene in a Korean Patient with May-Hegglin Anomaly

    JANG, MOON JU; Park, Hyun-Jeong; Chong, So Young; Huh, Ji Young; Kim, In-Ho; Jang, Ja-Hyun; Kim, Hee-Jin; Oh, Doyeun

    2012-01-01

    In this report, we describe a Korean patient with May-Hegglin anomaly from a mutation of the MYH9 gene. The proband was a 21-year-old man with thrombocytopenia. He did not have a bleeding tendency. His neutrophil count was normal at 7490/mm3; however, the neutrophils contained abnormal basophilic inclusions in their cytoplasm. The platelet count was decreased at 15000/mm3 with giant platelets. Coagulation test results were not remarkable. Direct sequencing of MYH9 revealed that he was heteroz...

  7. Molecular characterization of the argJ mutation in Neisseria gonorrhoeae strains with requirements for arginine, hypoxanthine, and uracil.

    Martin, P R; Mulks, M H

    1992-01-01

    Arginine auxotrophs are commonly encountered among clinical isolates of Neisseria gonorrhoeae. Arginine auxotrophs which also require hypoxanthine and uracil (AHU strains) compose a unique set of strains that are highly homogeneous and are believed to be clonally derived. The Arg- phenotype of these strains is due to a lesion in the argJ gene encoding ornithine acetyltransferase. We have cloned the mutant argJ gene from an AHU strain and compared the sequence of this gene to the wild-type arg...

  8. XRCC1 Arg194Trp and Arg399Gln polymorphisms and arsenic methylation capacity are associated with urothelial carcinoma

    The association between DNA repair gene polymorphisms and bladder cancer has been widely studied. However, few studies have examined the correlation between urothelial carcinoma (UC) and arsenic or its metabolites. The aim of this study was to examine the association between polymorphisms of the DNA repair genes, XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln, with urinary arsenic profiles and UC. To this end, we conducted a hospital-based case–control study with 324 UC patients and 647 age- and gender-matched non-cancer controls. Genomic DNA was used to examine the genotype of XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln by PCR-restriction fragment length polymorphism analysis (PCR-RFLP). Urinary arsenic profiles were measured by high performance liquid chromatography (HPLC) linked with hydride generator and atomic absorption spectrometry. The XRCC1 399 Gln/Gln and 194 Arg/Trp and Trp/Trp genotypes were significantly related to UC, and the odds ratio (OR) and 95% confidence interval (95%CI) were 1.68 (1.03–2.75) and 0.66 (0.48–0.90), respectively. Participants with higher total urinary arsenic levels, a higher percentage of inorganic arsenic (InAs%) and a lower percentage of dimethylarsinic acid (DMA%) had a higher OR of UC. Participants carrying XRCC1 risk diplotypes G-C/G-C, A-C/A-C, and A-T/G-T, and who had higher total arsenic levels, higher InAs%, or lower DMA% compared to those with other XRCC1 diplotypes had a higher OR of UC. Our results suggest that the XRCC1 399 Gln/Gln and 194 Arg/Arg DNA repair genes play an important role in poor arsenic methylation capacity, thereby increasing the risk of UC in non-obvious arsenic exposure areas. - Highlights: • The XRCC1 399Gln/Gln genotype was significantly associated with increased OR of UC. • The XRCC1 194 Arg/Trp and Trp/Trp genotype had a significantly decreased OR of UC. • Combined effect of the XRCC1 genotypes and poor arsenic methylation capacity on

  9. XRCC1 Arg194Trp and Arg399Gln polymorphisms and arsenic methylation capacity are associated with urothelial carcinoma

    Chiang, Chien-I [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Huang, Ya-Li [Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Chen, Wei-Jen [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Shiue, Horng-Sheng [Department of Chinese Medicine, Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan (China); Huang, Chao-Yuan; Pu, Yeong-Shiau [Department of Urology, National Taiwan University Hospital, College of Medicine National Taiwan University, Taipei, Taiwan (China); Lin, Ying-Chin [Department of Family Medicine, Shung Ho Hospital, Taipei Medical University, New Taipei, Taiwan (China); Department of Health Examination, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan (China); Division of Family Medicine, School of Medicine, Taipei Medical University, Taipei, Taiwan (China); Hsueh, Yu-Mei, E-mail: ymhsueh@tmu.edu.tw [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2014-09-15

    The association between DNA repair gene polymorphisms and bladder cancer has been widely studied. However, few studies have examined the correlation between urothelial carcinoma (UC) and arsenic or its metabolites. The aim of this study was to examine the association between polymorphisms of the DNA repair genes, XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln, with urinary arsenic profiles and UC. To this end, we conducted a hospital-based case–control study with 324 UC patients and 647 age- and gender-matched non-cancer controls. Genomic DNA was used to examine the genotype of XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln by PCR-restriction fragment length polymorphism analysis (PCR-RFLP). Urinary arsenic profiles were measured by high performance liquid chromatography (HPLC) linked with hydride generator and atomic absorption spectrometry. The XRCC1 399 Gln/Gln and 194 Arg/Trp and Trp/Trp genotypes were significantly related to UC, and the odds ratio (OR) and 95% confidence interval (95%CI) were 1.68 (1.03–2.75) and 0.66 (0.48–0.90), respectively. Participants with higher total urinary arsenic levels, a higher percentage of inorganic arsenic (InAs%) and a lower percentage of dimethylarsinic acid (DMA%) had a higher OR of UC. Participants carrying XRCC1 risk diplotypes G-C/G-C, A-C/A-C, and A-T/G-T, and who had higher total arsenic levels, higher InAs%, or lower DMA% compared to those with other XRCC1 diplotypes had a higher OR of UC. Our results suggest that the XRCC1 399 Gln/Gln and 194 Arg/Arg DNA repair genes play an important role in poor arsenic methylation capacity, thereby increasing the risk of UC in non-obvious arsenic exposure areas. - Highlights: • The XRCC1 399Gln/Gln genotype was significantly associated with increased OR of UC. • The XRCC1 194 Arg/Trp and Trp/Trp genotype had a significantly decreased OR of UC. • Combined effect of the XRCC1 genotypes and poor arsenic methylation capacity on

  10. Cloning and characterization of HbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals

    Li, Yan; Chen, Yue Yi; Yang, Shu Guang; Tian, Wei Min, E-mail: wmtian9110@126.com

    2015-05-22

    Metallothioneins (MTs) are of low molecular mass, cysteine-rich proteins. They play an important role in the detoxification of heavy metals and homeostasis of intracellular metal ions, and protecting against intracellular oxidative damages. In this study a full-length cDNA of type 2 plant metallothioneins, HbMT2a, was isolated from 25 mM Polyethyleneglycol (PEG) stressed leaves of Hevea brasiliensis by RACE. The HbMT2a was 372 bp in length and had a 237 bp open reading frame (ORF) encoding for a protein of 78 amino acid residues with molecular mass of 7.772 kDa. The expression of HbMT2a in the detached leaves of rubber tree clone RY7-33-97 was up-regulated by Me-JA, ABA, PEG, H{sub 2}O{sub 2}, Cu{sup 2+} and Zn{sup 2+}, but down-regulated by water. The role of HbMT2a protein in protecting against metal toxicity was demonstrated in vitro. PET-28a-HbMT2-beared Escherichia coli. Differential expression of HbMT2a upon treatment with 10 °C was observed in the detached leaves of rubber tree clone 93-114 which is cold-resistant and Reken501 which is cold-sensitive. The expression patterns of HbMT2a in the two rubber tree clones may be ascribed to a change in the level of endogenous H{sub 2}O{sub 2}. - Highlights: • Cloning an HbMT2a gene from rubber tree. • Analyzing expression patterns of HbMT2a upon abiotic stress and heavy metal stress. • Finding different expression patterns of HbMT2a among two Hevea germplasm. • The expressed protein of HbMT2a enhances copper and zinc tolerance in Escherichia coli.

  11. Cloning and characterization of HbMT2a, a metallothionein gene from Hevea brasiliensis Muell. Arg differently responds to abiotic stress and heavy metals

    Metallothioneins (MTs) are of low molecular mass, cysteine-rich proteins. They play an important role in the detoxification of heavy metals and homeostasis of intracellular metal ions, and protecting against intracellular oxidative damages. In this study a full-length cDNA of type 2 plant metallothioneins, HbMT2a, was isolated from 25 mM Polyethyleneglycol (PEG) stressed leaves of Hevea brasiliensis by RACE. The HbMT2a was 372 bp in length and had a 237 bp open reading frame (ORF) encoding for a protein of 78 amino acid residues with molecular mass of 7.772 kDa. The expression of HbMT2a in the detached leaves of rubber tree clone RY7-33-97 was up-regulated by Me-JA, ABA, PEG, H2O2, Cu2+ and Zn2+, but down-regulated by water. The role of HbMT2a protein in protecting against metal toxicity was demonstrated in vitro. PET-28a-HbMT2-beared Escherichia coli. Differential expression of HbMT2a upon treatment with 10 °C was observed in the detached leaves of rubber tree clone 93-114 which is cold-resistant and Reken501 which is cold-sensitive. The expression patterns of HbMT2a in the two rubber tree clones may be ascribed to a change in the level of endogenous H2O2. - Highlights: • Cloning an HbMT2a gene from rubber tree. • Analyzing expression patterns of HbMT2a upon abiotic stress and heavy metal stress. • Finding different expression patterns of HbMT2a among two Hevea germplasm. • The expressed protein of HbMT2a enhances copper and zinc tolerance in Escherichia coli

  12. Transcriptional cross-regulation between Gram-negative and gram-positive bacteria, demonstrated using ArgP-argO of Escherichia coli and LysG-lysE of Corynebacterium glutamicum.

    Marbaniang, Carmelita N; Gowrishankar, J

    2012-10-01

    The protein-gene pairs ArgP-argO of Escherichia coli and LysG-lysE of Corynebacterium glutamicum are orthologous, with the first member of each pair being a LysR-type transcriptional regulator and the second its target gene encoding a basic amino acid exporter. Whereas LysE is an exporter of arginine (Arg) and lysine (Lys) whose expression is induced by Arg, Lys, or histidine (His), ArgO exports Arg alone, and its expression is activated by Arg but not Lys or His. We have now reconstituted in E. coli the activation of lysE by LysG in the presence of its coeffectors and have shown that neither ArgP nor LysG can regulate expression of the noncognate orthologous target. Of several ArgP-dominant (ArgP(d)) variants that confer elevated Arg-independent argO expression, some (ArgP(d)-P274S, -S94L, and, to a lesser extent, -P108S) activated lysE expression in E. coli. However, the individual activating effects of LysG and ArgP(d) on lysE were mutually extinguished when both proteins were coexpressed in Arg- or His-supplemented cultures. In comparison with native ArgP, the active ArgP(d) variants exhibited higher affinity of binding to the lysE regulatory region and less DNA bending at both argO and lysE. We conclude that the transcription factor LysG from a Gram-positive bacterium, C. glutamicum, is able to engage appropriately with the RNA polymerase from a Gram-negative bacterium, E. coli, for activation of its cognate target lysE in vivo and that single-amino-acid-substitution variants of ArgP can also activate the distantly orthologous target lysE, but by a subtly different mechanism that renders them noninterchangeable with LysG. PMID:22904281

  13. Evidence for an Arginine Exporter Encoded by yggA (argO) That Is Regulated by the LysR-Type Transcriptional Regulator ArgP in Escherichia coli

    Nandineni, Madhusudan R.; J.Gowrishankar

    2004-01-01

    The anonymous open reading frame yggA of Escherichia coli was identified in this study as a gene that is under the transcriptional control of argP (previously called iciA), which encodes a LysR-type transcriptional regulator protein. Strains with null mutations in either yggA or argP were supersensitive to the arginine analog canavanine, and yggA-lac expression in vivo exhibited argP+-dependent induction by arginine. Lysine supplementation phenocopied the argP null mutation in that it virtual...

  14. Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis of the Gene Expression in Laticifers on the Basis of Latex Flow in Rubber Tree (Hevea brasiliensis Muell. Arg.)

    Chao, Jinquan; Yang, Shuguang; Chen, Yueyi; Tian, Wei-Min

    2016-01-01

    Latex exploitation-caused latex flow is effective in enhancing latex regeneration in laticifer cells of rubber tree. It should be suitable for screening appropriate reference gene for analysis of the expression of latex regeneration-related genes by quantitative real-time PCR (qRT-PCR). In the present study, the expression stability of 23 candidate reference genes was evaluated on the basis of latex flow by using geNorm and NormFinder algorithms. Ubiquitin-protein ligase 2a (UBC2a) and ubiquitin-protein ligase 2b (UBC2b) were the two most stable genes among the selected candidate references in rubber tree clones with differential duration of latex flow. The two genes were also high-ranked in previous reference gene screening across different tissues and experimental conditions. By contrast, the transcripts of latex regeneration-related genes fluctuated significantly during latex flow. The results suggest that screening reference gene during latex flow should be an efficient and effective clue for selection of reference genes in qRT-PCR. PMID:27524995

  15. Cloning and organization of seven arginine biosynthesis genes from Neisseria gonorrhoeae.

    Picard, F J; Dillon, J R

    1989-01-01

    A genomic library for Neisseria gonorrhoeae, constructed in the lambda cloning vector EMBL4, was screened for clones carrying arginine biosynthesis genes by complementation of Escherichia coli mutants. Clones complementing defects in argA, argB, argE, argG, argIF, carA, and carB were isolated. An E. coli defective in the acetylornithine deacetylase gene (argE) was complemented by the ornithine acetyltransferase gene (argJ) from N. gonorrhoeae. This heterologous complementation is reported for...

  16. The Effect of the Arg389Gly Beta-1 Adrenoceptor Polymorphism on Plasma Renin Activity and Heart Rate and the Genotype-Dependent Response to Metoprolol Treatment

    Petersen, Morten; Andersen, Jon T; Jimenez-Solem, Espen;

    2012-01-01

    A gene-drug interaction has been indicated between beta-1 selective beta-blockers and the Arg389Gly polymorphism (rs1801253) in the adrenergic beta-1 receptor gene (ADRB1). We studied the effect of the ADRB1 Arg389Gly polymorphism on plasma renin activity (PRA) and heart rate (HR) and the genotype...

  17. Expression of the benign HEXA mutations, Arg247Trp and Arg249Trp, associated with beta-hexosaminidase A pseudodeficiency

    Cao, Z.; Petroulakis, E.; Salo, T. [Univ. of Manitoba (Canada)] [and others

    1994-09-01

    {beta}-Hexosaminidase (Hex A) is a heterodimer of {alpha} and {beta} subunits encoded by the HEXA and HEXB genes, respectively. Mutations in the HEXA gene typically cause Tay-Sachs disease or less severe forms of G{sub M2} gangliosidosis. However, two benign mutations (Arg247Trp and Arg249Trp) in the {alpha}-subunit of Hex A account for Hex A deficiency in {approximately}36% of non-Jewish enzyme-defined Tay-Sachs disease carriers. These mutations do not result in any apparent clinical phenotype in individuals who are genetic compounds with a second disease-causing mutation. We expressed the {alpha}-subunit harboring each of the benign mutations separately to study activity toward the synthetic substrate, 4-MUGS, for comparison to activity from enzymes containing mutations associated with other forms of G{sub M2} gangliosidosis. The C739T (Arg247Trp;benign), C745T (Arg 249Trp; benign), G805A (Gly269Ser; adult-onset), G749A (Gly250Asp; juvenile), and C508T (Arg170Trp; infantile) mutations were introduced into the {alpha}-subunit cDNA. These were transfected alone, or with the {beta}-subunit cDNA, to generate Hex S ({alpha}{alpha}) or Hex A ({alpha}{beta}), respectively. The activities were monitored using 4-MUGS, and the levels of {alpha}-subunit protein were assessed by Western blotting. Repeated experiments show that the benign mutations produce approximately 35% of normal Hex S and 40% of normal Hex A activity. This level is much higher than that of Hex A harbouring the Gly169Ser adult-onset mutation (12%). A sequential decrease in expressed Hex A activity is observed as mutations associated with more severe phenotypes are expressed. The benign mutations also result in lower levels of mature {alpha}-subunit protein compared to normal, and slightly reduced levels of {alpha}-subunit precursor protein. The Hex A deficiency resulting from benign mutations is not as great as that associated with disease-causing mutations.

  18. Homozygosity for Pro of p53 Arg72Pro as a potential risk factor for hepatocellular carcinoma in Chinese population

    Zhong-Zheng Zhu; Wen-Ming Cong; Shu-Fang Liu; Hui Dong; Guan-Shan Zhu; Meng-Chao Wu

    2005-01-01

    AIM: Codon 72 exon 4 polymorphism (Arg72Pro) of the p53 gene has been implicated in cancer risk. Our objective was to investigate the possible association between p53Arg72Pro polymorphism and susceptibility to hepatocellular carcinoma (HCC) among Chinese population.METHODS: The p53 Arg72Pro genotypes were determined by PCR-based restriction fragment length polymorphism (RFLP) analysis in 507 HCC cases and 541 controls. Odds ratios (ORs) for HCC and 95% confidence intervals (CIs)from unconditional logistic regression models were used to evaluate relative risks. Potential risk factors were included in the logistic regression models as covariates in the multivariate analyses on genotype and HCC.RESULTS: The frequencies for Pro and Arg alleles were 44.5%, 55.5% in HCC cases, and 40.3% and 59.7% in controls, respectively. The Pro allele was significantly associated with the presence of HCC (P = 0.05) and had a higher risk for HCC (OR = 1.19, 95% CI 1.00-1.41) as compared with the Arg allele. After adjusted for potential risk factors, Arg/Pro heterozygotes had an 1.21-fold increased risk (95% CI 0.82-1.78, P = 0.34) of HCC compared with Arg homozygotes, whereas the risk for Pro homozygotes was 1.79 (95% CI 1.06-3.01, P = 0.03) times higher than that for Arg homozygotes. Pro-allele carriers had a higher relative risk of HCC than the Arg-only carriers (adjusted OR = 1.33, 95% CI 0.92-1.92, P = 0.13), although the difference was not statistically significant.CONCLUSION: Homozygosity for Pro of p53 Arg72Pro is potentially one of the genetic risk factors for HCC in Chinese population. The p53 Arg72Pro polymorphism may be used as a stratification marker in screening individuals at a high risk of HCC.

  19. ARG portable neutron radiography. Final report

    In this report all available neutron radiographic data, including results of tests run at LANL, McClellan AFB, and University of Virginia, will be combined to outline specific transportable neutron radiography systems that could achieve the desired results as a complement to x-radiography capabilities for the Accident Response Group (ARG)

  20. Effects of lifestyle intervention on weight and metabolic parameters in patients with impaired glucose tolerance related to beta-3 adrenergic receptor gene polymorphism Trp64Arg(C/T): Results from the Japan Diabetes Prevention Program.

    Sakane, Naoki; Sato, Juichi; Tsushita, Kazuyo; Tsujii, Satoru; Kotani, Kazuhiko; Tominaga, Makoto; Kawazu, Shoji; Sato, Yuzo; Usui, Takeshi; Kamae, Isao; Yoshida, Toshihide; Kiyohara, Yutaka; Sato, Shigeaki; Tsuzaki, Kokoro; Takahashi, Kaoru; Kuzuya, Hideshi

    2016-05-01

    The beta-3 adrenergic receptor (ADRB3), primarily expressed in adipose tissue, is involved in the regulation of energy metabolism. The present study hypothesized that ADRB3 (Trp64Arg, rs4994) polymorphisms modulate the effects of lifestyle intervention on weight and metabolic parameters in patients with impaired glucose tolerance. Data were analyzed from 112 patients with impaired glucose tolerance in the Japan Diabetes Prevention Program, a lifestyle intervention trial, randomized to either an intensive lifestyle intervention group or usual care group. Changes in weight and metabolic parameters were measured after the 6-month intervention. The ADRB3 polymorphisms were determined using the polymerase chain reaction restriction fragment length polymorphism method. Non-carriers showed a greater weight reduction compared with the carriers in both the lifestyle intervention group and usual care group, and a greater increase of high-density lipoprotein cholesterol levels than the carriers only in the lifestyle intervention group. ADRB3 polymorphisms could influence the effects of lifestyle interventions on weight and lipid parameters in impaired glucose tolerance patients. PMID:27330719

  1. Promoter polymorphism -119C/G in MYG1 (C12orf10) gene is related to vitiligo susceptibility and Arg4Gln affects mitochondrial entrance of Myg1

    Philips, Mari-Anne; Kingo, Külli; Karelson, Maire;

    2010-01-01

    MYG1 (Melanocyte proliferating gene 1, also C12orf10 in human) is a ubiquitous nucleo-mitochondrial protein, involved in early developmental processes and in adult stress/illness conditions. We recently showed that MYG1 mRNA expression is elevated in the skin of vitiligo patients. Our aim was to...... examine nine known polymorphisms in the MYG1 gene, to investigate their functionality, and to study their association with vitiligo susceptibility....

  2. Abnormal dentin structure in two novel gene mutations [COL1A1, Arg134Cys] and [ADAMTS2, Trp795-to-ter] causing rare type I collagen disorders.

    De Coster, P J; Cornelissen, M; De Paepe, A; Martens, L C; Vral, A

    2007-02-01

    Histological and ultrastructural observations of dentin of two patients affected with rare types of type I collagen disorders are presented. In the first case, a homozygous nonsense mutation in ADAMTS2 (substitution of a codon for tryptophan by a stopcodon) causes type VIIC Ehlers-Danlos syndrome (EDS) with multiple tooth agenesis and focal dysplastic dentin defects. In the second case, a missense mutation in COL1A1 (substitution of arginine by cysteine) results in a type I EDS phenotype with clinically normal-appearing dentition. Tooth samples are investigated by using light microscopy (LM), transmission electron microscopy (TEM) and immunostaining for types I and III collagen, and tenascin. These are compared with samples from patients with types III and IV osteogenesis imperfecta (OI) in association with dentinogenesis imperfecta (DI), showing a consistently abnormal appearance of the dentin in all specimens, with variations being primarily those of degree of change. Similarities in histological changes include the alternating presence of normal and severe pathological areas in primary and secondary dentin, the latter being characterized by large canal-like structures in atubular areas. Ultrastructural evidence of pathological dentinogenesis include abnormal distribution, size and organization of collagen fibers, which may also be found in clinically unaffected teeth. The histological and ultrastructural changes seen can be explained on the basis of odontoblast dysfunction which may be secondary to the collagen defect, interfering with different levels of odontoblast cell function and intercellular communication. These observations on (ultra)structural dentin defects associated with the two novel gene mutations are the first ever reported. PMID:17118335

  3. Variant Toll-Like Receptor4 (Asp299Gly and Thr399Ile Alleles) and Toll-Like Receptor2 (Arg753Gln and Arg677Trp Alleles) in Colorectal Cancer

    Homa Davoodi; Fong Seow

    2011-01-01

    The innate immune system recognizes the presence of bacterial products through the expression of a family of membrane receptors known as Toll-like receptors (TLRs). Polymorphisms in TLRs have been shown to be associated with increased susceptibility to diseases such as inflammatory bowel disease.The aim of this study was to determine whether there was a correlation between polymorphisms of TLR4 (Asp299Gly; Thr399Ile) and TLR2 (Arg677Trp; Arg753Gln) genes and risk of colorectal cancer. DNA fro...

  4. Variant Toll-Like Receptor4 (Asp299Gly and Thr399Ile Alleles) and Toll-Like Receptor2 (Arg753Gln and Arg677Trp Alleles) in Colorectal Cancer

    Fong Seow; Homa Davoodi

    2011-01-01

    The innate immune system recognizes the presence of bacterial products through the expression of a family of membrane receptors known as Toll-like receptors (TLRs). Polymorphisms in TLRs have been shown to be associated with increased susceptibility to diseases such as inflammatory bowel disease. The aim of this study was to determine whether there was a correlation between polymorphisms of TLR4 (Asp299Gly; Thr399Ile) and TLR2 (Arg677Trp; Arg753Gln) genes and risk of colorectal cancer. DNA fr...

  5. Draft Genome Sequence of Methylobacterium sp. Strain ARG-1 Isolated from the White-Rot Fungus Armillaria gallica

    Collins, Caitlin; Kowalski, Caitlin; Zebrowski, Jessica; Tulchinskaya, Yevgeniya; Tai, Albert K.; James-Pederson, Magdalena

    2016-01-01

    Methylobacterium sp. strain ARG-1 was isolated from a cell culture of hyphal tips of the white-rot fungus Armillaria gallica. We describe here the sequencing, assembly, and annotation of its genome, confirming the presence of genes involved in methylotrophy. This is the first genome announcement of a strain of Methylobacterium associated with A. gallica. PMID:27257212

  6. Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function

    Aprile-Garcia, Fernando; Metzger, Michael W.; Paez-Pereda, Marcelo; Stadler, Herbert; Acuña, Matías; Liberman, Ana C.; Senin, Sergio A.; Gerez, Juan; Hoijman, Esteban; Refojo, Damian; Mitkovski, Mišo; Panhuysen, Markus; Stühmer, Walter; Holsboer, Florian; Deussing, Jan M.; Arzt, Eduardo

    2016-01-01

    The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain) has been described for this SNP so far. Here we show that although the P2X7R-Gln460Arg variant per se is not compromised in its function, co-expression of wild-type P2X7R with P2X7R-Gln460Arg impairs receptor function with respect to calcium influx, channel currents and intracellular signaling in vitro. Moreover, co-immunoprecipitation and FRET studies show that the P2X7R-Gln460Arg variant physically interacts with P2X7R-WT. Specific silencing of either the normal or polymorphic variant rescues the heterozygous loss of function phenotype and restores normal function. The described loss of function due to co-expression, unique for mutations in the P2RX7 gene so far, explains the mechanism by which the P2X7R-Gln460Arg variant affects the normal function of the channel and may represent a mechanism of action for other mutations. PMID:26986975

  7. One isoform of Arg/Abl2 tyrosine kinase is nuclear and the other seven cytosolic isoforms differently modulate cell morphology, motility and the cytoskeleton

    Bianchi, Cristina; Torsello, Barbara; Di Stefano, Vitalba; Zipeto, Maria A.; Facchetti, Rita; Bombelli, Silvia; Perego, Roberto A., E-mail: roberto.perego@unimib.it

    2013-08-01

    The non-receptor tyrosine kinase Abelson related gene (Arg/Abl2) regulates cell migration and morphogenesis by modulating the cytoskeleton. Arg promotes actin-based cell protrusions and spreading, and inhibits cell migration by attenuating stress fiber formation and contractility via activation of the RhoA inhibitor, p190RhoGAP, and by regulating focal adhesion dynamics also via CrkII phosphorylation. Eight full-length Arg isoforms with different N- and C-termini are endogenously expressed in human cells. In this paper, the eight Arg isoforms, subcloned in the pFLAG-CMV2 vector, were transfected in COS-7 cells in order to study their subcellular distribution and role in cell morphology, migration and cytoskeletal modulation. The transfected 1BSCTS Arg isoform has a nuclear distribution and phosphorylates CrkII in the nucleus, whilst the other isoforms are detected in the cytoplasm. The 1BLCTL, 1BSCTL, 1ASCTS isoforms were able to significantly decrease stress fibers, induce cell shrinkage and filopodia-like protrusions with a significant increase in p190RhoGAP phosphorylation. In contrast, 1ALCTL, 1ALCTS, 1ASCTL and 1BLCTS isoforms do not significantly decrease stress fibers and induce the formation of retraction tail-like protrusions. The 1BLCTL and 1ALCTL isoforms have different effects on cell migration and focal adhesions. All these data may open new perspectives to study the mechanisms of cell invasiveness. -Highlights: • Each of the eight Arg isoforms was transfected in COS-7 cells. • Only the 1BSCTS Arg isoform has a nuclear distribution in transfected cells. • The cytoplasmic isoforms and F-actin colocalize cortically and in cell protrusions. • Arg isoforms differently phosphorylate p190RhoGAP and CrkII. • Arg isoforms differently modulate stress fibers, cell protrusions and motility.

  8. Leptin receptor polymorphism Gln223Arg (rs1137101) in oral squamous cell carcinoma and potentially malignant oral lesions.

    Domingos, Patrícia Luciana Batista; Farias, Lucyana Conceição; Pereira, Camila Santos; das Graças Pena, Geórgia; Reis, Tatiana Carvalho; Silva, Rosângela Ramos Veloso; Fraga, Carlos Alberto de Carvalho; de Souza, Marcela Gonçalves; Soares, Mariana Batista; Jones, Kimberly Marie; Menezes, Elytania Veiga; Nobre, Sérgio Avelino Mota; Rodrigues Neto, João Felício; de Paula, Alfredo Maurício Batista; Velásquez-Meléndez, Jorge Gustavo; Sena Guimarães, André Luiz

    2014-01-01

    The purpose of this study was to assess the LEPR gene Gln223Arg polymorphism (rs1137101) in oral squamous cell carcinoma (OSCC) and in potentially malignant oral lesions (PMOL) in comparison to normal oral mucosa in a Brazilian population. Smokers (n = 89) were selected from a representative sample of 471 individuals from the general population of Montes Claros, Brazil. Participants were age and gender matched to patients with OSCC (n = 25) and oral epithelial dysplasia (n = 25). We investigated the LEPR Gln223Arg polymorphism (A>G; rs1137101) in these groups. Genotype variants were assessed by RFLP-PCR, using MspI (HPAII) restriction endonuclease. The institutional review board of the Universidade Estadual de Montes Claros approved the study (process number 2667/2011). Written informed consent for this study was obtained from all participants. The GG genotype (Arg223Arg) appears to be the more relevant polymorphic variant in OSCC. It occurred, approximately, twice as frequently in OSCC patients than in the general population. In contrast, the A allele in its homozygosis form (Gln223Gln) is significantly associated with the development of PMOL; 80% of the samples from the PMOL group exhibit AA genotype. Our findings suggest new insights regarding LEPR gene variations in the development of OSCC and PMOL. PMID:26034683

  9. A Multi-Scale Settlement Matching Algorithm Based on ARG

    Yue, Han; Zhu, Xinyan; Chen, Di; Liu, Lingjia

    2016-06-01

    Homonymous entity matching is an important part of multi-source spatial data integration, automatic updating and change detection. Considering the low accuracy of existing matching methods in dealing with matching multi-scale settlement data, an algorithm based on Attributed Relational Graph (ARG) is proposed. The algorithm firstly divides two settlement scenes at different scales into blocks by small-scale road network and constructs local ARGs in each block. Then, ascertains candidate sets by merging procedures and obtains the optimal matching pairs by comparing the similarity of ARGs iteratively. Finally, the corresponding relations between settlements at large and small scales are identified. At the end of this article, a demonstration is presented and the results indicate that the proposed algorithm is capable of handling sophisticated cases.

  10. Molecular basis for modulated regulation of gene expression in the arginine regulon of Escherichia coli K-12.

    Cunin, R; T. Eckhardt; Piette, J.; Boyen, A; Piérard, A; Glansdorff, N

    1983-01-01

    We compare the nucleotide sequences of the regulatory regions of five genes or groups of genes of the arginine regulon of Escherichia coli K-12: argF, argI, argR, the bipolar argECBH operon and the carAB operon. All these regions harbour one or two copies of a conserved 18 bp sequence which appears to constitute the basic arginine operator sequence (ARG box). We discuss the influence of ARG box copy number, degree of dyad symmetry, base composition, and position relative to the cognate promot...

  11. Leptin receptor Gln223Arg polymorphism and breast cancer risk in Nigerian women: A case control study

    Leptin, a 16 kDa polypeptide hormone, implicated in various physiological processes, exerts its action through the leptin receptor, a member of the class I cytokine receptor family. Both leptin and leptin receptor have recently been implicated in processes leading to breast cancer initiation and progression in animal models and humans. An A to G transition mutation in codon 223 in exon 6 of the leptin receptor gene, resulting in glutamine to arginine substitution (Gln223Arg), lies within the first of two putative leptin-binding regions and may be associated with impaired signaling capacity of the leptin receptor. This study was designed to assess the role of this polymorphism in breast cancer susceptibility in Nigerian women. We utilized a polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) assay to evaluate the association between the Gln223Arg polymorphism of the leptin receptor gene and breast risk in Nigeria in a case control study involving 209 women with breast cancer and 209 controls without the disease. Study participants were recruited from surgical outpatient clinics and surgical wards of four University Teaching Hospitals located in Midwestern and southeastern Nigeria between September 2002 and April 2004. Premenopausal women carrying at least one LEPR 223Arg allele were at a modestly increased risk of breast cancer after adjusting for confounders (OR = 1.8, 95% confidence interval [CI] 1.0–3.2, p = 0.07). There was no association with postmenopausal breast cancer risk (OR = 0.9, 95% CI 0.4–1.8, p = 0.68). Our results suggest that the LEPR Gln223Arg polymorphism in the extracellular domain of the LEPR receptor gene is associated with a modestly increased risk of premenopausal breast cancer in Nigerian women

  12. Association of p53 codon72 Arg>Pro polymorphism with susceptibility to nasopharyngeal carcinoma: evidence from a case-control study and meta-analysis.

    Sahu, S K; Chakrabarti, S; Roy, S D; Baishya, N; Reddy, R R; Suklabaidya, S; Kumar, A; Mohanty, S; Maji, S; Suryanwanshi, A; Rajasubramaniam, S; Asthana, M; Panda, A K; Singh, S P; Ganguly, S; Shaw, O P; Bichhwalia, A K; Sahoo, P K; Chattopadhyay, N R; Chatterjee, K; Kundu, C N; Das, A K; Kannan, R; Zorenpuii; Zomawia, E; Sema, S A; Singh, Y I; Ghosh, S K; Sharma, K; Das, B S; Choudhuri, T

    2016-01-01

    Tumor suppressor p53 is a critical player in the fight against cancer as it controls the cell cycle check point, apoptotic pathways and genomic stability. It is known to be the most frequently mutated gene in a wide variety of human cancers. Single-nucleotide polymorphism of p53 at codon72 leading to substitution of proline (Pro) in place of arginine (Arg) has been identified as a risk factor for development of many cancers, including nasopharyngeal carcinoma (NPC). However, the association of this polymorphism with NPC across the published literature has shown conflicting results. We aimed to conduct a case-control study for a possible relation of p53 codon72 Arg>Pro polymorphism with NPC risk in underdeveloped states of India, combine the result with previously available records from different databases and perform a meta-analysis to draw a more definitive conclusion. A total of 70 NPC patients and 70 healthy controls were enrolled from different hospitals of north-eastern India. The p53 codon72 Arg>Pro polymorphism was typed by polymerase chain reaction, which showed an association with NPC risk. In the meta-analysis consisting of 1842 cases and 2330 controls, it was found that individuals carrying the Pro allele and the ProPro genotype were at a significantly higher risk for NPC as compared with those with the Arg allele and the ArgArg genotype, respectively. Individuals with a ProPro genotype and a combined Pro genotype (ProPro+ArgPro) also showed a significantly higher risk for NPC over a wild homozygote ArgArg genotype. Additionally, the strength of each study was tested by power analysis and genotype distribution by Hardy-Weinberg equilibrium. The outcome of the study indicated that both allele frequency and genotype distribution of p53 codon72 Arg>Pro polymorphism were significantly associated with NPC risk. Stratified analyses based on ethnicity and source of samples supported the above result. PMID:27159678

  13. Variant Toll-Like Receptor4 (Asp299Gly and Thr399Ile Alleles and Toll-Like Receptor2 (Arg753Gln and Arg677Trp Alleles in Colorectal Cancer

    Homa Davoodi

    2011-06-01

    Full Text Available The innate immune system recognizes the presence of bacterial products through the expression of a family of membrane receptors known as Toll-like receptors (TLRs. Polymorphisms in TLRs have been shown to be associated with increased susceptibility to diseases such as inflammatory bowel disease.The aim of this study was to determine whether there was a correlation between polymorphisms of TLR4 (Asp299Gly; Thr399Ile and TLR2 (Arg677Trp; Arg753Gln genes and risk of colorectal cancer. DNA from 60 colorectal carcinoma patients from 3 major races in Malaysia  (22  Malays,  20  Chinese  and  18  Indians  and  blood  from  50  apparently  healthy individuals were evaluated. Control group were matched to study group by race and age. The polymorphisms were determined by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP.Genotyping results showed two out of sixty tumor specimens (3.3% harbored both variant TLR4 Asp299Gly and Thr399Ile alleles. In contrast, DNA isolated from blood cells of 50 apparently healthy individuals harbored wild type TLR4. In the case of TLR2 Arg753Gln genotyping, all of the fifty normal and 60 tumors were of the wild type genotype. TLR2 Arg677Trp genotyping showed a heterozygous pattern in all samples. However, this may not be a true polymorphism of the TLR2 gene as it is likely due to a variation of a duplicated (pseudogene region. There was only a low incidence (2/60; 3.3% of TLR4 polymorphism at the Asp299Gly and Thr399Ile alleles in colorectal cancer patients. All normal and tumor samples harbored the wild type TLR2 Arg753 allele.Our study suggests that variant TLR4 (Asp299Gly and Thr399Ile alleles as well as TLR2 (Arg753Gln allele are not associated with risk of colorectal cancer.

  14. Occurrence of sulfonamide-, tetracycline-, plasmid-mediated quinolone- and macrolide-resistance genes in livestock feedlots in Northern China.

    Mu, Quanhua; Li, Jin; Sun, Yingxue; Mao, Daqing; Wang, Qing; Luo, Yi

    2015-05-01

    Antibiotic resistance genes (ARGs) in livestock feedlots deserve attention because they are prone to transfer to human pathogens and thus pose threats to human health. In this study, the occurrence of 21 ARGs, including tetracycline (tet)-, sulfonamide (sul)-, plasmid-mediated quinolone (PMQR)- and macrolide-resistance (erm) genes were investigated in feces and adjacent soils from chicken, swine, and cattle feedlots in Northern China. PMQR and sul ARGs were the most prevalent and account for over 90.0 % of the total ARGs in fecal samples. Specifically, PMQR genes were the most prevalent, accounting for 59.6 % of the total ARGs, followed by sul ARGs (34.2 %). The percentage of tet ARGs was 3.4 %, and erm ARGs accounted for only 1.9 %. Prevalence of PMQR and sul ARGs was also found in swine and cattle feces. The overall trend of ARG concentrations in feces of different feeding animals was chicken > swine > beef cattle in the studied area. In soils, sul ARGs had the highest concentration and account for 71.1 to 80.2 % of the total ARGs, which is possibly due to the widely distributed molecular carriers (i.e., class one integrons), facilitating sul ARG propagation. Overall, this study provides integrated profiles of various types of ARGs in livestock feedlots and thus provides a reference for the management of antibiotic use in livestock farming. PMID:25475616

  15. Positive relationship detected between soil bioaccessible organic pollutants and antibiotic resistance genes at dairy farms in Nanjing, Eastern China.

    Sun, Mingming; Ye, Mao; Wu, Jun; Feng, Yanfang; Wan, Jinzhong; Tian, Da; Shen, Fangyuan; Liu, Kuan; Hu, Feng; Li, Huixin; Jiang, Xin; Yang, Linzhang; Kengara, Fredrick Orori

    2015-11-01

    Co-contaminated soils by organic pollutants (OPs), antibiotics and antibiotic resistance genes (ARGs) have been becoming an emerging problem. However, it is unclear if an interaction exists between mixed pollutants and ARG abundance. Therefore, the potential relationship between OP contents and ARG and class 1 integron-integrase gene (intI1) abundance was investigated from seven dairy farms in Nanjing, Eastern China. Phenanthrene, pentachlorophenol, sulfadiazine, roxithromycin, associated ARG genes, and intI1 had the highest detection frequencies. Correlation analysis suggested a stronger positive relationship between the ARG abundance and the bioaccessible OP content than the total OP content. Additionally, the significant correlation between the bioaccessible mixed pollutant contents and ARG/intI1 abundance suggested a direct/indirect impact of the bioaccessible mixed pollutants on soil ARG dissemination. This study provided a preliminary understanding of the interaction between mixed pollutants and ARGs in co-contaminated soils. PMID:26256145

  16. Quantitative assessment of the association between XRCC1 Arg399Gln and Arg194Trp polymorphisms and risk of cutaneous melanoma.

    Jiang, Huayong; Xu, Weidong; Zhang, Fuli; Wei, Li; Wang, Yajie; Wang, Yadi; Liu, Chuan

    2016-06-01

    Accumulating evidence has suggested that the XRCC1 Arg399Gln and Arg194Trp polymorphisms might be related to cutaneous melanoma susceptibility. However, epidemiologic findings have been inconsistent. We have assessed reported studies by meta-analysis to perform a more precise estimation of the association between the XRCC1 two polymorphisms (Arg399Gln, Arg194Trp) and risk of cutaneous melanoma. A total of seven eligible articles were selected for this meta-analysis, including 3454 cases and 3811 controls for the XRCC1 Arg399Gln polymorphism and 1256 cases and 1575 controls for the XRCC1 Arg194Trp polymorphism. Overall, no significant associations were found in all genetic models when the studies were pooled into the meta-analysis for the Arg399Gln and Arg194Trp polymorphisms. When stratified by source of control, significant associations were found for the Arg399Gln polymorphism in the population-based subgroup under AA versus GG [odds ratio (OR)=1.43, 95% confidence interval (CI)=1.08-1.88]; the dominant model AA/GA versus GG (OR=1.25, 95% CI=1.04-1.51); and the recessive model AA versus GA/GG (OR=1.31, 95% CI=1.01-1.68). No significant associations were found for the Arg194Trp polymorphism in the subgroup analysis. This meta-analysis suggested that the XRCC1 Arg399Gln polymorphism was a risk factor for cutaneous melanoma in population-based subgroup. PMID:26967970

  17. Characterization of broad-spectrum antibiotic resistance genes in wastewater treatment reactors through metagenomic approaches

    Yang, Ying; 楊穎

    2014-01-01

    The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have attracted great concerns worldwide. Wastewater treatment plants (WWTPs) are reservoirs of ARGs while wastewater/sludge treatment processes are considered as important means to control these emerging biological pollutants. However, the full profiles of ARGs in WWTPs or the removal efficiency of ARGs by wastewater/sludge treatment process was not well characterized yet. Thus, the major tasks in this st...

  18. Replacement of Lys Linker with Arg Linker Resulting in Improved Melanoma Uptake and Reduced Renal Uptake of Tc-99m-Labeled Arg-Gly-Asp-Conjugated Alpha-Melanocyte Stimulating Hormone Hybrid Peptide

    Yang, Jianquan; Guo, Haixun; Padilla, R. Steve; Berwick, Marianne; Miao, Yubin

    2010-01-01

    The purpose of this study was to reduce the non-specific renal uptake of Arg-Gly-Asp (RGD)-conjugated alpha-melanocyte stimulating hormone (α-MSH) hybrid peptide through structural modification or L-lysine co-injection. The RGD motif {cyclic(Arg-Gly-Asp-dTyr-Asp)} was coupled to [Cys3,4,10, d-Phe7, Arg11]α-MSH3-13 {(Arg11)CCMSH} through the Arg linker (substituting the Lys linker) to generate a novel RGD-Arg-(Arg11)CCMSH hybrid peptide. The melanoma targeting and pharmacokinetic properties of...

  19. Arg-Gly-Asp (RGD) Modified Biomimetic Polymeric Materials

    Xufeng NIU; Yuanliang WANG; Yanfeng LUO; Juan XIN; Yonggang LI

    2005-01-01

    The new generation of biomaterials focuses on the design of biomimetic polymeric materials that are capable of eliciting specific cellular responses and directing new tissue formation. Since Arg-Gly-Asp (RGD) sequences have been found to promote cell adhesion in 1984, numerous polymers have been functionalized with RGD peptides for tissue engineering applications. This review gave the advance in RGD modified biomimetic polymeric materials,focusing on the mechanism of RGD, the surface and bulk modification of polymer with RGD peptides and the evaluation in vitro and in vivo of the modified biomimetic materials.

  20. Morfoanatomia foliar de Rauvolfia sellowii Müll. Arg., Apocynaceae

    Baratto, Leopoldo; Duarte, Márcia do Rocio; de Moraes Santos, Cid Aimbiré

    2010-01-01

    Rauvolfia sellowii Müll. Arg. (Apocynaceae), uma árvore nativa brasileira rica em alcaloides indólicos, é conhecida como "pau-pra-tudo" e utilizada popularmente como hipocolesterolêmica, hipoglicêmica e anti-hipertensiva. O presente trabalho teve como objetivo caracterizar morfoanatomicamente a folha desta planta medicinal, a fim de contribuir para o controle de qualidade e autenticidade desta espécie. O material vegetal foi fixado e submetido às microtécnicas usuais. Como caracte...

  1. ARG1 and ARL2 contribute to gravity signal transduction in the statocytes of Arabidopsis thaliana roots and hypocotyls

    Masson, Patrick; Harrison, Benjamin; Stanga, John; Otegui, Marisa; Sedbrook, John

    Gravity is an important cue that plant organs use to guide their growth. Each organ is characterized by a defined gravity set point angle that dictates its optimal orientation within the gravity field. Specialized cells, named statocytes, enable this directional growth response by perceiving gravity via the sedimentation of, and/or tension/pressure exerted by, starch-filled plastids within their cytoplasm. Located in the columella region of the cap in roots and in the endodermis of hypocotyls and stems, these cells modulate the lateral transport of auxin across the corresponding organ in a gravistimulus-dependent manner. Upon plant reorientation within the gravity field, a gravity signal transduction pathway is activated within those cells, which in roots leads to a relocalization of the PIN3 auxin efflux carrier toward the lower membrane and an alkalinization of the cytoplasm. In turn, these events appear to promote a lateral transport of auxin toward the bottom side of the stimulated organ, which promotes a curvature. We previously uncovered ARG1 and ARL2 as essential contributors to these cellular processes. Mutations in these genes result in altered root and hypocotyl gravitropism. In roots, this abnormal growth behavior is associated with a lack of PIN3 relocalization within the statocytes and an absence of preferential downward auxin transport upon gravistimulation. These two genes encode paralogous J-domain proteins that are associated with the plasma membrane and other membranes of the vesicular trafficking pathway, and appear to modulate protein trafficking within the statocytes. An analysis of the root gravitropic phenotypes associated with different double mutant configurations affecting ARG1, ARL2 and PIN3 suggest that all three proteins function in a common gravity-signaling pathway. Surprisingly, when a mutation that affects starch biosynthesis (pgm) is introgressed into an arg1-2 mutant, the gravitropic defects are dramatically enhanced relative to

  2. Leptin receptor polymorphism Gln223Arg (rs1137101) in oral squamous cell carcinoma and potentially malignant oral lesions

    Domingos, Patrícia Luciana Batista; Farias, Lucyana Conceição; Pereira, Camila Santos; das Graças Pena, Geórgia; Reis, Tatiana Carvalho; Silva, Rosângela Ramos Veloso; Fraga, Carlos Alberto de Carvalho; de Souza, Marcela Gonçalves; Soares, Mariana Batista; Jones, Kimberly Marie; Menezes, Elytania Veiga; Nobre, Sérgio Avelino Mota; Rodrigues Neto, João Felício; de Paula, Alfredo Maurício Batista; Velásquez-Meléndez, Jorge Gustavo

    2014-01-01

    The purpose of this study was to assess the LEPR gene Gln223Arg polymorphism (rs1137101) in oral squamous cell carcinoma (OSCC) and in potentially malignant oral lesions (PMOL) in comparison to normal oral mucosa in a Brazilian population. Smokers (n = 89) were selected from a representative sample of 471 individuals from the general population of Montes Claros, Brazil. Participants were age and gender matched to patients with OSCC (n = 25) and oral epithelial dysplasia (n = 25). We investiga...

  3. IL-4 regulates specific Arg-1(+) macrophage sFlt-1-mediated inhibition of angiogenesis.

    Wu, Wei-Kang; Georgiadis, Anastasios; Copland, David A; Liyanage, Sidath; Luhmann, Ulrich F O; Robbie, Scott J; Liu, Jian; Wu, Jiahui; Bainbridge, James W; Bates, David O; Ali, Robin R; Nicholson, Lindsay B; Dick, Andrew D

    2015-08-01

    One of the main drivers for neovascularization in age-related macular degeneration is activation of innate immunity in the presence of macrophages. Here, we demonstrate that T helper cell type 2 cytokines and, in particular, IL-4 condition human and murine monocyte phenotype toward Arg-1(+), and their subsequent behavior limits angiogenesis by increasing soluble fms-like tyrosine kinase 1 (sFlt-1) gene expression. We document that T helper cell type 2 cytokine-conditioned murine macrophages neutralize vascular endothelial growth factor-mediated endothelial cell proliferation (human umbilical vein endothelial cell and choroidal vasculature) in a sFlt-1-dependent manner. We demonstrate that in vivo intravitreal administration of IL-4 attenuates laser-induced choroidal neovascularization (L-CNV) due to specific IL-4 conditioning of macrophages. IL-4 induces the expression of sFlt-1 by resident CD11b(+) retinal microglia and infiltrating myeloid cells but not from retinal pigment epithelium. IL-4-induced suppression of L-CNV is not prevented when sFlt-1 expression is attenuated in retinal pigment epithelium. IL-4-mediated suppression of L-CNV was abrogated in IL-4R-deficient mice and in bone marrow chimeras reconstituted with myeloid cells that had undergone lentiviral-mediated shRNA silencing of sFlt-1, demonstrating the critical role of this cell population. Together, these data establish how lL-4 directly drives macrophage sFlt-1 production expressing an Arg-1(+) phenotype and support the therapeutic potential of targeted IL-4 conditioning within the tissue to regulate disease conditions such as neovascular age-related macular degeneration. PMID:26079814

  4. ARGE DFC - A European approach to MCFC commercialization

    Kraus, P.; Huppmann, G. [MTU Friedrichshafen GmbH, Muenchen (Germany); Heiming, A. [Ruhgras AG, Dorsten (Germany)] [and others

    1996-12-31

    The largest European program for the commercialization of the molten carbonate fuel cell technology is carried out by the European Direct Fuel Cell Consortium (ARGE DFC). The consortium consists of the following companies: (1) MTU Friedrichshafen GmbH (Germany), within the DaimlerBenz Group responsible for off-road propulsion and decentralized energy systems. (2) Haldor Topsoe A/S (Denmark), a plant engineering company and specialist for catalytic processes (3) Elkraft A.m.b.A. (Denmark), one of the countries two utility companies (4) Ruhrgas AG (Germany), the largest German gas company (5) RWE AG (Germany), the largest German electrical utility company MTU acts as a consortium leader. The company shares a license and technology exchange agreement with Energy Research Corporation of Danbury, Connecticut.

  5. Arginine induces GH gene expression by activating NOS/NO signaling in rat isolated hemi-pituitaries

    S.C.F. Olinto

    2012-11-01

    Full Text Available The amino acid arginine (Arg is a recognized secretagogue of growth hormone (GH, and has been shown to induce GH gene expression. Arg is the natural precursor of nitric oxide (NO, which is known to mediate many of the effects of Arg, such as GH secretion. Arg was also shown to increase calcium influx in pituitary cells, which might contribute to its effects on GH secretion. Although the mechanisms involved in the effects of Arg on GH secretion are well established, little is known about them regarding the control of GH gene expression. We investigated whether the NO pathway and/or calcium are involved in the effects of Arg on GH gene expression in rat isolated pituitaries. To this end, pituitaries from approximately 170 male Wistar rats (~250 g were removed, divided into two halves, pooled (three hemi-pituitaries and incubated or not with Arg, as well as with different pharmacological agents. Arg (71 mM, the NO donor sodium nitroprusside (SNP, 1 and 0.1 mM and a cyclic guanosine monophosphate (cGMP analogue (8-Br-cGMP, 1 mM increased GH mRNA expression 60 min later. The NO acceptor hemoglobin (0.3 µM blunted the effect of SNP, and the combined treatment with Arg and L-NAME (a NO synthase (NOS inhibitor, 55 mM abolished the stimulatory effect of Arg on GH gene expression. The calcium channel inhibitor nifedipine (3 µM also abolished Arg-induced GH gene expression. The present study shows that Arg directly induces GH gene expression in hemi-pituitaries isolated from rats, excluding interference from somatostatinergic neurons, which are supposed to be inhibited by Arg. Moreover, the data demonstrate that the NOS/NO signaling pathway and calcium mediate the Arg effects on GH gene expression.

  6. Arginine induces GH gene expression by activating NOS/NO signaling in rat isolated hemi-pituitaries

    The amino acid arginine (Arg) is a recognized secretagogue of growth hormone (GH), and has been shown to induce GH gene expression. Arg is the natural precursor of nitric oxide (NO), which is known to mediate many of the effects of Arg, such as GH secretion. Arg was also shown to increase calcium influx in pituitary cells, which might contribute to its effects on GH secretion. Although the mechanisms involved in the effects of Arg on GH secretion are well established, little is known about them regarding the control of GH gene expression. We investigated whether the NO pathway and/or calcium are involved in the effects of Arg on GH gene expression in rat isolated pituitaries. To this end, pituitaries from approximately 170 male Wistar rats (∼250 g) were removed, divided into two halves, pooled (three hemi-pituitaries) and incubated or not with Arg, as well as with different pharmacological agents. Arg (71 mM), the NO donor sodium nitroprusside (SNP, 1 and 0.1 mM) and a cyclic guanosine monophosphate (cGMP) analogue (8-Br-cGMP, 1 mM) increased GH mRNA expression 60 min later. The NO acceptor hemoglobin (0.3 µM) blunted the effect of SNP, and the combined treatment with Arg and L-NAME (an NO synthase (NOS) inhibitor, 55 mM) abolished the stimulatory effect of Arg on GH gene expression. The calcium channel inhibitor nifedipine (3 µM) also abolished Arg-induced GH gene expression. The present study shows that Arg directly induces GH gene expression in hemi-pituitaries isolated from rats, excluding interference from somatostatinergic neurons, which are supposed to be inhibited by Arg. Moreover, the data demonstrate that the NOS/NO signaling pathway and calcium mediate the Arg effects on GH gene expression

  7. Arginine induces GH gene expression by activating NOS/NO signaling in rat isolated hemi-pituitaries

    Olinto, S.C.F. [Faculdade de Ciências Integradas do Pontal, Universidade Federal de Uberlândia, Ituiutaba, MG (Brazil); Adrião, M.G. [Departamento de Morfologia e Fisiologia, Universidade Federal Rural de Pernambuco, Recife, PE (Brazil); Castro-Barbosa, T.; Goulart-Silva, F.; Nunes, M.T. [Departamento de Fisiologia e Biofísica, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP (Brazil)

    2012-06-01

    The amino acid arginine (Arg) is a recognized secretagogue of growth hormone (GH), and has been shown to induce GH gene expression. Arg is the natural precursor of nitric oxide (NO), which is known to mediate many of the effects of Arg, such as GH secretion. Arg was also shown to increase calcium influx in pituitary cells, which might contribute to its effects on GH secretion. Although the mechanisms involved in the effects of Arg on GH secretion are well established, little is known about them regarding the control of GH gene expression. We investigated whether the NO pathway and/or calcium are involved in the effects of Arg on GH gene expression in rat isolated pituitaries. To this end, pituitaries from approximately 170 male Wistar rats (∼250 g) were removed, divided into two halves, pooled (three hemi-pituitaries) and incubated or not with Arg, as well as with different pharmacological agents. Arg (71 mM), the NO donor sodium nitroprusside (SNP, 1 and 0.1 mM) and a cyclic guanosine monophosphate (cGMP) analogue (8-Br-cGMP, 1 mM) increased GH mRNA expression 60 min later. The NO acceptor hemoglobin (0.3 µM) blunted the effect of SNP, and the combined treatment with Arg and L-NAME (an NO synthase (NOS) inhibitor, 55 mM) abolished the stimulatory effect of Arg on GH gene expression. The calcium channel inhibitor nifedipine (3 µM) also abolished Arg-induced GH gene expression. The present study shows that Arg directly induces GH gene expression in hemi-pituitaries isolated from rats, excluding interference from somatostatinergic neurons, which are supposed to be inhibited by Arg. Moreover, the data demonstrate that the NOS/NO signaling pathway and calcium mediate the Arg effects on GH gene expression.

  8. An ARGS-aggrecan assay for analysis in blood and synovial fluid

    Larsson, S; Lohmander, Stefan; Struglics, A

    2014-01-01

    the Meso Scale Discovery (MSD) platform for detection of ARGS-aggrecan was validated, using a standard made from recombinant human aggrecan. Matched samples of SF, serum, plasma, and urine were obtained from 36 subjects at different time points after knee injury, and analysed for ARGS-aggrecan content...

  9. Aminopeptidase activity in rat brain synaptosomes - 2-mercaptoethanol stimulation and Arg-vasopressin degradation

    Rat brain synaptic plasma membranes contain an amastatin-inhibited aminopeptidase activity which degrades Arg-vaso-pressin (AVP). The pH optimum for AVP cleavage was found to be 6.8, similar to that reported for oxytocin. The ability of other peptides and arylamides such as oxytocin, Tyr-Phe-Met-Arg-Phe-NH2 and Arg-Arg-βNA to inhibit cleavage of [3H-Tyr2]-AVP suggests that the enzyme may not be specific for AVP. The AVP-cleaving activity has been solubilized and partially characterized. Synaptosomes were lysed with hypotonic buffer, washed, and extracted with 1% Nonidet P-40 detergent. The solubilized protein was chromatographed by gel filtration HPLC on Superose 6. A single peak of activity was found with a M.W. = 117,000 which could hydrolyze 1mM Ala-βNA, Arg-βNA, Arg-Arg-βNA, Phe-Met and Phe-Arg as well as slowly cleave AVP with the ultimate release of 3H-Tyr. 2-Mercaptoethanol (3.9mM) (ME) stimulated activity 3.6 to 6.6-fold for arylamide and dipeptide substrates, but 35-fold for labelled AVP, possibly owing to reduction of the AVP disulfide bond. All activities in the presence of ME were completely inhibited by 0.2mM amastatin

  10. Aminopeptidase activity in rat brain synaptosomes - 2-mercaptoethanol stimulation and Arg-vasopressin degradation

    Simmons, W.H.; Orawski, A.T.

    1986-03-05

    Rat brain synaptic plasma membranes contain an amastatin-inhibited aminopeptidase activity which degrades Arg-vaso-pressin (AVP). The pH optimum for AVP cleavage was found to be 6.8, similar to that reported for oxytocin. The ability of other peptides and arylamides such as oxytocin, Tyr-Phe-Met-Arg-Phe-NH/sub 2/ and Arg-Arg-..beta..NA to inhibit cleavage of (/sup 3/H-Tyr/sup 2/)-AVP suggests that the enzyme may not be specific for AVP. The AVP-cleaving activity has been solubilized and partially characterized. Synaptosomes were lysed with hypotonic buffer, washed, and extracted with 1% Nonidet P-40 detergent. The solubilized protein was chromatographed by gel filtration HPLC on Superose 6. A single peak of activity was found with a M.W. = 117,000 which could hydrolyze 1mM Ala-..beta..NA, Arg-..beta..NA, Arg-Arg-..beta..NA, Phe-Met and Phe-Arg as well as slowly cleave AVP with the ultimate release of /sup 3/H-Tyr. 2-Mercaptoethanol (3.9mM) (ME) stimulated activity 3.6 to 6.6-fold for arylamide and dipeptide substrates, but 35-fold for labelled AVP, possibly owing to reduction of the AVP disulfide bond. All activities in the presence of ME were completely inhibited by 0.2mM amastatin.

  11. Relationships between plasma leptin levels, leptin G2548A, leptin receptor Gln223Arg polymorphisms and gestational diabetes mellitus in Chinese population

    Mei Yang; Songxu Peng; Wei Li; Zhihua Wan; Linlin Fan; Yukai Du

    2016-01-01

    The purposes of this study were to examine concentrations of leptin and biochemical parameters in gestational diabetes mellitus (GDM) patients and normal glucose tolerance (NGT) individuals, and also to explore the links of leptin (LEP) G2548A and leptin receptor (LEPR) Gln223Arg polymorphisms with leptin levels and GDM risk among Chinese. Our study included 357 GDM and 355 NGT individuals who were at 24~30 gestational weeks. Plasma leptin and insulin levels were analyzed by ELISA. Gene polym...

  12. [Fate of ARB and ARGs During Wastewater Treatment Process of Spiramycin Production].

    Qin, Cai-xia; Tong, Juan; Shen, Pei-hong; Wei, Yuan-song

    2015-09-01

    Antibiotic resistant bacteria (ARB) and antibiotic resistance gene(ARG) pose great risk to both environment and human health. This study aimed to investigate the fate of macrolide resistant bacteria, six macrolide resistance genes ermB, ermF, ermX, mefA, ereA, mphB and three transfer elements ISCR1, intIl and Tn916/1545 during wastewater biological treatment processes of spiramycin production. Samples were collected from an antibiotic wastewater treatment station in different seasons. Results showed that the total heterotrophs and Enterococci were mostly removed during wastewater biological treatment, with the reduction of 1. 6-2. 1 logs for total heterotrophs and of 3. 7 logs for Enterococci, respectively. For 94 antibiotic resistant Enterococci individually isolated from four different treatment units including adjusting tank, anaerobic tank, anoxic tank, and aerobic tank, all of these strains showed resistance to spiramycin, azithromycin, erythromycin, and clarithromycin; moreover, the antibiotic resistance rates was not reduced in the effluent. Results of PCR and quantitative PCR showed that 80% of antibiotic resistant Enterococci were positive for PCR amplification of erAB, but negative for PCR amplification of other genes. Concentrations of ermB and ermF were peaked in the spring and autumn samples. Resistance genes of ermB, ermF, mefA, ereA, mphB and transfer element of Tn916/1545 were reduced to some degree during antibiotic production wastewater treatment, but concentrations of ermX, intIl, ISCRl in the effluent were higher than those in the influent. The abundance of mefA, ereA and Tn916/1545 were reduced during wastewater treatment process, and the better removal performance for mefA, ereA, Tn916/1545 occurred in spring than in autumn; however, the abundance of ermX, intI1 and ISCR1 were increased. PMID:26717693

  13. Cleavage at both Arg306 and Arg506 is required and sufficient for timely and efficient inactivation of factor Va by activated protein C*

    Barhoover, Melissa A.; Kalafatis, Michael

    2011-01-01

    Background Activated protein C (APC) inactivates membrane-bound factor Va following cleavages of the heavy chain at Arg306, Arg506, and Arg679. Objectives To examine which cleavage is most important for inactivation, recombinant factor V molecules were constructed as follows: factor V306Q (R306→Q), factor V506Q (R506→Q), and factor V306Q/506Q (R306→Q and R506→Q). Methods The recombinant molecules were expressed in mammalian cells, purified, and assayed prior and after incubation with APC and lipids for 30 min (factor Vai) in clotting assays and in an assay using purified reagents and saturating concentrations of factor Va. Results Clotting assays demonstrated that factor VaiWT, factor Vai306Q and factor Vai506Q were devoid of activity, while factor Vai306Q/506Q maintained ~70% activity following a 30 minute incubation with APC. Prothrombinase assembled with all mutant cofactor molecules before and after treatment with APC had Km values similar to values found with prothrombinase assembled with factor VaWT. Prothrombinase assembled with factor VaiWT demonstrated 20-fold reduction in kcat, while prothrombinase assembled with factor Vai506Q had a 2-fold reduction in kcat as compared to prothrombinase assembled with factor VaWT. In contrast, factor Vai306Q and factor Vai306Q/506Q didn’t show any loss in kcat under similar experimental conditions. Conclusion Our data demonstrate that: 1) the activity of an APC-treated factor Va molecule bearing a single mutation at Arg306 or Arg506 depends on the assay used; and 2) regardless of the assay employed, in the absence of the APC-cleavage sites at Arg306 and Arg506, the active cofactor is unable to be significantly inactivated by APC in the presence of a membrane surface. PMID:21467919

  14. Identification of Genes Important for Growth of Asymptomatic Bacteriuria Escherichia coli in Urine

    Vejborg, Rebecca Munk; de Evgrafov, Mari Cristina Rodriguez; Phan, Minh Duy;

    2012-01-01

    clinical ABU E. coli strain VR89. Six genes involved in the biosynthesis of various amino acids and nucleobases were identified (carB, argE, argC, purA, metE, and ilvC), and site-specific mutants were subsequently constructed in E. coli 83972 and E. coli VR89 for each of these genes. In all cases, these...

  15. Seasonal variations in antibiotic resistance gene transport in the Almendares River, Havana, Cuba

    Charles W Knapp

    2012-11-01

    Full Text Available Numerous studies have quantified antibiotic resistance genes (ARG in rivers and streams around the world, and significant relationships have been shown that relate different pollutant outputs and increased local ARG levels. However, most studies have not considered ambient flow conditions, which can vary dramatically especially in tropical countries. Here, ARG were quantified in water-column and sediment samples during the dry-and wet-seasons to assess how seasonal and other factors influence ARG transport down the Almendares River (Havana, Cuba. Eight locations were sampled and stream flow estimated during both seasons; qPCR was used to quantify four tetracycline, two erythromycin, and three beta-lactam resistance genes. ARG concentrations were higher in wet-season versus dry-season samples, which combined with higher flows, indicated greater ARG transport downstream during the wet season. Water-column ARG levels were more spatially variable in the dry-season than the wet-season, with the proximity of waste outfalls strongly influencing local ARG levels. Results confirm that dry-season sampling provides a useful picture of the impact of individual waste inputs on local stream ARG levels, whereas, the majority of ARGs in this tropical river were transported downstream during the wet season, possibly due to re-entrainment of ARG from sediments.

  16. Mallotus philippinensis Muell. Arg (Euphorbiaceae: Ethnopharmacology and Phytochemistry Review

    Mayank Gangwar

    2014-01-01

    Full Text Available Mallotus philippinensis Muell. Arg (Euphorbiaceae are widely distributed perennial shrub or small tree in tropical and subtropical region in outer Himalayas regions with an altitude below 1,000 m and are reported to have wide range of pharmacological activities. Mallotus philippinensis species are known to contain different natural compounds, mainly phenols, diterpenoids, steroids, flavonoids, cardenolides, triterpenoids, coumarins, isocoumarins, and many more especially phenols; that is, bergenin, mallotophilippinens, rottlerin, and isorottlerin have been isolated, identified, and reported interesting biological activities such as antimicrobial, antioxidant, antiviral, cytotoxicity, antioxidant, anti-inflammatory, immunoregulatory activity protein inhibition against cancer cell. We have selected all the pharmacological aspects and toxicological and all its biological related studies. The present review reveals that Mallotus philippinensis is a valuable source of medicinally important natural molecules and provides convincing support for its future use in modern medicine. However, the existing knowledge is very limited about Mallotus philippinensis and its different parts like steam, leaf, and fruit. Further, more detailed safety data pertaining to the acute and subacute toxicity and cardio- and immunotoxicity also needs to be generated for crude extracts or its pure isolated compounds. This review underlines the interest to continue the study of this genus of the Euphorbiaceae.

  17. Hemocytes of the Rose Sawfly Arge ochropus (Gmelin) (Hymenoptera: Argidae).

    Khosravi, R; Sendi, J J; Brayner, F A; Alves, L C; Feitosa, A P S

    2016-02-01

    We characterized individual morphological types of the rose sawfly, Arge ochropus (Gmelin) (Hymenoptera: Argidae), hemocytes for the first time by means of light and differential interference contrast microscopy and scanning and transmission electron microscopy. Four types of hemocytes were identified in the hemolymph of larvae and pupae of A. ochropus: prohemocytes, plasmatocytes, granulocytes, and oenocytoids. Prohemocytes are the smallest type of hemocytes, rounded to ovoid cells with large nuclei. Plasmatocytes are polymorphic and variable in size. Granulocytes are oval and spherical cells variable in size, with variable number of rough endoplasmic reticulum, mitochondria, and microtubules in the cytoplasm. Oenocytoids contain eccentric nucleus and cytoplasm with small mitochondria and few rough endoplasmic reticula. Differential hemocyte counts indicated that plasmatocytes are the most abundant hemocyte type during early instars while granulocytes are the most abundant hemocyte type in the last instar. The pattern of total hemocyte count changed during rose sawfly development and reached its peak in prepupae and then declined slowly in the pupal stage. PMID:26429582

  18. Influence of Glu/Arg, Asp/Arg, and Glu/Lys Salt Bridges on α-Helical Stability and Folding Kinetics.

    Meuzelaar, Heleen; Vreede, Jocelyne; Woutersen, Sander

    2016-06-01

    Using a combination of ultraviolet circular dichroism, temperature-jump transient-infrared spectroscopy, and molecular dynamics simulations, we investigate the effect of salt bridges between different types of charged amino-acid residue pairs on α-helix folding. We determine the stability and the folding and unfolding rates of 12 alanine-based α-helical peptides, each of which has a nearly identical composition containing three pairs of positively and negatively charged residues (either Glu(-)/Arg(+), Asp(-)/Arg(+), or Glu(-)/Lys(+)). Within each set of peptides, the distance and order of the oppositely charged residues in the peptide sequence differ, such that they have different capabilities of forming salt bridges. Our results indicate that stabilizing salt bridges (in which the interacting residues are spaced and ordered such that they favor helix formation) speed up α-helix formation by up to 50% and slow down the unfolding of the α-helix, whereas salt bridges with an unfavorable geometry have the opposite effect. Comparing the peptides with different types of charge pairs, we observe that salt bridges between side chains of Glu(-) and Arg(+) are most favorable for the speed of folding, probably because of the larger conformational space of the salt-bridging Glu(-)/Arg(+) rotamer pairs compared to Asp(-)/Arg(+) and Glu(-)/Lys(+). We speculate that the observed impact of salt bridges on the folding kinetics might explain why some proteins contain salt bridges that do not stabilize the final, folded conformation. PMID:27276251

  19. Role of Arg-72 of pharaonis Phoborhodopsin (Sensory Rhodopsin II) on its Photochemistry

    Ikeura, Yukako; Shimono, Kazumi; Iwamoto, Masayuki; Sudo, Yuki; Kamo, Naoki

    2004-01-01

    Pharaonis phoborhodopsin (ppR, or pharaonis sensory rhodopsin II, NpsRII) is a sensor for the negative phototaxis of Natronomonas (Natronobacterium) pharaonis. Arginine 72 of ppR corresponds to Arg-82 of bacteriorhodopsin, which is a highly conserved residue among microbial rhodopsins. Using various Arg-72 ppR mutants, we obtained the following results: 1), Arg-72ppR together possibly with Asp-193 influenced the pKa of the counterion of the protonated Schiff base. 2), The M-rise became approx...

  20. WOOD ANATOMY OF Phyllanthus sellowianus Müll. Arg. (PHYLLANTHACEAE

    Luciano Denardi

    2013-06-01

    Full Text Available http://dx.doi.org/10.5902/198050989294The present study aimed to describe the wood anatomy of Phyllanthus sellowianus Müll. Arg.(Phyllanthaceae, as well as the research of anatomical characters associated with the flexibility of their stems, desirable characteristic for bioengineering works. Belonging to the group of  reophyllous and with up to three  meters in height, the species occurs naturally in the south of Brazil, Uruguay and Argentina. The exsicat and wood sample from a P. sellowianus, located in the municipality of Santa Maria, Rio Grande do Sul state, Brazi, were deposited in the University Federal de Santa Maria under the numbers HDCF 5587 e 5588. Standard technique was used to set the histological lamina of wood. For macerations used the method of Jeffrey. The Wood anatomy description followed the IAWA Committee (1989. The microscopic analysis revealed the presence vassels of small diameter (< 60µm, radial multiples of 2 to 4 units, and simple perforation plates, obliquely, axial parenchyma absent, heterogeneous rays, with 1-3 cells large, and gelatinous fibers. The absence of axial parenchyma and laticifer tubes is compatible to the Phyllanthaceae group. The presence of perforated ray cells is, probably, the main anatomic component for differentiation of P. sellowianus from the other species of the genera. The abundant presence of gelatinous fibers certainly is one of the main anatomical characteristics related to the high flexibility of its stem.

  1. Polimorfisme Gly972Arg Gen IRS-1 dan Cys981Tyr Gen PTPN1 sebagai Faktor Risiko pada Sindrom Metabolik dengan Riwayat Berat Bayi Lahir Rendah

    Hikmat Permana

    2012-09-01

    Full Text Available Increasing prevalence of obesity occurs worldwide and can affect all ages levels. Risk factors of increasing metabolic syndrome were associated with changes in lifestyle, obesity and low birth weight (LBW. This study was aimed to analyze the genotype IRS-1, genotype PTPN1 and LBW in metabolic syndrome risk factors. In 2009, this restrospective cohort study was comprised of 97 LBW and 100 normal birth weight (NBW, aged 20–21 years old, from the previous cohort study in District Tanjung Sari since 1989. The role of IRS-1 gene, PTPN1 gene and LBW in the metabolic syndrome factors were analyzed using multiple logistic regressions. The result showed there were no significant relationship between IRS-1 and PTPN1 genotype with metabolic syndrome factors but only IRS-1 genotype was significantly associated with trygliceride (p=0.006. Gly972Arg IRS-1 gene polymorphism in LBW has predictive value of increasing diastolic blood pressure, waist circumference and trygliceride. Cys981 Tyr PTPN1 gene polymorphism showed a risk factor for systolic blood pressure and high density lipoprotein (HDL cholesterol. Gly972Arg IRS-1 gene polymorphism was a dominant factor to increase diastolic blood pressure, waist circumference and trygliceride. In conclusions, Cys981 Tyr PTPN1 gene polymorphism is a risk factor of increased systolic blood pressure and HDL cholesterol, and LBW is the risk factor increasing fasting glucose level.

  2. Arg deficiency does not influence the course of Myelin Oligodendrocyte Glycoprotein (MOG35-55)-induced experimental autoimmune encephalomyelitis

    Jacobsen, Freja Aksel; Hulst, Camilla; Bäckström, Thomas;

    2016-01-01

    Background: Inhibition of Abl kinases has an ameliorating effect on the rodent model for multiple sclerosis, experimental autoimmune encephalomyelitis, and arrests lymphocyte activation. The family of Abl kinases consists of the Abl1/Abl and Abl2/Arg tyrosine kinases. While the Abl kinase has bee...... encephalomyelitis is not dependent on Arg, but Arg plays a role for the number of B cells in immunized mice. This might suggest a novel role for the Arg kinase in B-cell trafficking or regulation. Furthermore, the results suggest that Arg is important for normal embryonic development....

  3. N- and C-terminal isoforms of Arg quantified by real-time PCR are specifically expressed in human normal and neoplastic cells, in neoplastic cell lines, and in HL-60 cell differentiation.

    Perego, Roberto A; Corizzato, Matteo; Bianchi, Cristina; Eroini, Barbara; Bosari, Silvano

    2005-04-01

    The human ABL2 (or ARG) gene codes for a nonreceptor tyrosine kinase is involved in translocation with the ETV6 gene in human leukemia and has an altered expression in several human carcinomas. Two isoforms of Arg with different N-termini (1A and 1B) have been described. The C-terminal domain of Arg contains two F-actin-binding sequences that perform a number of actions related to cell morphology and motility by interacting with actin filaments. We have identified different-sized specific cDNAs in hematopoietic, epithelial, nervous, and fibroblastic cells by means of the reverse transcription (RT)-polymerase chain reaction (PCR) analysis of human Arg mRNA. Some of these cDNAs showed an adjunctive alternative splice event involving the 63 bp sequence of exon II, thus leading to four cDNA types with different N-termini: 1A long and short, and 1B long and short. Other cDNAs lacked a 309 bp sequence in the last exon involving one of the C-terminal F-actin binding domains, thus giving rise to two cDNA types: C-termini long and short. Quantified by real-time PCR-quantitative RT-PCR-these Arg transcript isoforms have specific expression patterns not only in different normal and tumor cell types, but also during cell differentiation and growth arrest. These isoforms maintained the open reading frames, and eight putative proteins were predicted. The different C-termini isoforms seem to retain the same quantitative reciprocal ratio of their respective transcripts. The Arg protein isoforms with different C-terminal actin-binding domains and different N-termini might have specific cellular localizations/concentrations, and differently regulated catalytic activity with different implications in normal and neoplastic cells. PMID:15765532

  4. Dominant Retinitis Pigmentosa, p.Gly56Arg Mutation in NR2E3: Phenotype in a Large Cohort of 24 Cases

    Lopez Martinez, Miguel Angel; Lopez-Molina, Maria Isabel; Riveiro-Alvarez, Rosa; Fernandez-San Jose, Patricia; Avila-Fernandez, Almudena; Corton, Marta; Millan, Jose M.; García Sandoval, Blanca; Ayuso, Carmen

    2016-01-01

    Importance This research is the single largest NR2E3 genotype-phenotype correlation study performed to date in autosomal dominant Retinitis Pigmentosa. Objective The aim of this study is to analyse the frequency of the p.Gly56Arg mutation in NR2E3 for the largest cohort of autosomal dominant Retinitis Pigmentosa patients to date and its associated phenotype. Patients and Methods A cohort of 201 unrelated Spanish families affected by autosomal dominant Retinitis Pigmentosa. The p.Gly56Arg mutation in the NR2E3 (NM_014249.2) gene was analysed in 201 families. In the 24 cases where the mutation had been detected, a haplotype analysis linked to the p.Gly56Arg families was performed, using four extragenic polymorphic markers D15S967, D15S1050, D15S204 and D15S188. Phenotype study included presence and age of onset of night blindness, visual field loss and cataracts; and an ophthalmoscopic examination after pupillary dilation and electroretinogram for the 24 cases. Results Seven of the 201 analyzed families were positive for the p.Gly56Arg, leading to a prevalence of 3.5%. Clinical data were available for 24 subjects. Night blindness was the first noticeable symptom (mean 15.9 years). Visual field loss onset was variable (23.3 ± 11.9 years). Loss of visual acuity appeared late in the disease´s evolution. Most of the patients with cataracts (50%) presented it from the third decade of life. Fundus changes showed inter and intrafamiliar variability, but most of the patients showed typical RP changes and it was common to find macular affectation (47.4%). Electroretinogram was impaired from the beginning of the disease. Two families shared a common haplotype. Additionally, all patients shared a 104Kb region between D15S1050 and the NR2E3 gene. Conclusions This study highlights the importance of p.Gly56Arg in the NR2E3 gene as a common mutation associated with adRP, and provides new clues to its phenotype, which can allow for a better clinical management and genetic

  5. Real-Time Imaging Reveals Properties of Glutamate-Induced Arc/Arg 3.1 Translation in Neuronal Dendrites.

    Na, Youn; Park, Sungjin; Lee, Changhee; Kim, Dong-Kyu; Park, Joo Min; Sockanathan, Shanthini; Huganir, Richard L; Worley, Paul F

    2016-08-01

    The immediate early gene Arc (also Arg3.1) produces rapid changes in synaptic properties that are linked to de novo translation. Here we develop a novel translation reporter that exploits the rapid maturation and "flash" kinetics of Gaussia luciferase (Gluc) to visualize Arc translation. Following glutamate stimulation, discrete Arc-Gluc bioluminescent flashes representing sites of de novo translation are detected within 15 s at distributed sites in dendrites, but not spines. Flashes are episodic, lasting ∼20 s, and may be unitary or repeated at ∼minute intervals at the same sites. Analysis of flash amplitudes suggests they represent the quantal product of one or more polyribosomes, while inter-flash intervals appear random, suggesting they arise from a stochastic process. Surprisingly, glutamate-induced translation is dependent on Arc open reading frame. Combined observations support a model in which stalled ribosomes are reactivated to rapidly generate Arc protein. PMID:27397520

  6. Apert Syndrome: Molecularly Confirmed C.758C>G (P.Pro253Arg) in FGFR2

    A 5-day-old girl was referred to our clinic for evaluation of congenital malformations. She was identified with a pathogenic mutation c.758C>G (p.Pro253Arg) in FGFR2 gene using targeted exome sequencing. The de novo mutation was confirmed with Sanger sequencing in the patient and her parents. She showed occipital plagiocephaly with frontal bossing (Figure A and B). Skull frontal and lateral radiography revealed fusion of most of the sutures except coronal suture, with convolutional markings (Figure D and E). She had complete cleft palate (Figure C). Her fused bilateral hands showed type II syndactyly with complete syndactyly between the ring and the little fingers (Figure F1-F3). Both toes were simple syndactyly with side-to-side fusion of skin (Figure G1-)

  7. Apert Syndrome: Molecularly Confirmed C.758C>G (P.Pro253Arg) in FGFR2

    Cha Gon, Lee, E-mail: leechagon@eulji.ac.kr [Department of Pediatrics, Eulji General Hospital, College of Medicine, Eulji University, 68 Hangeulbiseok-ro, Nowon-gu, Seoul 139-711 (Korea, Republic of)

    2016-03-21

    A 5-day-old girl was referred to our clinic for evaluation of congenital malformations. She was identified with a pathogenic mutation c.758C>G (p.Pro253Arg) in FGFR2 gene using targeted exome sequencing. The de novo mutation was confirmed with Sanger sequencing in the patient and her parents. She showed occipital plagiocephaly with frontal bossing (Figure A and B). Skull frontal and lateral radiography revealed fusion of most of the sutures except coronal suture, with convolutional markings (Figure D and E). She had complete cleft palate (Figure C). Her fused bilateral hands showed type II syndactyly with complete syndactyly between the ring and the little fingers (Figure F1-F3). Both toes were simple syndactyly with side-to-side fusion of skin (Figure G1-)

  8. Functional analysis of the p.(Leu15Pro) and p.(Gly20Arg) sequence changes in the signal sequence of LDL receptor.

    Pavloušková, Jana; Réblová, Kamila; Tichý, Lukáš; Freiberger, Tomáš; Fajkusová, Lenka

    2016-07-01

    The low density lipoprotein receptor (LDLR) is a transmembrane protein that plays a key role in cholesterol metabolism. It contains 860 amino acids including a 21 amino acid long signal sequence, which directs the protein into the endoplasmic reticulum. Mutations in the LDLR gene lead to cholesterol accumulation in the plasma and results in familial hypercholesterolemia (FH). Knowledge of the impact of a mutation on the LDLR protein structure and function is very important for the diagnosis and management of FH. Unfortunately, for a large proportion of mutations this information is still missing. In this study, we focused on the LDLR signal sequence and carried out functional and in silico analyses of two sequence changes, p.(Gly20Arg) and p.(Leu15Pro), localized in this part of the LDLR. Our results revealed that the p.(Gly20Arg) change, previously described as disease causing, has no detrimental effect on protein expression or LDL particle binding. In silico analysis supports this observation, showing that both the wt and p.(Gly20Arg) signal sequences adopt an expected α-helix structure. In contrast, the mutation p.(Leu15Pro) is not associated with functional protein expression and exhibits a structure with disrupted a α-helical arrangement in the signal sequence, which most likely affects protein folding in the endoplasmic reticulum. PMID:27175606

  9. Meta analysis of association between XRCC1 Arg399Gln and nasopharyngeal carcinoma susceptibility%XRCC1 Arg399Gln 基因多态性与鼻咽癌易感性的 Meta 分析

    吴晓红; 于祥远; 王程强

    2015-01-01

    was I2 = 32% ,PHet = 0 .18 .Under the recessive and co‐dominant models ,combined OR and 95% CI were 1 .30(1 .04 - 1 .63) and 1 .37(1 .09 - 1 .72) respectively ,and with no significant heterogeneity was observed (I2 = 0 ,PHet = 1 .00) and (I2 = 0 ,PHet = 0 .96) .Conclusion XRCC1 gene Arg399Gln polymorphism is closely related to the genetic susceptibility of NPC ,399Gln allele may be a risk of genetic factors in NPC incidence in asians .

  10. Site-directed mutagenesis from Arg195 to His of a microalgal chloroplastidial glycerol-3-phosphate acyltransferase causes an increase in phospholipid levels in yeast

    Long-Ling eOuyang

    2016-03-01

    Full Text Available To analyze the contribution of glycerol-3-phosphate acyltransferase (GPAT to the first acylation of glycerol-3-phosphate (G-3-P, the present study focused on a functional analysis of the GPAT gene from Lobosphaera incisa (designated as LiGPAT and the subcellular localization of the encoded protein LiGPAT. A full-length cDNA of LiGPAT consisting of a 1,305-bp ORF, a 1,652-bp 5′-UTR, and a 354-bp 3′-UTR, was cloned. The ORF encoded a 434-amino acid peptide, of which 63 residues at the N-terminus defined a chloroplast transit peptide. LiGPAT was exclusively localized to chloroplasts, which was shown by co-expression of LiGPAT with eGFP in Chlamydomonas reinhardtii and by immunogold labeling in L. incisa. Considering the conservation of His among the G-3-P binding sites from chloroplastidial GPATs and the substitution of His by Arg at position 195 in the LiGPAT mature protein (designated mLiGPAT, we established the heterologous expression of either mLiGPAT or its mutant (Arg195His (sdmLiGPAT in the GPAT-deficient yeast mutant gat1Δ. Lipid profile analyses of these transgenic yeasts not only validated the acylation function of LiGPAT but also indicated that the site-directed mutagenesis from Arg195 to His led to an increase in the phospholipid level in yeast. Semi-quantitative analysis of mLiGPAT and sdmLiGPAT, together with the structural superimposition of their G-3-P binding sites, indicated that the increased enzymatic activity was caused by the enlarged accessible surface of the phosphate group binding pocket when Arg195 was mutated to His. Thus, the potential of genetic manipulation of GPAT to increase the glycerolipid level in L. incisa and other microalgae would be of great interest.

  11. Housefly Larva Vermicomposting Efficiently Attenuates Antibiotic Resistance Genes in Swine Manure, with Concomitant Bacterial Population Changes

    Wang, Hang; Li, Hongyi; Gilbert, Jack A.; Li, Haibo; Wu, Longhua; Liu, Meng; Wang, Liling; Zhou, Qiansheng; Yuan, Junxiang; Zhang, Zhijian

    2015-01-01

    Manure from swine treated with antimicrobials as feed additives is a major source for the expansion of the antibiotic resistance gene (ARG) reservoir in the environment. Vermicomposting via housefly larvae (Musca domestica) can be efficiently used to treat manure and regenerate biofertilizer, but few studies have investigated its effect on ARG attenuation. Here, we tracked the abundances of 9 ARGs and the composition and structure of the bacterial communities in manure samples across 6 days o...

  12. Genotype-phenotype correlations of TGFBI p.Leu509Pro, p.Leu509Arg, p.Val613Gly, and the allelic association of p.Met502Val-p.Arg555Gln mutations

    Niel-Butschi, Florence; Kantelip, Bernadette; Iwaszkiewicz, Justyna; Zoete, Vincent; Boimard, Mathieu; Delpech, Marc; Bourges, Jean-Louis; Renard, Gilles; D’Hermies, François; Pisella, Pierre-Jean; Hamel, Christian; Delbosc, Bernard

    2011-01-01

    Purpose Investigate the genotype-phenotype correlations for five TGFBI (transforming growth factor, beta-induced) mutations including one novel pathogenic variant and one complex allele affecting the fourth FAS1 domain of keratoepithelin, and their potential effects on the protein’s structure. Methods Three unrelated families were clinically diagnosed with lattice corneal dystrophy (CD) and one with an unclassified CD of Bowman’s layer. Mutations in the TGFBI gene were detected by direct sequencing, and the functional impact of each variant was predicted using in silico algorithms. Corneal phenotypes, including histological examinations, were compared with the literature data. Furthermore, molecular modeling studies of these mutations were performed. Results Two distinct missense mutations affecting the same residue at position 509 of keratoepithelin: p.Leu509Pro (c.1526T>C) and p.Leu509Arg (c.1526T>G) were found to be associated with a lattice-type CD. The novel p.Val613Gly (c.1828T>G) TGFBI mutation was found in a sporadic case of an Algerian individual affected by lattice CD. Finally, the Bowman’s layer CD was linked to the association in cis of the p.Met502Val and p.Arg555Gln variants, leading to the reclassification of this CD as atypical Thiel-Behnke CD. Structural modeling of these TGFBI mutations argues in favor of these mutations being responsible for instability and/or incorrect folding of keratoepithelin, predictions that are compatible with the clinical diagnoses. Conclusions Description of a novel TGFBI mutation and a complex TGFBI allele further extends the mutational spectrum of TGFBI. Moreover, we show convincing evidence that TGFBI mutations affecting Leu509 are linked to the lattice phenotype in two unrelated French families, contrasting with findings previously reported. The p.Leu509Pro was reported to be associated with both amyloid and non-amyloid aggregates, whereas p.Leu509Arg has been described as being responsible for Epithelial

  13. Arg-Phe-amide-related peptides influence gonadotropin-releasing hormone neurons

    Haluk Kelestimur; Emine Kacar; Aysegul Uzun; Mete Ozcan; Selim Kutlu

    2013-01-01

    The hypothalamic Arg-Phe-amide-related peptides, gonadotropin-inhibitory hormone and orthologous mammalian peptides of Arg-Phe-amide, may be important regulators of the hypothalamus-pituitary-gonadal reproductive axis. These peptides may modulate the effects of kisspeptins because they are presently recognized as the most potent activators of the hypothalamus-pituitary-gonadal axis. However, their effects on gonadotropin-releasing hormone neurons have not been investigated. In the current study, the GT1–7 cell line-expressing gonadotropin-releasing hormone was used as a model to explore the effects of Arg-Phe- amide-related peptides on kisspeptin activation. Intracellular calcium concentration was quantified using the calcium-sensitive dye, fura-2 acetoxymethyl ester. Gonadotropin-releasing hormone released into the medium was detected via enzyme-linked immunosorbent assay. Results showed that 100 nmol/L kisspeptin-10 significantly increased gonadotropin-releasing hormone levels (at 120 minutes of exposure) and intracellular calcium concentrations. Co-treatment of kisspeptin with 1 μmol/L gonadotropin-inhibitory hormone or 1 μmol/L Arg-Phe-amide-related peptide-1 significantly attenuated levels of kisspeptin-induced gonadotropin-releasing hormone but did not affect kisspeptin-induced elevations of intracellular calcium concentration. Overall, the results suggest that gonadotropin-inhibitory hormone and Arg-Phe-amide-related peptide-1 may have inhibitory effects on kisspeptin-activated gonadotropin-releasing hormone neurons independent of the calcium signaling pathway.

  14. Cloning and Bioinformatical Analysis of Phospholipase Dα1 Gene and Its Promoter in the Latex of Rubber Tree (Hevea brasiliensis Muell.Arg.)%橡胶树胶乳HbPLDα1基因及其启动子的克隆与生物信息学分析

    夏可灿; 康桂娟; 黎瑜; 聂智毅; 代龙军; 段翠芳; 曾日中

    2012-01-01

    植物磷脂酶PLDα1与伤害信号转导密切相关,是伤害诱导内源茉莉酸(Jasmonic acid,JA)生物合成的关键酶之一.橡胶树胶乳PLDα1基因(HbPLDα1)表达的研究将有助于揭示橡胶树乳管细胞JA信号转导及其调控橡胶生物合成的机制.在EST序列的基础上,通过RACE和Genome Walking方法分别克隆了橡胶树胶乳的HbPLDα1基因及其启动子序列.HbPLDα1基因的cDNA全长为2 870 bp,包含长度为2427 bp的完整开放阅读框(ORF),具有典型的植物PLDα蛋白保守功能域,与同属大戟科的蓖麻和麻风树的PLDα1基因亲缘关系最近.HbPLDα1基因启动子区域长为1 559 bp,除含有TATA box和CAAT box等基本顺式作用元件外,还存在JA和脱落酸等激素响应元件以及干旱胁迫等环境信号响应元件,这表明HbPLDα1基因的表达可能受激素和环境信号的调控,在橡胶树乳管细胞对激素和环境信号的响应过程中发挥重要作用.%Phospholipase D alpha 1(PLDαl) is involved in wound signal transduction in plant cells, and is a key component for wound -induced jasmonic acid (JA) production. The investigation of PLDαl expression would facilitate unraveling the signaling pathway of JA and its regulation of the rubber biosynthesis in Hevea brasilienais (rubber tree). The PLDal gene from the Hevea latex, HbPLDal, and its promoter were cloned based on the EST sequences using RACE and Genome Walking. The full-length cDNA of HbPLDal gene was 2 870 bp with a complete ORF of 2 427 bp. HbPLDal had the similar typical conserved domains of plant PLD with those of other plants, and showed the highest identity with those of PLDal from Ricinus communis and Jatropha curcas. The promoter of HbPLDal was 1 559 bp long, including not only the ordinary cis-acting elements of TATA and CAAT box, but also such cis-acting elements responsive to phytohormones (JA, ABA, GB) and environmental stresses. The data demonstrated that the expressed of HbPLDal gene could be

  15. Retinoic acid promotes the development of Arg1-expressing dendritic cells for the regulation of T-cell differentiation

    Chang, Jinsam; Thangamani, Shankar; Kim, Myung H.; Ulrich, Benjamin; Morris, Sidney M.; Chang H Kim

    2013-01-01

    Arginase I (Arg1), an enzyme expressed by many cell types including myeloid cells, can regulate immune responses. Expression of Arg1 in myeloid cells is regulated by a number of cytokines and tissue factors that influence cell development and activation. Retinoic acid, produced from vitamin A, regulates the homing and differentiation of lymphocytes and plays important roles in the regulation of immunity and immune tolerance. We report here that optimal expression of Arg1 in dendritic cells re...

  16. CD and UV Resonance Raman Indicate Little arg-glu Side Chain α-helix Peptide Stabilization

    Hong, Zhenmin; Ahmed, Zeeshan; Asher, Sanford A.

    2011-01-01

    Electrostatic interactions between side chains can control the conformation and folding of peptides and proteins. We used CD and UV resonance Raman spectroscopy (UVRR) to examine the impact of side chain charge on the conformations of two 21 residue mainly polyala peptides with a few arg and glu residues. We expected that attractions between arg-10 and glu-14 side chains would stabilize the α-helix conformation compared to a peptide with an arg-14. Surprisingly, CD suggests that the peptide w...

  17. Biosynthesis of homoarginine (hArg) and asymmetric dimethylarginine (ADMA) from acutely and chronically administered free L-arginine in humans.

    Kayacelebi, Arslan Arinc; Langen, Jennifer; Weigt-Usinger, Katharina; Chobanyan-Jürgens, Kristine; Mariotti, François; Schneider, Jessica Y; Rothmann, Sabine; Frölich, Jürgen C; Atzler, Dorothee; Choe, Chi-Un; Schwedhelm, Edzard; Huneau, Jean François; Lücke, Thomas; Tsikas, Dimitrios

    2015-09-01

    Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthesis, whereas L-arginine (Arg) and L-homoarginine (hArg) serve as substrates for NO synthesis. ADMA and other methylated arginines are generally believed to exclusively derive from guanidine (N (G))-methylated arginine residues in proteins by protein arginine methyltransferases (PRMTs) that use S-adenosylmethionine (SAM) as the methyl donor. L-Lysine is known for decades as a precursor for hArg, but only recent studies indicate that arginine:glycine amidinotransferase (AGAT) is responsible for the synthesis of hArg. AGAT catalyzes the formation of guanidinoacetate (GAA) that is methylated to creatine by guanidinoacetate methyltransferase (GAMT) which also uses SAM. The aim of the present study was to learn more about the mechanisms of ADMA and hArg formation in humans. Especially, we hypothesized that ADMA is produced by N (G)-methylation of free Arg in addition to the known PRMTs-involving mechanism. In knockout mouse models of AGAT- and GAMT-deficiency, we investigated the contribution of these enzymes to hArg synthesis. Arg infusion (0.5 g/kg, 30 min) in children (n = 11) and ingestion of high-fat protein meals by overweight men (n = 10) were used to study acute effects on ADMA and hArg synthesis. Daily Arg ingestion (10 g) or placebo for 3 or 6 months by patients suffering from peripheral arterial occlusive disease (PAOD, n = 20) or coronary artery disease (CAD, n = 30) was used to study chronic effects of Arg on ADMA synthesis. Mass spectrometric methods were used to measure all biochemical parameters in plasma and urine samples. In mice, AGAT but not GAMT was found to contribute to plasma hArg, while ADMA synthesis was independent of AGAT and GAMT. Arg infusion acutely increased plasma Arg, hArg and ADMA concentrations, but decreased the plasma hArg/ADMA ratio. High-fat protein meals acutely increased plasma Arg, hArg, ADMA concentrations, as well as the plasma hArg

  18. Polymorphisms in α- and β-Adrenergic Receptor Genes, Hypertension, and Obstructive Sleep Apnea: The Skaraborg Sleep Study

    Kristina Bengtsson Boström

    2010-01-01

    Full Text Available The sympathetic nervous system and the adrenergic receptors play an important role in regulation of blood pressure. This study explored the associations between functional polymorphisms of the α2B-, β1-, and β2-adrenergic receptor genes and obstructive sleep apnea (OSA in hypertensive patients and hypertension in patients with OSA in a populationbased sample of 157 hypertensive patients and 181 healthy control subjects. Only the Arg389Gly polymorphism of the β1-adrenergic receptor gene was associated with increased risk for mild OSA in hypertensive patients (Arg/Arg versus Gly/Arg/Gly/Gly, 2.1, 95% CI, 1.02–4.7. Hypertensive men carrying the Arg389Arg genotype had higher crude and age-adjusted AHI than carriers of the Arg389Gly/Gly389Gly genotypes. When adjusted also for BMI this difference became borderline significant. This difference was not observed in women. The risk of hypertension in mild OSA was associated with increasing number of Arg-alleles (Arg/Arg OR 5.4, 95% CI 1.4–21.2.

  19. Insulin Receptor Substrate-1 (IRS-1 Gly927Arg: Correlation with Gestational Diabetes Mellitus in Saudi Women

    Khalid Khalaf Alharbi

    2014-01-01

    Full Text Available Pregnant women with gestational diabetes mellitus (GDM and type 2 diabetes mellitus (T2DM share a common pathophysiology associated with similar risk factors. Genetic variants used to determine the risk of developing T2DM might also be associated with the prevalence of GDM. The aim of the present study was to scrutinize the relationship between the G972R polymorphism of the insulin receptor substrate-1 (IRS-1 gene with GDM in the Saudi female population. This is a case-control study that monitored 500 Saudi women. Subjects with GDM (n=200 were compared with non-GDM (n=300 controls. We opted to evaluate rs1801278 polymorphism in the IRS1 gene, which plays a critical role in the insulin-signaling pathway. Genotyping was performed with the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP method. The frequency of the rs1801278 polymorphism was significantly higher in women with GDM than in women with non-GDM (for TT + CT versus CC: P=0.02. Additionally, there was a significant increase in the frequency of the Arg-encoding mutant allele from GDM to non-GDM (for T versus C: P=0.01. Our results suggest that the rs1801278 polymorphism in the IRS-1 gene is involved in the occurrence of GDM in the Saudi population.

  20. Contribution of Arg288 of Escherichia coli elongation factor Tu to translational functionality

    Rattenborg, Thomas; Nautrup-Pedersen, Gitte; Clark, Brian F. C.;

    1997-01-01

    network is disrupted upon formation of the ternary complex. Arg288 was replaced by alanine, isoleucine, lysine or glutamic acid, and the resulting mutants have been subjected to an in vitro characterisation with the aim of clarifying the function of Arg288. Unexpectedly, the mutants behaved like the wild......-type factor with regard to the association and dissociation of guanine nucleotides, and the intrinsic GTPase activities are unchanged. Furthermore, the mutants were as efficient as the wild-type factor in carrying out protein synthesis in vitro in the presence of an excess of aminoacyl-tRNA. However, the...

  1. Microsomal epoxide hydrolase (EPHX1), slow (exon 3, 113His) and fast (exon 4, 139Arg) alleles confer susceptibility to squamous cell esophageal cancer

    Genetic polymorphisms in xenobiotic metabolizing enzymes may alter risk of various cancers. Present case-control study evaluated the influence of EPHX1 genetic variations on squamous cell esophageal cancer (ESCC) susceptibility in 107 patients and 320 controls. EPHX1 polymorphic alleles were genotyped by direct sequencing (exon 3, Tyr113His) or PCR-RFLP (exon 4, His139Arg). Patients with exon 3 genotypes (Tyr113His, His113His) and 113His allele were at risk of ESCC (ORTyr113His 2.0, 95% CI = 1.2-3.4, p = 0.007; ORHis113His 2.3 95% CI = 1.0-5.2, p = 0.03 and ORHis 1.5, 95% CI = 1.0-2.1, p = 0.01). In contrast, individuals with exon 4, 139Arg allele were at low risk of cancer (OR 0.34, 95% CI = 0.20-0.56, p = 0.001). However, none of haplotype combinations of exon 3 (Tyr113His) and exon 4 (His139Arg) polymorphisms showed modulation of risk for ESCC. Sub-grouping of patients based on anatomical location of tumor predicted that patients with exon 3, His113His and Tyr113His genotypes were at higher risk for developing ESCC tumor at upper and middle third locations (OR 4.4, 95% CI = 1.0-18.5, p = 0.04; OR 2.5, 95% CI = 1.3-5.0, p = 0.005 respectively). The frequency of exon 4, His139Arg genotype was significantly lower in ESCC patients with lower third tumor location as compared to controls (14.8% vs. 36.3%, p = 0.02). In case-only study, gene-environment interaction of EPHX1 genotypes with tobacco, alcohol and occupational exposures did not appear to modulate the cancer susceptibility. In conclusion, exon 3, Tyr113His genotype was associated with higher risk of ESCC particularly at upper and middle-third anatomical locations of tumor. However, His139Arg genotype of exon 4, exhibited low risk for ESCC as well as its clinical characteristics

  2. Fibroblast growth factor receptor 4 Gly388Arg polymorphism in Chinese gastric cancer patients

    Yan-Ying Shen

    2013-01-01

    Full Text Available AIM: To investigate the contribution of the fibroblast growth factor receptor 4 (FGFR4 Gly388Arg polymorphism as a genetic risk factor for gastric cancer (GC and to investigate any associations between this polymorphism and clinicopathological parameters and survival. METHODS: Tumors and matched adjacent non-cancer tissues were collected from 304 GC patients, and 5 mL of venous blood was collected from 62 GC patients and 392 age- and sex-matched healthy controls without cancer history from the same ethnic population. DNA was extracted, and direct sequencing analyses were performed to genotype the FGFR4 Gly388Arg polymorphism in all the samples. Differences in the genotype frequencies of the FGFR4 Gly388Arg polymorphism between GC patients and healthy controls were estimated using the χ2 test. Binary logistic regression was used for all analysis variables to estimate risk as the ORs with 95%CIs. The relationships between the FGFR4 genotype and clinicopathological parameters were tested with the χ2 test. The Kaplan-Meier product-limit method, the log-rank test, and the Cox regression model were applied to evaluate the effect of the FGFR4 genotype on the overall survival of patients with GC. RESULTS: In the present GC cohort, 118 patients (38.8% were homozygous for the Gly388 allele, 124 patients (40.8% were heterozygous, and 62 patients (20.4% were homozygous for the Arg388 allele. The frequencies of the Gly/Gly, Gly/Arg, and Arg/Arg genotypes in the healthy controls were 33.6%, 48.0%, and 18.4%, respectively. The distributions of genotypes (χ2 = 3.589, P = 0.166 and alleles (χ2 = 0.342, P = 0.559 of the FGFR4 Gly388Arg polymorphism were not different between the GC patients and the healthy controls. Although we observed no correlation between the FGFR4 Gly388Arg polymorphism and clinicopathological parameters or survival in the total cohort of GC patients, the presence of the Arg388 allele was associated with shorter survival time in patients

  3. GABAB R活性水平对致痫大鼠认知功能及Arc/Arg3.1表达的影响%Effects of GABAB receptor expression level on cognitive impairment and Arc/Arg3.1 expression in induced epileptic rats model

    兰彦平; 孙涛; 张春; 袁聪聪; 杨征; 王峰

    2016-01-01

    目的 探讨GABABR活性变化对癫痫大鼠认知功能及Arc/Arg3.1的影响.方法 建立氯化锂-匹罗卡品致痫模型,随机分成正常组、巴氯酚组、CGP组、单纯点燃组.避暗、水迷宫实验观察大鼠认知情况,免疫组化、荧光定量PCR、免疫印迹检测海马组织内GABABR(GB1、GB2)、Arc/Arg3.1蛋白及mRNA表达情况.结果 避暗实验:4组大鼠穿梭次数为:6.8±0.6、1.2土0.2、5.4±0.5及3.6±0.3,潜伏期为:26.1 ±3.9、152.2±12.9、65.8 ±7.0、91.2±9.1,与水迷宫行为学变化趋势一致,显示致痫大鼠认知功能减退,巴氯酚进一步抑制致痫大鼠学习和记忆获取能力,CGP35348可改善致痫大鼠认知功能.Arc/Arg3.1及GB1、GB2相对表达量检测显:致痫大鼠较正常大鼠Arc/Arg3.1及GB1、GB2表达量明显增高,致痫组大鼠相对比,巴氯酚组Arc/Arg3.1表达量下降,GB1、GB2增高;而CGP35348组Arc/Arg3.1表达量增高,GB1、GB2降低.结论 GABABR活性水平可以调控Arc/Arg3.1表达,并影响致痫大鼠认知功能.%Objective To investigate the effects of GABAB receptor on cognitive impairment by using pilocarpine induced kindled rats model and also to check early gene (Arc/Arg3.1) expression.Methods Pilocarpine induced kindled rats were divided into four groups (Group normal,Baclofen,CGP and Kindled) randomly,and every group included 20 rats.We checked their cognitive impairment by using passive avoidance test and water maze test.The expression of GABAB receptor (GB1,GB2) and Arc/Arg3.1 was tested by immunohistochemical staining,RT-PCR and Western blot.Results Passive avoidance test showed four Group rats shuttle times were 6.8 ± 0.6,1.2 ± 0.2,5.4 ± 0.5,3.6 ± 0.3,incubation period were 26.1 ±3.9,152.2 ± 12.9,65.8 ±7.0,91.2 ±9.1,and water maze test had the same trend,with values in epilepsy groups significantly lower than the normal group of rats,which meant cognitive dysfunction.The above results also showed Baclofen further inhibited the learning

  4. Occurrence and distribution of antibiotic resistance genes in the coastal area of the Bohai Bay, China.

    Niu, Zhi-Guang; Zhang, Kai; Zhang, Ying

    2016-06-15

    Considering the abuse of antibiotics worldwide, we investigated the abundance of three classes of antibiotic resistance genes (ARGs) and the concentrations of corresponding antibiotics in water and sediments of Bohai Bay. The results showed that sulI and sulII were detected in all samples, and their abundance range was 10(-5)-10(-2)/16S gene copies. The abundance of tetM and ermB were relatively higher than the other genes of tet-ARGs and erm-ARGs. Sulfonamides were the most prevalent antibiotics, and the concentrations of antibiotic in sediments were higher than those in water. The correlation analysis revealed that antibiotics had pertinence with corresponding ARGs, indicating that antibiotics play an important role in the creation and transfer of ARGs. The results of regression analysis indicated that the propagation and maintenance of sulI and sulII were facilitated by class I integrons. PMID:27107623

  5. Fate and removal of various antibiotic resistance genes in typical pharmaceutical wastewater treatment systems.

    Zhai, Wenchao; Yang, Fengxia; Mao, Daqing; Luo, Yi

    2016-06-01

    The high levels of antibiotic residues in pharmaceutical wastewater treatment plants (PWWTPs) make these plants the hotspots for the proliferation of antibiotic resistance genes (ARGs). This study investigated the fate and removal of 11 ARG subtypes for sulfonamide, tetracycline, β-lactam, and macrolide resistance in each processing stage of two full-scale PWWTPs in northern China. The levels of typical ARG subtypes in the final effluents ranged from (2.56 ± 0.13) × 10(1) to (2.36 ± 0.11) × 10(7) copies/ml. The absolute abundance of ARGs in effluents accounted for only 0.03-78.1 % of influents of the two PWWTPs, while the majority of the ARGs were transported to the dewatered sludge with concentrations from (2.65 ± 0.43) × 10(5) to (4.27 ± 0.03) × 10(10) copies/g dry weight (dw). The total loads of ARGs discharged through dewatered sludge plus effluent was 1.01-14.09-fold higher than that in the raw influents, suggesting the proliferation of ARGs occurred in the wastewater treatment. The proliferation of ARGs mainly occurs in biological treatment process, such as aeration tank, anoxic tank, sequencing batch reactor (SBR), and bio-contact oxidation, facilitates the proliferation of various ARGs, implying significant replication of certain ARG subtypes may be attributable to microbial growth. Chemical oxidation seems promising to remove ARGs, with removal efficiency ranged from 29.3 to 85.7 %, while the partial correlation analysis showed significant correlations between antibiotic concentration and ARG removal. Thus, the high antibiotic residues within the PWWTPs may have an influence on the proliferation, fate, and removal of the associated ARG subtypes. PMID:26961534

  6. High-throughput profiling of antibiotic resistance genes in drinking water treatment plants and distribution systems.

    Xu, Like; Ouyang, Weiying; Qian, Yanyun; Su, Chao; Su, Jianqiang; Chen, Hong

    2016-06-01

    Antibiotic resistance genes (ARGs) are present in surface water and often cannot be completely eliminated by drinking water treatment plants (DWTPs). Improper elimination of the ARG-harboring microorganisms contaminates the water supply and would lead to animal and human disease. Therefore, it is of utmost importance to determine the most effective ways by which DWTPs can eliminate ARGs. Here, we tested water samples from two DWTPs and distribution systems and detected the presence of 285 ARGs, 8 transposases, and intI-1 by utilizing high-throughput qPCR. The prevalence of ARGs differed in the two DWTPs, one of which employed conventional water treatments while the other had advanced treatment processes. The relative abundance of ARGs increased significantly after the treatment with biological activated carbon (BAC), raising the number of detected ARGs from 76 to 150. Furthermore, the final chlorination step enhanced the relative abundance of ARGs in the finished water generated from both DWTPs. The total enrichment of ARGs varied from 6.4-to 109.2-fold in tap water compared to finished water, among which beta-lactam resistance genes displayed the highest enrichment. Six transposase genes were detected in tap water samples, with the transposase gene TnpA-04 showing the greatest enrichment (up to 124.9-fold). We observed significant positive correlations between ARGs and mobile genetic elements (MGEs) during the distribution systems, indicating that transposases and intI-1 may contribute to antibiotic resistance in drinking water. To our knowledge, this is the first study to investigate the diversity and abundance of ARGs in drinking water treatment systems utilizing high-throughput qPCR techniques in China. PMID:26890482

  7. Protease-catalyzed Synthesis of Bz-Arg-Gly-Asp-OMe in Full Aqueous Medium

    HOU Rui-zhen; LI Hong-mei; LIU Yun-jia; ZHANG Long; XU Li; ZHANG Xue-zhong

    2007-01-01

    Synthesis of N-benzoyl-argininylglycylasparagine methyl ester(Bz-Arg-Gly-Asp-OMe), a precursor tripeptide of Arg-Gly-Asp) was catalyzed by papain under kinetic control, at alkaline pH, in a full aqueous medium. The substrates were N-benzoyl-argininylglycine ethyl ester and asparagine dimethyl ester. An aqueous solution of 0. 1 mol/L KCl/NaOH containing 8 mmol/L EDTA and 2 mmol/L DTT was selected as the reaction medium. The synthesized hydrophilic tripeptide was soluble in the reaction medium during the reaction process, however, the secondary hydrolysis of the tripeptide product was not considerable. The effects of different factors, including water content, temperature, reaction time, and molar ratio of the substrates, on the yield of Bz-Arg-Gly-Asp-OMe were examined. The optimal reaction conditions were 0.05 mol/L Bz-Arg-Gly-OEt and 0.15 mol/L Asp(-OMe)2 · HCl in 0.1 mol/L KCl/NaOH solution(pH 8.5), at 40 ℃, and a reaction time of 60 min, with a maximum conversion yield of 62.4%.

  8. Molecular Interactions Between the Active Sites of RGD (Arg-Gly-Asp with its Receptor (Integrine

    E. Jauregui

    2000-03-01

    Full Text Available A study of the molecular interactions between the active sites of RGD (Arg-Gly-Asp with it Receptor using simultaions is reported. Our calculations indicate that the guanidine-carboxylate complex is energetically favourd with respect to the guanidine-methyl tetrazole complex.

  9. Increased risk of cutaneous melanoma associated with p53 Arg72Pro polymorphism.

    Peiliang Geng

    Full Text Available The objective of this study was to test the hypothesis that p53 Arg72Pro polymorphism may contribute to an increased risk of cutaneous melanoma (CM.By searching the databases of PubMed, EMBASE, and Web of Science, a total of 8 eligible case-control studies with 1,957 CM cases and 2,887 controls were included in this meta-analysis. Stata software was used to analyze all the statistical data.The pooled data by a fixed-effects model suggested an increased risk of CM associated with p53 Arg72Pro polymorphism under the genetic model of Arg/Pro vs. Pro/Pro without heterogeneity (ORArg/Pro vs. Pro/Pro = 1.76, 95% CI = 1.55-1.99, Pheterogeneity = 0.075. A similar trend was seen in subgroups of hospital-based studies and population-based studies.Our meta-analysis based on all studies shows that the p53 Arg72Pro polymorphism may increase individual susceptibility to CM, particularly in Caucasians and could serve as a biomarker to predict the population at high risk of CM.

  10. Effect of temperature on removal of antibiotic resistance genes by anaerobic digestion of activated sludge revealed by metagenomic approach.

    Zhang, Tong; Yang, Ying; Pruden, Amy

    2015-09-01

    As antibiotic resistance continues to spread globally, there is growing interest in the potential to limit the spread of antibiotic resistance genes (ARGs) from wastewater sources. In particular, operational conditions during sludge digestion may serve to discourage selection of resistant bacteria, reduce horizontal transfer of ARGs, and aid in hydrolysis of DNA. This study applied metagenomic analysis to examine the removal efficiency of ARGs through thermophilic and mesophilic anaerobic digestion using bench-scale reactors. Although the relative abundance of various ARGs shifted from influent to effluent sludge, there was no measureable change in the abundance of total ARGs or their diversity in either the thermophilic or mesophilic treatment. Among the 35 major ARG subtypes detected in feed sludge, substantial reductions (removal efficiency >90%) of 8 and 13 ARGs were achieved by thermophilic and mesophilic digestion, respectively. However, resistance genes of aadA, macB, and sul1 were enriched during the thermophilic anaerobic digestion, while resistance genes of erythromycin esterase type I, sul1, and tetM were enriched during the mesophilic anaerobic digestion. Efflux pump remained to be the major antibiotic resistance mechanism in sludge samples, but the portion of ARGs encoding resistance via target modification increased in the anaerobically digested sludge relative to the feed. Metagenomic analysis provided insight into the potential for anaerobic digestion to mitigate a broad array of ARGs. PMID:25994259

  11. Selective proteolysis of apolipoprotein B-100 by Arg-gingipain mediates atherosclerosis progression accelerated by bacterial exposure.

    Hashimoto, Munetaka; Kadowaki, Tomoko; Tsukuba, Takayuki; Yamamoto, Kenji

    2006-11-01

    Epidemiological studies suggest the association of periodontal infections with atherosclerosis, however, the mechanism underlying this association remains poorly understood. Porphyromonas gingivalis is the primary etiologic agent of adult periodontitis and produces a unique class of cysteine proteinases consisting of Arg-gingipain (Rgp) and Lys-gingipain (Kgp). To elucidate key mechanisms for progression of atherosclerosis by P. gingivalis infection, we tested the effects of the disruption of genes encoding Rgp and/or Kgp and inhibitors specific for the respective enzymes on atherosclerosis progression in apolipoprotein E-knockout mice. Repeated intravenous injection of wild-type P. gingivalis resulted in an increase in atherosclerotic lesions as well as an increase in the serum LDL cholesterol and a decrease of HDL cholesterol in these animals. LDL particles in P. gingivalis-injected animals were modified as a result of selective proteolysis of apoB-100 in LDL particles. This modification of LDL by P. gingivalis resulted in an increase in LDL uptake by macrophages and consequent foam cell formation in vitro. The atherosclerotic changes induced by P. gingivalis infection were attenuated by disruption of Rgp-encoding genes or by an Rgp-specific inhibitor. Our results indicate that degradation of apoB-100 by Rgp plays a crucial role in the promotion of atherosclerosis by P. gingivalis infection. PMID:17030507

  12. ARG1 Is a Novel Bronchodilator Response Gene: Screening and Replication in Four Asthma Cohorts

    Litonjua, Augusto A.; Lasky-Su, Jessica; Schneiter, Kady; Tantisira, Kelan G.; Lazarus, Ross; Klanderman, Barbara; Lima, John J.; Irvin, Charles G.; Peters, Stephen P; Hanrahan, John P.; Liggett, Stephen B.; Hawkins, Gregory A.; Meyers, Deborah A; Bleecker, Eugene R.; Lange, Christoph

    2008-01-01

    Rationale: Inhaled β-agonists are one of the most widely used classes of drugs for the treatment of asthma. However, a substantial proportion of patients with asthma do not have a favorable response to these drugs, and identifying genetic determinants of drug response may aid in tailoring treatment for individual patients.

  13. Influence of ADRB2 Gln27Glu and ADRB3 Trp64Arg polymorphisms on body weight and body composition changes after a controlled weight-loss intervention.

    Szendrei, Barbara; González-Lamuño, Domingo; Amigo, Teresa; Wang, Guan; Pitsiladis, Yannis; Benito, Pedro J; Gomez-Candela, Carmen; Calderón, Francisco J; Cupeiro, Rocío

    2016-03-01

    The β-2 and β-3 adrenergic receptors (ADRB2 and ADRB3) are thought to play a role in energy expenditure and lipolysis. However, the effects of the ADRB2 glutamine (Gln) 27 glutamic acid (glutamate) (Glu) and ADRB3 tryptophan (Trp) 64 arginine (Arg) polymorphisms on weight loss remain controversial. The aim of this study was to investigate the effect of these polymorphisms on changes in weight and body composition during a controlled weight-loss program. One hundred seventy-three healthy overweight and obese participants (91 women, 82 men) aged 18-50 years participated in a 22-week-long intervention based on a hypocaloric diet and exercise. They were randomly assigned to 1 of 4 groups: strength, endurance, strength and endurance combined, and physical activity recommendations only. Body weight, body mass index (BMI), and body composition variables were assessed before and after the intervention. Genetic analysis was carried out according to standard protocols. No effect of the ADRB2 gene was shown on final weight, BMI, or body composition, although in the supervised male group, Glu27 carriers tended to have greater weight (p = 0.019, 2.5 kg) and BMI (p = 0.019, 0.88 kg/m(2)) reductions than did noncarriers. There seems to be an individual effect of the ADRB3 polymorphism on fat mass (p = 0.004) and fat percentage (p = 0.036), in addition to an interaction with exercise for fat mass (p = 0.038). After the intervention, carriers of the Arg64 allele had a greater fat mass and fat percentage than did noncarriers (p = 0.004, 2.8 kg). In conclusion, the ADRB2 Gln27Glu and ADRB3 Trp64Arg polymorphisms may influence weight loss and body composition, although the current evidence is weak; however, further studies are necessary to clarify their roles. PMID:26888112

  14. ARG1 (altered response to gravity) encodes a DnaJ-like protein that potentially interacts with the cytoskeleton

    Sedbrook, J. C.; Chen, R.; Masson, P. H.

    1999-01-01

    Gravitropism allows plant organs to direct their growth at a specific angle from the gravity vector, promoting upward growth for shoots and downward growth for roots. Little is known about the mechanisms underlying gravitropic signal transduction. We found that mutations in the ARG1 locus of Arabidopsis thaliana alter root and hypocotyl gravitropism without affecting phototropism, root growth responses to phytohormones or inhibitors of auxin transport, or starch accumulation. The positional cloning of ARG1 revealed a DnaJ-like protein containing a coiled-coil region homologous to coiled coils found in cytoskeleton-interacting proteins. These data suggest that ARG1 participates in a gravity-signaling process involving the cytoskeleton. A combination of Northern blot studies and analysis of ARG1-GUS fusion-reporter expression in transgenic plants demonstrated that ARG1 is expressed in all organs. Ubiquitous ARG1 expression in Arabidopsis and the identification of an ortholog in Caenorhabditis elegans suggest that ARG1 is involved in other essential processes.

  15. Point mutation Gln121-Arg increased temperature optima of Bacillus lipase (1.4 subfamily) by fifteen degrees.

    Goomber, Shelly; Kumar, Rakesh; Singh, Ranvir; Mishra, Neelima; Kaur, Jagdeep

    2016-07-01

    Small molecular weight Bacillus lipases are industrially attractive because of its alkaline optimum pH, broad substrate specificity and production in high yield by overexpression both in Escherichia coli and Bacillus subtilis. Its major limitation of being mesophilic in nature is constantly targeted by laboratory evolution studies. Herein metagenomically isolated Bacillus LipJ was randomly evolved by error prone PCR and library of variants were screened for enhanced thermostability. Point mutant Gln121Arg was extensively characterized and it showed dramatic shift of Temp. opt to 50°C compared to 37°C for parent enzyme. Thermostability studies at 45°C and 50°C determined six fold increase in half life for point variant Gln121Arg compared to LipJ. Circular dichroism (CD) and tryptophan fluorescence study established enhanced thermostability of Gln121Arg. Specific activity of point variant Gln121Arg was comparable to wild type with increased substrate affinity (Km reduced). Reduced kcat for variant Gln121Arg infer that kinetic and catalytic efficiency of mutant was compromised. Structural implications by homolog modelling predicted Gln121 to be placed within longest loop of the structure at surface. Localization of loop due to additional polar interactions by Arg121 to protein core defines molecular basis of enhanced thermostability of random point variant Gln121Arg. PMID:27083848

  16. Prevalence of antibiotic resistance genes of wastewater and surface water in livestock farms of Jiangsu Province, China.

    Chen, Biao; Hao, Lijun; Guo, Xinyan; Wang, Na; Ye, Boping

    2015-09-01

    The overuse of antibiotics in livestock farms is general, leading to a wide distribution of antibiotic resistance genes (ARGs) in aquatic environment adjacent to livestock farms. However, researches of the distribution and types of ARGs in aquatic environment of China are still in the initial stage. In this study, wastewater and surface water samples were collected from 12 livestock farms (four pig farms, four cattle farms, and four chicken farms) in Jiangsu Province of China. The prevalence, abundance, and distribution of 22 ARGs were investigated, which were categorized into six groups, including nine tetracyclin resistance genes, three sulfonamides resistance genes, three quinolone resistance genes, two macrolide resistance genes, three aminoglycoside resistance genes, and two multidrug resistance genes, employing quantitative real-time PCR (qPCR). The results suggested that all of the 22 ARGs were detected in samples. Sul1, sul2, and tetM were the most abundant with the average concentration of 3.84 × 10(1) copies/16S recombinant RNA (rRNA) gene copies, 1.62 × 10(1) copies/16S rRNA gene copies, 2.33 × 10(1) copies/16S rRNA gene copies, respectively. Principle component analysis revealed that the comprehensive pollution of ARGs in northern Jiangsu was more serious. ARGs in wastewater were more abundant when compared to that in surface water. A preliminary study regarding the fate of ARGs after an aerobiotic process showed that tetA, tetC, sul1, sul2, oqxB, and qnrS were significantly increased. And, among the tetracycline resistance genes, the efflux pump genes were enriched while the ribosomal protection protein encoding genes were decreased in the aerobiotic process. The prevalance of ARGs in water environment is of concern; more surveillance is required to determine the pollution level and pattern of antibiotic resistance genes. PMID:25948386

  17. Three new species of the genus Arge (Hymenoptera: Symphyta: Argidae from South Korea with key to species of the subfamily Arginae

    Jin-Kyung Choi

    2016-06-01

    Full Text Available Three new species, Arge koreana Wei & Lee sp. nov. from South Korea, Arge pseudorejecta Wei & Lee sp. nov., and Arge shengi Wei & Lee sp. nov. from South Korea and China are described. Keys to known genera of Argidae and known species of Arginae from South Korea are provided.

  18. Binding of ArgTX-636 in the NMDA receptor ion channel

    Poulsen, Mette H; Andersen, Jacob; Christensen, Rune;

    2015-01-01

    N1 and GluN2A where mutation influences ArgTX-636 potency suggests differential contribution of the M2-loops of GluN1 and GluN2A to binding of ArgTX-636. The results of the mutational analysis are highly relevant for the future structure-based development of argiotoxin-derived NMDAR channel blockers.......The N-methyl-d-aspartate receptors (NMDARs) constitute an important class of ligand-gated cation channels that are involved in the majority of excitatory neurotransmission in the human brain. Compounds that bind in the NMDAR ion channel and act as blockers are use- and voltage-dependent inhibitors...... of NMDAR activity and have therapeutic potential for treatment of a variety of brain diseases or as pharmacological tools for studies of the neurobiological role of NMDARs. We have performed a kinetic analysis of the blocking mechanism of the prototypical polyamine toxin NMDAR ion channel blocker...

  19. Persistence of naturally occurring antibiotic resistance genes in the bacteria and bacteriophage fractions of wastewater.

    Calero-Cáceres, William; Muniesa, Maite

    2016-05-15

    The emergence and prevalence of antibiotic resistance genes (ARGs) in the environment is a serious global health concern. ARGs from bacteria can be mobilized by mobile genetic elements, and recent studies indicate that phages and phage-derived particles, among others, could play a role in the spread of ARGs through the environment. ARGs are abundant in the bacterial and bacteriophage fractions of water bodies and for successful transfer of the ARGs, their persistence in these environments is crucial. In this study, three ARGs (blaTEM, blaCTX-M and sul1) that naturally occur in the bacterial and phage fractions of raw wastewater were used to evaluate the persistence of ARGs at different temperatures (4 °C, 22 °C and 37 °C) and pH values (3, 7 and 9), as well as after various disinfection treatments (thermal treatment, chlorination and UV) and natural inactivation in a mesocosm. Gene copies (GC) were quantified by qPCR; then the logarithmic reduction and significance of the differences between their numbers were evaluated. The ARGs persisted for a long time with minimal reductions after all the treatments. In general, they showed greater persistence in the bacteriophage fraction than in the bacterial fraction. Comparisons showed that the ARGs persisted under conditions that reduced culturable Escherichia coli and infectious coliphages below the limit of detection. The prevalence of ARGs, particularly in the bacteriophage fraction, poses the threat of the spread of ARGs and their incorporation into a new bacterial background that could lead to the emergence of new resistant clones. PMID:26978717

  20. A sustainable urban development and crises management: Empirical investigation on redesigning Tehran Arg square

    Mohammad Rahim Rahnama; Hana Yazdanfar

    2014-01-01

    This paper presents an empirical investigation on redesigning Tehran Arge square based on principles of sustainable urban development and crises management. The proposed study performs the study based on three perspectives including quality improvement, sustainability and crisis management. In terms of city quality improvement, the study detects six factors including social interaction, cleanliness, attractiveness, comfort, vitality and security. In terms of sustainability, the study discusse...

  1. Tracking and Monitoring with Dosimeter-Enabled ARG-US RFID System - 12009

    Automated monitoring and tracking of materials with radio frequency identification (RFID) technology can significantly improve both the operating efficiency of radiological facilities and the application of the ALARA (as low as reasonably achievable) principle in them. One such system, called ARG-US, has been developed by Argonne National Laboratory for the U.S. Department of Energy (DOE) Packaging and Certification Program to use in managing sensitive nuclear and radioactive materials. Several ARG-US systems are in various stages of deployment and advanced testing across DOE sites. ARG-US utilizes sensors in the tags to continuously monitor the state of health of the packaging and promptly disseminates alarms to authorized users. In conjunction with global positioning system (GPS) tracking provided by TRANSCOM, the system can also monitor and track packages during transport. A compact dosimeter has been incorporated in the ARG-US tags via an onboard universal asynchronous receiver/transmitter interface. The detector has a wide measurement range for gamma radiation - from 0.1 mSv/h to 8 Sv/h. The detector is able to generate alarms for both high and low radiation and for a high cumulative dose. In a large installation, strategically located dosimeter-enabled tags can yield an accurate, real-time, 2D or 3D dose field map that can be used to enhance facility safety, security, and safeguards. This implementation can also lead to a reduced need for manned surveillance and reduced exposure of personnel to radiation, consistent with the ALARA principle at workplaces. (authors)

  2. The common germline Arg72Pro polymorphism of p53 and increased longevity in humans

    Bojesen, S.E.; Nordestgaard, Børge

    2008-01-01

    Arg72Pro substitution in the p53 protein has important influence on cell death via increased apoptosis. Thus, the increased longevity may be due to a generally increased robustness after a diagnosis of any life-threatening disease. In contrast to widespread skepticism on the importance of SNPs in...... humans, this gain-of-function p53 SNP of importance for cell repair mechanisms has a profound influence on longevity Udgivelsesdato: 2008/1/15...

  3. Atypical distyly in Psychotria goyazensis Mull. Arg. (Rubiaceae), an intramorph self-compatible species

    Ebenezer Barbosa Rodrigues; Hélder Consolaro

    2013-01-01

    Distyly is a genetically controlled floral dimorphism, characterized by the reciprocal positioning of pin and thrum morphs, a heteromorphic incompatibility system and a balanced morph ratio (isoplethy). This study aimed to investigate the phenology, features of distyly, reproductive biology, and floral visitors of Psychotria goyazensis Mull. Arg. (Rubiaceae), an intramorph self-compatible species, in a tropical semideciduous forest in the municipality of Catalão, located in the state of Goiás...

  4. Tracking and Monitoring with Dosimeter-Enabled ARG-US RFID System - 12009

    Anderson, J.; Lee, H.; De Lurgio, P.; Kearney, C.M.; Craig, B.; Soos, I.H.; Tsai, H.; Liu, Y. [Argonne National Laboratory, Argonne, IL 60439 (United States); Shuler, J. [U.S. Department of Energy, Washington, D.C. 20585 (United States)

    2012-07-01

    Automated monitoring and tracking of materials with radio frequency identification (RFID) technology can significantly improve both the operating efficiency of radiological facilities and the application of the ALARA (as low as reasonably achievable) principle in them. One such system, called ARG-US, has been developed by Argonne National Laboratory for the U.S. Department of Energy (DOE) Packaging and Certification Program to use in managing sensitive nuclear and radioactive materials. Several ARG-US systems are in various stages of deployment and advanced testing across DOE sites. ARG-US utilizes sensors in the tags to continuously monitor the state of health of the packaging and promptly disseminates alarms to authorized users. In conjunction with global positioning system (GPS) tracking provided by TRANSCOM, the system can also monitor and track packages during transport. A compact dosimeter has been incorporated in the ARG-US tags via an onboard universal asynchronous receiver/transmitter interface. The detector has a wide measurement range for gamma radiation - from 0.1 mSv/h to 8 Sv/h. The detector is able to generate alarms for both high and low radiation and for a high cumulative dose. In a large installation, strategically located dosimeter-enabled tags can yield an accurate, real-time, 2D or 3D dose field map that can be used to enhance facility safety, security, and safeguards. This implementation can also lead to a reduced need for manned surveillance and reduced exposure of personnel to radiation, consistent with the ALARA principle at workplaces. (authors)

  5. PHARMACOGNOSTICAL EVALUATION OF FRUITS OF MALLOTUS PHILIPPENENSIS (LAM). MUELL- ARG (EUPHORBIACEAE)

    Kadam Prasad Vijay; Bhingare Chandrashekhar Laxman; Soni Surajkumar Bansilal; Rathi Sandesh Ashok; Patil Manohar Janardhan

    2013-01-01

    Correct identification of starting material is necessary to ensure the quality of herbal medicines. Pharmacognostic evaluation is first step towards establishing identity and purity of crude drug. With this aim the present paper deals with study of pharmacognostic and physicochemical characteristics of fruits of Mallotus philippinensis (lam) Muell. Arg. (Euphorbiaceae). The powder of Mallotus philippinensis was used for detection of various chemical constituents. The transverse section and th...

  6. The Possibility of Energy Recovery from Waste Material in Arges County, Romania

    Nordström, Emma; Enochsson, Evelina

    2009-01-01

    Abstract Waste disposal is a global problem contributing to the ongoing climate change by large emissions of greenhouse gases. By using waste material as a resource instead of landfilling, the greenhouse gas emissions from landfills are reduced. Waste material can be used for waste incineration with energy recovery, thus decreasing the greenhouse gas emission from energy utilization by changing from fossil fuels to a partly renewable fuel. Arges County in Romania has severe problems with its ...

  7. A cell surface receptor complex for collagen type I recognizes the Arg- Gly-Asp sequence

    1987-01-01

    To isolate collagen-binding cell surface proteins, detergent extracts of surface-iodinated MG-63 human osteosarcoma cells were chromatographed on affinity matrices of either type I collagen- Sepharose or Sepharose carrying a collagen-like triple-helical peptide. The peptide was designed to be triple helical and to contain the sequence Arg-Gly-Asp, which has been implicated as the cell attachment site of fibronectin, vitronectin, fibrinogen, and von Willebrand factor, and is also present in ty...

  8. Gene

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  9. Multifactorial likelihood assessment of BRCA1 and BRCA2 missense variants confirms that BRCA1:c.122A>G(p.His41Arg is a pathogenic mutation.

    Phillip J Whiley

    Full Text Available Rare exonic, non-truncating variants in known cancer susceptibility genes such as BRCA1 and BRCA2 are problematic for genetic counseling and clinical management of relevant families. This study used multifactorial likelihood analysis and/or bioinformatically-directed mRNA assays to assess pathogenicity of 19 BRCA1 or BRCA2 variants identified following patient referral to clinical genetic services. Two variants were considered to be pathogenic (Class 5. BRCA1:c.4484G> C(p.Arg1495Thr was shown to result in aberrant mRNA transcripts predicted to encode truncated proteins. The BRCA1:c.122A>G(p.His41Arg RING-domain variant was found from multifactorial likelihood analysis to have a posterior probability of pathogenicity of 0.995, a result consistent with existing protein functional assay data indicating lost BARD1 binding and ubiquitin ligase activity. Of the remaining variants, seven were determined to be not clinically significant (Class 1, nine were likely not pathogenic (Class 2, and one was uncertain (Class 3.These results have implications for genetic counseling and medical management of families carrying these specific variants. They also provide additional multifactorial likelihood variant classifications as reference to evaluate the sensitivity and specificity of bioinformatic prediction tools and/or functional assay data in future studies.

  10. Molecular mechanisms mediating the beneficial metabolic effects of [Arg4]tigerinin-1R in mice with diet-induced obesity and insulin resistance.

    Ojo, Opeolu O; Srinivasan, Dinesh K; Owolabi, Bosede O; McGahon, Mary K; Moffett, R Charlotte; Curtis, Tim M; Conlon, J Michael; Flatt, Peter R; Abdel-Wahab, Yasser H A

    2016-08-01

    The frog skin host-defense peptide tigerinin-1R stimulates insulin release in vitro and improves glucose tolerance and insulin sensitivity in animal models of type 2 diabetes. This study extends these observations by investigating the molecular mechanisms of action underlying the beneficial metabolic effects of the analogue [Arg4]tigerinin-1R in mice with diet-induced obesity, glucose intolerance and insulin resistance. The study also investigates the electrophysiological effects of the peptide on KATP and L-type Ca2+ channels in BRIN-BD11 clonal β cells. Non-fasting plasma glucose and glucagon concentrations were significantly (pinsulin increased by twice daily treatment with [Arg4]tigerinin-1R (75 nmol/kg body weight) for 28 days. Oral and intraperitoneal glucose tolerance were significantly (pinsulin. The peptide blocked KATP channels and, consistent with this, improved beta cell responses of isolated islets to a range of secretagogues. Peptide administration resulted in up-regulation of key functional genes in islets involved insulin secretion (Abcc8, Kcnj11, Cacna1c and Slc2a2) and in skeletal muscle involved with insulin action (Insr, Irs1, Pdk1, Pik3ca, and Slc2a4). These observations encourage further development of tigerinin-1R analogues for the treatment of patients with type 2 diabetes. PMID:26966929

  11. Hippocampal Arc (Arg3.1) expression is induced by memory recall and required for memory reconsolidation in trace fear conditioning.

    Chia, Chester; Otto, Tim

    2013-11-01

    Mounting evidence suggests that long-lasting, protein synthesis-dependent changes in synaptic strength accompany both the initial acquisition and subsequent recall of specific memories. Within brain areas thought to be important for learning and memory, including the hippocampus, learning-related plasticity is likely mediated in part by NMDA receptor activation and experience-dependent changes in gene expression. In the present study, we examined the role of activity-regulated cytoskeletal-associated protein (Arc/Arg3.1) expression in the acquisition, recall, and reconsolidation of memory in a trace fear conditioning paradigm. First, we show that the expression of Arc protein in ventral hippocampus (VH) is dramatically enhanced by memory recall 24h after the acquisition of trace fear conditioning, and that both memory recall and the associated recall-induced enhancement of Arc expression are blocked by pre-training administration of 2-amino-5-phosphonovaleric acid (APV). Next, we show that while infusion of Arc antisense oligodeoxynucleotides (ODNs) into VH prior to testing had little effect on memory recall, it significantly reduced both Arc protein expression and freezing behavior during subsequent testing sessions. Collectively, these results suggest that Arc/Arg3.1 protein plays an important functional role in both the initial acquisition of hippocampal-dependent memory and the reconsolidation of these memories after recall. PMID:23872190

  12. Who Possesses Drug Resistance Genes in the Aquatic Environment? : Sulfamethoxazole (SMX) Resistance Genes among the Bacterial Community in Water Environment of Metro-Manila, Philippines

    Satoru eSuzuki; Mitsuko eOgo; Miller, Todd W.; Akiko eShimizu; Hideshige eTakada; Maria Auxilia eSiringan

    2013-01-01

    Recent evidence has shown that antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are ubiquitous in natural environments, including sites considered pristine. To understand the origin of ARGs and their dynamics, we must first define their actual presence in the natural bacterial assemblage. Here we found varying distribution profiles of sul genes in “colony forming bacterial assemblages” and “natural bacterial assemblages.” Our monitoring for antibiotic contamination r...

  13. Constitutive presence of antibiotic resistance genes within the bacterial community of a large subalpine lake.

    Di Cesare, Andrea; Eckert, Ester M; Teruggi, Alessia; Fontaneto, Diego; Bertoni, Roberto; Callieri, Cristiana; Corno, Gianluca

    2015-08-01

    The fate of antibiotic resistance genes (ARGs) in environmental microbial communities is of primary concern as prodromal of a potential transfer to pathogenic bacteria. Although of diverse origin, the persistence of ARGs in aquatic environments is highly influenced by anthropic activities, allowing potential control actions in well-studied environments. However, knowledge of abundance and space-time distribution of ARGs in ecosystems is still scarce. Using quantitative real-time PCR, we investigated the presence and the abundance of twelve ARGs (against tetracyclines, β-lactams, aminoglycosides, quinolones and sulphonamides) at different sampling sites, depths and seasons, in Lake Maggiore, a large subalpine lake, and in the area of its watershed. We then evaluated the correlation between each ARG and a number of ecological parameters in the water column in the deepest part of the lake. Our results suggest the constitutive presence of at least four ARGs within the bacterial community with a high proportion of bacteria potentially resistant to tetracyclines and sulphonamides. The presence of these ARGs was independent of the total bacterial density and temperature. The dynamics of tet(A) and sulII genes were, however, positively correlated with dissolved oxygen and negatively to chlorophyll a, suggesting that the resistant microbes inhabit specific niches. These observations indicate that the lake is a reservoir of antibiotic resistances, highlighting the need of a deeper understanding of the sources of ARGs and the factors allowing their persistence in waters. PMID:26118321

  14. Klasik Dönem İslam felsefesinde kozmolojik argüman ve modern fizikteki gelişmeler

    EVLİ, Mehmet Mustafa

    2014-01-01

    Kozmolojik argüman, âlem hakkındaki bazı gerçekliklerden yola çıkarak Tanrı'ya ulaşmaya çalışan a posteriori bir argümandır. Felsefe tarihinde belli bazı tenkitlere maruz kalan argüman, dayandığı ana doktrinlere bağlı olarak değişen çeşitli formlarda inşa edilmiştir. Bu formlar, İlk Muharrik Delili, İlk Neden Delili, İmkan Delili, Yeter Sebep Delili, Hudûs Delili şeklinde sınıflandırılabilir. Bu çalışmanın birinci bölümü, kozmolojik argüman hakkında genel bilgiler vermeni...

  15. Chapa aglomerada de cimento-madeira de Hevea brasiliensis Müll. Arg. Cement-bonded particleboard of Hevea brasiliensis Müll. Arg.

    Esmeralda Yoshico Arakaki Okino; Mário Rabelo de Souza; Marcos Antonio Eduardo Santana; Maria Eliete de Sousa; Divino Eterno Teixeira

    2004-01-01

    Chapas de partículas de cimento-madeira foram confeccionadas com a madeira de quatro clones de Hevea brasiliensis Müll. Arg. (seringueira): IAN 717, IAN 873, GT 711 e AVROS 1301. Confeccionaram-se as chapas na proporção de 1:4:1 (madeira:cimento:água) por peso e nas dimensões de 450 x 450 x 13 mm e densidade nominal de 1,4 g/cm³, com a adição de 4% de cloreto de cálcio di-hidratado (CaCl2.2H2O) como acelerador. Foram testadas partículas fervidas e não-fervidas dos quatro clones, totaliza...

  16. Association of β1 and β3 adrenergic receptors gene polymorphisms with insulin resistance and high lipid profiles related to type 2 diabetes and metabolic syndrome

    Ana I. Burguete-García

    2014-06-01

    Full Text Available Background: Among the diverse genes associated to type 2 diabetes (T2D, the β-adrenergic receptors are an excellent candidate to study in Mexican population. The objective of this work was to analyze the association of polymorphisms in ADRB1 (rs1801253 (Arg389Gly and ADRB3 (Trp64Arg genes with T2D and metabolic syndrome (MS. Methods: We studied 445 MS patients, 502 with T2D and 552 healthy controls. Anthropometric features and complete biochemical profile were evaluated, and Arg389Gly and Trp64Arg SNPs were determined by TaqMan assays. Data analysis was adjusted by African, Caucasian and Amerindian ancestral percentage. Results: The variant Arg389Gly of ADRB1 was statistically associated with an increase of LDL levels (P < 0.008, and the variant ADRB3 Trp64Arg was associated to larger HOMA-IR (P < 0.018 and with an increase of insulin levels (P < 0.001. A multiple logistic regression analysis was made in three grouping models: For ADRB3 in the codominant model Trp/Arg genotype, there was an OR of 1.53 (1.09-2.13, P < 0.003 which was increased up to OR 2.99 (1.44-6.22, P < 0.003 for the Arg/Arg genotype. Similar risk association was found under the dominant model Trp/Arg-Arg/Arg genotype with OR 1.67 (1.21-2.30; P < 0.002. In the recessive model (Arg/Arg genotype, there was also a high association OR 2.56 (1.24-5.26, P < 0.01. Conclusions: The ADRB3 Trp64Arg variant is a susceptibility gene polymorphism for T2D and the ADRB1 Gly389Arg for lipid metabolism disruption. These results show that these variants are potential biomarkers for predicting metabolic alterations and evolution in diabetic and metabolic syndrome patients.

  17. Fatal accident due to anti-personnel ARGES EM01 rifle grenade explosion

    Pavlos Pavlidis; Valeria Karakasi; Theodossios A.Birbilis

    2016-01-01

    During the process of unsealing an old ammunition box in order to destroy it,a 42-year-old ammunition technician was fatally injured due to an anti-personnel ARGES EM01-type rifle grenade detonation.The explosion took place in the victim's hands,in point-blank range.This report aimed to show the anatomical position,the severity and the dispersion extent of the multiple injuries in the human body due to the detonation,and draw firm conclusions regarding the position of the human body and the circumstances prevailing at the moment of the explosion.

  18. Opening the Arg-Glu salt bridge in myosin: computational study.

    Kaliman, Ilya; Grigorenko, Bella; Shadrina, Maria; Nemukhin, Alexander

    2009-06-28

    Opening the Arg-Glu salt bridge in myosin, which presumably succeeds the myosin-catalyzed hydrolysis of adenosine triphosphate, was modeled computationally on the basis of the structures corresponding to the enzyme-substrate and enzyme-product complexes found in the quantum mechanics-molecular mechanics simulations. According to the calculations of the potential of mean force, opening the bridge is considerably facilitated upon termination of the chemical reaction, but does not promote egress of inorganic phosphate by the back-door mechanism. PMID:19506754

  19. Estructura territorial y estado en la cultura argárica

    Legarra Herrero, B.

    2014-01-01

    La cultura de El Argar es habitualmente presentada como una formación estatal en la que varias comunidades ejercieron un control social, económico y político sobre amplios territorios. Sin embargo esta configuración geo-política de las sociedades argáricas ha comenzado a ser reconocida como problemática y nuevas investigaciones han revelado discrepancias entre la evidencia y este modelo territorial clásico. Este artículo explora la organización territorial de El Argar a través de ...

  20. Rational design and characterization of D-Phe-Pro-D-Arg-derived direct thrombin inhibitors.

    Figueiredo, Ana C; Clement, Cristina C; Zakia, Sheuli; Gingold, Julian; Philipp, Manfred; Pereira, Pedro J B

    2012-01-01

    The tremendous social and economic impact of thrombotic disorders, together with the considerable risks associated to the currently available therapies, prompt for the development of more efficient and safer anticoagulants. Novel peptide-based thrombin inhibitors were identified using in silico structure-based design and further validated in vitro. The best candidate compounds contained both L- and D-amino acids, with the general sequence D-Phe(P3)-Pro(P2)-D-Arg(P1)-P1'-CONH₂. The P1' position was scanned with L- and D-isomers of natural or unnatural amino acids, covering the major chemical classes. The most potent non-covalent and proteolysis-resistant inhibitors contain small hydrophobic or polar amino acids (Gly, Ala, Ser, Cys, Thr) at the P1' position. The lead tetrapeptide, D-Phe-Pro-D-Arg-D-Thr-CONH₂, competitively inhibits α-thrombin's cleavage of the S2238 chromogenic substrate with a K(i) of 0.92 µM. In order to understand the molecular details of their inhibitory action, the three-dimensional structure of three peptides (with P1' L-isoleucine (fPrI), L-cysteine (fPrC) or D-threonine (fPrt)) in complex with human α-thrombin were determined by X-ray crystallography. All the inhibitors bind in a substrate-like orientation to the active site of the enzyme. The contacts established between the D-Arg residue in position P1 and thrombin are similar to those observed for the L-isomer in other substrates and inhibitors. However, fPrC and fPrt disrupt the active site His57-Ser195 hydrogen bond, while the combination of a P1 D-Arg and a bulkier P1' residue in fPrI induce an unfavorable geometry for the nucleophilic attack of the scissile bond by the catalytic serine. The experimental models explain the observed relative potency of the inhibitors, as well as their stability to proteolysis. Moreover, the newly identified direct thrombin inhibitors provide a novel pharmacophore platform for developing antithrombotic agents by exploring the conformational

  1. Rational design and characterization of D-Phe-Pro-D-Arg-derived direct thrombin inhibitors.

    Ana C Figueiredo

    Full Text Available The tremendous social and economic impact of thrombotic disorders, together with the considerable risks associated to the currently available therapies, prompt for the development of more efficient and safer anticoagulants. Novel peptide-based thrombin inhibitors were identified using in silico structure-based design and further validated in vitro. The best candidate compounds contained both L- and D-amino acids, with the general sequence D-Phe(P3-Pro(P2-D-Arg(P1-P1'-CONH₂. The P1' position was scanned with L- and D-isomers of natural or unnatural amino acids, covering the major chemical classes. The most potent non-covalent and proteolysis-resistant inhibitors contain small hydrophobic or polar amino acids (Gly, Ala, Ser, Cys, Thr at the P1' position. The lead tetrapeptide, D-Phe-Pro-D-Arg-D-Thr-CONH₂, competitively inhibits α-thrombin's cleavage of the S2238 chromogenic substrate with a K(i of 0.92 µM. In order to understand the molecular details of their inhibitory action, the three-dimensional structure of three peptides (with P1' L-isoleucine (fPrI, L-cysteine (fPrC or D-threonine (fPrt in complex with human α-thrombin were determined by X-ray crystallography. All the inhibitors bind in a substrate-like orientation to the active site of the enzyme. The contacts established between the D-Arg residue in position P1 and thrombin are similar to those observed for the L-isomer in other substrates and inhibitors. However, fPrC and fPrt disrupt the active site His57-Ser195 hydrogen bond, while the combination of a P1 D-Arg and a bulkier P1' residue in fPrI induce an unfavorable geometry for the nucleophilic attack of the scissile bond by the catalytic serine. The experimental models explain the observed relative potency of the inhibitors, as well as their stability to proteolysis. Moreover, the newly identified direct thrombin inhibitors provide a novel pharmacophore platform for developing antithrombotic agents by exploring the

  2. Fear extinction requires Arc/Arg3.1 expression in the basolateral amygdala

    Onoue, Kousuke; Nakayama, Daisuke; Ikegaya, Yuji; Matsuki, Norio; Nomura, Hiroshi

    2014-01-01

    Background Prolonged re-exposure to a fear-eliciting cue in the absence of an aversive event extinguishes the fear response to the cue, and has been clinically used as an exposure therapy. Arc (also known as Arg3.1) is implicated in synaptic and experience-dependent plasticity. Arc is regulated by the transcription factor cAMP response element binding protein, which is upregulated with and necessary for fear extinction. Because Arc expression is also activated with fear extinction, we hypothe...

  3. THE INCIDENCE OF HIV INFECTIONS IN ARGES POPULATION IN 2012 - 2013

    Ionica Deliu; Nicoleta Buică; Maria Ruxandra Gaisteanu

    2015-01-01

    Human immunodeficiency virus (HIV) from Retroviridae family was isolated in early 80´s but his presence in Africa is much older than we previously thought, before the pandemic spread because of the social changes of 70´s (Cernescu, 1998). The incidence of HIV infections is a constant concern for public health specialists, with inferences for whole population. In this paper we presented the incidence of HIV infections in 2012/2013 period in Arges County, Romania. Studies were made in Lab...

  4. Ecological effects of Arges River hydroelectric systems. The minimum flow problem

    The paper presents general observations of the ecological status of riverside areas, on Arges River between Capataneni and Bascov villages. These are affected by the presence of an important hydroelectric power system. The main problem generating negative impact on the environment is non-observance of Waters' Law no.107/1996 with reference to minimum flow. The minimum flow is defined as the flow through the cross section of a watercourse which ensures the living conditions for the existent aquatic ecosystems. It should be taken into account in order to protect the aquatic ecosystems. A methodology regarding the determination of minimum flow is proposed. (author)

  5. Melanoma imaging using 111In-, 86Y- and 68Ga-labeled CHX-A''-Re(Arg11)CCMSH

    Introduction: A novel alpha-melanocyte-stimulating hormone peptide analog CHX-A''-Re(Arg11)CCMSH, which targeted the melanocortin-1 receptor (MC1-R) overexpressed on melanoma cells, was investigated for its biodistribution and tumor imaging properties. Methods: The metal bifunctional chelator CHX-A'' was conjugated to the melanoma targeting peptide (Arg11)CCMSH and cyclized by Re incorporation to yield CHX-A''-Re(Arg11)CCMSH. CHX-A''-Re(Arg11)CCMSH was labeled with 111In, 86Y and 68Ga, and the radiolabeled peptides were examined in B16/F1 melanoma-bearing mice for their pharmacokinetic as well as their tumor targeting properties using small animal SPECT and PET. Results: The radiolabeling efficiencies of the 111In-, 86Y- and 68Ga-labeled CHX-A''-Re(Arg11)CCMSH peptides were >95%, resulting in specific activities of 4.44, 3.7 and 1.85 MBq/μg, respectively. Tumor uptake of the 111In-, 86Y- and 68Ga-labeled peptides was rapid with 4.17±0.94, 4.68±1.02 and 2.68±0.69 %ID/g present in the tumors 2 h postinjection, respectively. Disappearance of radioactivity from the normal organs and tissues was rapid with the exception of the kidneys. Melanoma tumors were imaged with all three radiolabeled peptides 2 h postinjection. MC1-R-specific uptake was confirmed by competitive receptor blocking studies. Conclusions: Melanoma tumor uptake and imaging was exhibited by the 111In-, 86Y- and 68Ga-labeled Re(Arg11)CCMSH peptides, although the tumor uptake was moderated by low specific activity. The facile radiolabeling properties of CHX-A''-Re(Arg11)CCMSH allow it to be employed as a melanoma imaging agent with little or no purification after 111In, 86Y and 68Ga labeling.

  6. Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

    Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

    2016-07-01

    Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

  7. Biologia reprodutiva de Psychotria poeppigiana Mull. Arg. (Rubiaceae em mata de galeria Reproductive biology of Psychotria poeppigiana Mull. Arg. (Rubiaceae in gallery forest

    Christiano Peres Coelho

    2004-09-01

    Full Text Available Psychotria L. (Rubiaceae é o gênero com mais espécies heterostílicas dentre as Angiospermas. O objetivo deste trabalho é descrever a biologia floral, fenologia, o sistema reprodutivo e os polinizadores de Psychotria poeppigiana Müll. Arg. As inflorescências são capitadas terminais, com brácteas vermelhas e flores amarelas apresentando dois morfos distintos: brevistiladas e longistiladas, caracterizadas por flores hermafroditas com diferentes comprimentos de estiletes e posicionamento oposto das anteras (hercogamia recíproca. Encontrou-se diferenças no tamanho das anteras, dos grãos de pólen e da superfície estigmática entre os morfotipos. Polinizações controladas mostraram que os morfotipos apresentam auto-incompatibilidade e intramorfo-incompatibilidade ao nível do estigma e do estilete. Os visitantes são pequenas vespas, abelhas, borboletas e beija-flores, que visitam as flores com maior freqüência no início da manhã. De acordo com a freqüência e eficiência no comportamento de transportar os grãos de pólen entre os morfos florais, o principal polinizador foi o beija-flor Thalurania furcata. O néctar é produzido em pequena quantidade (máximo de 8µl e durante a abertura das flores, que ocorre entre 5 e 7h. A espécie forma agrupamentos devido à existência de reprodução vegetativa.Psychotria L. (Rubiaceae is the genus with the most heterostylous species in the angiosperms. The objective of this study is to describe floral biology, phenology, reproductive system, and pollinators of Psychotria poeppigiana Müll. Arg. The terminal inflorescences are capitate with red bracts and yellow flowers, and presented two different morphs: pin and thrum, characterized by hermaphroditic flowers with different lengths of styles and opposed position of the anthers (reciprocal herkogamy. Dimorphism was also observed in the size of anthers, pollen grains and stigmatic surfaces. Controlled pollination revealed self

  8. Chapa aglomerada de cimento-madeira de Hevea brasiliensis Müll. Arg. Cement-bonded particleboard of Hevea brasiliensis Müll. Arg.

    Esmeralda Yoshico Arakaki Okino

    2004-06-01

    Full Text Available Chapas de partículas de cimento-madeira foram confeccionadas com a madeira de quatro clones de Hevea brasiliensis Müll. Arg. (seringueira: IAN 717, IAN 873, GT 711 e AVROS 1301. Confeccionaram-se as chapas na proporção de 1:4:1 (madeira:cimento:água por peso e nas dimensões de 450 x 450 x 13 mm e densidade nominal de 1,4 g/cm³, com a adição de 4% de cloreto de cálcio di-hidratado (CaCl2.2H2O como acelerador. Foram testadas partículas fervidas e não-fervidas dos quatro clones, totalizando oito tratamentos, sendo em cada um destes, com quatro repetições, avaliadas as propriedades mecânicas e físicas das chapas, segundo a norma ASTM D 1037 - 96a. De forma geral, os melhores resultados de propriedades físicas e mecânicas foram obtidos nas chapas com partículas do clone AVROS 1301. No teste de hidratação do cimento, a madeira de seringueira in natura foi classificada como de "inibição extrema", porém com a adição de CaCl2 o foi como de "baixa inibição". Essa madeira se mostrou tecnicamente viável à produção de chapas de cimento-madeira, independentemente do clone.Cement-bonded particleboards of rubberwood were manufactured with four clones of Hevea brasiliensis Müll. Arg. (rubberwood: IAN 717, IAN 873, GT 711 and AVROS 1301. Boards of 450 x 450 x 13 mm were manufactured in a ratio of 1:4:1 (wood/cement/water, weight basis, with 1.4 g/cm³ density and 4% calcium chloride dihydrated - CaCl2.2H2O as accelerator. The particles of four clones were tested in treated and untreated conditions, totaling eight treatments. In each treatment with four replicates, the physical and mechanical properties were evaluated according to ASTM D 1037 - 96a standard. Overall, the best mechanical and physical results were obtained with the cement-bonded particleboard made with particles from clone AVROS 1301. Rubberwood has shown to be "highly inhibitory" in the hydration test, however when CaCl2 was added the inhibition index decreased and

  9. Fortuitous description of haemoglobin A2' [δ16 (A13) Gly→Arg (GGC→CGC)] in a Tunisian family: study of the molecular defect and its origin.

    Sahli, Chaima Abdelhafidh; Gritli, Sami; Dabboubi, Rym; Omar, Souheil; Siala, Hajer; Kaabachi, Naziha; Bibi, Amina; Messaoud, Taieb

    2015-01-01

    The most common inherited haemoglobin disorders encountered in Tunisia are β-thalassemia and sickle cell disease, which result from mutations in the β-globin gene. Few studies focused on δ-globin gene variations responsible for δ-thalassemia or HbA2 variants. HbA2' [δ16 (A13) Gly→Arg (GGC→CGC)] is a δ-chain variant that has been identified in several populations of African origin. We report herein for the first time the description of HbA2' in the Tunisian population. Identification of HbA2' in the studied family was carried out by high-performance liquid chromatography and confirmed by sequencing analyses of the whole δ-globin gene. Haplotypes of the β-globin gene cluster were constructed by mapping the restriction sites using polymerase chain reaction followed by enzymatic digestion. Compound heterozygosity of HbA2' with HbO-Arab was identified in the proband. The mother and two other siblings showed heterozygous HbA2' whereas the father showed heterozygous HbO-Arab. The sum of HbA2 and HbA2' in all cases was less than 4%, thus excluding β-thalassemia. β-cluster haplotype analysis revealed that this mutation was associated with the F haplotype (-+--+++). The unique origin of this mutation in Africa is likely since the linked β-cluster haplotype is one of the major haplotypes found in African populations. PMID:25858298

  10. Occurrence of antimicrobials and antimicrobial resistance genes in beef cattle storage ponds and swine treatment lagoons

    Zhang, Yuping; Zhang, Chiqian [Department of Civil Engineering, University of Nebraska-Lincoln, Lincoln, NE (United States); Parker, David B. [USDA Meat Animal Research Center, Clay Center, NE (United States); Snow, Daniel D. [Water Sciences Laboratory, University of Nebraska-Lincoln, Lincoln, NE (United States); Zhou, Zhi [Department of Civil and Environmental Engineering, National University of Singapore (Singapore); Li, Xu, E-mail: xuli@unl.edu [Department of Civil Engineering, University of Nebraska-Lincoln, Lincoln, NE (United States)

    2013-10-01

    Livestock manure treatment and storage structures are potential environmental sources of antimicrobials and antimicrobial resistance genes (ARGs). In this study, the occurrence of antimicrobials and ARGs was investigated in the water and the sludge compartments of beef cattle storage ponds and swine lagoons. Analysis was focused on two families of antimicrobials (sulfonamide and tetracycline) and the corresponding ARGs (sul1, sul2, tetO, tetQ and tetX). Results showed that the pseudo-partitioning coefficients of tetracyclines were higher than those of sulfonamides, suggesting different distributions of these two classes of antimicrobials between water and sludge. The ARGs tested were detected in nearly all ponds and lagoons, with the highest relative abundance in sul2 at 6.3 × 10{sup −1} copies per 16S rRNA gene. A positive correlation was observed between total sul genes and total sulfonamides in water while the correlation was negative in sludge. No significant correlation was found between total tet genes and total tetracyclines in either water or sludge, but significant correlations were observed for certain individual tet genes. Ammonia concentrations strongly correlated with all ARGs except tetX. This study provided quantitative information on the occurrence of antimicrobials and ARGs in the liquid and solid compartments of typical manure treatment and storage structures. - Highlights: • Partitioning of antimicrobials between water and sludge is compound specific. • Antimicrobial resistance genes occurred in both water and sludge. • The ARG abundance varied more substantially in swine lagoons than in cattle ponds. • Correlations between ARGs and antimicrobials are system dependent.

  11. RESEARCH ON ARGES RIVER FISH FAUNA IN BUDEASA-GOLESTI AREA

    Alina-Mihaela Truţă

    2015-12-01

    Full Text Available Arges River was subject to periodic ichthyologic, hydrobiological and hydrological research. By its content and approach the present paper shows a series of research on fish fauna in Budeasa-Golesti area of Arges River, Pitesti. By research presented in the study we sought to evaluate the state and evolution of fish fauna in the city reservoirs, Pitesti area, over the last 30 years, trying to highlight the causes that led to the current situation and to propose measures for the conservation of natural fish fauna in the future. Fish fauna in Pitesti area currently consists of 14 species belonging to four families: Cyprinidae (9 species, Cobitidae (1 species, Esocidae (1 species and Percidae (3 species. Most species live naturally in lakes studied except for one species Pseudorasbora parva which was introduced accidentally. The research undertaken to reflect changes in the fish fauna in the last 30 years, indicates an increase in the number of species, either through deliberate stocking for sport fishing purposes or due to changes in biotope favouring the development of certain species which were accidental in the past.

  12. Characterization of Aspartate Kinase from Corynebacterium pekinense and the Critical Site of Arg169

    Weihong Min

    2015-11-01

    Full Text Available Aspartate kinase (AK is the key enzyme in the biosynthesis of aspartate-derived amino acids. Recombinant AK was efficiently purified and systematically characterized through analysis under optimal conditions combined with steady-state kinetics study. Homogeneous AK was predicted as a decamer with a molecular weight of ~48 kDa and a half-life of 4.5 h. The enzymatic activity was enhanced by ethanol and Ni2+. Moreover, steady-state kinetic study confirmed that AK is an allosteric enzyme, and its activity was inhibited by allosteric inhibitors, such as Lys, Met, and Thr. Theoretical results indicated the binding mode of AK and showed that Arg169 is an important residue in substrate binding, catalytic domain, and inhibitor binding. The values of the kinetic parameter Vmax of R169 mutants, namely, R169Y, R169P, R169D, and R169H AK, with l-aspartate as the substrate, were 4.71-, 2.25-, 2.57-, and 2.13-fold higher, respectively, than that of the wild-type AK. Furthermore, experimental and theoretical data showed that Arg169 formed a hydrogen bond with Glu92, which functions as the entrance gate. This study provides a basis to develop new enzymes and elucidate the corresponding amino acid production.

  13. THE INCIDENCE OF HIV INFECTIONS IN ARGES POPULATION IN 2012 - 2013

    Ionica Deliu

    2015-12-01

    Full Text Available Human immunodeficiency virus (HIV from Retroviridae family was isolated in early 80´s but his presence in Africa is much older than we previously thought, before the pandemic spread because of the social changes of 70´s (Cernescu, 1998. The incidence of HIV infections is a constant concern for public health specialists, with inferences for whole population. In this paper we presented the incidence of HIV infections in 2012/2013 period in Arges County, Romania. Studies were made in Laboratories of Public Health Department Arges. Immunoenzymatic methods were used for anti-HIV antibodies detection (Genscreen ULTRA HIV Ag-Ab kit from BIO RAD. The results of the immunoenzymatic tests were presented in order to age and gender of the tested people. New cases of HIV infection were 1.73% in 2012 and 1.10% in 2013 from all investigated samples. The positive samples were frequent in male population, and for 25 -34 years and 35-44 years age groups. Prophylaxis is the best weapon in fight against HIV/AIDS. But the properly way to stop the spread of HIV infections may be the development of vaccine or drugs to stop progression to AIDS (Al-Jabri, 2007.

  14. Prevalence of the apolipoprotein E Arg145Cys dyslipidemia at-risk polymorphism in African-derived populations.

    Abou Ziki, Maen D; Strulovici-Barel, Yael; Hackett, Neil R; Rodriguez-Flores, Juan L; Mezey, Jason G; Salit, Jacqueline; Radisch, Sharon; Hollmann, Charleen; Chouchane, Lotfi; Malek, Joel; Zirie, Mahmoud A; Jayyuosi, Amin; Gotto, Antonio M; Crystal, Ronald G

    2014-01-15

    Apolipoprotein E, a protein component of blood lipid particles, plays an important role in lipid transport. Different mutations in the apolipoprotein E gene have been associated with various clinical phenotypes. In an initiated study of Qataris, we observed that 17% of the African-derived genetic subgroup were heterozygotes for a rare Arg145Cys (R145C) variant that functions as a dominant trait with incomplete penetrance associated with type III hyperlipoproteinemia. On the basis of this observation, we hypothesized that the R145C polymorphism might be common in African-derived populations. The prevalence of the R145C variant was assessed worldwide in the "1000 Genomes Project" and in 1,012 whites and 1,226 African-Americans in New York, New York. The 1000 Genomes Project data demonstrated that the R145C polymorphism is rare in non-African-derived populations but present in 5% to 12% of Sub-Saharan African-derived populations. The R145C polymorphism was also rare in New York whites (1 of 1,012, 0.1%); however, strikingly, 53 of the 1,226 New York African-Americans (4.3%) were R145C heterozygotes. The lipid profiles of the Qatari and New York R145C heterozygotes were compared with those of controls. The Qatari R145C subjects had higher triglyceride levels than the Qatari controls (p worldwide derived from Sub-Saharan Africans are apolipoprotein E R145C. In conclusion, although larger epidemiologic studies are necessary to determine the long-term consequences of this polymorphism, the available evidence suggests it is a common cause of a mild triglyceride dyslipidemia. PMID:24239320

  15. Comparative MD Simulations Indicate a Dual Role for Arg1323.50 in Dopamine-Dependent D2R Activation.

    Ralf C Kling

    Full Text Available Residue Arg3.50 belongs to the highly conserved DRY-motif of class A GPCRs, which is located at the bottom of TM3. On the one hand, Arg3.50 has been reported to help stabilize the inactive state of GPCRs, but on the other hand has also been shown to be crucial for stabilizing active receptor conformations and mediating receptor-G protein coupling. The combined results of these studies suggest that the exact function of Arg3.50 is likely to be receptor-dependent and must be characterized independently for every GPCR. Consequently, we now present comparative molecular-dynamics simulations that use our recently described inactive-state and Gα-bound active-state homology models of the dopamine D2 receptor (D2R, which are either bound to dopamine or ligand-free, performed to identify the function of Arg1323.50 in D2R. Our results are consistent with a dynamic model of D2R activation in which Arg1323.50 adopts a dual role, both by stabilizing the inactive-state receptor conformation and enhancing dopamine-dependent D2R-G protein coupling.

  16. Can chlorination co-select antibiotic-resistance genes?

    Lin, Wenfang; Zhang, Menglu; Zhang, Shenghua; Yu, Xin

    2016-08-01

    Selective pressures, such as chemical or heavy metal pollution, may co-select for bacterial antibiotic resistance in the environment. However, whether chlorination in water treatment can co-select antibiotic-resistant bacteria is controversial. In this study, high capacity quantitative polymerase chain reaction (qPCR) analysis was applied to target almost all known antibiotic-resistance genes (ARGs) (282 types) and 13 mobile genetic elements (MGEs) in bacteria detected in secondary effluents from a municipal wastewater treatment plant after chlorination. The results revealed that 125 unique ARGs were detected in non-chlorinated samples, and the number decreased (79-91 types) as the chlorine concentration was increased. Moreover, 7.49 × 10(4)-3.92 × 10(7) copies/100 ml water reduction of ARGs occurred with 4 mg Cl2/l. Considering the relative abundance of ARGs (i.e., ARG copies normalized to 16S rRNA gene copies), 119 ARGs decreased in response to chlorination, whereas only six ARGs, such as dfrA1, tetPB-03, tetPA, ampC-04, tetA-02, and erm(36), were potentially enriched by 10.90-, 10.06-, 8.63-, 6.86-, 3.77-, and 1.09-fold, respectively. Furthermore, the relative abundance of 12 detected MGEs was lower after chlorination. Therefore, chlorination was effective in reducing ARGs and MGEs rather than co-selecting them. PMID:27192478

  17. Evidence of an association between the Arg72 allele of the peptide YY and increased risk of type 2 diabetes

    Torekov, Signe S; Larsen, Lesli H; Glümer, Charlotte; Borch-Johnsen, Knut; Jørgensen, Torben; Holst, Jens J; Madsen, Ole D; Hansen, Torben; Pedersen, Oluf

    2005-01-01

    1.05-1.35]). The same polymorphism associated with overweight (25 Arg allele was associated with an increased plasma glucose level 2 h after an oral glucose...... tolerance test (OGTT) (P = 0.03), an increased area under the curve for the post-OGTT plasma glucose level (P = 0.03), and a lower insulinogenic index (P = 0.01). In conclusion, the common Arg allele of the PYY Arg72Thr variant modestly associates with type 2 diabetes and with type 2 diabetes...

  18. Tet and sul antibiotic resistance genes in livestock lagoons of various operation type, configuration, and antibiotic occurrence

    McKinney, C.W.; Loftin, K.A.; Meyer, M.T.; Davis, J.G.; Pruden, A.

    2010-01-01

    Although livestock operations are known to harbor elevated levels of antibiotic resistant bacteria, few studies have examined the potential of livestock waste lagoons to reduce antibiotic resistance genes (ARGs). The purpose of this study was to determine the prevalence and examine the behavior of tetracycline [tet(O) and tet(W)] and sulfonamide [sul(I) and su/(II)] ARGsin a broad cross-section of livestock lagoons within the same semiarid western watershed. ARGs were monitored for one year in the water and the settled solids of eight lagoon systems by quantitative polymerase chain reaction. In addition, antibiotic residues and various bulk water quality constituents were analyzed. It was found that the lagoons of the chicken layer operation had the lowest concentrations of both tet and sul ARGs and low total antibiotic concentrations, whereas su ARGs were highest in the swine lagoons, which generally corresponded to the highest total antibiotic concentrations. A marginal benefit of organic and small dairy operations also was observed compared to conventional and large dairies, respectively. In all lagoons, su ARGs were observed to be generally more recalcitrant than tet ARGs. Also, positive correlations of various bulk water quality constituents were identified with tet ARGs but not sul ARGs. Significant positive correlations were identified between several metals and tet ARGs, but Pearson's correlation coefficients were mostly lower than those determined between antibiotic residues and ARGs. This study represents a quantitative characterization of ARGs in lagoons across a variety of livestock operations and provides insight into potential options for managing antibiotic resistance emanating from agricultural activities. ?? 2010 American Chemical Society.

  19. Fate of antibiotic resistance bacteria and genes during enhanced anaerobic digestion of sewage sludge by microwave pretreatment.

    Tong, Juan; Liu, Jibao; Zheng, Xiang; Zhang, Junya; Ni, Xiaotang; Chen, Meixue; Wei, Yuansong

    2016-10-01

    The fate of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) were investigated during the sludge anaerobic digestion (AD) with microwave-acid (MW-H), microwave (MW) and microwave-H2O2-alkaline (MW-H2O2) pretreatments. Results showed that combined MW pretreatment especially for the MW-H pretreatment could efficiently reduce the ARB concentration, and most ARG concentrations tended to attenuate during the pretreatment. The subsequent AD showed evident removal of the ARB, but most ARGs were enriched after AD. Only the concentration of tetX kept continuous declination during the whole sludge treatment. The total ARGs concentration showed significant correlation with 16S rRNA during the pretreatment and AD. Compared with unpretreated sludge, the AD of MW and MW-H2O2 pretreated sludge presented slightly better ARB and ARGs reduction efficiency. PMID:26970692

  20. Aquaculture changes the profile of antibiotic resistance and mobile genetic element associated genes in Baltic Sea sediments.

    Muziasari, Windi I; Pärnänen, Katariina; Johnson, Timothy A; Lyra, Christina; Karkman, Antti; Stedtfeld, Robert D; Tamminen, Manu; Tiedje, James M; Virta, Marko

    2016-04-01

    Antibiotics are commonly used in aquaculture and they can change the environmental resistome by increasing antibiotic resistance genes (ARGs). Sediment samples were collected from two fish farms located in the Northern Baltic Sea, Finland, and from a site outside the farms (control). The sediment resistome was assessed by using a highly parallel qPCR array containing 295 primer sets to detect ARGs, mobile genetic elements and the 16S rRNA gene. The fish farm resistomes were enriched in transposon and integron associated genes and in ARGs encoding resistance to antibiotics which had been used to treat fish at the farms. Aminoglycoside resistance genes were also enriched in the farm sediments despite the farms not having used aminoglycosides. In contrast, the total relative abundance values of ARGs were higher in the control sediment resistome and they were mainly genes encoding efflux pumps followed by beta-lactam resistance genes, which are found intrinsically in many bacteria. This suggests that there is a natural Baltic sediment resistome. The resistome associated with fish farms can be from native ARGs enriched by antibiotic use at the farms and/or from ARGs and mobile elements that have been introduced by fish farming. PMID:26976842

  1. Microsporogênese em clones normais e tetraplóides de Hevea brasiliensis Muell.-Arg Microsporo genesis in normal and tetraploid Hevea brasiliensis (Muell.-Arg.

    Cândida H. T. M. Conagin

    1971-01-01

    Full Text Available Pesquisas sôbre o efeito da colquicina em Hevea brasiliensis Muell.-Arg. realizadas anteriormente levaram à obtenção de clones com número duplicado de cromossomos; tais clones, atualmente em fase de amplas e detalhadas observações (6, floresceram em 1969, pela primeira vez. Foi então realizado um estudo citológico comparativo da microsporo-gènese de duas plantas, uma pertencente ao clone normal n.° 3064, com 2n = 36 cromossomos, e outra pertencente ao clone duplicado n.° 3065, com 2n=72 cromossomos. Ambos são considerados clones gêmeos, porque foram obtidos de uma mesma semente, por técnica especial (7. Na planta com 2n = 36 cromossomos, o processo meiótico é normal, dando tétrades perfeitas e grãos de pólen aparentemente funcionais. A planta 3065, com 2n=72 cromossomos, apresenta, além de células-mães de pólen que se dividem normalmente, outras que no final da meiose produzem tétrades anormais, com micrócitos excedentes e grãos de pólen vazios. Caracteriza-se também por grãos de pólen que não passam pelas divisões mitóticas, isto é, apresentam sempre um núcleo só, que não se divide. Em virtude destas primeiras observações pode-se formular uma hipótese de esterilidade masculina para o clone em estudo.Previous works on Hevea brasiliensis Muell.-Arg. produced several pairs of twin clones, one member having the normal chromosome number and the other the duplicated set after colchicine treatment. Plants of normal clone 3064 are fertile and have 32 chromosomes. Microsporogenesis is normal, producing only normal tetrads of four microsporocytes. The resulting pollen grains have three germinal pores. Grains in different stages of development could be noticed, from one-nucleated cytoplasm to the two-nucleated reproductive cell, which undoubtedly means normal game to genesis. On the other hand plants of the duplicated twin clone 3065, blossomed during the year of 1969 for the first time. Microsporogenesis studied

  2. PHARMACOGNOSTICAL EVALUATION OF FRUITS OF MALLOTUS PHILIPPENENSIS (LAM. MUELL- ARG (EUPHORBIACEAE

    Kadam Prasad Vijay

    2013-06-01

    Full Text Available Correct identification of starting material is necessary to ensure the quality of herbal medicines. Pharmacognostic evaluation is first step towards establishing identity and purity of crude drug. With this aim the present paper deals with study of pharmacognostic and physicochemical characteristics of fruits of Mallotus philippinensis (lam Muell. Arg. (Euphorbiaceae. The powder of Mallotus philippinensis was used for detection of various chemical constituents. The transverse section and the powder of fruit were studied for macroscopy, microscopy and physicochemical parameters. Fruit is globose, 3-lobed capsule with abundant orange or reddish glandular granules. The Microscopy showed the presence of epicarp, mesocarp endocarp. The cells also showed the presence of starch. From the pharmacognostical evaluation it was found that the fruit contains alkaloids, saponins, flavanoids etc. Physicochemical parameters were also within the limit. It can be concluded that Pharmacognostic evaluation can serve as tool for developing standards for identification, quality and purity of M. philippinensis.

  3. AVATAR: AdVanced Aerodynamic Tools for lArge Rotors

    Schepers, J.C.; Ceyhan, O.; Savenije, F.J.; Stettner, M.; Kooijman, H.J.; Chaviarapoulos, P.; Sieros, G.; Ferreira, C.S.; Sørensen, Niels N.; Wächter, M.; Stoevesandt, B.; Lutz, T.; Gonzalez, A.; Barakos, G.; Voutsinas, A.; Croce, A.; Madsen, J.

    An EERA (European Energy Research Alliance) consortium started an ambitious EU FP7 project AVATAR (AdVanced Aerodynamic Tools of lArge Rotors) in November 2013. The project lasts 4 years and is carried out in a consortium with 11 research institutes and two industry partners. The motivation for the...... AVATAR project lies in the fact that future 10 to 20 MW turbine design model analysis will importantly violate known validity limits of today’s aerodynamic and aero-elastic models in aspects like compressibility and Reynolds number effects, laminar/turbulent transition and separation effects, all in...... modelling improvements will be demonstrated on a new 10MW reference turbine design model description. The first results from the AVATAR project are presented in this paper....

  4. Monitoring and evaluation of antibiotic resistance genes in four municipal wastewater treatment plants in Harbin, Northeast China.

    Wen, Qinxue; Yang, Lian; Duan, Ruan; Chen, Zhiqiang

    2016-05-01

    The development and proliferation of antibiotic resistance in pathogenic and environmental microorganisms is of great concern for public health. In this study, the distribution and removal efficiency of intI1 and eight subtypes of antibiotic resistance genes (ARGs) for tetracycline, sulfonamides, beta-lactams resistance in four municipal wastewater treatment plants (WWTPs) in Harbin, which locates in Songhua River basin in cold areas of China, were monitored by real-time fluorescent quantitative PCR. The results showed that intI1 and 6 ARGs except for blaTEM and blaSHV were detected in wastewater and sludge samples and 0.3-2.7 orders of magnitude of ARGs removal efficiency in the four WWTPs were observed. The investigation on the removal of ARGs of different treatment units in one WWTP showed that the biological treatment unit played the most important role in ARGs removal (1.2-1.8 orders of magnitude), followed by UV disinfection, while primary physical treatment units can hardly remove any ARGs. Although all the WWTPs can remove ARGs effectively, ARGs concentrations are still relatively high in the effluent, their further attenuation should be investigated. PMID:26840514

  5. ARG (juegos de realidad alternativa. Contribuciones, limitaciones y potencialidades para la docencia universitaria

    Teresa Piñeiro Otero

    2015-01-01

    Full Text Available La ludificación de la educación ha representado una oportunidad para fomentar la interacción, la motivación y la participación del alumnado. Los ARG (las siglas inglesas de juegos de realidad alternativa ofrecen una nueva herramienta altamente inmersiva que puede implementarse en el logro de los objetivos docentes. Uno de sus puntos fuertes consiste en la suma de esfuerzos y recursos (la llamada inteligencia colectiva aplicada a la resolución de problemas. A esto se añade su combinación de plataformas en los entornos online y offline, lo que favorece el «realismo» de la experiencia. En este sentido, el presente trabajo pretende condensar las potencialidades, limitaciones y retos de los ARG al servicio de la educación universitaria. Basándose, a nivel metodológico, en la elaboración de un corpus teórico relevante y adecuado, analiza el potencial educativo de esta herramienta que, en ámbitos como el marketing o la comunicación corporativa ya ha despegado con éxito, pero que en el área educativa todavía no había sido abordada en profundidad. Recopila, además, ejemplos satisfactorios que se han desarrollado en diversas disciplinas académicas en otros países y que no resultan fácilmente localizables. Se concluye que, dados los antecedentes, potencialidades y análisis expuesto, debe valorarse la posibilidad de incorporar los juegos de realidad alternativa a la práctica de la docencia universitaria en el marco de una estrategia educativa que determine sus objetivos y sistema de evaluación más adecuado.

  6. Arg188 in rice sucrose transporter OsSUT1 is crucial for substrate transport

    Sun Ye

    2012-11-01

    Full Text Available Abstract Background Plant sucrose uptake transporters (SUTs are H+/sucrose symporters related to the major facilitator superfamily (MFS. SUTs are essential for plant growth but little is known about their transport mechanism. Recent work identified several conserved, charged amino acids within transmembrane spans (TMS in SUTs that are essential for transport activity. Here we further evaluated the role of one of these positions, R188 in the fourth TMS of OsSUT1, a type II SUT. Results The OsSUT1(R188K mutant, studied by expression in plants, yeast, and Xenopus oocytes, did not transport sucrose but showed a H+ leak that was blocked by sucrose. The H+ leak was also blocked by β-phenyl glucoside which is not translocated by OsSUT1. Replacing the corresponding Arg in type I and type III SUTs, AtSUC1(R163K and LjSUT4(R169K, respectively, also resulted in loss of sucrose transport activity. Fluorination at the glucosyl 3 and 4 positions of α-phenyl glucoside greatly decreased transport by wild type OsSUT1 but did not affect the ability to block H+ leak in the R188K mutant. Conclusion OsSUT1 R188 appears to be essential for sucrose translocation but not for substrate interaction that blocks H+ leak. Therefore, we propose that an additional binding site functions in the initial recognition of substrates. The corresponding Arg in type I and III SUTs are equally important. We propose that R188 interacts with glucosyl 3-OH and 4-OH during translocation.

  7. Association of ADHIB and ALDH2 gene polymorphisms with alcohol dependence: A pilot study from India

    Vaswani Meera; Prasad Pushplata; Kapur Suman

    2009-01-01

    Abstract Functional polymorphism in the genes encoding alcohol dehydrogenase (ADH) 1B and aldehyde dehydrogenase (ALDH) 2 are considered most important among several genetic determinants of alcohol dependence, a complex disorder. There is no report on the widely studied Arg47His and Glu487Lys polymorphisms from Indian alcoholdependent populations. In this paper, we report, for the first time, allelic and genotypic frequencies of Arg47His and Glu487Lys single nucleotide polymorphisms (SNPs) in...

  8. Excretion of Antibiotic Resistance Genes by Dairy Calves Fed Milk Replacers with Varying Doses of Antibiotics

    Thames, Callie H; Pruden, Amy; James, Robert E.; Ray, Partha P.; Knowlton, Katharine F.

    2012-01-01

    Elevated levels of antibiotic resistance genes (ARGs) in soil and water have been linked to livestock farms and in some cases feed antibiotics may select for antibiotic resistant gut microbiota. The purpose of this study was to examine the establishment of ARGs in the feces of calves receiving milk replacer containing no antibiotics versus subtherapeutic or therapeutic doses of tetracycline and neomycin. The effect of antibiotics on calf health was also of interest. Twenty-eight male and fema...

  9. Antibiotic resistance gene abundance in feces of calves fed pirlimycin-dosed whole milk

    Littier, Heather Melissa

    2015-01-01

    Exposure to antibiotics has the potential to increase the incidence and proliferation of antibiotic resistance genes (ARG) in the gut and fecal microbiome. Non-saleable, antibiotic-containing milk from cows treated with antibiotics (waste milk) is commonly fed to dairy calves but the effects of ingestion of antibiotics at an early age on the gut microbiome and the development of ARG in the naive gut are not well understood. Pirlimycin, a lincosamide antibiotic acting against Gram positive bac...

  10. A Comprehensive Analysis on Spread and Distribution Characteristic of Antibiotic Resistance Genes in Livestock Farms of Southeastern China

    Wang, Na; Guo, Xinyan; Yan, Zheng; Wang, Wei; Chen, Biao; Ge, Feng; Ye, Boping

    2016-01-01

    The pollution of antibiotic resistance genes (ARGs) in livestock farms is a problem which need to be paid more attention to, due to the severe resistance dissemination and the further human health risk. In this study, all the relevant exposure matrices (manure, soil and water) of sixteen animal farms in Southeastern China were sampled to determine twenty-two ARGs conferring resistance to five major classes of antibiotics including tetracyclines, sulfonamides, quinolones, aminoglycosides, and macrolides. The results showed that the spread property of sul genes was most extensive and strong, followed by tet and erm genes. The abundance of tet genes expressing ribosomal protection proteins (tetM, tetO, tetQ, tetT and tetW) was higher than that expressing efflux pump proteins (tetA, tetC, tetE and tetG) in each type of samples. The high abundance and frequency of ermB gene in the matrices should be paid more attention, because macrolides is a major medicine for human use. For manures, it was found that the similar ARGs distribution rules were existing in poultry manure or porcine manure samples, despite of the different origins of these two types of livestock farms. Meanwhile, it was interesting that the distribution rule of tet genes in animal manure was nearly the same as all the ARGs. For soils, the result of nonmetric multi-dimensional scaling (NMDS) analysis showed that the pollution of ARGs in the soils fertilized by poultry and cattle manures were more substantial in northern Jiangsu, but no significant ARGs diversity was observed among porcine manured soils of five different regions. Furthermore, most ARGs showed significant positive relationships with environmental variables such as concentration of sulfonamides, tetracyclines, Cu, Zn and total organic carbon (TOC). The pollution profile and characteristics of so many ARGs in livestock farms can provide significative foundation for the regulation and legislation of antibiotics in China. PMID:27388166

  11. A Comprehensive Analysis on Spread and Distribution Characteristic of Antibiotic Resistance Genes in Livestock Farms of Southeastern China.

    Wang, Na; Guo, Xinyan; Yan, Zheng; Wang, Wei; Chen, Biao; Ge, Feng; Ye, Boping

    2016-01-01

    The pollution of antibiotic resistance genes (ARGs) in livestock farms is a problem which need to be paid more attention to, due to the severe resistance dissemination and the further human health risk. In this study, all the relevant exposure matrices (manure, soil and water) of sixteen animal farms in Southeastern China were sampled to determine twenty-two ARGs conferring resistance to five major classes of antibiotics including tetracyclines, sulfonamides, quinolones, aminoglycosides, and macrolides. The results showed that the spread property of sul genes was most extensive and strong, followed by tet and erm genes. The abundance of tet genes expressing ribosomal protection proteins (tetM, tetO, tetQ, tetT and tetW) was higher than that expressing efflux pump proteins (tetA, tetC, tetE and tetG) in each type of samples. The high abundance and frequency of ermB gene in the matrices should be paid more attention, because macrolides is a major medicine for human use. For manures, it was found that the similar ARGs distribution rules were existing in poultry manure or porcine manure samples, despite of the different origins of these two types of livestock farms. Meanwhile, it was interesting that the distribution rule of tet genes in animal manure was nearly the same as all the ARGs. For soils, the result of nonmetric multi-dimensional scaling (NMDS) analysis showed that the pollution of ARGs in the soils fertilized by poultry and cattle manures were more substantial in northern Jiangsu, but no significant ARGs diversity was observed among porcine manured soils of five different regions. Furthermore, most ARGs showed significant positive relationships with environmental variables such as concentration of sulfonamides, tetracyclines, Cu, Zn and total organic carbon (TOC). The pollution profile and characteristics of so many ARGs in livestock farms can provide significative foundation for the regulation and legislation of antibiotics in China. PMID:27388166

  12. Association of β1 and β3 adrenergic receptors gene polymorphisms with insulin resistance and high lipid profiles related to type 2 diabetes and metabolic syndrome

    Burguete-García, Ana I; Gabriela A. Martínez-Nava; Adán Valladares-Salgado; V. H. Bermúdez; Bárbara Estrada-Velasco; Niels Wacher; Jesús Peralta-Romero; Jaime Garcia-Mena; Esteban Parra; Miguel Cruz

    2014-01-01

    Background: Among the diverse genes associated to type 2 diabetes (T2D), the β-adrenergic receptors are an excellent candidate to study in Mexican population. The objective of this work was to analyze the association of polymorphisms in ADRB1 (rs1801253) (Arg389Gly) and ADRB3 (Trp64Arg) genes with T2D and metabolic syndrome (MS). Methods: We studied 445 MS patients, 502 with T2D and 552 healthy controls. Anthropometric features and complete biochemical profile were evaluated, and Arg389Gly an...

  13. The architecture of ArgR-DNA complexes at the genome-scale in Escherichia coli

    Cho, Suhyung; Cho, Yoo-Bok; Kang, Taek Jin;

    2015-01-01

    DNA-binding motifs that are recognized by transcription factors (TFs) have been well studied; however, challenges remain in determining the in vivo architecture of TF-DNA complexes on a genome-scale. Here, we determined the in vivo architecture of Escherichia coli arginine repressor (ArgR)-DNA co...

  14. High constitutive activity of a virus-encoded seven transmembrane receptor in the absence of the conserved DRY motif (Asp-Arg-Tyr) in transmembrane helix 3

    Rosenkilde, Mette M; Kledal, Thomas N; Schwartz, Thue W

    2005-01-01

    The highly conserved Arg in the so-called DRY motif (Asp-Arg-Tyr) at the intracellular end of transmembrane helix 3 is in general considered as an essential residue for G protein coupling in rhodopsin-like seven transmembrane (7TM) receptors. In the open reading frame 74 (ORF74) receptor encoded by...

  15. Regional differences in distribution volume of I-123 IMP in the human brain. Effect on CBF calculated by ARG method

    Two methods of quantitating cerebral blood flow (CBF) with iodine-123-labeled N-isopropyl-p-iodoamphetamine (I-123 IMP) and a two-compartment model had been proposed; one is the table look-up (TLU) method and the other is the autoradiographic (ARG) method. The TLU method provides values of the cerebral blood flow (CBF) values and distribution volume of I-123 IMP (Vd) independently. In the ARG method, a fixed Vd is applied for the entire brain to calculate CBF. Our purpose was to evaluate regional differences in Vd in the human brain, or possible effects of regional differences in Vd on CBF calculated by the ARG method. In the present study, two SPECT scans were acquired from each of eight normal subjects (aged 44.0±16.7) at 40 min and 180 min of mid-scan-time after intravenous 1 min infusion of 111 MBq IMP. A single arterial blood sampling was performed 10 min after the IMP infusion. All images were anatomically normalized and analyzed with SPM99 and Matlab. We generated CBF and Vd images for each subject by the TLU method and evaluated differences in Vd among brain structures. We subsequently generated another set of CBF images by the ARG method and examined differences between CBF calculated by the TLU method and that by the ARG method. Significant main effects of subject and brain structure in Vd were observed (two-way ANOVA). Vd values were higher in the deep gray matter than in the cerebral cortical regions. Among the cerebral cortical regions, no significant difference in Vd was observed. In spite of the significant differences in Vd among the brain structures, the voxel-by-voxel analyses as well as the ROI analyses revealed no statistically significant difference between CBF calculated by the TLU method and that by the ARG method. Although regional differences in Vd were observed, the present results support the assumption that a fixed Vd does not cause significant error in the calculation of CBF by the ARG method. (author)

  16. Initial study of some methanogen genes: expression of Methanococcus voltae genes in Escherichia coli

    This report elates studies of gene transfer experiments aimed at elucidating the genetic organization of a methanogen, Methanococcus volta. Our initial cloning study, was concerned with complementation of hisA and argG mutations in E. coli by DNA derived from M. voltae and cloned into the PstI site of the plasmid vector pBR322. This complemetation was correlated with production in maxicells of methanogen-encoded polypeptides. We found that expression of the argG-complementing DNA was dependent upon orientation with respect to the β-lactamase promoters of pBR322. 17 refs., 3 figs

  17. Technetium-99m-labeled Arg-Gly-Asp-conjugated alpha-melanocyte stimulating hormone hybrid peptides for human melanoma imaging

    Introduction: The purpose of this study was to examine whether 99mTc-labeled Arg-Gly-Asp (RGD)-conjugated alpha-melanocyte stimulating hormone (α-MSH) hybrid peptide targeting both melanocortin-1 (MC1) and αvβ3 integrin receptors was superior in melanoma targeting to 99mTc-labeled α-MSH or RGD peptide targeting only the MC1 or αvβ3 integrin receptor. Methods: RGD-Lys-(Arg11)CCMSH, RAD-Lys-(Arg11)CCMSH and RGD-Lys-(Arg11)CCMSHscramble were designed to target both MC1 and αvβ3 integrin receptors, MC1 receptor only and αvβ3 integrin receptor only, respectively. The MC1 or αvβ3 integrin receptor binding affinities of three peptides were determined in M21 human melanoma cells. The melanoma targeting properties of 99mTc-labeled RGD-Lys-(Arg11)CCMSH, RAD-Lys-(Arg11)CCMSH and RGD-Lys-(Arg11)CCMSHscramble were determined in M21 human melanoma-xenografted nude mice. Meanwhile, the melanoma uptake of 99mTc-RGD-Lys-(Arg11)CCMSH was blocked with various non-radiolabeled peptides in M21 melanoma xenografts. Results: RGD-Lys-(Arg11)CCMSH displayed 2.0 and 403 nM binding affinities to both MC1 and αvβ3 integrin receptors, whereas RAD-Lys-(Arg11)CCMSH or RGD-Lys-(Arg11)CCMSHscramble lost their αvβ3 integrin receptor binding affinity by greater than 248-fold or MC1 receptor binding affinity by more than 100-fold, respectively. The melanoma uptake of 99mTc-RGD-Lys-(Arg11)CCMSH was 2.49 and 2.24 times (P 99mTc-RAD-Lys-(Arg11)CCMSH and 99mTc-RGD-Lys-(Arg11)CCMSHscramble at 2 h post-injection, respectively. Either RGD or (Arg11)CCMSH peptide co-injection could block 42% and 57% of the tumor uptake of 99mTc-RGD-Lys-(Arg11)CCMSH, whereas the coinjection of RGD+(Arg11)CCMSH peptide mixture could block 66% of the tumor uptake of 99mTc-RGD-Lys-(Arg11)CCMSH. Conclusions: Targeting both MC1 and αvβ3 integrin receptors enhanced the melanoma uptake of 99mTc-RGD-Lys-(Arg11)CCMSH in M21 human melanoma xenografts. Flank M21 human melanoma tumors were clearly visualized by single

  18. Relationships between plasma leptin levels, leptin G2548A, leptin receptor Gln223Arg polymorphisms and gestational diabetes mellitus in Chinese population

    Yang, Mei; Peng, Songxu; Li, Wei; Wan, Zhihua; Fan, Linlin; Du, Yukai

    2016-01-01

    The purposes of this study were to examine concentrations of leptin and biochemical parameters in gestational diabetes mellitus (GDM) patients and normal glucose tolerance (NGT) individuals, and also to explore the links of leptin (LEP) G2548A and leptin receptor (LEPR) Gln223Arg polymorphisms with leptin levels and GDM risk among Chinese. Our study included 357 GDM and 355 NGT individuals who were at 24~30 gestational weeks. Plasma leptin and insulin levels were analyzed by ELISA. Gene polymorphisms were genotyped using TaqMan real-time polymerase chain reaction assay. The results showed that plasma leptin levels were significantly higher in the impaired fasting glucose (IFG) group than NGT group (34.35 (26.54, 56.48) ng/mL vs 26.31 (17.99, 37.87) ng/mL, P  0.05). Our findings showed that high leptin level was associated with GDM. And larger and more rigorous researches were needed to further explore the association of LEP and LEPR gene polymorphisms and GDM among Chinese population. PMID:27034205

  19. [des-Arg(1)]-Proctolin: A novel NEP-like enzyme inhibitor identified in Tityus serrulatus venom.

    Duzzi, Bruno; Cajado-Carvalho, Daniela; Kuniyoshi, Alexandre Kazuo; Kodama, Roberto Tadashi; Gozzo, Fabio Cesar; Fioramonte, Mariana; Tambourgi, Denise Vilarinho; Portaro, Fernanda Vieira; Rioli, Vanessa

    2016-06-01

    The scorpion Tityus serrulatus venom comprises a complex mixture of molecules that paralyzes and kills preys, especially insects. However, venom components also interact with molecules in humans, causing clinic envenomation. This cross-interaction may result from homologous molecular targets in mammalians and insects, such as (NEP)-like enzymes. In face of these similarities, we searched for peptides in Tityus serrulatus venom using human NEP as a screening tool. We found a NEP-inhibiting peptide with the primary sequence YLPT, which is very similar to that of the insect neuropeptide proctolin (RYLPT). Thus, we named the new peptide [des-Arg(1)]-proctolin. Comparative NEP activity assays using natural substrates demonstrated that [des-Arg(1)]-proctolin has high specificity for NEP and better inhibitory activity than proctolin. To test the initial hypothesis that molecular homologies allow Tityus serrulatus venom to act on both mammal and insect targets, we investigated the presence of a NEP-like in cockroaches, the main scorpion prey, that could be likewise inhibited by [des-Arg(1)]-proctolin. Indeed, we detected a possible NEP-like in a homogenate of cockroach heads whose activity was blocked by thiorphan and also by [des-Arg(1)]-proctolin. Western blot analysis using a human NEP monoclonal antibody suggested a NEP-like enzyme in the homogenate of cockroach heads. Our study describes for the first time a proctolin-like peptide, named [des-Arg(1)]-proctolin, isolated from Tityus serrulatus venom. The tetrapeptide inhibits human NEP activity and a NEP-like activity in a cockroach head homogenate, thus it may play a role in human envenomation as well as in the paralysis and death of scorpion preys. PMID:26056922

  20. Seasonality of antibiotic prescriptions for outpatients and resistance genes in sewers and wastewater treatment plant outflow.

    Caucci, Serena; Karkman, Antti; Cacace, Damiano; Rybicki, Marcus; Timpel, Patrick; Voolaid, Veiko; Gurke, Robert; Virta, Marko; Berendonk, Thomas U

    2016-05-01

    To test the hypothesis of a seasonal relationship of antibiotic prescriptions for outpatients and the abundance of antibiotic resistance genes (ARGs) in the wastewater, we investigated the distribution of prescriptions and different ARGs in the Dresden sewer system and wastewater treatment plant during a two-year sampling campaign. Based on quantitative PCR (qPCR), our results show a clear seasonal pattern for relative ARGs abundances. The higher ARGs levels in autumn and winter coincide with the higher rates of overall antibiotic prescriptions. While no significant differences of relative abundances were observed before and after the wastewater treatment for most of the relative ARGs, the treatment clearly influenced the microbial community composition and abundance. This indicates that the ARGs are probably not part of the dominant bacterial taxa, which are mainly influenced by the wastewater treatment processes, or that plasmid carrying bacteria remain constant, while plasmid free bacteria decrease. An exception was vancomycin (vanA), showing higher relative abundance in treated wastewater. It is likely that a positive selection or community changes during wastewater treatment lead to an enrichment ofvanA. Our results demonstrate that in a medium-term study the combination of qPCR and next generation sequencing corroborated by drug-related health data is a suitable approach to characterize seasonal changes of ARGs in wastewater and treated wastewater. PMID:27073234

  1. Occurrence of Antibiotic Resistance Genes and Bacterial Markers in a Tropical River Receiving Hospital and Urban Wastewaters.

    Naresh Devarajan

    Full Text Available The occurrence of emerging biological contaminants including antibiotic resistance genes (ARGs and Faecal Indicator Bacteria (FIB is still little investigated in developing countries under tropical conditions. In this study, the total bacterial load, the abundance of FIB (E. coli and Enterococcus spp. (ENT, Pseudomonas spp. and ARGs (blaTEM, blaCTX-M, blaSHV, blaNDM and aadA were quantified using quantitative PCR in the total DNA extracted from the sediments recovered from hospital outlet pipes (HOP and the Cauvery River Basin (CRB, Tiruchirappalli, Tamil Nadu, India. The abundance of bacterial marker genes were 120, 104 and 89 fold higher for the E. coli, Enterococcus spp. and Pseudomonas spp., respectively at HOP when compared with CRB. The ARGs aadA and blaTEM were most frequently detected in higher concentration than other ARGs at all the sampling sites. The ARGs blaSHV and blaNDM were identified in CRB sediments contaminated by hospital and urban wastewaters. The ARGs abundance strongly correlated (r ≥ 0.36, p < 0.05, n = 45 with total bacterial load and E. coli in the sediments, indicating a common origin and extant source of contamination. Tropical aquatic ecosystems receiving wastewaters can act as reservoir of ARGs, which could potentially be transferred to susceptible bacterial pathogens at these sites.

  2. Occurrence of Antibiotic Resistance Genes and Bacterial Markers in a Tropical River Receiving Hospital and Urban Wastewaters

    Devarajan, Naresh; Laffite, Amandine; Mulaji, Crispin Kyela; Otamonga, Jean-Paul; Mpiana, Pius Tshimankinda; Mubedi, Josué Ilunga; Prabakar, Kandasamy; Ibelings, Bastiaan Willem; Poté, John

    2016-01-01

    The occurrence of emerging biological contaminants including antibiotic resistance genes (ARGs) and Faecal Indicator Bacteria (FIB) is still little investigated in developing countries under tropical conditions. In this study, the total bacterial load, the abundance of FIB (E. coli and Enterococcus spp. (ENT)), Pseudomonas spp. and ARGs (blaTEM, blaCTX-M, blaSHV, blaNDM and aadA) were quantified using quantitative PCR in the total DNA extracted from the sediments recovered from hospital outlet pipes (HOP) and the Cauvery River Basin (CRB), Tiruchirappalli, Tamil Nadu, India. The abundance of bacterial marker genes were 120, 104 and 89 fold higher for the E. coli, Enterococcus spp. and Pseudomonas spp., respectively at HOP when compared with CRB. The ARGs aadA and blaTEM were most frequently detected in higher concentration than other ARGs at all the sampling sites. The ARGs blaSHV and blaNDM were identified in CRB sediments contaminated by hospital and urban wastewaters. The ARGs abundance strongly correlated (r ≥ 0.36, p wastewaters can act as reservoir of ARGs, which could potentially be transferred to susceptible bacterial pathogens at these sites. PMID:26910062

  3. Hyperproinsulinemia in a three-generation Caucasian family due to mutant proinsulin (Arg{sup 65}{yields}His) not associated with impaired glucose tolerance: The contribution of mutant proinsulin to insulin bioactivity

    Roder, M.E.; Vissing, H. [Health Care Discovery, Bagsvaerd (Denmark); Nauck, M.A. [Ruhr-Univ. Bochum (Germany)

    1996-04-01

    Familial hyperproinsulinemia is a genetic abnormality characterized by an increased proportion of proinsulin immunoreactivity in the circulation due to mutations affecting the posttranslational processing of proinsulin. In affected Japanese families, this has been associated with noninsulin-dependent diabetes mellitus or impaired glucose tolerance. A three-generation Caucasian family with hyperproinsulinemia was identified through unexplained hyperinsulinemia in a normal volunteer participating in a metabolic study. High pressure liquid chromatography analysis of fasting plasma revealed a major peak eluting close to the position of proinsulin. Direct sequencing of the proinsulin gene exon 3 showed a heterozygous point mutation (CGT{yields}CAT) resulting in the substitution of Arg{yields}His in position 65 (corresponding to the AC cleavage site) in the index case, his mother, and his maternal grandmother. All affected subjects had normal oral glucose tolerance. In the basal state and after oral glucose administration, their proinsulin responses were slightly reduced. However, when calculating insulin bioactivity by assuming 9% activity for mutant Arg{sup 65}{yields}His proinsulin, responses in affected subjects were comparable to those in normal subjects. In conclusion, our data demonstrate hyperproinsulinemia in a three-generation Caucasian family due to heterozygous mutant Arg{sup 65}{yields}His proinsulin. This was not associated with impaired glucose tolerance. These results suggest that this mutation in the heterozygous state per se does not affect glucose tolerance and that the biological activity of mutant proinsulin contributes to glucose homeostasis in this family. The association of the same mutation with impaired glucose tolerance or diabetes in previous studies may be the result of selection bias or associated conditions (e.g. the genetic background of the kindreds examined). 29 refs., 5 figs., 3 tabs.

  4. Melanocortin-1 receptor gene variants in four Chinese ethnic populations

    2001-01-01

    There is strong relationship between melanocortin-1 receptor (MC1R) gene variants and human hair color and skin type.Based on a sequencing study of MC1R gene in 50 individuals from the Uygur,Tibetan,Wa and Dai ethnic populations,we discuss the occurrence of 7 mc1r variants consisting of 5 nonsynonymous sites (Val60Leu,Arg67Gln,Val92Met,Arg163Gln and Ala299Val) and 2 synonymous sites (C414T and A942G),among which C414T and Ala299Val were reported for the first time.Confirmation and analysis were also made of 122 individuals at three common point mutations (Val92Met,Arg163Gln,A942G) using PCR-SSCP.The frequency of Arg163Gln variant varies in the four ethnic populations,with percentage of 40%,85.0%,66.2% and 72.7%,respectively,while those of Val92Met and A942G are roughly similar in these four populations.The different environments,migration and admixture of various ethnic groups in China might have impact on the observed frequency of Arg163Gln.

  5. Sulfamethoxazole, tetracycline and oxytetracycline and related antibiotic resistance genes in a large-scale landfill, China.

    Song, Liyan; Li, Lei; Yang, Shu; Lan, Jiwu; He, Haijie; McElmurry, Shawn P; Zhao, Youcai

    2016-05-01

    Landfills are likely to be important reservoirs of antibiotics and antibiotic resistant genes (ARGs) as they receive unused and unwanted antibiotics and ARGs in municipal solid waste (MSW). The distribution, transportation and dynamics of antibiotics and ARGs in landfills remain largely unknown. In the present study, 3 antibiotics - sulfamethoxazole (SMX), tetracycline (TC), and oxytetracycline (OTC) - and their related ARGs (sulI and tetO) were quantified in 51 refuse samples from different depths at 8 locations within a large-scale landfill in central China. The average concentration of OTC was the highest, up to 100.9±141.81μg/kg (dw, n=48), followed by TC (63.8±37.7μg/kg, n=40), and SMX (47.9±8.1μg/kg, n=30). Both sulI and tetO were detected in all samples. Of the ARGs, sul1 (-3.06±1.18, n=51, log10 ARGs/16SrDNA) was more abundant than tetO (-4.37±0.97) in all refuse samples (p<0.05). Both sulI and tetO negatively correlated to refuse age, suggesting they are attenuated during landfill stabilization. OTC and TC positively correlated to tetO (r=0.41, p<0.01) and sulI (r=0.29, p=0.04), respectively. Chemical conditions (e.g. moisture content and nitrate concentrations) within the refuse correlated to antibiotics and ARGs suggesting environmental factors impact the distribution of antibiotics and ARGs in landfill matrix. This study is the first comprehensive in situ landfill study to connect the concentrations of antibiotic residues to ARGs. PMID:26874755

  6. Discharge of swine wastes risks water quality and food safety: Antibiotics and antibiotic resistance genes from swine sources to the receiving environments.

    He, Liang-Ying; Ying, Guang-Guo; Liu, You-Sheng; Su, Hao-Chang; Chen, Jun; Liu, Shuang-Shuang; Zhao, Jian-Liang

    2016-01-01

    Swine feedlots are widely considered as a potential hotspot for promoting the dissemination of antibiotic resistance genes (ARGs) in the environment. ARGs could enter the environment via discharge of animal wastes, thus resulting in contamination of soil, water, and food. We investigated the dissemination and diversification of 22 ARGs conferring resistance to sulfonamides, tetracyclines, chloramphenicols, and macrolides as well as the occurrence of 18 corresponding antibiotics from three swine feedlots to the receiving water, soil environments and vegetables. Most ARGs and antibiotics survived the on-farm waste treatment processes in the three swine farms. Elevated diversity of ARGs was observed in the receiving environments including river water and vegetable field soils when compared with respective controls. The variation of ARGs along the vertical soil profiles of vegetable fields indicated enrichment and migration of ARGs. Detection of various ARGs and antibiotic residues in vegetables fertilized by swine wastes could be of great concern to the general public. This research demonstrated the contribution of swine wastes to the occurrence and development of antibiotic resistance determinants in the receiving environments and potential risks to food safety and human health. PMID:27107226

  7. Effects of Arg26 and Lys27 mutation on the bioactivity of HNTX-Ⅳ

    XIONG Xia; XU Xia; LI Dongling; CHEN Ping; LIANG Songping

    2007-01-01

    Hainantoxin-Ⅳ (HNTX-Ⅳ)was isolated from the Chinese bird spider Ornithoctorcs hainana and identified as a novel antagonist of tetrodotoxin-sensitive (TTX-S)sodium channels.As revealed by the solution structure of HNTX-Ⅳ solved by two-dimensional nuclear magnetic resonance (2D-NMR),HNTX-Ⅳ adopts an inhibitor cystine knot motif.To check the role of basic residues during HNTX-Ⅳ's interaction with TTX-S sodium channels,R26A and K27A mutants of HNTX-Ⅳ were constructed by solid-phase chemical synthesis.The synthesized peptides were purified and refolded under optimized oxidation conditions.Correct synthesis and folding were confirmed by MALDI-TOF mass spectrometry and NMR spectroscopy,respectively.Using the whole-cell patch-clamp technique,Lys27 but not Arg26 was identified as a key residue for HNTX-Ⅳ's bioactivity against TTX-S sodium channels,because R26A-HNTX-Ⅳ showed slightly reduced activity and K27A-HNTX-Ⅳ showed almost no inhibition.

  8. CLONAL AND PLANTING DENSITY EFFECTS ON SOME PROPERTIES OF RUBBER WOOD (HEVEA BRASILIENSIS MUELL. ARG.

    Hamid Reza Naji,

    2011-10-01

    Full Text Available Inter-clonal and intra-clonal wood properties and their variations from pith to bark were evaluated for wood density and anatomical features on rubber wood (Hevea brasiliensis Muell. Arg from a 9-year-old plantation with planting densities of 500 and 2000 trees per hectare comprised of clones RRIM 2020 and RRIM 2025. Planting density had uneven effects on wood density and wood cell features. Intra-clonal and inter-clonal variations were significant for wood density in both clones and planting densities. Wood density demonstrated an increasing trend in the radial direction. However, at the lower planting density wood density near the bark decreased slightly. Fiber diameter, lumen diameter, and cell wall thickness showed an increasing trend from pith to bark. Best average fiber characteristics were observed at the lower planting density in clone RRIM 2025. Vessel frequency had a direct relationship with planting density in that it was higher in the higher planting density of 2000 trees per hectare. Overall, planting density had a significant effect on wood quality. The properties of clone RRIM 2025 were found to be comparatively better with longer fiber length and higher wood density than those of RRIM 2020.

  9. The tetrapeptide Arg-Leu-Tyr-Glu inhibits VEGF-induced angiogenesis

    Baek, Yi-Yong; Lee, Dong-Keon [Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); So, Ju-Hoon; Kim, Cheol-Hee [Department of Biology, Chungnam National University, Daejeon, 305-764 (Korea, Republic of); Jeoung, Dooil [Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); Lee, Hansoo [Department of Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); Choe, Jongseon [Department of Immunology, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); Won, Moo-Ho [Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); Ha, Kwon-Soo [Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of); Kwon, Young-Guen [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, 120-752 (Korea, Republic of); Kim, Young-Myeong, E-mail: ymkim@kangwon.ac.kr [Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, 200-702 (Korea, Republic of)

    2015-08-07

    Kringle 5, derived from plasminogen, is highly capable of inhibiting angiogenesis. Here, we have designed and synthesized 10 tetrapeptides, based on the amino acid properties of the core tetrapeptide Lys-Leu-Tyr-Asp (KLYD) originating from anti-angiogenic kringle 5 of human plasminogen. Of these, Arg-Leu-Tyr-Glu (RLYE) effectively inhibited vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation, migration and tube formation, with an IC{sub 50} of 0.06–0.08 nM, which was about ten-fold lower than that of the control peptide KLYD (0.79 nM), as well as suppressed developmental angiogenesis in a zebrafish model. Furthermore, this peptide effectively inhibited the cellular events that precede angiogenesis, such as ERK and eNOS phosphorylation and nitric oxide production, in endothelial cells stimulated with VEGF. Collectively, these data demonstrate that RLYE is a potent anti-angiogenic peptide that targets the VEGF signaling pathway. - Highlights: • The tetrapeptide RLYE inhibited VEGF-induced angiogenesis in vitro. • RLYE also suppressed neovascularization in a zebrafish model. • Its effect was correlated with inhibition of VEGF-induced ERK and eNOS activation. • RLYE may be used as a therapeutic drug for angiogenesis-related diseases.

  10. The tetrapeptide Arg-Leu-Tyr-Glu inhibits VEGF-induced angiogenesis

    Kringle 5, derived from plasminogen, is highly capable of inhibiting angiogenesis. Here, we have designed and synthesized 10 tetrapeptides, based on the amino acid properties of the core tetrapeptide Lys-Leu-Tyr-Asp (KLYD) originating from anti-angiogenic kringle 5 of human plasminogen. Of these, Arg-Leu-Tyr-Glu (RLYE) effectively inhibited vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation, migration and tube formation, with an IC50 of 0.06–0.08 nM, which was about ten-fold lower than that of the control peptide KLYD (0.79 nM), as well as suppressed developmental angiogenesis in a zebrafish model. Furthermore, this peptide effectively inhibited the cellular events that precede angiogenesis, such as ERK and eNOS phosphorylation and nitric oxide production, in endothelial cells stimulated with VEGF. Collectively, these data demonstrate that RLYE is a potent anti-angiogenic peptide that targets the VEGF signaling pathway. - Highlights: • The tetrapeptide RLYE inhibited VEGF-induced angiogenesis in vitro. • RLYE also suppressed neovascularization in a zebrafish model. • Its effect was correlated with inhibition of VEGF-induced ERK and eNOS activation. • RLYE may be used as a therapeutic drug for angiogenesis-related diseases

  11. Validation of Community Models: 2. Development of a Baseline, Using the Wang-Sheeley-Arge Model

    MacNeice, Peter

    2009-01-01

    This paper is the second in a series providing independent validation of community models of the outer corona and inner heliosphere. Here I present a comprehensive validation of the Wang-Sheeley-Arge (WSA) model. These results will serve as a baseline against which to compare the next generation of comparable forecasting models. The WSA model is used by a number of agencies to predict Solar wind conditions at Earth up to 4 days into the future. Given its importance to both the research and forecasting communities, it is essential that its performance be measured systematically and independently. I offer just such an independent and systematic validation. I report skill scores for the model's predictions of wind speed and interplanetary magnetic field (IMF) polarity for a large set of Carrington rotations. The model was run in all its routinely used configurations. It ingests synoptic line of sight magnetograms. For this study I generated model results for monthly magnetograms from multiple observatories, spanning the Carrington rotation range from 1650 to 2074. I compare the influence of the different magnetogram sources and performance at quiet and active times. I also consider the ability of the WSA model to forecast both sharp transitions in wind speed from slow to fast wind and reversals in the polarity of the radial component of the IMF. These results will serve as a baseline against which to compare future versions of the model as well as the current and future generation of magnetohydrodynamic models under development for forecasting use.

  12. Monitoring and evaluation of the water quality of Budeasa Reservoir-Arges River, Romania.

    Ion, Antoanela; Vladescu, Luminita; Badea, Irinel Adriana; Comanescu, Laura

    2016-09-01

    The purpose of this study was to monitor and record the specific characteristics and properties of the Arges River water in the Budeasa Reservoir (the principal water resources of municipal tap water of the big Romanian city Pitesti and surrounding area) for a period of 5 years (2005-2009). The monitored physical and chemical parameters were turbidity, pH, electrical conductivity, chemical oxygen demand, 5 days biochemical oxygen demand, free dissolved oxygen, nitrite, nitrate, ammonia nitrogen, chloride, total dissolved iron ions, sulfate, manganese, phosphate, total alkalinity, and total hardness. The results were discussed in correlation with the precipitation values during the study. Monthly and annual values of each parameter determined in the period January 2005-December 2009 were used as a basis for the classification of Budeasa Reservoir water, according to the European legislation, as well as for assessing its quality as a drinking water supply. Principal component analysis and Pearson correlation coefficients were used as statistical procedures in order to evaluate the data obtained during this study. PMID:27566323

  13. Primary structure of the precursor for the sea anemone neuropeptide Antho-RFamide (less than Glu-Gly-Arg-Phe-NH2)

    Darmer, D; Schmutzler, C; Diekhoff, D; Grimmelikhuijzen, C J

    1991-01-01

    Neuropeptides containing the carboxylterminal sequence Arg-Phe-NH2 are found throughout the animal kingdom and are important substances mediating neuronal communication. Here, we have cloned the cDNA coding for the precursor protein of the sea anemone neuropeptide (Antho-RFamide) less than Glu......-Gly-Arg-Phe-NH2. This precursor is 334 amino acids in length and contains 19 copies of unprocessed Antho-RFamide (Gln-Gly-Arg-Phe-Gly), which are tandemly arranged in the C-terminal part of the protein. Paired basic residues (Lys-Arg) or single basic residues (Arg) occur at the C-terminal side of each Antho...... glutamic acid residues. Such processing is, to our knowledge, hitherto unknown for peptidergic neurons. The Antho-RFamide precursor also contains two copies of the putative Antho-RFamide-related peptide Phe-Gln-Gly-Arg-Phe-NH2 and one copy of Tyr-Val-Pro-Gly-Arg-Tyr-NH2. In addition, the precursor protein...

  14. Floral biology and breeding system of Psychotria tenuinervis Muell. Arg. (Rubiaceae) in the Atlantic rain forest, SE Brazil Biologia floral e sistema reprodutivo de Psychotria tenuinervis Muell. Arg. (Rubiaceae) na Floresta Atlântica, SE Brasil

    Carolina Bernucci Virillo; Flavio Nunes Ramos; Cibele Cardoso de Castro; João Semir

    2007-01-01

    (Floral biology and breeding system oi Psychotria tenuinervis Muell. Arg. (Rubiaceae) in the Atlantic rain forest, SE Brazil). The aim of this study was to investigate pollination biology, floral morphometry, morph ratio and breeding system oiPsychotria tenuinervis in an area of Atlantic rain forest in southeastern Brazil. Pollination biology was studied based on focal observations and the breeding system was determined using controlled crosses; data on flower production and floral morphometr...

  15. Deletion of Genes Encoding Arginase Improves Use of "Heavy" Isotope-Labeled Arginine for Mass Spectrometry in Fission Yeast.

    Weronika E Borek

    Full Text Available The use of "heavy" isotope-labeled arginine for stable isotope labeling by amino acids in cell culture (SILAC mass spectrometry in the fission yeast Schizosaccharomyces pombe is hindered by the fact that under normal conditions, arginine is extensively catabolized in vivo, resulting in the appearance of "heavy"-isotope label in several other amino acids, most notably proline, but also glutamate, glutamine and lysine. This "arginine conversion problem" significantly impairs quantification of mass spectra. Previously, we developed a method to prevent arginine conversion in fission yeast SILAC, based on deletion of genes involved in arginine catabolism. Here we show that although this method is indeed successful when (13C6-arginine (Arg-6 is used for labeling, it is less successful when (13C6(15N4-arginine (Arg-10, a theoretically preferable label, is used. In particular, we find that with this method, "heavy"-isotope label derived from Arg-10 is observed in amino acids other than arginine, indicating metabolic conversion of Arg-10. Arg-10 conversion, which severely complicates both MS and MS/MS analysis, is further confirmed by the presence of (13C5(15N2-arginine (Arg-7 in arginine-containing peptides from Arg-10-labeled cells. We describe how all of the problems associated with the use of Arg-10 can be overcome by a simple modification of our original method. We show that simultaneous deletion of the fission yeast arginase genes car1+ and aru1+ prevents virtually all of the arginine conversion that would otherwise result from the use of Arg-10. This solution should enable a wider use of heavy isotope-labeled amino acids in fission yeast SILAC.

  16. Prevalence and proliferation of antibiotic resistance genes in two municipal wastewater treatment plants.

    Mao, Daqing; Yu, Shuai; Rysz, Michal; Luo, Yi; Yang, Fengxia; Li, Fengxiang; Hou, Jie; Mu, Quanhua; Alvarez, P J J

    2015-11-15

    The propagation of antibiotic resistance genes (ARGs) is an emerging health concern worldwide. Thus, it is important to understand and mitigate their occurrence in different systems. In this study, 30 ARGs that confer resistance to tetracyclines, sulfonamides, quinolones or macrolides were detected in two activated sludge wastewater treatment plants (WWTPs) in northern China. Bacteria harboring ARGs persisted through all treatment units, and survived disinfection by chlorination in greater percentages than total Bacteria (assessed by 16S rRNA genes). Although the absolute abundances of ARGs were reduced from the raw influent to the effluent by 89.0%-99.8%, considerable ARG levels [(1.0 ± 0.2) × 10(3) to (9.5 ± 1.8) × 10(5) copies/mL)] were found in WWTP effluent samples. ARGs were concentrated in the waste sludge (through settling of bacteria and sludge dewatering) at (1.5 ± 2.3) × 10(9) to (2.2 ± 2.8) × 10(11) copies/g dry weight. Twelve ARGs (tetA, tetB, tetE, tetG, tetH, tetS, tetT, tetX, sul1, sul2, qnrB, ermC) were discharged through the dewatered sludge and plant effluent at higher rates than influent values, indicating overall proliferation of resistant bacteria. Significant antibiotic concentrations (2%-50% of raw influent concentrations) remained throughout all treatment units. This apparently contributed selective pressure for ARG replication since the relative abundance of resistant bacteria (assessed by ARG/16S rRNA gene ratios) was significantly correlated to the corresponding effluent antibiotic concentrations. Similarly, the concentrations of various heavy metals (which induce a similar bacterial resistance mechanism as antibiotics - efflux pumps) were also correlated to the enrichment of some ARGs. Thus, curtailing the release of antibiotics and heavy metals to sewage systems (or enhancing their removal in pre-treatment units) may alleviate their selective pressure and mitigate ARG proliferation in WWTPs. PMID:26372743

  17. P53 codon 11, 72, and 248 gene polymorphisms in endometriosis

    Hsieh, Yao-Yuan; Lin, Chich-Sheng

    2006-01-01

    Objective: Mutated p53 gene is related to the instability of cell growth and cell cycle progression. We aimed to evaluate the association between endometriosis and p53 codon 11, 72 and 248 gene polymorphisms. Patients and methods: Women were divided into two groups: (1) moderate/severe endometriosis (n=148), and (2) non-endometriosis groups (n=150). P53 gene polymorphisms include codon11 Glu/Gln or Lys (GAG->CAG or AAG), codon 72 Arg/Pro (CGC->CCC), and codon 248 Arg/Thr (CGG->TCG). These gen...

  18. Housefly Larva Vermicomposting Efficiently Attenuates Antibiotic Resistance Genes in Swine Manure, with Concomitant Bacterial Population Changes.

    Wang, Hang; Li, Hongyi; Gilbert, Jack A; Li, Haibo; Wu, Longhua; Liu, Meng; Wang, Liling; Zhou, Qiansheng; Yuan, Junxiang; Zhang, Zhijian

    2015-11-01

    Manure from swine treated with antimicrobials as feed additives is a major source for the expansion of the antibiotic resistance gene (ARG) reservoir in the environment. Vermicomposting via housefly larvae (Musca domestica) can be efficiently used to treat manure and regenerate biofertilizer, but few studies have investigated its effect on ARG attenuation. Here, we tracked the abundances of 9 ARGs and the composition and structure of the bacterial communities in manure samples across 6 days of full-scale manure vermicomposting. On day 6, the abundances of genes encoding tetracycline resistance [tet(M), tet(O), tet(Q), and tet(W)] were reduced (P vermicomposting, accompanied by a 100 times increase in the relative abundance of Flavobacteriaceae spp. Variations in the abundances of ARGs were correlated with the changing microbial community structure and the relative abundances of the family Ruminococcaceae, class Bacilli, or phylum Proteobacteria. Vermicomposting, as a waste management practice, can reduce the overall abundance of ARGs. More research is warranted to assess the use of this waste management practice as a measure to attenuate the dissemination of antimicrobial residues and ARGs from livestock production before vermicompost can be safely used as biofertilizer in agroecosystems. PMID:26296728

  19. Does human activity impact the natural antibiotic resistance background? Abundance of antibiotic resistance genes in 21 Swiss lakes.

    Czekalski, Nadine; Sigdel, Radhika; Birtel, Julia; Matthews, Blake; Bürgmann, Helmut

    2015-08-01

    Antibiotic resistance genes (ARGs) are emerging environmental contaminants, known to be continuously discharged into the aquatic environment via human and animal waste. Freshwater aquatic environments represent potential reservoirs for ARG and potentially allow sewage-derived ARG to persist and spread in the environment. This may create increased opportunities for an eventual contact with, and gene transfer to, human and animal pathogens via the food chain or drinking water. However, assessment of this risk requires a better understanding of the level and variability of the natural resistance background and the extent of the human impact. We have analyzed water samples from 21 Swiss lakes, taken at sampling points that were not under the direct influence of local contamination sources and analyzed the relative abundance of ARG using quantitative real-time PCR. Copy numbers of genes mediating resistance to three different broad-spectrum antibiotic classes (sulfonamides: sul1, sul2, tetracyclines: tet(B), tet(M), tet(W) and fluoroquinolones: qnrA) were normalized to copy numbers of bacterial 16S rRNA genes. We used multiple linear regression to assess if ARG abundance is related to human activities in the catchment, microbial community composition and the eutrophication status of the lakes. Sul genes were detected in all sampled lakes, whereas only four lakes contained quantifiable numbers of tet genes, and qnrA remained below detection in all lakes. Our data indicate higher abundance of sul1 in lakes with increasing number and capacity of wastewater treatment plants (WWTPs) in the catchment. sul2 abundance was rather related to long water residence times and eutrophication status. Our study demonstrates the potential of freshwater lakes to preserve antibiotic resistance genes, and provides a reference for ARG abundance from lake systems with low human impact as a baseline for assessing ARG contamination in lake water. PMID:25913323

  20. Fate and proliferation of typical antibiotic resistance genes in five full-scale pharmaceutical wastewater treatment plants

    Wang, Jilu [College of Environmental Science and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Nankai University, Tianjin 300071 (China); Mao, Daqing, E-mail: mao@tju.edu.cn [School of Environmental Science and Engineering, Tianjin University, Tianjin 300072 (China); Mu, Quanhua [College of Environmental Science and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Nankai University, Tianjin 300071 (China); Luo, Yi, E-mail: luoy@nankai.edu.cn [College of Environmental Science and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Nankai University, Tianjin 300071 (China)

    2015-09-01

    This study investigated the characteristics of 10 subtypes of antibiotic resistance genes (ARGs) for sulfonamide, tetracycline, β-lactam and macrolide resistance and the class 1 integrase gene (intI1). In total, these genes were monitored in 24 samples across each stage of five full-scale pharmaceutical wastewater treatment plants (PWWTPs) using qualitative and real-time quantitative polymerase chain reactions (PCRs). The levels of typical ARG subtypes in the final effluents ranged from (2.08 ± 0.16) × 10{sup 3} to (3.68 ± 0.27) × 10{sup 6} copies/mL. The absolute abundance of ARGs in effluents accounted for only 0.6%–59.8% of influents of the five PWWTPs, while the majority of the ARGs were transported to the dewatered sludge with concentrations from (9.38 ± 0.73) × 10{sup 7} to (4.30 ± 0.81) × 10{sup 10} copies/g dry weight (dw). The total loads of ARGs discharged through dewatered sludge was 7–308 folds higher than that in the raw influents and 16–638 folds higher than that in the final effluents. The proliferation of ARGs mainly occurs in the biological treatment processes, such as conventional activated sludge, cyclic activated sludge system (CASS) and membrane bio-reactor (MBR), implying that significant replication of certain subtypes of ARGs may be attributable to microbial growth. High concentrations of antibiotic residues (ranging from 0.14 to 92.2 mg/L) were detected in the influents of selected wastewater treatment systems and they still remain high residues in the effluents. Partial correlation analysis showed significant correlations between the antibiotic concentrations and the associated relative abundance of ARG subtypes in the effluent. Although correlation does not prove causation, this study demonstrates that in addition to bacterial growth, the high antibiotic residues within the pharmaceutical WWTPs may influence the proliferation and fate of the associated ARG subtypes. - Highlights: • The ARGs in final discharges were 7

  1. Fate and proliferation of typical antibiotic resistance genes in five full-scale pharmaceutical wastewater treatment plants

    This study investigated the characteristics of 10 subtypes of antibiotic resistance genes (ARGs) for sulfonamide, tetracycline, β-lactam and macrolide resistance and the class 1 integrase gene (intI1). In total, these genes were monitored in 24 samples across each stage of five full-scale pharmaceutical wastewater treatment plants (PWWTPs) using qualitative and real-time quantitative polymerase chain reactions (PCRs). The levels of typical ARG subtypes in the final effluents ranged from (2.08 ± 0.16) × 103 to (3.68 ± 0.27) × 106 copies/mL. The absolute abundance of ARGs in effluents accounted for only 0.6%–59.8% of influents of the five PWWTPs, while the majority of the ARGs were transported to the dewatered sludge with concentrations from (9.38 ± 0.73) × 107 to (4.30 ± 0.81) × 1010 copies/g dry weight (dw). The total loads of ARGs discharged through dewatered sludge was 7–308 folds higher than that in the raw influents and 16–638 folds higher than that in the final effluents. The proliferation of ARGs mainly occurs in the biological treatment processes, such as conventional activated sludge, cyclic activated sludge system (CASS) and membrane bio-reactor (MBR), implying that significant replication of certain subtypes of ARGs may be attributable to microbial growth. High concentrations of antibiotic residues (ranging from 0.14 to 92.2 mg/L) were detected in the influents of selected wastewater treatment systems and they still remain high residues in the effluents. Partial correlation analysis showed significant correlations between the antibiotic concentrations and the associated relative abundance of ARG subtypes in the effluent. Although correlation does not prove causation, this study demonstrates that in addition to bacterial growth, the high antibiotic residues within the pharmaceutical WWTPs may influence the proliferation and fate of the associated ARG subtypes. - Highlights: • The ARGs in final discharges were 7–308 times higher than that

  2. Three novel and the common Arg677Ter RP1 protein truncating mutations causing autosomal dominant retinitis pigmentosa in a Spanish population

    Antiñolo Guillermo

    2006-04-01

    Full Text Available Abstract Background Retinitis pigmentosa (RP, a clinically and genetically heterogeneous group of retinal degeneration disorders affecting the photoreceptor cells, is one of the leading causes of genetic blindness. Mutations in the photoreceptor-specific gene RP1 account for 3–10% of cases of autosomal dominant RP (adRP. Most of these mutations are clustered in a 500 bp region of exon 4 of RP1. Methods Denaturing gradient gel electrophoresis (DGGE analysis and direct genomic sequencing were used to evaluate the 5' coding region of exon 4 of the RP1 gene for mutations in 150 unrelated index adRP patients. Ophthalmic and electrophysiological examination of RP patients and relatives according to pre-existing protocols were carried out. Results Three novel disease-causing mutations in RP1 were detected: Q686X, K705fsX712 and K722fsX737, predicting truncated proteins. One novel missense mutation, Thr752Met, was detected in one family but the mutation does not co-segregate in the family, thereby excluding this amino acid variation in the protein as a cause of the disease. We found the Arg677Ter mutation, previously reported in other populations, in two independent families, confirming that this mutation is also present in a Spanish population. Conclusion Most of the mutations reported in the RP1 gene associated with adRP are expected to encode mutant truncated proteins that are approximately one third or half of the size of wild type protein. Patients with mutations in RP1 showed mild RP with variability in phenotype severity. We also observed several cases of non-penetrant mutations.

  3. BDNF-induced LTP is associated with rapid Arc/Arg3.1-dependent enhancement in adult hippocampal neurogenesis.

    Kuipers, Sjoukje D; Trentani, Andrea; Tiron, Adrian; Mao, Xiaosong; Kuhl, Dietmar; Bramham, Clive R

    2016-01-01

    Adult neurogenesis in the hippocampus is a remarkable phenomenon involved in various aspects of learning and memory as well as disease pathophysiology. Brain-derived neurotrophic factor (BDNF) represents a major player in the regulation of this unique form of neuroplasticity, yet the mechanisms underlying its pro-neurogenic actions remain unclear. Here, we examined the effects associated with brief (25 min), unilateral infusion of BDNF in the rat dentate gyrus. Acute BDNF infusion induced long-term potentiation (LTP) of medial perforant path-evoked synaptic transmission and, concomitantly, enhanced hippocampal neurogenesis bilaterally, reflected by increased dentate gyrus BrdU + cell numbers. Importantly, inhibition of activity-regulated cytoskeleton-associated protein (Arc/Arg3.1) translation through local, unilateral infusion of anti-sense oligodeoxynucleotides (ArcAS) prior to BDNF infusion blocked both BDNF-LTP induction and the associated pro-neurogenic effects. Notably, basal rates of proliferation and newborn cell survival were unaltered in homozygous Arc/Arg3.1 knockout mice. Taken together these findings link the pro-neurogenic effects of acute BDNF infusion to induction of Arc/Arg3.1-dependent LTP in the adult rodent dentate gyrus. PMID:26888068

  4. Pharmacobotanic characterization of young stems and stem barks of Rauvolfia sellowii Müll. Arg., Apocynaceae

    Leopoldo Clemente Baratto

    2010-09-01

    Full Text Available Rauvolfia sellowii Müll. Arg. (Apocynaceae, a Brazilian native tree rich in indole alkaloids, is known as "pau-pra-tudo" and popularly used as hypocholesterolemic, hypoglycemic and antihypertensive. The aim of this work was to study the anatomy of the young stems and stem barks of this medicinal plant, in order to contribute to the identification of the species as a drug. The plant material was fixed and prepared according to standard microtechniques. The young stems have remaining epidermis, but a suberified peridermis is evident. The phellogen is located in the cortical region, forming suber externally. Underneath the phellogen, lies the phelloderm and collenchymatic region. In the cortex, there are numerous laticifers and some fibers. There is an incomplete sclerenchymatic sheath, consisting of several groups of fibers and stone cells. The stem has internal phloem ordered as isolated groups side by side. Numerous laticifers, calcium oxalate crystals, idioblasts and amyloplasts are found in the cortex, phloem, xylem and pith. The stem bark has many layers of suber and cortical parenchyma, a sheath composed of fibers and stone cells totally lignified, and external phloem. These anatomical characteristic, taken together, can be used as quality control parameters for this species.Rauvolfia sellowii Müll. Arg. (Apocynaceae, uma árvore nativa brasileira rica em alcaloides indólicos, é conhecida como "pau-pra-tudo" e utilizada popularmente como hipocolesterolêmica, hipoglicêmica e anti-hipertensiva. O presente trabalho teve como objetivo caracterizar anatomicamente o caule jovem e a casca caulinar dessa planta medicinal, a fim de contribuir para a identificação e autenticidade da droga. O material vegetal foi fixado e submetido às microtécnicas usuais. O caule jovem possui epiderme remanescente, porém uma periderme suberificada é observada. O felogênio instala-se na região cortical, formando súber externamente. Subjacentes ao felog

  5. Bioinformatic analysis of an unusual gene-enzyme relationship in the arginine biosynthetic pathway among marine gamma proteobacteria: implications concerning the formation of N-acetylated intermediates in prokaryotes.

    Labedan Bernard; Glansdorff Nicolas; Xu Ying

    2006-01-01

    Abstract Background The N-acetylation of L-glutamate is regarded as a universal metabolic strategy to commit glutamate towards arginine biosynthesis. Until recently, this reaction was thought to be catalyzed by either of two enzymes: (i) the classical N-acetylglutamate synthase (NAGS, gene argA) first characterized in Escherichia coli and Pseudomonas aeruginosa several decades ago and also present in vertebrates, or (ii) the bifunctional version of ornithine acetyltransferase (OAT, gene argJ)...

  6. Affinity capture of (Arg sup 8 )vasopressin-receptor complex using immobilized antisense peptide

    Feng Xian Lu; Aiyar, N.; Chaiken, I. (SmithKline Beecham, King of Prussia, PA (United States))

    1991-05-01

    Solubilized noncovalent complexes of (Arg{sup 8})-vasopressin (AVP) with receptor proteins from rat liver membranes were isolated by selective binding to silica-immobilized antisense (AS) peptide. The affinity chromatographic support was prepared with a chemically synthesized AS peptide whose sequence is encoded by the AS DNA corresponding to the 20 amino-terminal residues of the AVP bovine neurophysin II biosynthetic precursor (pro-AVP/BNPII-(20-1)), region that includes the AVP sequence at residues 1-9. The AS peptide-AVP interaction mechanism hypothesized, contact by hydropathic complementarity at multiple sites along the peptide chains, led to the prediction that AVP bound to its receptor would still have enough free surface to interact with immobilized AS peptide. To test this prediction of a three-way interaction, ({sup 3}H)AVP-receptor was obtained as a solubilized, partially purified fraction from rat liver membrane. Covalently crosslinked ({sup 3}H)AVP complex also was bound to the AS peptide column; binding was blocked by competition with unlabeled AVP in the elution buffer. Since the AVP-linked 31- and 38-kDa proteins have the same apparent molecular mass on SDS/PAGE as found previously by photoaffinity labeling, the authors conclude that the AS peptide column has affinity-captured AVP-receptor complexes. The 15-kDa protein appears to be an active AVP-receptor fragment of one or both of the larger proteins. It is generally concluded that immobilized AS peptides may be useful to isolate peptide and protein receptor complexes in other systems as well.

  7. Laxative activities of Mareya micrantha (Benth. Müll. Arg. (Euphorbiaceae leaf aqueous extract in rats

    Djaman Joseph A

    2010-02-01

    Full Text Available Abstract Background Mareya micrantha (Benth. Müll. Arg. (Euphorbiaceae is a shrub that is commonly used in Côte d'Ivoire (West Africa for the treatment of constipation and as an ocytocic drug. The present study was carried out to investigate the laxative activity of Mareya micrantha in albino's Wistar rats. Methods Rats were divided in 5 groups of 5 animals each, first group as control, second group served as standard (sodium picosulfate while group 3, 4 and 5 were treated with leaf aqueous extract of Mareya micrantha at doses of 100, 200 and 400 mg/kg body weight (b.w., per os respectively. The laxative activity was determined based on the weight of the faeces matter. The effects of the leaves aqueous extract of Mareya micrantha and castor oil were also evaluated on intestinal transit, intestinal fluid accumulation and ions secretion. Results Phytochemicals screening of the extract revealed the presence of flavonoids, alkaloids, tannins, polyphenols, sterols and polyterpenes. The aqueous extract of Mareya micrantha applied orally (100, 200 and 400 mg/kg; p.o., produced significant laxative activity and reduced loperamide induced constipation in dose dependant manner. The effect of the extract at 200 and 400 mg/kg (p.o. was similar to that of reference drug sodium picosulfate (5 mg/kg, p.o. The same doses of the extract (200 and 400 mg/kg, p.o. produced a significant increase (p -, Na+, K+ and Ca2+ in the intestinal fluid (p Conclusions The results showed that the aqueous extract of Mareya micrantha has a significant laxative activity and supports its traditional use in herbal medicine.

  8. Changes of resistome, mobilome and potential hosts of antibiotic resistance genes during the transformation of anaerobic digestion from mesophilic to thermophilic.

    Tian, Zhe; Zhang, Yu; Yu, Bo; Yang, Min

    2016-07-01

    This study aimed to reveal how antibiotic resistance genes (ARGs) and their horizontal and vertical transfer-related items (mobilome and bacterial hosts) respond to the transformation of anaerobic digestion (AD) from mesophilic to thermophilic using one-step temperature increase. The resistomes and mobilomes of mesophilic and thermophilic sludge were investigated using metagenome sequencing, and the changes in 24 representative ARGs belonging to three categories, class 1 integron and bacterial genera during the transition period were further followed using quantitative PCR and 454-pyrosequencing. After the temperature increase, resistome abundance in the digested sludge decreased from 125.97 ppm (day 0, mesophilic) to 50.65 ppm (day 57, thermophilic) with the reduction of most ARG types except for the aminoglycoside resistance genes. Thermophilic sludge also had a smaller mobilome, including plasmids, insertion sequences and integrons, than that of mesophilic sludge, suggesting the lower horizontal transfer potential of ARGs under thermophilic conditions. On the other hand, the total abundance of 18 bacterial genera, which were suggested as the possible hosts for 13 ARGs through network analysis, decreased from 23.27% in mesophilic sludge to 11.92% in thermophilic sludge, indicating fewer hosts for the vertical expansion of ARGs after the increase in temperature. These results indicate that the better reduction of resistome abundance by thermophilic AD might be associated with the decrease of both the horizontal and vertical transferability of ARGs. PMID:27108212

  9. Impact of GNB3-C825T, ADRB3-Trp64Arg, UCP2-3′UTR 45 bp del/ins, and PPARγ-Pro12Ala Polymorphisms on Bofutsushosan Response in Obese Subjects: A Randomized, Double-Blind, Placebo-Controlled Trial

    Park, Junghyun; Bose, Shambhunath; Hong, Sun-Woo; Lee, Dong-Ki; Yoo, Jae-Wook; Lim, Chi-Yeon; Lee, Myeongjong; Kim, Hojun

    2014-01-01

    Obesity is known to be influenced by a number of genes, including the β3 subunit of G protein (GNB3), β3-adrenergic receptor (ADRB3), uncoupling protein 2 (UCP2), and peroxisome proliferator activated receptor gamma (PPARγ). The single nucleotide polymorphisms (SNPs) of the above genes, such as GNB3-C825T, ADRB3-Trp64Arg, UCP2-3′UTR 45 bp del/ins, and PPARγ-Pro12Ala, are associated with obesity and body mass index. The present study evaluates the impact of Bofutsushosan, a traditional Eastern...

  10. Polymorphisms in the Melanocortin-1 Receptor (Mc1R Gene in Vitiligo

    Eylem Acar

    2012-03-01

    Full Text Available Objective: Vitiligo is a progressive skin pigmentation disorder, which may be acquired or hereditary, frequently seen, and may influence every age group. The melanocortin 1 receptor (MC1R gene is a major determinant of human pigmentation. In our study, polymorphic differences of the MC1R gene at the DNA level has been investigated in patients with vitiligo.Materials and Methods: In our study, polymorphic differences of the MC1R gene at the DNA level has been investigated in vitiligo 60 patients, whose families had resided in the Thrace region of Turkey for at least three generations. The 60 volunteer healthy individuals have no other systemic and dermatological disease.Results: Totally, five types of Single Nucleotide Polymorphism (SNP were found in each case and control groups: Val60Leu (G178T, Val92Met (G274A, Arg151Cys (C451T, Arg160Trp (C478T, and Arg163Gln (G488A. Comparing both groups in terms of genotype frequencies, no statistically meaningful difference was detected (p>0.05. However, assessing in terms of allele frequencies, a meaningful difference was found in the Arg163Gln (G488A allele statistically in favor of the control group (p<0.05.Conclusion: It has been found in our study population that the MC1R gene Arg163Gln (G488A allele may be a protective factor for vitiligo.

  11. The association between changes in synovial fluid levels of ARGS-aggrecan fragments, progression of radiographic osteoarthritis and self-reported outcomes: a cohort study

    Larsson, S; Englund, M; Struglics, A; Lohmander, L S

    2012-01-01

    subjects at two time points after meniscectomy. Time point A was on average 18years after meniscectomy, time point B was on average 7.5years later; 74 subjects had SF available from both examinations. We measured SF ARGS by an electrochemiluminescence immunoassay, graded radiographic features of...... between examinations was increased 6- and 4-fold respectively compared to those increasing in SF ARGS (odds ratio (OR) 5.72; 95% confidence interval (CI) 1.53-21.4 and 3.66; 1.01-13.2, respectively). No significant associations were seen between decreasing SF ARGS and progression of osteophytes (OR 0.......88; 0.28-2.78), or for patient-reported outcomes other than KOOS pain. CONCLUSION: Having decreasing levels of SF ARGS over time was associated with an increased risk of loss of joint space and pain worsening, but showed no association with other patient-reported outcomes or osteophyte progression....

  12. STUDY ON INVOLVEMENT OF L-ARG-NO IN THE PROTECTIVE ACTION OF ELECTROACUPUNCTURE ON CEREBRAL ISCHEMIC INJURY IN THE RAT

    MaYang; XuNenggui; XuGuansun; ZhuShunli; SunYong; ZhangDaoqin; MaQinyun

    2000-01-01

    In the present study, the involvement of L-arginine(L-Arg) NO on ihe protective action of electroacupuncture(EA) on cerebral ischemic injury was observed in acute ischemia-reperfusion(IR) rat model by taking regional cerebral blood flow(r-CBF), cerebral water content(CWC), and blood nitric oxide(NO) contents as indexes. Results showed that 1) EA could cause r-CBF and serum NO content to increase and CWC to lower, suggesting an protective action of EA on IR cerebral injury; 2) intravenous injection of L-Arg also had an protective effect on cerebral IR cerebral injury, while L-NNA had no this effect; and 3) pre-treatment with L-Arg might strengthen the effect of EA further, while pretreatment with L-NNA could weaken its effect. It indicates that L-Arg-NO may be involved in the effect of EA in protecting the brain from ischemic injury.

  13. STUDY ON INVOLVEMENT OF L-ARG-NO IN THE PROTECTIVE ACTION OF ELECTROACUPUNCTURE ON CEREBRAL ISCHEMIC INJURY IN THE RAT

    马扬; 许能贵; 许冠荪; 朱舜丽; 孙勇; 张道芹; 马勤耘

    2000-01-01

    In the present study, the involvement of L-arginine(L-Arg) NO on he protective action of electroacupuncture(EA) on cerebral ischemic injury was observed in acute ischemia-reperfusion(IR)rat model by taking regional cerebral blood flow(r-CBF), cerebral water content(CWC), and blood nitric oxide(NO) contents as indexes. Results showed that 1) EA could cause r-CBF and serum NO content to increase and CWC to lower, suggesting an protective action of EA on IR cerebral injury; 2) intravenous injection of L-Arg also had an protective effect on cerebral IR cerebral injury, while L-NNA had no this effect; and 3) pre-treatment with L-Arg might strengthen the effect of EA further, while pretreatment with L-NNA could weaken its effect. It indicates that L-Arg-NO may be involved in the effect of EA in protecting the brain from ischemic injury.

  14. Impacts of human activities on distribution of sulfate-reducing prokaryotes and antibiotic resistance genes in marine coastal sediments of Hong Kong.

    Guo, Feng; Li, Bing; Yang, Ying; Deng, Yu; Qiu, Jian-Wen; Li, Xiangdong; Leung, Kenneth My; Zhang, Tong

    2016-09-01

    Sulfate-reducing prokaryotes (SRPs) and antibiotic resistance genes (ARGs) in sediments could be biomarkers for evaluating the environmental impacts of human activities, although factors governing their distribution are not clear yet. By using metagenomic approach, this study investigated the distributions of SRPs and ARGs in marine sediments collected from 12 different coastal locations of Hong Kong, which exhibited different pollution levels and were classified into two groups based on sediment parameters. Our results showed that relative abundances of major SRP genera to total prokaryotes were consistently lower in the more seriously polluted sediments (P-value niche differentiation of SRPs and behavior of ARGs in marine coastal sediment. PMID:27297722

  15. Proteomic analysis of the effects of aged garlic extract and its FruArg component on lipopolysaccharide-induced neuroinflammatory response in microglial cells.

    Hui Zhou

    Full Text Available Aged garlic extract (AGE is widely used as a dietary supplement, and is claimed to promote human health through anti-oxidant/anti-inflammatory activities with hypolipidemic, antiplatelet and neuroprotective effects. Prior studies of AGE have mainly focused on its organosulfur compounds, with little attention paid to its carbohydrate derivatives, such as N-α-(1-deoxy-D-fructos-1-yl-L-arginine (FruArg. The goal of this study is to investigate actions of AGE and FruArg on antioxidative and neuroinflammatory responses in lipopolysaccharide (LPS-activated murine BV-2 microglial cells using a proteomic approach. Our data show that both AGE and FruArg can significantly inhibit LPS-induced nitric oxide (NO production in BV-2 cells. Quantitative proteomic analysis by combining two dimensional differential in-gel electrophoresis (2D-DIGE with mass spectrometry revealed that expressions of 26 proteins were significantly altered upon LPS exposure, while levels of 20 and 21 proteins exhibited significant changes in response to AGE and FruArg treatments, respectively, in LPS-stimulated BV-2 cells. Notably, approximate 78% of the proteins responding to AGE and FruArg treatments are in common, suggesting that FruArg is a major active component of AGE. MULTICOM-PDCN and Ingenuity Pathway Analyses indicate that the proteins differentially affected by treatment with AGE and FruArg are involved in inflammatory responses and the Nrf2-mediated oxidative stress response. Collectively, these results suggest that AGE and FruArg attenuate neuroinflammatory responses and promote resilience in LPS-activated BV-2 cells by suppressing NO production and by regulating expression of multiple protein targets associated with oxidative stress.

  16. Inactivation of antibiotic resistance genes in municipal wastewater effluent by chlorination and sequential UV/chlorination disinfection

    This study investigated disinfection methods including chlorination, ultraviolet (UV) irradiation and sequential UV/chlorination treatment on the inactivation of antibiotic resistance genes (ARGs). ARGs including sul1, tetX, tetG, intI1, and 16S rRNA genes in municipal wastewater treatment plant (MWTP) effluent were examined. The results indicated a positive correlation between the removal of ARGs and chlorine dosage (p = 0.007–0.014, n = 6),as well as contact time (p = 0.0001, n = 10). Greater free chlorine (FC) dosage leads to higher removal for all the genes and the maximum removal (1.30–1.49 logs) could be achieved at FC dosage of 30 mg L−1. The transformation kinetic data for ARGs removal (log C0 / C) followed the second-order reaction kinetic model with FC dosage (R2 = 0.6829–0.9999) and contact time (R2 = 0.7353–8634), respectively. Higher ammonia nitrogen (NH3–N) concentration was found to lead to lower removal of ARGs at the same chlorine dosage. When the applied Cl2:NH3–N ratio was over 7.6:1, a significant reduction of ARGs (1.20–1.49 logs) was achieved. By using single UV irradiation, the log removal values of tetX and 16Ss rRNA genes were 0.58 and 0.60, respectively, while other genes were 0.36–0.40 at a fluence of 249.5 mJ cm−2, which was observed to be less effective than chlorination. With sequential UV/chlorination treatment, 0.006 to 0.31 log synergy values of target genes were observed under different operation parameters. - Highlights: • Chlorine is more effective than UV irradiation in removing ARGs from MWTP effluent. • The chlorination reaction followed the second-order reaction kinetic model. • Higher NH3–N contents result in lower ARGs removal in the chlorination process. • FC is more effective than CC on the inactivation of ARGs. • UV irradiation followed by chlorination shows high efficiency in removing ARGs

  17. Inactivation of antibiotic resistance genes in municipal wastewater effluent by chlorination and sequential UV/chlorination disinfection

    Zhang, Yingying; Zhuang, Yao; Geng, Jinju, E-mail: jjgeng@nju.edu.cn; Ren, Hongqiang, E-mail: hqren@nju.edu.cn; Zhang, Yan; Ding, Lili; Xu, Ke

    2015-04-15

    This study investigated disinfection methods including chlorination, ultraviolet (UV) irradiation and sequential UV/chlorination treatment on the inactivation of antibiotic resistance genes (ARGs). ARGs including sul1, tetX, tetG, intI1, and 16S rRNA genes in municipal wastewater treatment plant (MWTP) effluent were examined. The results indicated a positive correlation between the removal of ARGs and chlorine dosage (p = 0.007–0.014, n = 6),as well as contact time (p = 0.0001, n = 10). Greater free chlorine (FC) dosage leads to higher removal for all the genes and the maximum removal (1.30–1.49 logs) could be achieved at FC dosage of 30 mg L{sup −1}. The transformation kinetic data for ARGs removal (log C{sub 0} / C) followed the second-order reaction kinetic model with FC dosage (R{sup 2} = 0.6829–0.9999) and contact time (R{sup 2} = 0.7353–8634), respectively. Higher ammonia nitrogen (NH{sub 3}–N) concentration was found to lead to lower removal of ARGs at the same chlorine dosage. When the applied Cl{sub 2}:NH{sub 3}–N ratio was over 7.6:1, a significant reduction of ARGs (1.20–1.49 logs) was achieved. By using single UV irradiation, the log removal values of tetX and 16Ss rRNA genes were 0.58 and 0.60, respectively, while other genes were 0.36–0.40 at a fluence of 249.5 mJ cm{sup −2}, which was observed to be less effective than chlorination. With sequential UV/chlorination treatment, 0.006 to 0.31 log synergy values of target genes were observed under different operation parameters. - Highlights: • Chlorine is more effective than UV irradiation in removing ARGs from MWTP effluent. • The chlorination reaction followed the second-order reaction kinetic model. • Higher NH{sub 3}–N contents result in lower ARGs removal in the chlorination process. • FC is more effective than CC on the inactivation of ARGs. • UV irradiation followed by chlorination shows high efficiency in removing ARGs.

  18. Dissemination of veterinary antibiotics and corresponding resistance genes from a concentrated swine feedlot along the waste treatment paths.

    Wang, Jian; Ben, Weiwei; Yang, Min; Zhang, Yu; Qiang, Zhimin

    2016-01-01

    Swine feedlots are an important pollution source of antibiotics and antibiotic resistance genes (ARGs) to the environment. This study investigated the dissemination of two classes of commonly-used veterinary antibiotics, namely, tetracyclines (TCs) and sulfonamides (SAs), and their corresponding ARGs along the waste treatment paths from a concentrated swine feedlot located in Beijing, China. The highest total TC and total SA concentrations detected were 166.7mgkg(-1) and 64.5μgkg(-1) in swine manure as well as 388.7 and 7.56μgL(-1) in swine wastewater, respectively. Fourteen tetracycline resistance genes (TRGs) encoding ribosomal protection proteins (RPP), efflux proteins (EFP) and enzymatic inactivation proteins, three sulfonamide resistance genes (SRGs), and two integrase genes were detected along the waste treatment paths with detection frequencies of 33.3-75.0%. The relative abundances of target ARGs ranged from 2.74×10(-6) to 1.19. The antibiotics and ARGs generally declined along both waste treatment paths, but their degree of reduction was more significant along the manure treatment path. The RPP TRGs dominated in the upstream samples and then decreased continuously along both waste treatment paths, whilst the EFP TRGs and SRGs maintained relatively stable. Strong correlations between antibiotic concentrations and ARGs were observed among both manure and wastewater samples. In addition, seasonal temperature, and integrase genes, moisture content and nutrient level of tested samples could all impact the relative abundances of ARGs along the swine waste treatment paths. This study helps understand the evolution and spread of ARGs from swine feedlots to the environment as well as assess the environmental risk arising from swine waste treatment. PMID:27128716

  19. Structural and population-based evaluations of TBC1D1 p.Arg125Trp.

    Tom G Richardson

    Full Text Available Obesity is now a leading cause of preventable death in the industrialised world. Understanding its genetic influences can enhance insight into molecular pathogenesis and potential therapeutic targets. A non-synonymous polymorphism (rs35859249, p.Arg125Trp in the N-terminal TBC1D1 phosphotyrosine-binding (PTB domain has shown a replicated association with familial obesity in women. We investigated these findings in the Avon Longitudinal Study of Parents and Children (ALSPAC, a large European birth cohort of mothers and offspring, and by generating a predicted model of the structure of this domain. Structural prediction involved the use of three separate algorithms; Robetta, HHpred/MODELLER and I-TASSER. We used the transmission disequilibrium test (TDT to investigate familial association in the ALSPAC study cohort (N = 2,292 mother-offspring pairs. Linear regression models were used to examine the association of genotype with mean measurements of adiposity (Body Mass Index (BMI, waist circumference and Dual-energy X-ray absorptiometry (DXA assessed fat mass, and logistic regression was used to examine the association with odds of obesity. Modelling showed that the R125W mutation occurs in a location of the TBC1D1 PTB domain that is predicted to have a function in a putative protein:protein interaction. We did not detect an association between R125W and BMI (mean per allele difference 0.27 kg/m(2 (95% Confidence Interval: 0.00, 0.53 P = 0.05 or obesity (odds ratio 1.01 (95% Confidence Interval: 0.77, 1.31, P = 0.96 in offspring after adjusting for multiple comparisons. Furthermore, there was no evidence to suggest that there was familial association between R125W and obesity (χ(2 = 0.06, P = 0.80. Our analysis suggests that R125W in TBC1D1 plays a role in the binding of an effector protein, but we find no evidence that the R125W variant is related to mean BMI or odds of obesity in a general population sample.

  20. Metagenomic profiling of antibiotic resistance genes and mobile genetic elements in a tannery wastewater treatment plant.

    Zhu Wang

    Full Text Available Antibiotics are often used to prevent sickness and improve production in animal agriculture, and the residues in animal bodies may enter tannery wastewater during leather production. This study aimed to use Illumina high-throughput sequencing to investigate the occurrence, diversity and abundance of antibiotic resistance genes (ARGs and mobile genetic elements (MGEs in aerobic and anaerobic sludge of a full-scale tannery wastewater treatment plant (WWTP. Metagenomic analysis showed that Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria dominated in the WWTP, but the relative abundance of archaea in anaerobic sludge was higher than in aerobic sludge. Sequencing reads from aerobic and anaerobic sludge revealed differences in the abundance of functional genes between both microbial communities. Genes coding for antibiotic resistance were identified in both communities. BLAST analysis against Antibiotic Resistance Genes Database (ARDB further revealed that aerobic and anaerobic sludge contained various ARGs with high abundance, among which sulfonamide resistance gene sul1 had the highest abundance, occupying over 20% of the total ARGs reads. Tetracycline resistance genes (tet were highly rich in the anaerobic sludge, among which tet33 had the highest abundance, but was absent in aerobic sludge. Over 70 types of insertion sequences were detected in each sludge sample, and class 1 integrase genes were prevalent in the WWTP. The results highlighted prevalence of ARGs and MGEs in tannery WWTPs, which may deserve more public health concerns.

  1. Dihydropteroate Synthase and Novel Dihydrofolate Reductase Gene Mutations in Strains of Pneumocystis jirovecii from South Africa

    Robberts, F. J. L.; Chalkley, L J; Weyer, K.; Goussard, P.; Liebowitz, L. D.

    2005-01-01

    Dihydropteroate synthase (DHPS) gene mutations have raised concerns about emerging sulfonamide resistance in Pneumocystis jirovecii. DHPS and dihydrofolate reductase (DHFR) gene products were amplified in clinical specimens from South African patients. One of 53 DHPS genes sequenced contained the double mutation Thr55Ala Pro57Ser. DHFR gene mutations detected were Ala67Val and the new mutations Arg59Gly and C278T.

  2. Long-term field application of sewage sludge increases the abundance of antibiotic resistance genes in soil.

    Chen, Qinglin; An, Xinli; Li, Hu; Su, Jianqiang; Ma, Yibing; Zhu, Yong-Guan

    2016-01-01

    Sewage sludge and manure are common soil amendments in crop production; however, their impact on the abundance and diversity of the antibiotic resistome in soil remains elusive. In this study, by using high-throughput sequencing and high-throughput quantitative PCR, the patterns of bacterial community and antibiotic resistance genes (ARGs) in a long-term field experiment were investigated to gain insights into these impacts. A total of 130 unique ARGs and 5 mobile genetic elements (MGEs) were detected and the long-term application of sewage sludge and chicken manure significantly increased the abundance and diversity of ARGs in the soil. Genes conferring resistance to beta-lactams, tetracyclines, and multiple drugs were dominant in the samples. Sewage sludge or chicken manure applications caused significant enrichment of 108 unique ARGs and MGEs with a maximum enrichment of up to 3845 folds for mexF. The enrichment of MGEs suggested that the application of sewage sludge or manure may accelerate the dissemination of ARGs in soil through horizontal gene transfer (HGT). Based on the co-occurrence pattern of ARGs subtypes revealed by network analysis, aacC, oprD and mphA-02, were proposed to be potential indicators for quantitative estimation of the co-occurring ARGs subtypes abundance by power functions. The application of sewage sludge and manure resulted in significant increase of bacterial diversity in soil, Proteobacteria, Acidobacteria, Actinobacteria and Chloroflexi were the dominant phyla (>10% in each sample). Five bacterial phyla (Chloroflexi, Planctomycetes, Firmicutes, Gemmatimonadetes and Bacteroidetes) were found to be significantly correlated with the ARGs in soil. Mantel test and variation partitioning analysis (VPA) suggested that bacterial community shifts, rather than MGEs, is the major driver shaping the antibiotic resistome. Additionally, the co-occurrence pattern between ARGs and microbial taxa revealed by network analysis indicated that four

  3. Monitoring Antibiotic Residues and Corresponding Antibiotic Resistance Genes in an Agroecosystem

    Yasser M. Awad

    2015-01-01

    Full Text Available Antibiotic resistance genes (ARGs have been commonly reported due to the overuse worldwide of antibiotics. Antibiotic overuse disturbs the environment and threatens public human health. The objective of this study was to measure the residual concentrations of veterinary antibiotics in the tetracycline group (TCs, including tetracycline (TC and chlortetracycline (CTC, as well as those in the sulfonamide group (SAs, including sulfamethazine (SMT, sulfamethoxazole (SMX, and sulfathiazole (STZ. We also isolated the corresponding ARGs in the agroecosystem. Four sediment samples and two rice paddy soil samples were collected from sites near a swine composting facility along the Naerincheon River in Hongcheon, Korea. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS was employed with a solid-phase extraction method to measure the concentration of each antibiotic. ARGs were identified by the qualitative polymerase chain-reaction using synthetic primers. SAs and their corresponding ARGs were highly detected in sediment samples whereas TCs were not detected except for sediments sample #1. ARGs for TCs and SAs were detected in rice paddy soils, while ARGs for TCs were only found in sediment #2 and #4. Continuous monitoring of antibiotic residue and its comprehensive impact on the environment is needed to ensure environmental health.

  4. [Hemoglobin Boumerdès alpha 2(37) (C2) Pro----Arg beta 2: a new variant of the alpha chain associated with hemoglobin S in an Algerian family].

    Dahmane-Arbane, M; Blouquit, Y; Arous, N; Bardakdjian, J; Benamani, M; Riou, J; Benabadji, M; Rosa, J; Galacteros, F

    1987-01-01

    We report the first case of Hb Boumerdes, an alpha chain variant alpha 2(37) (C2) Pro----Arg beta 2, in an Algerian family. The propositus was also homozygous for the sickle cell gene. The abnormal hybrid Hb alpha 2Boum. beta 2S had an electrophoretic mobility on cellulose acetate pH 8.7 electrophoresis between those of Hb S and Hb A2. Its expression was about 16%. The alpha 2Boum. beta 2A fraction has a mobility between those of Hb F and Hb S. The effects of this mutation on Hb oxygen affinity and deoxy Hb S polymer formation were not studied. The propositus' sickle cell phenotype was benign. PMID:3438164

  5. E-Selectin Gene Polymorphisms in Iranian Chronic Hepatitis B Patients

    Mehrdad Hajilooi; Amir-Houshang Mohammad Alizade; Mitra Ranjbar; Farahnaz Fallahian; Seyed-Moayed Alavian

    2007-01-01

    Background and Aims: The aim of this study is to detect the substitutions Ser128Arg (A128C) and Leu554Phe (T554C) which responsible for E-selectin polymorphisms in patients with chronic hepatitis B and healthy controls. We investigated association of the Ser128Arg, Leu554Phe gene polymorphisms in the E-selectin gene as prototypical inflammatory molecules for susceptibility to chronic hepatitis B.Methods: Sixty-three patients with chronic hepatitis B virus infection and 150 healthy subjects we...

  6. Synthesis of Fluorine-18 Labeled Glucose-Lys-Arg-Gly-Asp-D-Phe as a Potential Tumor Imaging Agent

    The αvβ3 integrin is an important receptor affecting tumor growth, metastatic potential on proliferating endothelial cells as well as on tumor cells of various origin, tumor-induced angiogenesis could be blocked by antagonizing the αvβ3 integrin with RGD. Therefore, αvβ3 integrin is a target for angiogenesis imaging that might be useful in assessing tumor-induced angiogenesis and identifying tumor metastasis. To design potent radiotracer for imaging angiogenesis containing a cRGD moiety should include low hepatic uptake in vivo. Tripeptide Arg-Gly-Asp (RGD), naturally existed in extracellular matrix proteins, is known to be the primary binding site of the αvβ3 integrin. The imaging of αvβ3 receptor expression will give the information of the metastatic ability of the tumor which is not available by [18F]FDG. Our interest in developing new radiopharmaceuticals for in vivo visualization of angiogenesis has led us to synthesize derivatives of cRGD (cyclic arginineglycine-aspartic acid) that contains glucose moiety. Because sugar-protein interaction is a key step in metastasis and angiogenesis, it has also been proposed to play an intriguing role in imaging of tumor. We designed and synthesized two fluorine-18 labeled RGD glycopeptides . N-fluorobenzyl-diaminobutane-N'-glucose-Lys-Arg-Gly-Asp-D-Phe ([18F]fluorobenzyl-glucose-KRGDf, and Nfluorobenzoyl- diaminobutane-N'-glucose-Lys-Arg-Gly-Asp-D-Phe ([18F]fluorobenzoyl-glucose-KRGDf, from same precursor as a diagnostic tumor imaging agent for positron emission tomography (PET). Fluorine-18 labeled cRGD glycopeptides were prepared using two different simple labeling methods: one is reductive alkylation of an amine with [18F]fluorobenzaldehyde and the other is amide condensation with [18F]fluorobenzoic acid

  7. ARGE Weser (Working Group for Water Pollution Abatement in the Weser River). Weser water quality report 1996

    The ARGE Weser (Arbeitsgemeinschaft zur Reinhaltung der Weser), which comprises representatives of the German states of Bremen, Hessen, Niedersachsen, Nordrhein-Westfalen and Thueringen, publishes a yearly water quality report based on the measuring programmes for the Weser river and for the Werra/Ulster rivers (the latter is a joint project of the states of Hessen and Thrueringen). The Weser river today has a water quality of II-III after the successful implementation of the 1989 action programme. Chloride concentrations were reduced as well, but efforts will still be made by communal authorities, agriculture, industry and administrative bodies if the goals set for 2000 are to be reached. (orig/AJ)

  8. Identification of Minor Secondary Metabolites from the Latex of Croton lechleri (Muell-Arg) and Evaluation of Their Antioxidant Activity

    Maria Iorizzi; Francesco Visioli; Gelsomina Fico; Sara Vitalini; Franco Zollo; Fulvio Gala; Simona De Marino

    2008-01-01

    Dragon’s blood (Sangre de drago), a viscous red sap derived from Croton lechleri Muell-Arg (Euphorbiaceae), is extensively used by indigenous cultures of the Amazonian basin for its wound healing properties. The aim of this study was to identify the minor secondary metabolites and test the antioxidant activity of this sustance. A bioguided fractionation of the n-hexane, chloroform, n-butanol, and aqueous extracts led to the isolation of 15 compounds: three megastigmanes, four flavan-3-ol...

  9. Direct Assessment of the Effect of the Gly380Arg Achondroplasia Mutation on FGFR3 Dimerization Using Quantitative Imaging FRET

    Placone, Jesse; Hristova, Kalina

    2012-01-01

    The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH), the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase...

  10. Genes, enzymes and regulation of arginine biosynthesis in plants.

    Slocum, Robert D

    2005-08-01

    Arabidopsis genes encoding enzymes for each of the eight steps in L-arginine (Arg) synthesis were identified, based upon sequence homologies with orthologs from other organisms. Except for N-acetylglutamate synthase (NAGS; EC 2.3.1.1), which is encoded by two genes, all remaining enzymes are encoded by single genes. Targeting predictions for these enzymes, based upon their deduced sequences, and subcellular fractionation studies, suggest that most enzymes of Arg synthesis reside within the plastid. Synthesis of the L-ornthine (Orn) intermediate in this pathway from L-glutamate occurs as a series of acetylated intermediates, as in most other organisms. An N-acetylornithine:glutamate acetyltransferase (NAOGAcT; EC 2.3.1.35) facilitates recycling of the acetyl moiety during Orn formation (cyclic pathway). A putative N-acetylornithine deacetylase (NAOD; EC 3.5.1.16), which participates in the "linear" pathway for Orn synthesis in some organisms, was also identified. Previous biochemical studies have indicated that allosteric regulation of the first and, especially, the second steps in Orn synthesis (NAGS; N-acetylglutamate kinase (NAGK), EC 2.7.2.8) by the Arg end-product are the major sites of metabolic control of the pathway in organisms using the cyclic pathway. Gene expression profiling for pathway enzymes further suggests that NAGS, NAGK, NAOGAcT and NAOD are coordinately regulated in response to changes in Arg demand during plant growth and development. Synthesis of Arg from Orn is further coordinated with pyrimidine nucleotide synthesis, at the level of allocation of the common carbamoyl-P intermediate. PMID:16122935

  11. Evaluation Of ARG-1 Samples Prepared By Cesium Carbonate Dissolution During The Isolok SME Acceptability Testing

    issued that recommended that the mixed acid method be replaced by the Cs2CO3 method for the measurement of magnesium (Mg), sodium (Na), and zirconium (Zr) with additional testing of the method by DWPF Laboratory being needed before further implementation of the Cs2CO3 method at that laboratory. While the SME acceptability testing of the Isolok does not address any of the open issues remaining after the publication of the recommendation for the replacement of the mixed acid method by the Cs2CO3 method (since those issues are to be addressed by the DWPF Laboratory), the Cs2CO3 testing associated with the Isolok testing does provide additional insight into the performance of the method as conducted by SRNL. The performance is to be investigated by looking to the composition measurement data generated by the samples of a standard glass, the Analytical Reference Glass - 1 (ARG-1), that were prepared by the Cs2CO3 method and included in the SME acceptability testing of the Isolok. The measurements of these samples were presented as part of the study results, but no statistical analysis of these measurements was conducted as part of those results. It is the purpose of this report to provide that analysis, which was supported using JMP Version 7.0.2.

  12. EVALUATION OF ARG-1 SAMPLES PREPARED BY CESIUM CARBONATE DISSOLUTION DURING THE ISOLOK SME ACCEPTABILITY TESTING

    Edwards, T.; Hera, K.; Coleman, C.

    2011-12-05

    from this work was issued that recommended that the mixed acid method be replaced by the Cs{sub 2}CO{sub 3} method for the measurement of magnesium (Mg), sodium (Na), and zirconium (Zr) with additional testing of the method by DWPF Laboratory being needed before further implementation of the Cs{sub 2}CO{sub 3} method at that laboratory. While the SME acceptability testing of the Isolok does not address any of the open issues remaining after the publication of the recommendation for the replacement of the mixed acid method by the Cs{sub 2}CO{sub 3} method (since those issues are to be addressed by the DWPF Laboratory), the Cs{sub 2}CO{sub 3} testing associated with the Isolok testing does provide additional insight into the performance of the method as conducted by SRNL. The performance is to be investigated by looking to the composition measurement data generated by the samples of a standard glass, the Analytical Reference Glass - 1 (ARG-1), that were prepared by the Cs{sub 2}CO{sub 3} method and included in the SME acceptability testing of the Isolok. The measurements of these samples were presented as part of the study results, but no statistical analysis of these measurements was conducted as part of those results. It is the purpose of this report to provide that analysis, which was supported using JMP Version 7.0.2.

  13. In vitro and in vivo pharmacological profile of PL-3994, a novel cyclic peptide (Hept-cyclo(Cys-His-Phe-d-Ala-Gly-Arg-d-Nle-Asp-Arg-Ile-Ser-Cys)-Tyr-[Arg mimetic]-NH2) natriuretic peptide receptor-A agonist that is resistant to neutral endopeptidase and acts as a bronchodilator

    Edelson, Jeffrey D.; Makhlina, Marie; Silvester, Kevin R.; Vengurlekar, Shailesh S.; Chen, Xiaomei; Zhang, Jie; Koziol-White, Cynthia J.; Cooper, Philip R.; Hallam, Trevor J.; Hay, Douglas W.P.; Panettieri, Reynold A.

    2014-01-01

    The pharmacological and airways relaxant profiles of PL-3994 (Hept-cyclo(Cys-His-Phe-d-Ala-Gly-Arg-dNle-Asp-Arg-Ile-Ser-Cys)-Tyr-[Arg mimetic]-NH2), a novel natriuretic peptide receptor-A (NPR-A) agonist, were evaluated. PL-3994, a full agonist, has high affinity for recombinant human (h), dog, or rat NPR-As (Kis of 1, 41, and 10 nm, respectively), and produced concentration-dependent cGMP generation in human, dog and rat NPR-As (respective EC50s of 2, 3 and 14 nm). PL-3994 has a Ki of 7 nm for hNPR-C but was without effect on cGMP generation in hNPR-B. PL-3994 (1 µm) was without significant effect against 75 diverse molecular targets. PL-3994 or BNP, a natural NPR ligand, produced concentration-dependent relaxation of pre-contracted guinea-pig trachea (IC50s of 42.7 and 10.7 nm, respectively). PL-3994, and also BNP, (0.1 nm–100 µm) elicited a potent, concentration-dependent but small relaxation of pre-contracted human precision-cut lung slices (hPCLS). Intratracheal PL-3994 (1–1000 µg/kg) produced a dose-dependent inhibition of the bronchoconstrictor response evoked by aerosolized methacholine, but was without significant effect on cardiovascular parameters. PL-3994 was resistant to degradation by human neutral endopeptidase (hNEP) (92% remaining after 2 h), whereas the natural ligands, ANP and CNP, were rapidly metabolized (≤1% remaining after 2 h). PL-3994 is a potent, selective NPR agonist, resistant to NEP, with relaxant effects in guinea-pig and human airway smooth muscle systems. PL-3994 has the profile predictive of longer clinical bronchodilator activity than observed previously with ANP, and suggests its potential utility in the treatment of asthma, in addition to being a useful research tool to evaluate NPR biology. PMID:23154072

  14. A Comprehensive Insight into Tetracycline Resistant Bacteria and Antibiotic Resistance Genes in Activated Sludge Using Next-Generation Sequencing

    Kailong Huang

    2014-06-01

    Full Text Available In order to comprehensively investigate tetracycline resistance in activated sludge of sewage treatment plants, 454 pyrosequencing and Illumina high-throughput sequencing were used to detect potential tetracycline resistant bacteria (TRB and antibiotic resistance genes (ARGs in sludge cultured with different concentrations of tetracycline. Pyrosequencing of 16S rRNA gene revealed that tetracycline treatment greatly affected the bacterial community structure of the sludge. Nine genera consisting of Sulfuritalea, Armatimonas, Prosthecobacter, Hyphomicrobium, Azonexus, Longilinea, Paracoccus, Novosphingobium and Rhodobacter were identified as potential TRB in the sludge. Results of qPCR, molecular cloning and metagenomic analysis consistently indicated that tetracycline treatment could increase both the abundance and diversity of the tet genes, but decreased the occurrence and diversity of non-tetracycline ARG, especially sulfonamide resistance gene sul2. Cluster analysis showed that tetracycline treatment at subinhibitory concentrations (5 mg/L was found to pose greater effects on the bacterial community composition, which may be responsible for the variations of the ARGs abundance. This study indicated that joint use of 454 pyrosequencing and Illumina high-throughput sequencing can be effectively used to explore ARB and ARGs in the environment, and future studies should include an in-depth investigation of the relationship between microbial community, ARGs and antibiotics in sewage treatment plant (STP sludge.

  15. Living with high putrescine: expression of ornithine and arginine biosynthetic pathway genes in high and low putrescine producing poplar cells.

    Page, Andrew F; Minocha, Rakesh; Minocha, Subhash C

    2012-01-01

    Arginine (Arg) and ornithine (Orn), both derived from glutamate (Glu), are the primary substrates for polyamine (PA) biosynthesis, and also play important roles as substrates and intermediates of overall N metabolism in plants. Their cellular homeostasis is subject to multiple levels of regulation. Using reverse transcription quantitative PCR (RT-qPCR), we studied changes in the expression of all genes of the Orn/Arg biosynthetic pathway in response to up-regulation [via transgenic expression of mouse Orn decarboxylase (mODC)] of PA biosynthesis in poplar (Populus nigra × maximowiczii) cells grown in culture. Cloning and sequencing of poplar genes involved in the Orn/Arg biosynthetic pathway showed that they have high homology with similar genes in other plants. The expression of the genes of Orn, Arg and PA biosynthetic pathway fell into two hierarchical clusters; expression of one did not change in response to high putrescine, while members of the other cluster showed a shift in expression pattern during the 7-day culture cycle. Gene expression of branch point enzymes (N-acetyl-Glu synthase, Orn aminotransferase, Arg decarboxylase, and spermidine synthase) in the sub-pathways, constituted a separate cluster from those involved in intermediary reactions of the pathway (N-acetyl-Glu kinase, N-acetyl-Glu-5-P reductase, N-acetyl-Orn aminotransferase, N (2)-acetylOrn:N-acetyl-Glu acetyltransferase, N (2)-acetyl-Orn deacetylase, Orn transcarbamylase, argininosuccinate synthase, carbamoylphosphate synthetase, argininosuccinate lyase, S-adenosylmethionine decarboxylase, spermine synthase). We postulate that expression of all genes of the Glu-Orn-Arg pathway is constitutively coordinated and is not influenced by the increase in flux rate through this pathway in response to increased utilization of Orn by mODC; thus the pathway involves mostly biochemical regulation rather than changes in gene expression. We further suggest that Orn itself plays a major role in the

  16. High frequencies of antibiotic resistance genes in infants’ meconium and early fecal samples

    Gosalbes, M. J.; Vallès, Y.; Jiménez-Hernández, N.;

    2016-01-01

    before birth. We have used culture-independent genetic screenings to explore whether ARGs are already present in the meconium accumulated in the GIT during fetal life and in feces of 1-week-old infants. We have analyzed resistance to β-lactam antibiotics (BLr) and tetracycline (Tcr), screening for a......The gastrointestinal tract (GIT) microbiota has been identified as an important reservoir of antibiotic resistance genes (ARGs) that can be horizontally transferred to pathogenic species. Maternal GIT microbes can be transmitted to the offspring, and recent work indicates that such transfer starts...... variety of genes conferring each. To evaluate whether ARGs could have been inherited by maternal transmission, we have screened perinatal fecal samples of the 1-week-old babies’ mothers, as well as a mother–infant series including meconium, fecal samples collected through the infant’s 1st year, maternal...

  17. Conformational study of Z-Glu-OH and Z-Arg-OH: dispersion interactions versus conventional hydrogen bonding.

    Jaeqx, Sander; Du, Weina; Meijer, Evert Jan; Oomens, Jos; Rijs, Anouk M

    2013-02-14

    The gas-phase conformational preferences of the model dipeptides Z-Glu-OH and Z-Arg-OH have been studied in the low-temperature environment of a supersonic jet. IR-UV ion-dip spectra obtained using the free electron laser FELIX provide conformation-specific IR spectra, which in combination with density functional theory (DFT) allow us to determine the conformational structures of the peptides. Molecular dynamics modeling using simulated annealing generates a variety of low-energy structures, for which geometry optimization and frequency calculations are then performed using the B3LYP functional with the 6-311+G(d,p) basis set. By comparing experimental and theoretical IR spectra, three conformations for Z-Glu-OH and two for Z-Arg-OH have been identified. For three of the five structures, the dispersion interaction provides an important contribution to the stabilization, emphasizing the importance of these forces in small peptides. Therefore, dispersion-corrected DFT functionals (M05-2X and B97D) have also been employed in our theoretical analysis. Second-order Møller-Plesset perturbation theory (MP2) has been used as benchmark for the relative energies of the different conformational structures. Finally, we address the ongoing debate on the gas-phase structure of arginine by elucidating whether isolated arginine is canonical, tautomeric, or zwitterionic. PMID:23095122

  18. Multimodal approach to explore the pathogenicity of BARD1, ARG 658 CYS, and ILE 738 VAL mutants.

    Choudhary, Rajan Kumar; Vikrant; Siddiqui, Quadir M; Thapa, Pankaj S; Raikundalia, Sweta; Gadewal, Nikhil; Kumar, Nachimuthu Senthil; Hosur, M V; Varma, Ashok K

    2016-07-01

    BARD1-BRCA1 complex plays an important role in DNA damage repair, apoptosis, chromatin remodeling, and other important processes required for cell survival. BRCA1 and BARD1 heterodimer possess E3 ligase activity and is involved in genome maintenance, by functioning in surveillance for DNA damage, thereby regulating multiple pathways including tumor suppression. BRCT domains are evolutionary conserved domains present in different proteins such as BRCA1, BARD1, XRCC, and MDC1 regulating damage response and cell-cycle control through protein-protein interactions. Nonetheless, the role of BARD1BRCT in the recruitment of DNA repair mechanism and structural integrity with BRCA1 complex is still implicit. To explicate the role of BARD1BRCT in the DNA repair mechanism, in silico, in vitro, and biophysical approach were applied to characterize BARD1 BRCT wild-type and Arg658Cys and Ile738Val mutants. However, no drastic secondary and tertiary structural changes in the mutant proteins were observed. Thermal and chemical denaturation studies revealed that mutants Arg658Cys and Ile738Val have a decrease in Tm and ∆G than the wild type. In silico studies of BARD1 BRCT (568-777) and mutant protein indicate loss in structural compactness on the Ile738Val mutant. Comparative studies of wild-type and mutants will thus be helpful in understanding the basic role of BARD1BRCT in DNA damage repair. PMID:26307947

  19. A Review on the Salt Bridge Between ASP177 and ARG163 of Wild-Type Rabbit Prion Protein

    Zhang, Jiapu

    2014-01-01

    Prion diseases are invariably fatal and highly infectious neurodegenerative diseases that affect a wide variety of mammalian species such as sheep and goats, cattle, deer, elks, humans and mice etc., but rabbits have a low susceptibility to be infected by prion diseases with respect to other species. The stability of rabbit prion protein is due to its highly ordered beta2-alpha2 loop [PLoS One 5 (10) e13273 (2010); Journal of Biological Chemistry 285 (41) 31682-31693 (2010)] and a helix-capping motif within this loop [PLoS One 8 (5) e63047 (2013)]. The beta2-alpha2 loop has been a focus in prion studies. For this loop we found a salt bridge linkage ASP177-ARG163 (O-N) [Journal of Theoretical Biology 342 (7 February 2014) 70-82 (2014)]. Some scientists said on the 2FJ3.pdb NMR file of the rabbit prion protein, the distance of ASP177-ARG163 (O-N) gives the salt bridge of about 10 angstroms which is nearly null in terms of energy thus think our result is wrong. This opinion is clearly wrong simply due to the 3O7...

  20. Cationic amino acid transporters and Salmonella Typhimurium ArgT collectively regulate arginine availability towards intracellular Salmonella growth.

    Priyanka Das

    Full Text Available Cationic amino acid transporters (mCAT1 and mCAT2B regulate the arginine availability in macrophages. How in the infected cell a pathogen can alter the arginine metabolism of the host remains to be understood. We reveal here a novel mechanism by which Salmonella exploit mCAT1 and mCAT2B to acquire host arginine towards its own intracellular growth within antigen presenting cells. We demonstrate that Salmonella infected bone marrow derived macrophages and dendritic cells show enhanced arginine uptake and increased expression of mCAT1 and mCAT2B. We show that the mCAT1 transporter is in close proximity to Salmonella containing vacuole (SCV specifically by live intracellular Salmonella in order to access the macrophage cytosolic arginine pool. Further, Lysosome associated membrane protein 1, a marker of SCV, also was found to colocalize with mCAT1 in the Salmonella infected cell. The intra vacuolar Salmonella then acquire the host arginine via its own arginine transporter, ArgT for growth. The argT knockout strain was unable to acquire host arginine and was attenuated in growth in both macrophages and in mice model of infection. Together, these data reveal survival strategies by which virulent Salmonella adapt to the harsh conditions prevailing in the infected host cells.

  1. Arc/Arg3.1 mRNA global expression patterns elicited by memory recall in cerebral cortex differ for remote versus recent spatial memories

    Pavel A Gusev

    2010-05-01

    Full Text Available The neocortex plays a critical role in the gradual formation and storage of remote declarative memories. Because the circuitry mechanisms of systems-level consolidation are not well understood, the precise cortical sites for memory storage and the nature of enduring memory correlates (mnemonic plasticity are largely unknown. Detailed maps of neuronal activity underlying recent and remote memory recall highlight brain regions that participate in systems consolidation and constitute putative storage sites, and thus may facilitate detection of mnemonic plasticity. To localize cortical regions involved in the recall of a spatial memory task, we trained rats in a water maze and then mapped mRNA expression patterns of a neuronal activity marker Arc/Arg3.1 (Arc upon recall of recent (24 hours after training or remote (one month after training memories and compared them with swimming and naive controls. Arc gene expression was significantly more robust 24 hours after training compared to one month after training. Arc expression diminished in the parietal, cingulate and visual areas, but select segments in the prefrontal, retrosplenial, somatosensory and motor cortical showed similar robust increases in the Arc expression. When Arc expression was compared across select segments of sensory, motor and associative regions within recent and remote memory groups, the overall magnitude and cortical laminar patterns of task-specific Arc expression were similar (stereotypical. Arc mRNA fractions expressed in the upper cortical layers (2/3, 4 increased after both recent and remote recall, while layer 6 fractions decreased only after the recent recall. The data suggest that robust recall of remote memory requires an overall smaller increase in neuronal activity within fewer cortical segments. This activity trend highlights the difficulty in detecting the storage sites and plasticity underlying remote memory. Application of the Arc maps may ameliorate this

  2. The LMNA mutation p.Arg321Ter associated with dilated cardiomyopathy leads to reduced expression and a skewed ratio of lamin A and lamin C proteins

    Dilated cardiomyopathy (DCM) is a disease of the heart muscle characterized by cardiac chamber enlargement and reduced systolic function of the left ventricle. Mutations in the LMNA gene represent the most frequent known genetic cause of DCM associated with disease of the conduction systems. The LMNA gene generates two major transcripts encoding the nuclear lamina major components lamin A and lamin C by alternative splicing. Both haploinsuffiency and dominant negative effects have been proposed as disease mechanism for premature termination codon (PTC) mutations in LMNA. These mechanisms however are still not clearly established. In this study, we used a representative LMNA nonsense mutation, p.Arg321Ter, to shed light on the molecular disease mechanisms. Cultured fibroblasts from three DCM patients carrying this mutation were analyzed. Quantitative reverse transcriptase PCR and sequencing of these PCR products indicated that transcripts from the mutant allele were degraded by the nonsense-mediated mRNA decay (NMD) mechanism. The fact that no truncated mutant protein was detectable in western blot (WB) analysis strengthens the notion that the mutant transcript is efficiently degraded. Furthermore, WB analysis showed that the expression of lamin C protein was reduced by the expected approximately 50%. Clearly decreased lamin A and lamin C levels were also observed by immunofluorescence microscopy analysis. However, results from both WB and nano-liquid chromatography/mass spectrometry demonstrated that the levels of lamin A protein were more reduced suggesting an effect on expression of lamin A from the wild type allele. PCR analysis of the ratio of lamin A to lamin C transcripts showed unchanged relative amounts of lamin A transcript suggesting that the effect on the wild type allele was operative at the protein level. Immunofluorescence microscopy analysis showed no abnormal nuclear morphology of patient fibroblast cells. Based on these data, we propose that

  3. The LMNA mutation p.Arg321Ter associated with dilated cardiomyopathy leads to reduced expression and a skewed ratio of lamin A and lamin C proteins

    Al-Saaidi, Rasha [Research Unit for Molecular Medicine, Aarhus University and Aarhus University Hospital, Aarhus (Denmark); Rasmussen, Torsten B. [Department of Cardiology, Aarhus University Hospital, Aarhus (Denmark); Palmfeldt, Johan [Research Unit for Molecular Medicine, Aarhus University and Aarhus University Hospital, Aarhus (Denmark); Nissen, Peter H. [Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus (Denmark); Beqqali, Abdelaziz [Heart Failure Research Center, Academic Medical Center, Amsterdam (Netherlands); Hansen, Jakob [Department of Forensic Medicine, Bioanalytical Unit, University of Aarhus (Denmark); Pinto, Yigal M. [Heart Failure Research Center, Academic Medical Center, Amsterdam (Netherlands); Boesen, Thomas [Department of Molecular Biology and Genetics, University of Aarhus (Denmark); Mogensen, Jens [Department of Cardiology, Odense University Hospital, Odense (Denmark); Bross, Peter, E-mail: peter.bross@ki.au.dk [Research Unit for Molecular Medicine, Aarhus University and Aarhus University Hospital, Aarhus (Denmark)

    2013-11-15

    Dilated cardiomyopathy (DCM) is a disease of the heart muscle characterized by cardiac chamber enlargement and reduced systolic function of the left ventricle. Mutations in the LMNA gene represent the most frequent known genetic cause of DCM associated with disease of the conduction systems. The LMNA gene generates two major transcripts encoding the nuclear lamina major components lamin A and lamin C by alternative splicing. Both haploinsuffiency and dominant negative effects have been proposed as disease mechanism for premature termination codon (PTC) mutations in LMNA. These mechanisms however are still not clearly established. In this study, we used a representative LMNA nonsense mutation, p.Arg321Ter, to shed light on the molecular disease mechanisms. Cultured fibroblasts from three DCM patients carrying this mutation were analyzed. Quantitative reverse transcriptase PCR and sequencing of these PCR products indicated that transcripts from the mutant allele were degraded by the nonsense-mediated mRNA decay (NMD) mechanism. The fact that no truncated mutant protein was detectable in western blot (WB) analysis strengthens the notion that the mutant transcript is efficiently degraded. Furthermore, WB analysis showed that the expression of lamin C protein was reduced by the expected approximately 50%. Clearly decreased lamin A and lamin C levels were also observed by immunofluorescence microscopy analysis. However, results from both WB and nano-liquid chromatography/mass spectrometry demonstrated that the levels of lamin A protein were more reduced suggesting an effect on expression of lamin A from the wild type allele. PCR analysis of the ratio of lamin A to lamin C transcripts showed unchanged relative amounts of lamin A transcript suggesting that the effect on the wild type allele was operative at the protein level. Immunofluorescence microscopy analysis showed no abnormal nuclear morphology of patient fibroblast cells. Based on these data, we propose that

  4. Study on the Recombining Conditions Between A and B Chains of "Mini-C" Human Proinsulin Analog and Human Proinsulin%小C肽人胰岛素原类似物(B-Arg-Arg-A)和人胰岛素原(B-C-A)A、B链重组条件的研究

    陈来同; 张明军; 胡美浩

    2000-01-01

    目的:获得更佳的变复性条件和提高人胰岛素原的重组率.方法:采用特有的重组方法,将小C肽人胰岛素原类似物(B-Arg-Arg-A)与人胰岛素原(B-C-A)进行重组条件的比较.结果:发现二者在pH10.6左右处重组率达到最大,且在相同的条件下B-Arg-Arg-A蛋白质的重组率(94%) 较B-C-A蛋白质(82%)的高.结论:进一步验证了"胰岛素的A、B链含有足够使二硫键正确配对的结构信息"的论点.

  5. Reclaimed water as a reservoir of antibiotic resistance genes: distribution system and irrigation implications

    AmyPruden

    2013-05-01

    Full Text Available Treated wastewater is increasingly being reused to achieve sustainable water management in arid regions. The objective of this study was to quantify the distribution of antibiotic resistance genes (ARGs in recycled water, particularly after it has passed through the distribution system, and to consider point-of-use implications for soil irrigation. Three separate reclaimed wastewater distribution systems in the western U.S. were examined. Quantitative polymerase chain reaction (qPCR was used to quantify ARGs corresponding to resistance to sulfonamides (sul1, sul2, macrolides (ermF, tetracycline (tet(A, tet(O, glycopeptides (vanA, and methicillin (mecA, in addition to genes present in waterborne pathogens Legionella pneumophila (Lmip, Escherichia coli (gadAB, and Pseudomonas aeruginosa (ecfx, gyrB. In a parallel lab study, the effect of irrigating an agricultural soil with secondary, chlorinated, or dechlorinated wastewater effluent was examined in batch microcosms. A broader range of ARGs were detected after the reclaimed water passed through the distribution systems, highlighting the importance of considering bacterial re-growth and the overall water quality at the point of use. Screening for pathogens with qPCR indicated presence of Lmip and gadAB genes, but not ecfx or gyrB. In the lab study, chlorination was observed to reduce 16S rRNA and sul2 gene copies in the wastewater effluent, while dechlorination had no apparent effect. ARGs levels did not change with time in soil slurries incubated after a single irrigation event with any of the effluents. However, when irrigated repeatedly with secondary wastewater effluent (not chlorinated or dechlorinated, elevated levels of sul1 and sul2 were observed. This study suggests that reclaimed water may be an important reservoir of ARGs, especially at the point of use, and that attention should be directed towards the fate of ARGs in irrigation water and the implications for human health.

  6. Environmental factors responsible for the incidence of antibiotic resistance genes in pristine Crassostrea virginica reefs

    Highlights: ► Estuary was the major source of antibiotic resistance genes (ARG) for tidal creeks. ► Watersheds were the secondary source of ARG for tidal creeks. ► Watershed contribution corresponded to the degree of its anthropogenic disturbance. ► ARG in tidal creeks were carried by native hosts preferring low termohaline niches. ► ARG incidence was the highest in oysters implying ARG bioaccumulation/proliferation. - Abstract: The occurrence of tetracycline resistance (TRG) and integrase (INT) genes were monitored in Crassostrea virginica oyster reefs of three pristine creeks (SINERR, Georgia, USA). Their profiles revealed 85% similarity with the TRG/INT profiles observed in the adjacent to the SINERR and contaminated Altamaha River estuary (Barkovskii et al., 2010). The TRG/INT spectra and incidence frequencies corresponded to the source of oceanic input and to run-offs from creeks’ watersheds. The highest incidence frequencies and concentrations were observed in oysters. TRG/INT incidences correlated positively (Spearman Rank = 0.88), and negatively correlated (−0.63 to −0.79) with creek salinity, conductivity, dissolved solids, and temperature. Coliform incidence positively correlated with temperature, and not with the TRG/INT incidence. The Altamaha River estuary was the primary TRG/INT source for the reefs with contributions from creek’s watersheds. TRG/INT were carried by non-coliforms with a preference for low-to-temperate thermohaline environments coupled with bioaccumulation by oysters.

  7. A review of the influence of treatment strategies on antibiotic resistant bacteria and antibiotic resistance genes.

    Sharma, Virender K; Johnson, Natalie; Cizmas, Leslie; McDonald, Thomas J; Kim, Hyunook

    2016-05-01

    Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG) in the aquatic environment have become an emerging contaminant issue, which has implications for human and ecological health. This review begins with an introduction to the occurrence of ARB and ARG in different environmental systems such as natural environments and drinking water resources. For example, ARG or ARB with resistance to ciprofloxacin, sulfamethoxazole, trimethoprim, quinolone, vancomycin, or tetracycline (e.g., tet(A), tet(B), tet(C), tet(G), tet(O), tet(M), tet(W), sul I, and sul II) have been detected in the environment. The development of resistance may be intrinsic, may be acquired through spontaneous mutations (de novo), or may occur due to horizontal gene transfer from donor bacteria, phages, or free DNA to recipient bacteria. An overview is also provided of the current knowledge regarding inactivation of ARB and ARG, and the mechanism of the effects of different disinfection processes in water and wastewater (chlorination, UV irradiation, Fenton reaction, ozonation, and photocatalytic oxidation). The effects of constructed wetlands and nanotechnology on ARB and ARG are also summarized. PMID:26775188

  8. EDA Gene Mutations Underlie Non-syndromic Oligodontia

    Song, S.; Han, D.; Qu, H.; Gong, Y.; Wu, H.; Zhang, X.; Zhong, N.; Feng, H.

    2009-01-01

    Recent studies have detected mutations in the EDA gene, previously identified as causing X-linked hypohidrotic ectodermal dysplasia (XLHED), in two families with X-linked non-syndromic hypodontia. Notably, all affected males in both families exhibited isolated oligodontia, while almost all female carriers showed a milder or normal phenotype. We hypothesized that the EDA gene could be responsible for sporadic non-syndromic oligodontia in affected males. In this study, we examined 15 unrelated males with non-syndromic oligodontia. Three novel EDA mutations (p.Ala259Glu, p.Arg289Cys, and p.Arg334His) were identified in four individuals (27%). A genetic defect in the EDA gene could result in non-syndromic oligodontia in affected males. PMID:19278982

  9. Fate of antibiotic resistance genes and its drivers during anaerobic co-digestion of food waste and sewage sludge based on microwave pretreatment.

    Zhang, Junya; Chen, Meixue; Sui, Qianwen; Wang, Rui; Tong, Juan; Wei, Yuansong

    2016-10-01

    In this study, anaerobic digestion of mono-SS, MW-SS:FW and SS:MW-FW was investigated to understand the fate of ARGs and its drivers. Anaerobic digestion was effective for the reduction of metal resistance genes (MRGs), and could reduce the abundance of blaOXA-1, sulI and tetG, while sulII in co-digestion and blaTEM and ereA only in MW-SS. ARGs reduction could be partly attributed to the reduction of co-selective pressure from heavy metals reflected by MRGs. However, the abundance of mefA/E, ermB, ermF, tetM and tetX increased significantly. Anaerobic co-digestion, especially for MW-SS, could reduce total ARGs abundance compared with mono-SS, and evolution of bacterial community was the main driver for the fate of ARGs. PMID:26988135

  10. Variable effects of oxytetracycline on antibiotic resistance gene abundance and the bacterial community during aerobic composting of cow manure.

    Qian, Xun; Sun, Wei; Gu, Jie; Wang, Xiao-Juan; Sun, Jia-Jun; Yin, Ya-Nan; Duan, Man-Li

    2016-09-01

    Livestock manure is often subjected to aerobic composting but little is known about the variation in antibiotic resistance genes (ARGs) during the composting process under different concentrations of antibiotics. This study compared the effects of three concentrations of oxytetracycline (OTC; 10, 60, and 200mg/kg) on ARGs and the succession of the bacterial community during composting. Very similar trends were observed in the relative abundances (RAs) of each ARG among the OTC treatments and the control during composting. After composting, the RAs of tetC, tetX, sul1, sul2, and intI1 increased 2-43 times, whereas those of tetQ, tetM, and tetW declined by 44-99%. OTC addition significantly increased the absolute abundances and RAs of tetC and intI1, while 200mg/kg OTC also enhanced those of tetM, tetQ, and drfA7. The bacterial community could be grouped according to the composting time under different treatments. The highest concentration of OTC had a more persistent effect on the bacterial community. In the present study, the succession of the bacterial community appeared to have a greater influence on the variation of ARGs during composting than the presence of antibiotics. Aerobic composting was not effective in reducing most of the ARGs, and thus the compost product should be considered as an important reservoir for ARGs. PMID:27179201

  11. Appearance of β-lactam Resistance Genes in Agricultural Soils and Clinical Isolates over the 20th Century

    Graham, David W.; Knapp, Charles W.; Christensen, Bent Tolstrup; McCluskey, Seánin; Dolfing, Jan

    archived since 1923 at Askov Experimental Station in Denmark, we quantified four broad-spectrum β-lactam AR genes (ARG; blaTEM, blaSHV, blaOXA and blaCTX-M) and class-1 integron genes (int1) in soils from manured (M) versus inorganic fertilised (IF) fields. “Total” β-lactam ARG levels were significantly...... higher in M versus IF in soils post-1940 (paired-t test; p < 0.001). However, dominant individual ARGs varied over time; blaTEM and blaSHV between 1963 and 1974, blaOXA slightly later, and blaCTX-M since 1988. These dates roughly parallel first reporting of these genes in clinical isolates, suggesting...... ARGs in animal manure and humans are historically interconnected. Archive data further show when non-therapeutic antibiotic use was banned in Denmark, blaCTX-M levels declined in M soils, suggesting accumulated soil ARGs can be reduced by prudent antibiotic stewardship. Conversely, int1 levels have...

  12. Recombinant Brucella abortus gene expressing immunogenic protein

    Mayfield, J.E.; Tabatabai, L.B.

    1991-06-11

    This patent describes a synthetic recombinant DNA molecule containing a DNA sequence. It comprises a gene of Brucella abortus encoding an immunogenic protein having a molecular weight of approximately 31,000 daltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under denaturing conditions, the protein having an isoelectric point around 4.9, and containing a twenty-five amino acid sequence from its amino terminal end consisting of Gln-Ala-Pro-Thr-Phe-Phe-Arg-Ile-Gly-Thr-Gly-Gly-Thr-Ala-Gly-Thr-Tyr-Tyr-Pro-Ile-Gly-Gly-Leu-Ile-Ala, wherein Gln, Ala, Pro, Thr, Phe, Arg, Ile, Gly, Tyr, and Leu, respectively, represent glutamine, alanine, proline, threonine, phenylalanine, arginine, isolecuine, glycine, tyrosine, and leucine.

  13. The Gln223Arg polymorphism of the leptin receptor in Pima Indians: influence on energy expenditure, physical activity and lipid metabolism

    Stefan, N; Vozarova, B; Del Parigi, A;

    2002-01-01

    Leptin regulates body weight by its receptor-mediated anorectic, thermogenic and antisteatotic effects. Recently, lower leptin binding to the soluble form of the leptin receptor (LEPR) was shown in carriers of the Arg223-encoding allele of the Gln223Arg polymorphism of the LEPR. To investigate wh...... who had undergone percutaneous fat biopsies from the periumbilical region (n=184). Genotype was not associated with percent body fat (P>0.39), but was associated with 24 h EE, PAL and mean subcutaneous abdominal adipocyte size (SAAS all P...

  14. Association of the beta3-adrenergic receptor Trp64Arg polymorphism with common metabolic traits: studies of 7605 middle-aged white people

    Gjesing, A. P.; Andersen, G; Borch-Johnsen, K; Jørgensen, T; Hansen, T; Pedersen, Oluf

    2008-01-01

    cases and 6049 controls) and type 2 diabetes (1373 cases and 4742 controls) and quantitative trait analyses among 5822 individuals. Furthermore, the association of Trp64Arg with type 2 diabetes was examined in a meta-analysis. RESULTS: The Trp64Arg polymorphism was not associated with obesity. However...... does not confer an increased risk of obesity among Danes. Yet, in the present study of 7605 Danes the variant is associated with type 2 diabetes and quantitative traits related to type 2 diabetes....

  15. Metabolism of the broad-spectrum neuropeptide growth factor antagonist: [D-Arg1, D-Phe5, D-Trp7,9, Leu11]-substance P.

    Jones, D. A.; Cummings, J.; Langdon, S. P.; Maclellan, A. J.; Higgins, T; Rozengurt, E; Smyth, J. F.

    1996-01-01

    Broad-spectrum neuropeptide growth factor antagonists, such as [D-Arg1, D-Phe5, D-Trp7,9, Leu11]substance P (antagonist D) and [Arg6, D-Trp7,9, NmePhe8]substance P(6-11) (antagonist G), are currently being investigated as possible anti-tumour agents. These compounds are hoped to be effective against neuropeptide-driven cancers such as small-cell lung cancer. Antagonist D possesses a broader antagonistic spectrum than antagonist G and hence may be of greater therapeutic use. The in vitro metab...

  16. Genotyping and meta-analysis of KIF6 Trp719Arg polymorphism in South Indian Coronary Artery Disease patients: A case–control study

    Durairajpandian Vishnuprabu

    2015-09-01

    Full Text Available The KIF6 719Arg allele is an interesting genomic variant widely screened in various populations and is reported to be associated with the risk of Coronary Artery Disease (CAD and statin treatment outcome. Recent population based clinical studies and large-scale meta-analyses pondered over the role of 719Arg variant in CAD risk and treatment response. We screened the KIF6 Trp719Arg polymorphism (rs20455 in south Indian CAD patients in a case–control approach. A total of 1042 samples (510 CAD patients and 532 controls were screened for the KIF6 Trp719Arg SNP by TaqMan SNP genotyping assay, followed by meta-analysis of the genotype data of non-Europeans reports. The 719Arg risk genotype (GG was observed in 29.6% of CAD cases and in 30.1% of controls with an odds ratio (OR of 1.07 (95% CI: 0.76–1.50, p value = 0.709. No significant difference in the genotype frequency was observed between CAD and controls in both dominant model (AG + GG vs AA and allelic model (719Arg vs 719Trp with an OR of 1.11 (p = 0.491 and 1.03 (p = 0.767, respectively. The covariate analysis indicated that smoking & alcohol consumption increased the risk for MI among CAD patients. Meta-analysis showed that the KIF6 719Arg allele is not associated with CAD risk in both fixed effect (p = 0.515, OR = 1.023, 95% CI = 0.956–1.094 and random effect (p = 0.547, OR = 1.022, 95% CI = 0.953–1.096. The symmetrical shape of the Egger's funnel plots revealed that there is no publication bias. These results suggest that there is no association of KIF6 719Arg allele with CAD risk in South Indian population and the meta-analysis confirms the same among non-European population.

  17. COMPARATIVE STUDY OF TOURIST TRAFFIC AT THE CITY, COUNTY AND HOUSE IN CÂMPULUNG MUSCEL ARGES

    Adelaida Cristina HONTUŞ

    2013-01-01

    Full Text Available This paper is based on a comparative analysis of tourism demand and supply and the development of mountain tourism tendinţelorde plannaţional and internationally, the degree of capitalization of the mountain tourism in Romania, focusing on assessment of tourism in the tourist resort Campulung Muscel County Arges. The objective of this study consists of a comparative analysis of tourist traffic at the county level tourist resort and tourist board level. To perform this study we used a series of statistical data provided by the representatives of the City of Delhi Muscel on tourism and tourist traffic statistics on the county level and at the boarding house.

  18. Three different prohormones yield a variety of Hydra-RFamide (Arg-Phe-NH2) neuropeptides in Hydra magnipapillata

    Darmer, D; Hauser, F; Nothacker, H P; Bosch, T C; Williamson, M; Grimmelikhuijzen, C J

    The freshwater polyp Hydra is the most frequently used model for the study of development in cnidarians. Recently we isolated four novel Arg-Phe-NH2 (RFamide) neuropeptides, the Hydra-RFamides I-IV, from Hydra magnipapillata. Here we describe the molecular cloning of three different preprohormones...... from H. magnipapillata, each of which gives rise to a variety of RFamide neuropeptides. Preprohormone A contains one copy of unprocessed Hydra-RFamide I (QWLGGRFG), II (QWFNGRFG), III/IV [(KP)HLRGRFG] and two putative neuropeptide sequences (QLMSGRFG and QLMRGRFG). Preprohormone B has the same general...... organization as preprohormone A, but instead of unprocessed Hydra-RFamide III/IV it contains a slightly different neuropeptide sequence [(KP)HYRGRFG]. Preprohormone C contains one copy of unprocessed Hydra-RFamide I and seven additional putative neuropeptide sequences (with the common N-terminal sequence QWF...

  19. Direct assessment of the effect of the Gly380Arg achondroplasia mutation on FGFR3 dimerization using quantitative imaging FRET.

    Jesse Placone

    Full Text Available The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH, the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand.

  20. In vitro and in vivo pharmacological profile of PL-3994, a novel cyclic peptide (Hept-cyclo(Cys-His-Phe-d-Ala-Gly-Arg-d-Nle-Asp-Arg-Ile-Ser-Cys)-Tyr-[Arg mimetic]-NH(2)) natriuretic peptide receptor-A agonist that is resistant to neutral endopeptidase and acts as a bronchodilator.

    Edelson, Jeffrey D; Makhlina, Marie; Silvester, Kevin R; Vengurlekar, Shailesh S; Chen, Xiaomei; Zhang, Jie; Koziol-White, Cynthia J; Cooper, Philip R; Hallam, Trevor J; Hay, Douglas W P; Panettieri, Reynold A

    2013-04-01

    The pharmacological and airways relaxant profiles of PL-3994 (Hept-cyclo(Cys-His-Phe-d-Ala-Gly-Arg-d-Nle-Asp-Arg-Ile-Ser-Cys)-Tyr-[Arg mimetic]-NH(2)), a novel natriuretic peptide receptor-A (NPR-A) agonist, were evaluated. PL-3994, a full agonist, has high affinity for recombinant human (h), dog, or rat NPR-As (K(i)s of 1, 41, and 10 nm, respectively), and produced concentration-dependent cGMP generation in human, dog and rat NPR-As (respective EC(50)s of 2, 3 and 14 nm). PL-3994 has a K(i) of 7 nm for hNPR-C but was without effect on cGMP generation in hNPR-B. PL-3994 (1 μm) was without significant effect against 75 diverse molecular targets. PL-3994 or BNP, a natural NPR ligand, produced concentration-dependent relaxation of pre-contracted guinea-pig trachea (IC(50)s of 42.7 and 10.7 nm, respectively). PL-3994, and also BNP, (0.1 nm-100 μm) elicited a potent, concentration-dependent but small relaxation of pre-contracted human precision-cut lung slices (hPCLS). Intratracheal PL-3994 (1-1000 μg/kg) produced a dose-dependent inhibition of the bronchoconstrictor response evoked by aerosolized methacholine, but was without significant effect on cardiovascular parameters. PL-3994 was resistant to degradation by human neutral endopeptidase (hNEP) (92% remaining after 2 h), whereas the natural ligands, ANP and CNP, were rapidly metabolized (≤1% remaining after 2 h). PL-3994 is a potent, selective NPR agonist, resistant to NEP, with relaxant effects in guinea-pig and human airway smooth muscle systems. PL-3994 has the profile predictive of longer clinical bronchodilator activity than observed previously with ANP, and suggests its potential utility in the treatment of asthma, in addition to being a useful research tool to evaluate NPR biology. PMID:23154072

  1. Dynamic properties of the native free antithrombin from molecular dynamics simulations: computational evidence for solvent- exposed Arg393 side chain.

    Tóth, László; Fekete, Attila; Balogh, Gábor; Bereczky, Zsuzsanna; Komáromi, István

    2015-09-01

    While antithrombin (AT) has small basal inhibitory activity, it reaches its full inhibitory potential against activated blood coagulation factors, FXa, FIXa, and FIIa (thrombin), via an allosteric and/or template (bridging) mechanism by the action of heparin, heparan sulfate, or heparin-mimetic pentasaccharides (PS). From the numerous X-ray structures available for different conformational states of AT, only indirect and incomplete conclusions can be drawn on the inherently dynamic properties of AT. As a typical example, the basal inhibitory activity of AT cannot be interpreted on the basis of "non-activated" free antithrombin X-ray structures since the Arg393 side chain, playing crucial role in antithrombin-proteinase interaction, is not exposed. In order to reveal the intrinsic dynamic properties and the reason of basal inhibitory activity of antithrombin, 2 μs molecular dynamics simulations were carried out on its native free-forms. It was shown from the simulation trajectories that the reactive center loop which is functioning as "bait" for proteases, even without any biasing potential can populate conformational state in which the Arg393 side chain is solvent exposed. It is revealed from the trajectory analysis that the peptide sequences correspond to the helix D extension, and new helix P formation can be featured with especially large root-mean-square fluctuations. Mutual information analyses of the trajectory showed remarkable (generalized) correlation between those regions of antithrombin which changed their conformations as the consequence of AT-PS complex formation. This suggests that allosteric information propagation pathways are present even in the non-activated native form of AT. PMID:25483839

  2. Homozygous tyrosinase gene mutation in an American black with tyrosinase-negative (type IA) oculocutaneous albinism.

    Spritz, R.A.; Strunk, K M; Hsieh, C L; G.S. Sekhon; Francke, U.

    1991-01-01

    We have identified a tyrosinase gene mutation in an American black with classic, tyrosinase-negative oculocutaneous albinism. This mutation results in an amino acid substitution (Cys----Arg) at codon 89 of the tyrosinase polypeptide. The proband is homozygous for the substitution, suggesting that this mutation may be frequently associated with tyrosinase-negative oculocutaneous albinism in blacks.

  3. Relationships between antibiotics and antibiotic resistance gene levels in municipal solid waste leachates in Shanghai, China.

    Wu, Dong; Huang, Zhiting; Yang, Kai; Graham, David; Xie, Bing

    2015-04-01

    Many studies have quantified antibiotics and antibiotic resistance gene (ARG) levels in soils, surface waters, and waste treatment plants (WTPs). However, similar work on municipal solid waste (MSW) landfill leachates is limited, which is concerning because antibiotics disposal is often in the MSW stream. Here we quantified 20 sulfonamide (SA), quinolone (FQ), tetracycline (TC), macrolide (ML), and chloramphenicol (CP) antibiotics, and six ARGs (sul1, sul2, tetQ, tetM, ermB, and mefA) in MSW leachates from two Shanghai transfer stations (TS; sites Hulin (HL) and Xupu (XP)) and one landfill reservoir (LR) in April and July 2014. Antibiotic levels were higher in TS than LR leachates (985 ± 1965 ng/L vs 345 ± 932 ng/L, n = 40), which was because of very high levels in the HL leachates (averaging at 1676 ± 5175 ng/L, n = 40). The mean MLs (3561 ± 8377 ng/L, n = 12), FQs (975 ± 1608 ng/L, n = 24), and SAs (402 ± 704 ng/L, n = 42) classes of antibiotics were highest across all samples. ARGs were detected in all leachate samples with normalized sul2 and ermB levels being especially elevated (-1.37 ± 1.2 and -1.76 ± 1.6 log (copies/16S-rDNA), respectively). However, ARG abundances did not correlate with detected antibiotic levels, except for tetW and tetQ with TC levels (r = 0.88 and 0.81, respectively). In contrast, most measured ARGs did significantly correlate with heavy metal levels (p < 0.05), especially with Cd and Cr. This study shows high levels of ARGs and antibiotics can prevail in MSW leachates and landfills may be an underappreciated as a source of antibiotics and ARGs to the environment. PMID:25760223

  4. Polymorphisms of XRCC1 genes and risk of nasopharyngeal carcinoma in the Cantonese population

    Lin Dong-Xin

    2006-06-01

    Full Text Available Abstract Background Nasopharyngeal carcinoma (NPC is one of the most common cancers in southern China. In addition to environmental factors such as Epstein-Barr virus infection and diet, genetic susceptibility has been reported to play a key role in the development of this disease. The x-ray repair cross-complementing group 1 (XRCC1 gene is important in DNA base excision repair. We hypothesized that two common single nucleotide polymorphisms of XRCC1 (codons 194 Arg→Trp and 399 Arg→Gln are related to the risk of NPC and interact with tobacco smoking. Methods We sought to determine whether these genetic variants of the XRCC1 gene were associated with the risk of NPC among the Cantonese population in a hospital-based case control study using polymerase chain reaction-restriction fragment length polymorphism analysis. We conducted this study in 462 NPC patients and 511 healthy controls. Results After adjustment for sex and age, we found a reduced risk of developing NPC in individuals with the Trp194Trp genotype (OR = 0.48; 95% CI, 0.27–0.86 and the Arg194Trp genotype (OR = 0.79; 95% CI, 0.60–1.05 compared with those with the Arg194Arg genotype. Compared with those with the Arg399Arg genotype, the risk for NPC was not significantly different in individuals with the Arg399Gln genotype (OR = 0.82; 95% CI, 0.62–1.08 and the Gln399Gln genotype (OR = 1.20; 95% CI, 0.69–2.06. Further analyses stratified by gender and smoking status revealed a significantly reduced risk of NPC among males (OR = 0.32; 95% CI, 0.14–0.70 and smokers (OR = 0.34; 95% CI, 0.14–0.82 carrying the XRCC1 194Trp/Trp genotype compared with those carrying the Arg/Arg genotype. No association was observed between Arg399Gln variant genotypes and the risk of NPC combined with smoking and gender. Conclusion Our findings suggest that the XRCC1 Trp194Trp variant genotype is associated with a reduced risk of developing NPC in Cantonese population, particularly in males and

  5. Polymorphisms of XRCC1 genes and risk of nasopharyngeal carcinoma in the Cantonese population

    Nasopharyngeal carcinoma (NPC) is one of the most common cancers in southern China. In addition to environmental factors such as Epstein-Barr virus infection and diet, genetic susceptibility has been reported to play a key role in the development of this disease. The x-ray repair cross-complementing group 1 (XRCC1) gene is important in DNA base excision repair. We hypothesized that two common single nucleotide polymorphisms of XRCC1 (codons 194 Arg→Trp and 399 Arg→Gln) are related to the risk of NPC and interact with tobacco smoking. We sought to determine whether these genetic variants of the XRCC1 gene were associated with the risk of NPC among the Cantonese population in a hospital-based case control study using polymerase chain reaction-restriction fragment length polymorphism analysis. We conducted this study in 462 NPC patients and 511 healthy controls. After adjustment for sex and age, we found a reduced risk of developing NPC in individuals with the Trp194Trp genotype (OR = 0.48; 95% CI, 0.27–0.86) and the Arg194Trp genotype (OR = 0.79; 95% CI, 0.60–1.05) compared with those with the Arg194Arg genotype. Compared with those with the Arg399Arg genotype, the risk for NPC was not significantly different in individuals with the Arg399Gln genotype (OR = 0.82; 95% CI, 0.62–1.08) and the Gln399Gln genotype (OR = 1.20; 95% CI, 0.69–2.06). Further analyses stratified by gender and smoking status revealed a significantly reduced risk of NPC among males (OR = 0.32; 95% CI, 0.14–0.70) and smokers (OR = 0.34; 95% CI, 0.14–0.82) carrying the XRCC1 194Trp/Trp genotype compared with those carrying the Arg/Arg genotype. No association was observed between Arg399Gln variant genotypes and the risk of NPC combined with smoking and gender. Our findings suggest that the XRCC1 Trp194Trp variant genotype is associated with a reduced risk of developing NPC in Cantonese population, particularly in males and smokers. Larger studies are needed to confirm our findings

  6. TP53 gene polymorphisms at codons 11, 72, and 248 and association with endometriosis in a Brazilian population.

    Camargo-Kosugi, C M; D'Amora, P; Kleine, J P F O; Carvalho, C V; Sato, H; Schor, E; Silva, I D C G

    2014-01-01

    We evaluated the association between TP53 gene polymorphisms and endometriosis in Brazilian women. Genomic DNA was extracted from swabs of buccal cells collected from hospital patients. TP53 gene polymorphisms were investigated at three codons: TP53 11 Glu/Gln or Lys (GAG->CAG or AAG), TP53 72 Arg/Pro (CCG->CCC), and TP53 248 Arg/Thr (CGG->TCG) using the polymerase chain reaction-restriction fragment length polymorphism method. TP53 11 presented the following genotypic distribution: the control group was 98.28% homozygous wild-type (Glu) and 1.72% homozygous variant (Gln/Lys), and the heterozygous genotype was not identified. The genotypic distribution in the endometriosis group was 96% homozygous wild-type (Glu) and 4% heterozygous (Glu-Gln/Lys); the homozygous variant genotype was not identified (P = 0.02). TP53 72 showed the following genotypic distribution: the control group was 29.75% homozygous wild-type (Arg), 47.11% heterozygous (Arg-Pro), and 23.14% homozygous variant (Pro). The genotypic distribution in the endometriosis group was 16.15% homozygous wild-type (Arg), 51.54% heterozygous (Arg-Pro), and 32.31% homozygous variant (Pro) (odds ratio = 2.26; 95% confidence interval = 1.19-4.03; P = 0.02). Only one patient had the homozygous TP53 248 genotype (Arg-Trp/Gln); all other patients were homozygous wild-type in both the control and endometriosis groups (P = 0.51; NS). We found that TP53 72 polymorphism may be associated with susceptibility to endometriosis; the presence of at least 1 polymorphic allele increased the chance of disease development by 2.26-fold. Hence, this genetic variant is a potential candidate marker for endometriosis. PMID:25177931

  7. Excretion of antibiotic resistance genes by dairy calves fed milk replacers with varying doses of antibiotics

    Callie H. Thames

    2012-04-01

    Full Text Available Elevated levels of antibiotic resistance genes (ARGs in soil and water have been linked to livestock farms and in some cases feed antibiotics may select for antibiotic resistant gut microbiota. The purpose of this study was to examine the establishment of ARGs in the feces of calves receiving milk replacer containing no antibiotics versus subtherapeutic or therapeutic doses of tetracycline and neomycin. The effect of antibiotics on calf health was also of interest. Twenty-eight male and female dairy calves were assigned to one of the three antibiotic treatment groups at birth and fecal samples were collected at weeks 6, 7 (prior to weaning, and 12 (5 weeks after weaning. ARGs corresponding to the tetracycline (tetC, tetG, tetO, tetW, and tetX, macrolide (ermB, ermF, and sulfonamide (sul1, sul2 classes of antibiotics along with the class I integron gene, intI1, were monitored by quantitative polymerase chain reaction as potential indicators of direct selection, co-selection, or horizontal gene transfer of ARGs. Surprisingly, there was no significant effect of antibiotic treatment on the absolute abundance (gene copies/ g wet manure of any of the ARGs except ermF, which was lower in the antibiotic-treated calf manure, presumably because a significant portion of host bacterial cells carrying ermF were not resistant to tetracycline or neomycin. However, relative abundance (gene copies normalized to 16S rRNA genes of tetO was higher in calves fed the highest dose of antibiotic than in the other treatments. All genes, except tetC and intI1, were detectable in feces from 6 weeks onwards, and tetW and tetG significantly increased (P<0.10, even in control calves. Overall, the results provide new insight into the colonization of calf gut flora with ARGs in the early weeks. Although feed antibiotics exerted little effect on the ARGs monitored in this study, the fact that they also provided no health benefit suggests that the greater than conventional

  8. Assessment of anaerobic bacterial diversity and its effects on anaerobic system stability and the occurrence of antibiotic resistance genes.

    Aydin, Sevcan; Ince, Bahar; Ince, Orhan

    2016-05-01

    This study evaluated the link between anaerobic bacterial diversity and, the biodegradation of antibiotic combinations and assessed how amending antibiotic combination and increasing concentration of antibiotics in a stepwise fashion influences the development of resistance genes in anaerobic reactors. The biodegradation, sorption and occurrence of the known antibiotic resistance genes (ARGs) of erythromycin and tetracycline were investigated using the processes of UV-HPLC and qPCR analysis respectively. Ion Torrent sequencing was used to detect microbial community changes in response to the addition of antibiotics. The overall results indicated that changes in the structure of a microbial community lead to changes in biodegradation capacity, sorption of antibiotics combinations and occurrence of ARGs. The enhanced biodegradation efficiency appeared to generate variations in the structure of the bacterial community. The results suggested that controlling the ultimate Gram-negative bacterial community, especially Acinetobacter-related populations, may promote the successful biodegradation of antibiotic combinations and reduce the occurrence of ARGs. PMID:26897411

  9. Survival of Antibiotic Resistant Bacteria and Horizontal Gene Transfer Control Antibiotic Resistance Gene Content in Anaerobic Digesters

    Miller, Jennifer H.; Novak, John T.; Knocke, William R.; Pruden, Amy

    2016-01-01

    Understanding fate of antibiotic resistant bacteria (ARB) vs. their antibiotic resistance genes (ARGs) during wastewater sludge treatment is critical in order to reduce the spread of antibiotic resistance through process optimization. Here, we spiked high concentrations of tetracycline-resistant bacteria, isolated from mesophilic (Iso M1-1—a Pseudomonas sp.) and thermophilic (Iso T10—a Bacillus sp.) anaerobic digested sludge, into batch digesters and monitored their fate by plate counts and quantitative polymerase chain reaction (QPCR) of their corresponding tetracycline ARGs. In batch studies, spiked ARB plate counts returned to baseline (thermophilic) or 1-log above baseline (mesophilic) while levels of the ARG present in the spiked isolate [tet(G)] remained high in mesophilic batch reactors. To compare results under semi-continuous flow conditions with natural influent variation, tet(O), tet(W), and sul1 ARGs, along with the intI1 integrase gene, were monitored over a 9-month period in the raw feed sludge and effluent sludge of lab-scale thermophilic and mesophilic anaerobic digesters. sul1 and intI1 in mesophilic and thermophilic digesters correlated positively (Spearman rho = 0.457–0.829, P < 0.05) with the raw feed sludge. There was no correlation in tet(O) or tet(W) ratios in raw sludge and mesophilic digested sludge or thermophilic digested sludge (Spearman rho = 0.130–0.486, P = 0.075–0.612). However, in the thermophilic digester, the tet(O) and tet(W) ratios remained consistently low over the entire monitoring period. We conclude that the influent sludge microbial composition can influence the ARG content of a digester, apparently as a result of differential survival or death of ARBs or horizontal gene transfer of genes between raw sludge ARBs and the digester microbial community. Notably, mesophilic digestion was more susceptible to ARG intrusion than thermophilic digestion, which may be attributed to a higher rate of ARB survival and

  10. Survival of Antibiotic Resistant Bacteria and Horizontal Gene Transfer Control Antibiotic Resistance Gene Content in Anaerobic Digesters.

    Miller, Jennifer H; Novak, John T; Knocke, William R; Pruden, Amy

    2016-01-01

    Understanding fate of antibiotic resistant bacteria (ARB) vs. their antibiotic resistance genes (ARGs) during wastewater sludge treatment is critical in order to reduce the spread of antibiotic resistance through process optimization. Here, we spiked high concentrations of tetracycline-resistant bacteria, isolated from mesophilic (Iso M1-1-a Pseudomonas sp.) and thermophilic (Iso T10-a Bacillus sp.) anaerobic digested sludge, into batch digesters and monitored their fate by plate counts and quantitative polymerase chain reaction (QPCR) of their corresponding tetracycline ARGs. In batch studies, spiked ARB plate counts returned to baseline (thermophilic) or 1-log above baseline (mesophilic) while levels of the ARG present in the spiked isolate [tet(G)] remained high in mesophilic batch reactors. To compare results under semi-continuous flow conditions with natural influent variation, tet(O), tet(W), and sul1 ARGs, along with the intI1 integrase gene, were monitored over a 9-month period in the raw feed sludge and effluent sludge of lab-scale thermophilic and mesophilic anaerobic digesters. sul1 and intI1 in mesophilic and thermophilic digesters correlated positively (Spearman rho = 0.457-0.829, P < 0.05) with the raw feed sludge. There was no correlation in tet(O) or tet(W) ratios in raw sludge and mesophilic digested sludge or thermophilic digested sludge (Spearman rho = 0.130-0.486, P = 0.075-0.612). However, in the thermophilic digester, the tet(O) and tet(W) ratios remained consistently low over the entire monitoring period. We conclude that the influent sludge microbial composition can influence the ARG content of a digester, apparently as a result of differential survival or death of ARBs or horizontal gene transfer of genes between raw sludge ARBs and the digester microbial community. Notably, mesophilic digestion was more susceptible to ARG intrusion than thermophilic digestion, which may be attributed to a higher rate of ARB survival and/or horizontal gene

  11. Site-directed mutagenesis of Arg58 and Asp86 of elongation factor Tu from Escherichia coli: effects on the GTPase reaction and aminoacyl-tRNA binding

    Knudsen, Charlotte Rohde; Clark, Brian F. C.

    1996-01-01

    Elongation factor Tu from Escherichia coli was mutated separately at positions Asp86 and Arg58, in order to shed light both on the GTPase mechanism of elongation factor Tu and on the binding of aminoacyl-tRNA. In addition, the binding of guanine nucleotides was investigated by determination of th...

  12. IL-10 and ARG-1 Concentrations in Bone Marrow and Peripheral Blood of Metastatic Neuroblastoma Patients Do Not Associate with Clinical Outcome

    Fabio Morandi

    2015-01-01

    Full Text Available The expression of the immunosuppressive molecules IL-10 and arginase 1 (ARG-1, and of FOXP3 and CD163, as markers of regulatory T cells (Treg and macrophages, respectively, was evaluated in bone marrow (BM and peripheral blood (PB samples collected at diagnosis from patients with metastatic neuroblastoma (NB. IL-10 and ARG-1 plasma concentrations were measured and the association of each parameter with patients’ outcome was tested. The percentages of immunosuppressive Treg and type-1 regulatory (Tr1 cells were also determined. In both BM and PB samples, IL-10 mRNA expression was higher in metastatic NB patients than in controls. IL-10 plasma concentration was higher in patients with NB regardless of stage. Neither IL-10 expression nor IL-10 plasma concentration significantly associated with patient survival. In PB samples from metastatic NB patients, ARG-1 and CD163 expression was higher than in controls but their expression did not associate with survival. Moreover, ARG-1 plasma concentration was lower than in controls, and no association with patient outcome was found. Finally, in metastatic NB patients, the percentage of circulating Treg was higher than in controls, whereas that of Tr1 cells was lower. In conclusion, although IL-10 concentration and Treg percentage were increased, their contribution to the natural history of metastatic NB appears uncertain.

  13. Cell adhesion to fibrillin-1: identification of an Arg-Gly-Asp-dependent synergy region and a heparin-binding site that regulates focal adhesion formation

    Bax, Daniel V; Mahalingam, Yashithra; Cain, Stuart; Mellody, Kieran; Freeman, Lyle; Younger, Kerri; Shuttleworth, C Adrian; Humphries, Martin J; Couchman, John R; Kielty, Cay M

    We have defined the molecular basis of cell adhesion to fibrillin-1, the major structural component of extracellular microfibrils that are associated with elastic fibres. Using human dermal fibroblasts, and recombinant domain swap fragments containing the Arg-Gly-Asp motif, we have demonstrated a...

  14. High Incidence of Noonan Syndrome Features Including Short Stature and Pulmonic Stenosis in Patients carrying NF1 Missense Mutations Affecting p.Arg1809: Genotype-Phenotype Correlation.

    Rojnueangnit, Kitiwan; Xie, Jing; Gomes, Alicia; Sharp, Angela; Callens, Tom; Chen, Yunjia; Liu, Ying; Cochran, Meagan; Abbott, Mary-Alice; Atkin, Joan; Babovic-Vuksanovic, Dusica; Barnett, Christopher P; Crenshaw, Melissa; Bartholomew, Dennis W; Basel, Lina; Bellus, Gary; Ben-Shachar, Shay; Bialer, Martin G; Bick, David; Blumberg, Bruce; Cortes, Fanny; David, Karen L; Destree, Anne; Duat-Rodriguez, Anna; Earl, Dawn; Escobar, Luis; Eswara, Marthanda; Ezquieta, Begona; Frayling, Ian M; Frydman, Moshe; Gardner, Kathy; Gripp, Karen W; Hernández-Chico, Concepcion; Heyrman, Kurt; Ibrahim, Jennifer; Janssens, Sandra; Keena, Beth A; Llano-Rivas, Isabel; Leppig, Kathy; McDonald, Marie; Misra, Vinod K; Mulbury, Jennifer; Narayanan, Vinodh; Orenstein, Naama; Galvin-Parton, Patricia; Pedro, Helio; Pivnick, Eniko K; Powell, Cynthia M; Randolph, Linda; Raskin, Salmo; Rosell, Jordi; Rubin, Karol; Seashore, Margretta; Schaaf, Christian P; Scheuerle, Angela; Schultz, Meredith; Schorry, Elizabeth; Schnur, Rhonda; Siqveland, Elizabeth; Tkachuk, Amanda; Tonsgard, James; Upadhyaya, Meena; Verma, Ishwar C; Wallace, Stephanie; Williams, Charles; Zackai, Elaine; Zonana, Jonathan; Lazaro, Conxi; Claes, Kathleen; Korf, Bruce; Martin, Yolanda; Legius, Eric; Messiaen, Ludwine

    2015-11-01

    Neurofibromatosis type 1 (NF1) is one of the most frequent genetic disorders, affecting 1:3,000 worldwide. Identification of genotype-phenotype correlations is challenging because of the wide range clinical variability, the progressive nature of the disorder, and extreme diversity of the mutational spectrum. We report 136 individuals with a distinct phenotype carrying one of five different NF1 missense mutations affecting p.Arg1809. Patients presented with multiple café-au-lait macules (CALM) with or without freckling and Lisch nodules, but no externally visible plexiform neurofibromas or clear cutaneous neurofibromas were found. About 25% of the individuals had Noonan-like features. Pulmonic stenosis and short stature were significantly more prevalent compared with classic cohorts (P < 0.0001). Developmental delays and/or learning disabilities were reported in over 50% of patients. Melanocytes cultured from a CALM in a segmental NF1-patient showed two different somatic NF1 mutations, p.Arg1809Cys and a multi-exon deletion, providing genetic evidence that p.Arg1809Cys is a loss-of-function mutation in the melanocytes and causes a pigmentary phenotype. Constitutional missense mutations at p.Arg1809 affect 1.23% of unrelated NF1 probands in the UAB cohort, therefore this specific NF1 genotype-phenotype correlation will affect counseling and management of a significant number of patients. PMID:26178382

  15. Sulfonamide and tetracycline resistance genes in total- and culturable-bacterial assemblages in South African aquatic environments

    Suzuki, Satoru; Ogo, Mitsuko; Koike, Tatsuya; Takada, Hideshige; Newman, Brent

    2015-01-01

    Antibiotic resistant bacteria are ubiquitous in the natural environment. The introduction of effluent derived antibiotic resistance genes (ARGs) into aquatic environments is of concern in the spreading of genetic risk. This study showed the prevalence of sulfonamide and tetracycline resistance genes, sul1, sul2, sul3, and tet(M), in the total bacterial assemblage and colony forming bacterial assemblage in river and estuarine water and sewage treatment plants (STP) in South Africa. There was no correlation between antibiotic concentrations and ARGs, suggesting the targeted ARGs are spread in a wide area without connection to selection pressure. Among sul genes, sul1 and sul2 were major genes in the total (over 10-2 copies/16S) and colony forming bacteria assemblages (∼10-1 copies/16S). In urban waters, the sul3 gene was mostly not detectable in total and culturable assemblages, suggesting sul3 is not abundant. tet(M) was found in natural assemblages with 10-3 copies/16S level in STP, but was not detected in colony forming bacteria, suggesting the non-culturable (yet-to-be cultured) bacterial community in urban surface waters and STP effluent possess the tet(M) gene. Sulfamethoxazole (SMX) resistant (SMXr) and oxytetracycline (OTC) resistant (OTCr) bacterial communities in urban waters possessed not only sul1 and sul2 but also sul3 and tet(M) genes. These genes are widely distributed in SMXr and OTCr bacteria. In conclusion, urban river and estuarine water and STP effluent in the Durban area were highly contaminated with ARGs, and the yet-to-be cultured bacterial community may act as a non-visible ARG reservoir in certain situations. PMID:26300864

  16. Effect of p53 Arg72Pro polymorphism on the induction of micronucleus by aflatoxin B1 in in vitro in human blood lymphocytes.

    Bayram, Süleyman; Rencüzoğulları, Eyyüp; Almas, Abdullah Muttalip; Genç, Ahmet

    2016-07-01

    Aflatoxin B1 (AFB1) is a class 1 carcinogen produced by Aspergillus flavus and Aspergillus parasiticus that can contaminate a variety of food substances, the liver being its target organ. A common p53 Arg72Pro polymorphism resulting in the substitution of an arginine amino acid by proline amino acid in the transactivating domain has been demonstrated to affect p53 function. The aim of this study is to investigate association between p53 Arg72Pro polymorphism and the frequencies of spontaneous and AFB1-induced DNA damage in peripheral blood lymphocytes from 100 healthy individuals in Turkish population. In vitro cytokinesis-blocked micronucleus (CBMN) assay was used to detect the spontaneous and AFB1-induced DNA damage whereas, genotyping of p53 Arg72Pro polymorphism was carried out by using a polymerase chain reaction restriction fragment length polymorphism assay. During 68 h incubation time, lymphocytes treated with AFB1 (1.56 μg/mL) and S9 mix for a total of 3 h (48-51 h). Treatment of the lymphocytes with AFB1significantly increased the overall frequencies of micronucleus (MN) when compared to untreated cultures (1.23 ± 0.05 versus 0.55 ± 0.02; p < 0.001). Moreover, genotype analysis revealed a statistically significant association between Pro/Pro genotype of p53 Arg72Pro polymorphism and increased frequencies of MN both spontaneous and AFB1-induced cultures when compared Arg/Arg genotype (0.69 ± 0.19 versus 0.46 ± 0.13, p < 0.001; 1.59 ± 0.65 versus 1.01 ± 0.41 p < 0.001; respectively). Our data indicate that p53 Arg72Pro polymorphism plays a significant role in human sensitivity to the genotoxic effects of AFB1. Further investigations in larger sample size and with different ethnic origins as well as including more functional single nucleotide polymorphisms might lead to the identification of novel genetic factors responsible for susceptibility to human carcinogens such as AFB1. PMID:26738694

  17. Characterization of Antibiotics and Antibiotic Resistance Genes on an Ecological Farm System

    Songhe Zhang

    2015-01-01

    Full Text Available There is a growing concern worldwide about the prevalence of antibiotics and antibiotic resistance genes (ARGs on the farm. In this study, we investigated the distribution of seven antibiotics and ten ARGs in fresh and dried pig feces, in biogas slurry, and in grape-planting soil from an ecological farm. Antibiotics including sulfamethazine, norfloxacin, ofloxacin, tetracycline, oxytetracycline, and chlortetracycline were detected in these samples (except for sulfamethoxazole in dried feces. In general, antibiotics levels in samples were in the sequence: biogas slurry > fresh feces > soil or dried feces. Results of ecological risk assessments revealed that among the seven antibiotics chlortetracycline showed the highest ecological risk. Among the ten ARGs, sulI and tetO were the most prevalent on this ecological farm. There were positive correlations between certain ARGs and the corresponding antibiotics on this ecological farm. Therefore, continuous monitoring of antibiotics and their corresponding ARGs should be conducted in the agroecosystem near the concentrated animal farming operation systems.

  18. Distributive characteristics of Ser49Gly and Gly389Arg genetic polymorphisms of β1-adrenoceptor in Chinese Han and Dai populations

    Zhao-qian LIU; Hong-hao ZHOU; Jie LIU; Zhi-hua XIANG; Min-yu HU; Wei MO; Lian-sheng WANG; Dong-sheng OU-YANG; Nan HE; Dan WANG

    2006-01-01

    Aim: Genetic polymorphisms causing Ser49Gly and Gly389Arg mutants of β1-adrenoceptor may result in significant changes in the function of this receptor.The aim of the present study was to investigate the frequencies of the Ser49Gly and Gly389Arg mutant alleles in healthy Chinese populations and to investigate the differences between 2 Chinese ethnic groups (Han and Dai populations) with respect to the frequencies of these alleles. Methods: A total of 225 Han Chinese and 175 Dai Chinese unrelated healthy volunteers were recruited for this study.Genomic DNA was extracted from peripheral blood leukocytes by using a standard manual chloroform-phenol extraction. Fragments spanning the 2 polymorphisms were amplified by using polymerase chain reaction with template genomic DNA and relevant primers. The DNA products including the polymorphic loci were subjected to restriction endonuclease digestion with Eco0l09I and BcgI.Digested fragments were detected with an ultraviolet detector after electrophoresis (100 V for approximately 1.5 h). Results: The frequencies of the Gly49 and Arg389 alleles were, respectively, 16.2% and 76.4% in the Han population and 14.6%and 75.7% in the Dai population. Conclusion: The polymorphisms causing the Ser49Gly and Gly389Arg mutations of the β1-adrenoceptor existed in both healthy Han and Dai Chinese populations. The frequencies of the Ser49Gly and Gly389Arg mutant alleles were not significantly different in the Han and Dai populations.However, the frequency of the Gly389 variant seems to be significantly lower in these 2 populations than in an African-American population.

  19. Fate of antibiotic resistant cultivable heterotrophic bacteria and antibiotic resistance genes in wastewater treatment processes.

    Zhang, Songhe; Han, Bing; Gu, Ju; Wang, Chao; Wang, Peifang; Ma, Yanyan; Cao, Jiashun; He, Zhenli

    2015-09-01

    Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are emerging contaminants of environmental concern. Heterotrophic bacteria in activated sludge have an important role in wastewater treatment plants (WWTPs). However, the fate of cultivable heterotrophic ARB and ARGs in WWPTs process remains unclear. In the present study, we investigated the antibiotic-resistant phenotypes of cultivable heterotrophic bacteria from influent and effluent water of three WWTPs and analysed thirteen ARGs in ARB and in activated sludge from anoxic, anaerobic and aerobic compartments. From each influent or effluent sample of the three plants, 200 isolates were randomly tested for susceptibility to 12 antibiotics. In these samples, between 5% and 64% isolates showed resistance to >9 antibiotics and the proportion of >9-drug-resistant bacteria was lower in isolates from effluent than from influent. Eighteen genera were identified in 188 isolates from influent (n=94) and effluent (n=94) of one WWTP. Six genera (Aeromonas, Bacillus, Lysinibacillus, Microbacterium, Providencia, and Staphylococcus) were detected in both influent and effluent samples. Gram-negative and -positive isolates dominated in influent and effluent, respectively. The 13 tetracycline-, sulphonamide-, streptomycin- and β-lactam-resistance genes were detected at a higher frequency in ARB from influent than from effluent, except for sulA and CTX-M, while in general, the abundances of ARGs in activated sludge from two of the three plants were higher in aerobic compartments than in anoxic ones, indicating abundant ARGs exit in the excess sledges and/or in uncultivable bacteria. These findings may be useful for elucidating the effect of WWTP on ARB and ARGs. PMID:25950407

  20. Ozone treatment of conditioned wastewater selects antibiotic resistance genes, opportunistic bacteria, and induce strong population shifts.

    Alexander, Johannes; Knopp, Gregor; Dötsch, Andreas; Wieland, Arne; Schwartz, Thomas

    2016-07-15

    An ozone treatment system was investigated to analyze its impact on clinically relevant antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs). A concentration of 0.9±0.1g ozone per 1g DOC was used to treat conventional clarified wastewater. PCR, qPCR analyses, Illumina 16S Amplicon Sequencing, and PCR-DGGE revealed diverse patterns of resistances and susceptibilities of opportunistic bacteria and accumulations of some ARGs after ozone treatment. Molecular marker genes for enterococci indicated a high susceptibility to ozone. Although they were reduced by almost 99%, they were still present in the bacterial population after ozone treatment. In contrast to this, Pseudomonas aeruginosa displayed only minor changes in abundance after ozone treatment. This indicated different mechanisms of microorganisms to cope with the bactericidal effects of ozone. The investigated ARGs demonstrated an even more diverse pattern. After ozone treatment, the erythromycin resistance gene (ermB) was reduced by 2 orders of magnitude, but simultaneously, the abundance of two other clinically relevant ARGs increased within the surviving wastewater population (vanA, blaVIM). PCR-DGGE analysis and 16S-Amplicon-Sequencing confirmed a selection-like process in combination with a substantial diversity loss within the vital wastewater population after ozone treatment. Especially the PCR-DGGE results demonstrated the survival of GC-rich bacteria after ozone treatment. PMID:27058129

  1. Environmental dissemination of antibiotic resistance genes and correlation to anthropogenic contamination with antibiotics

    Björn Berglund

    2015-09-01

    Full Text Available Antibiotic resistance is a growing problem which threatens modern healthcare globally. Resistance has traditionally been viewed as a clinical problem, but recently non-clinical environments have been highlighted as an important factor in the dissemination of antibiotic resistance genes (ARGs. Horizontal gene transfer (HGT events are likely to be common in aquatic environments; integrons in particular are well suited for mediating environmental dissemination of ARGs. A growing body of evidence suggests that ARGs are ubiquitous in natural environments. Particularly, elevated levels of ARGs and integrons in aquatic environments are correlated to proximity to anthropogenic activities. The source of this increase is likely to be routine discharge of antibiotics and resistance genes, for example, via wastewater or run-off from livestock facilities and agriculture. While very high levels of antibiotic contamination are likely to select for resistant bacteria directly, the role of sub-inhibitory concentrations of antibiotics in environmental antibiotic resistance dissemination remains unclear. In vitro studies have shown that low levels of antibiotics can select for resistant mutants and also facilitate HGT, indicating the need for caution. Overall, it is becoming increasingly clear that the environment plays an important role in dissemination of antibiotic resistance; further studies are needed to elucidate key aspects of this process. Importantly, the levels of environmental antibiotic contamination at which resistant bacteria are selected for and HGT is facilitated at should be determined. This would enable better risk analyses and facilitate measures for preventing dissemination and development of antibiotic resistance in the environment.

  2. TWO ETERNAL WORKS DEFYING TIME –THE MONASTERY OF ARGES AND THE BUILDING OF THE FORTRESS ON THE BOJANA

    Marina Puja-Badesku

    2012-12-01

    Full Text Available This paper proposes to analyze several motives from Serbian and Romanian folk literature. It identifies the main themes and motives from the local folklore, similar situations from the folklore of the two peoples, the Romanian and the Serbian. The common Balkan history and, especially, the struggle against the Ottoman Turks, are the starting point of both the Serbian and the Romanian legend (The Monastery of Arges and the Building of the Fortress on the River Bojana. This paper attempts to present several motives and their presence in the legends of the two neighboring peoples, stressing not only the similarities, but also the differences between them. What should be pointed out are previous, detailed studies about local and Balkan folklore, starting from names of references and the work they have done in this field. One of the main differences, and one that will mark the overall meaning of the two legends, is related to the ethnic, religious and traditional characteristics of each of the two nations. If with the Romanian the image of creation, the arch spreading over generations and generations - is a church, a religious temple, then with the Serbian it is a tower built as a defense against Turks. Their continuity in space and time is considered from the point of view of the building of the Arges Temple and the Fortress on the Bojana, respectively; in his play about Manole the Craftsman, Lucian Blaga also speaks of mythical time. In the Romanian version, Manole sacrifices himself, which is an idea of creation that defies time, and in the popular mind it implies the necessity of complete sacrifice, and death is transformed into immortality through the acts of building an edifice and a fountain. Common motives in Serbian and Romanian folk literature are the result of good neighborly relations in this part of the Balkans and the legends of Manole the Craftsman and the Building of the Fortress on the Bojana are the best example of this. These two

  3. Report on a 2009 mini-demonstration of the ARG-US Radio Frequency Identification (RFID) system in transportation.

    Tsai, H.; Chen, K.; Jusko, M.; Craig, B.; Liu, Y.; Decision and Information Sciences

    2009-11-23

    The Packaging Certification Program (PCP) of the U.S. Department of Energy (DOE) Environmental Management (EM), Office of Packaging and Transportation (EM-14), has developed a radio frequency identification (RFID) tracking and monitoring system for the management of nuclear materials during storage and transportation. The system, developed by the PCP team at Argonne National Laboratory, consists of hardware (Mk-series sensor tags, fixed and handheld readers, form factor for multiple drum types, seal integrity sensors, and enhanced battery management), software (application programming interface, ARG-US software for local and remote/web applications, secure server and database management), and cellular/satellite communication interfaces for vehicle tracking and item monitoring during transport. The ability of the above system to provide accurate, real-time tracking and monitoring of the status of multiple, certified containers of nuclear materials has been successfully demonstrated in a week-long, 1,700-mile DEMO performed in April 2008. While the feedback from the approximately fifty (50) stakeholders who participated in and/or observed the DEMO progression were very positive and encouraging, two major areas of further improvements - system integration and web application enhancement - were identified in the post-DEMO evaluation. The principal purpose of the MiniDemo described in this report was to verify these two specific improvements. The MiniDemo was conducted on August 28, 2009. In terms of system integration, a hybrid communication interface - combining the RFID item-monitoring features and a commercial vehicle tracking system by Qualcomm - was developed and implemented. In the MiniDemo, the new integrated system worked well in reporting tag status and vehicle location accurately and promptly. There was no incompatibility of components. The robust commercial communication gear, as expected, helped improve system reliability. The MiniDemo confirmed that system

  4. Report on a 2009 mini-demonstration of the ARG-US Radio Frequency Identification (RFID) system in transportation

    The Packaging Certification Program (PCP) of the U.S. Department of Energy (DOE) Environmental Management (EM), Office of Packaging and Transportation (EM-14), has developed a radio frequency identification (RFID) tracking and monitoring system for the management of nuclear materials during storage and transportation. The system, developed by the PCP team at Argonne National Laboratory, consists of hardware (Mk-series sensor tags, fixed and handheld readers, form factor for multiple drum types, seal integrity sensors, and enhanced battery management), software (application programming interface, ARG-US software for local and remote/web applications, secure server and database management), and cellular/satellite communication interfaces for vehicle tracking and item monitoring during transport. The ability of the above system to provide accurate, real-time tracking and monitoring of the status of multiple, certified containers of nuclear materials has been successfully demonstrated in a week-long, 1,700-mile DEMO performed in April 2008. While the feedback from the approximately fifty (50) stakeholders who participated in and/or observed the DEMO progression were very positive and encouraging, two major areas of further improvements - system integration and web application enhancement - were identified in the post-DEMO evaluation. The principal purpose of the MiniDemo described in this report was to verify these two specific improvements. The MiniDemo was conducted on August 28, 2009. In terms of system integration, a hybrid communication interface - combining the RFID item-monitoring features and a commercial vehicle tracking system by Qualcomm - was developed and implemented. In the MiniDemo, the new integrated system worked well in reporting tag status and vehicle location accurately and promptly. There was no incompatibility of components. The robust commercial communication gear, as expected, helped improve system reliability. The MiniDemo confirmed that system

  5. Obesity-related gene ADRB2, ADRB3 and GHRL polymorphisms and the response to a weight loss diet intervention in adult women

    Saliba, Louise F.; Reis, Rodrigo S; Brownson, Ross C.; Hino, Adriano A.; Tureck, Luciane V.; Cheryl Valko; Ricardo L.R. Souza; Lupe Furtado-Alle

    2014-01-01

    The individual response to diet may be influenced by gene polymorphisms. This study hypothesized that ADRB2 (Gln27Glu, rs1042714 and Arg16Gly, rs1042713), ADRB3 (Trp64Arg, rs4994) and GHRL (Leu72Met, rs696217) polymorphisms moderate weight loss. The study was a seven weeks dietary weight loss intervention with Brazilian adult obese women (n = 109). The body mass index (BMI) was calculated and polymorphisms in these genes were assessed by real-time PCR assays. Two-way repeated-measures ANOVA (...

  6. High-throughput quantification of antibiotic resistance genes from an urban wastewater treatment plant.

    Karkman, Antti; Johnson, Timothy A; Lyra, Christina; Stedtfeld, Robert D; Tamminen, Manu; Tiedje, James M; Virta, Marko

    2016-03-01

    Antibiotic resistance among bacteria is a growing problem worldwide, and wastewater treatment plants have been considered as one of the major contributors to the dissemination of antibiotic resistance to the environment. There is a lack of comprehensive quantitative molecular data on extensive numbers of antibiotic resistance genes (ARGs) in different seasons with a sampling strategy that would cover both incoming and outgoing water together with the excess sludge that is removed from the process. In order to fill that gap we present a highly parallel quantitative analysis of ARGs and horizontal gene transfer potential over four seasons at an urban wastewater treatment plant using a high-throughput qPCR array. All analysed transposases and two-thirds of primer sets targeting ARGs were detected in the wastewater. The relative abundance of most of the genes was highest in influent and lower in effluent water and sludge. The resistance profiles of the samples cluster by sample location with a shift from raw influent through the final effluents and dried sludge to the sediments. Wastewater discharge enriched only a few genes, namely Tn25 type transposase gene and clinical class 1 integrons, in the sediment near the discharge pipe, but those enriched genes may indicate a potential for horizontal gene transfer. PMID:26832203

  7. Class 1 integrase, sulfonamide and tetracycline resistance genes in wastewater treatment plant and surface water.

    Makowska, Nicoletta; Koczura, Ryszard; Mokracka, Joanna

    2016-02-01

    Wastewater treatment plants are considered hot spots for multiplication and dissemination of antibiotic-resistant bacteria and resistance genes. In this study, we determined the presence of class 1 integron integrase and genes conferring resistance to tetracyclines and sulfonamides in the genomes of culturable bacteria isolated from a wastewater treatment plant and the river that receives the treated wastewater. Moreover, using PCR-based metagenomic approach, we quantified intI1, tet and sul genes. Wastewater treatment caused the decrease in the total number of culturable heterotrophs and bacteria resistant to tetracycline and sulfonamides, along with the decrease in the number of intI1, sul and tet gene copies per ml, with significant reduction of tet(B). On the other hand, the treatment process increased both the frequency of tetracycline- and sulfonamide-resistant bacteria and intI1-positive strains, and the relative abundance of all quantified antibiotic resistance genes (ARGs) and intI1 gene; in the case of tet(A) and sul2 significantly. The discharge of treated wastewater increased the number of intI1, tet and sul genes in the receiving river water both in terms of copy number per ml and relative abundance. Hence, despite the reduction of the number of ARGs and ARBs, wastewater treatment selects for bacteria with ARGs in effluent. PMID:26519797

  8. Genes and Gene Therapy

    ... correctly, a child can have a genetic disorder. Gene therapy is an experimental technique that uses genes to ... or prevent disease. The most common form of gene therapy involves inserting a normal gene to replace an ...

  9. Gly71Arg UGT1A1 polymorphism is associated with breast cancer susceptibility in Han Chinese women.

    Shi, J; Li, L H; Duan, X Y; Liu, Q; Sun, L L; Tian, Y T

    2016-01-01

    Breast cancer is among the most common causes of cancer-related death in women worldwide. Previous studies have demonstrated an association between prolonged estrogen exposure and increased risk of breast cancer. Uridine 5'-diphospho-glucuronosyltransferase 1-1 (UGT1A1) plays a significant role in the detoxification of estrogens. Two major genetic polymorphisms have been identified in the UGT1A1 locus. UGT1A1*28 has been previously linked to increased risk of breast cancer. The aim of this study was to elucidate the possible correlation between UGT1A1*6, a single nucleotide polymorphism causing a Gly71Arg substitution, and breast cancer susceptibility. Forty-six women diagnosed with breast cancer, 15 patients with gastrointestinal cancer, and 13 healthy women were recruited to this study. The genotype in the polymorphic UGT1A1 locus was determined by DNA sequencing. The frequency of each genotype was compared among the three groups. The frequency of the UGT1A1*6 allele was significantly higher in breast cancer and gastrointestinal cancer patients than that in healthy females (both P 0.05). Therefore, the UGT1A1*6 polymorphism was deduced to be a risk factor for breast cancer in women of Han Chinese ethnicity. UGT1A1 may serve as a therapeutic target for the prevention and treatment of breast cancer and other estrogen-related diseases. PMID:27525948

  10. The role of Arg(78) in the metabotropic glutamate receptor mGlu(1) for agonist binding and selectivity

    Jensen, Anders A.; Sheppard, P O; O'Hara, P J; Krogsgaard-Larsen, P; Bräuner-Osborne, H

    The metabotropic glutamate receptors belong to family C of the G-protein coupled receptor superfamily. These receptors all possess large extracellular amino terminal domains, where agonist binding takes place. We have previously constructed a molecular model of the amino terminal domain of the m...... leucine or glutamate completely knocks out [3H]quisqualic acid binding to the receptor. The constructed mutants, R78L and R78E, have also been characterized in a inositol phosphate assay. Here, the potency of (S)-glutamic acid and (S)-quisqualic acid was reduced 1000- and 100-fold, respectively, on R78L...... compared to the wild type (WT) receptor. (S)-Quisqualic acid was as potent on mutant R78E as it was on R78L, whereas (S)-glutamic acid was unable to activate R78E significantly at concentrations up to 10 mM. In conclusion, Arg(78) appears to be essential for agonist binding to the mGlu(1) receptor, most...

  11. Identification of Minor Secondary Metabolites from the Latex of Croton lechleri (Muell-Arg and Evaluation of Their Antioxidant Activity

    Maria Iorizzi

    2008-06-01

    Full Text Available Dragon’s blood (Sangre de drago, a viscous red sap derived from Croton lechleri Muell-Arg (Euphorbiaceae, is extensively used by indigenous cultures of the Amazonian basin for its wound healing properties. The aim of this study was to identify the minor secondary metabolites and test the antioxidant activity of this sustance. A bioguided fractionation of the n-hexane, chloroform, n-butanol, and aqueous extracts led to the isolation of 15 compounds: three megastigmanes, four flavan-3-ols, three phenylpropanoids, three lignans, a clerodane, and the alkaloid taspine. In addition to these known molecules, six compounds were isolated and identified for the first time in the latex: blumenol B, blumenol C, 4,5-dihydroblumenol A, erythro-guaiacyl-glyceryl-β-O-4’- dihydroconiferyl ether, 2-[4-(3-hydroxypropyl-2-methoxyphenoxy]-propane-1,3-diol and floribundic acid glucoside. Combinations of spectroscopic methods (1H-, 13C- NMR and 2D-NMR experiments, ESI-MS, and literature comparisons were used for compound identification. In vitro antioxidant activities were assessed by DPPH, total antioxidant capacity and lipid peroxidation assays. Flavan-3-ols derivatives (as major phenolic compounds in the latex exhibited the highest antioxidant activity.

  12. Mothers of autistic children: lower plasma levels of oxytocin and Arg-vasopressin and a higher level of testosterone.

    Xin-Jie Xu

    Full Text Available BACKGROUND: Autism is a pervasive neurodevelopmental disorder,thought to be caused by a combination of genetic heritability and environmental risk factors. Some autistic-like traits have been reported in mothers of autistic children. We hypothesized that dysregulation of oxytocin (OXT, Arg-vasopressin (AVP and sex hormones, found in autistic children, may also exist in their mothers. METHODS: We determined plasma levels of OXT (40 in autism vs. 26 in control group, AVP (40 vs. 17 and sex hormones (61 vs. 47 in mothers of autistic and normal children by enzyme immunoassay and radioimmunoassay, respectively and investigated their relationships with the children's autistic behavior scores (Childhood Autism Rating Scale (CARS and Autism Behavior Checklist (ABC. RESULTS: Significantly lower plasma concentrations of OXT (p<0.001 and AVP (p<0.001, as well as a higher level of plasma testosterone (p<0.05, were found in mothers of autistic children vs. those of control. The children's autistic behavior scores were negatively associated with maternal plasma levels of OXT and AVP. CONCLUSIONS: These results suggest that dysregulation of OXT, AVP and/or testosterone systems exist in mothers of autistic children, which may impact children's susceptibility to autism.

  13. Blood pressure levels in male carriers of Arg82Cys in CD300LG

    Støy, Julie; Grarup, Niels; Hørlyck, Arne;

    2014-01-01

    UNLABELLED: The genetics of hypertension has been scrutinized in large-scale genome-wide association studies (GWAS) with a large number of common genetic variants identified, each exerting subtle effects on disease susceptibility. An amino acid polymorphism, p.Arg82Cys, in CD300LG was recently fo...... found to be associated with fasting HDL-cholesterol and triglyceride levels. The polymorphism has not been detected in hypertension GWAS potentially due to its low frequency, but CD300LG has been linked to blood pressure as CD300LG knockout mice have changes in blood pressure. Twenty...... (122 mmHg versus 115; p = 0.01) and diastolic blood pressure (77 mmHg versus 72; p<0.01) compared with CC-carriers. There were no differences in CIMT between the two groups. Metalloproteinase-9 level was higher in CT-carriers than in CC-carriers (P<0.01). However, no association between office blood...... pressure and CD300LG genotype was detected in the genetic-epidemiological study. CONCLUSIONS: Although 24-hour blood pressure, measured with a sensitive method, in a small sample of CD300LG rs72836561 CT-carriers was higher than in CC-carriers, this did not translate into significant differences in office...

  14. The Variant p.(Arg183Trp) in SPTLC2 Causes Late-Onset Hereditary Sensory Neuropathy.

    Suriyanarayanan, Saranya; Auranen, Mari; Toppila, Jussi; Paetau, Anders; Shcherbii, Maria; Palin, Eino; Wei, Yu; Lohioja, Tarja; Schlotter-Weigel, Beate; Schön, Ulrike; Abicht, Angela; Rautenstrauss, Bernd; Tyynismaa, Henna; Walter, Maggie C; Hornemann, Thorsten; Ylikallio, Emil

    2016-03-01

    Hereditary sensory and autonomic neuropathy 1 (HSAN1) is an autosomal dominant disorder that can be caused by variants in SPTLC1 or SPTLC2, encoding subunits of serine palmitoyl-CoA transferase. Disease variants alter the enzyme's substrate specificity and lead to accumulation of neurotoxic 1-deoxysphingolipids. We describe two families with autosomal dominant HSAN1C caused by a new variant in SPTLC2, c.547C>T, p.(Arg183Trp). The variant changed a conserved amino acid and was not found in public variant databases. All patients had a relatively mild progressive distal sensory impairment, with onset after age 50. Small fibers were affected early, leading to abnormalities on quantitative sensory testing. Sural biopsy revealed a severe chronic axonal neuropathy with subtotal loss of myelinated axons, relatively preserved number of non-myelinated fibers and no signs for regeneration. Skin biopsy with PGP9.5 labeling showed lack of intraepidermal nerve endings early in the disease. Motor manifestations developed later in the disease course, but there was no evidence of autonomic involvement. Patients had elevated serum 1-deoxysphingolipids, and the variant protein produced elevated amounts of 1-deoxysphingolipids in vitro, which proved the pathogenicity of the variant. Our results expand the genetic spectrum of HSAN1C and provide further detail about the clinical characteristics. Sequencing of SPTLC2 should be considered in all patients presenting with mild late-onset sensory-predominant small or large fiber neuropathy. PMID:26573920

  15. P53 codon 11, 72, and 248 gene polymorphisms in endometriosis

    Yao-Yuan Hsieh , Chich-Sheng Lin

    2006-01-01

    Full Text Available Objective: Mutated p53 gene is related to the instability of cell growth and cell cycle progression. We aimed to evaluate the association between endometriosis and p53 codon 11, 72 and 248 gene polymorphisms. Patients and methods: Women were divided into two groups: (1 moderate/severe endometriosis (n=148, and (2 non-endometriosis groups (n=150. P53 gene polymorphisms include codon11 Glu/Gln or Lys (GAG->CAG or AAG, codon 72 Arg/Pro (CGC->CCC, and codon 248 Arg/Thr (CGG->TCG. These gene polymorphisms were amplified by polymerase chain reaction and detected by electrophoresis after restriction enzyme (Taq I, BstU I, Hap II digestions. Associations between the endometriosis and p53 polymorphisms were evaluated. Results: The distributions of p53 codon 72 polymorphisms in both groups were significantly different. The proportions of Arg homozygotes/heterozygotes/Pro homozygotes in both groups were 9.5/66.2/24.3% and 30.7/50/19.3%. The proportions of Arg/Pro alleles were 42.6/57.4% and 56/44%. The distributions of p53 codon 11 and 248 polymorphisms in both groups were non-significantly different. All individuals appeared the wild genotypes (Glu11 and Arg248 homozygotes. Conclusion: Association between endometriosis and p53 codon 72 polymorphism exists. P53 codon 72*Pro-related genotype and allele are related with higher susceptibility of endometriosis. P53 codon 11 and 248 polymorphisms are not related with endometriosis susceptibility.

  16. Codon 201Gly Polymorphic Type of the DCC Gene is Related to Disseminated Neuroblastoma

    Xiao-Tang Kong

    2001-01-01

    Full Text Available The deleted in colorectal carcinoma (DCC gene is a potential tumor- suppressor gene on chromosome 18821.3. The relatively high frequency of loss of heterozygosity (LOH and loss of expression of this gene in neuroblastoma, especially in the advanced stages, imply the possibility of involvement of the DCC gene in progression of neuroblastoma. However, only few typical mutations have been identified in this gene, indicating that other possible mechanisms for the inactivation of this gene may exist. A polymorphic change (Arg to Gly at DCC codon 201 is related to advanced colorectal carcinoma and increases in the tumors with absent DCC protein expression. In order to understand whether this change is associated with the development or progression of neuroblastoma, we investigated codon 201 polymorphism of the DCC gene in 102 primary neuroblastomas by polymerase chain reaction single-strand conformation polymorphism. We found no missense or nonsense mutations, but a polymorphic change from CGA (Arg to GGA (Gly at codon 201 resulting in three types of polymorphism: codon 201Gly type, codon 201Arg/Gly type, and codon 201Arg type. The codon 201Gly type occurred more frequently in disseminated (stages IV and IVs neuroblastomas (72% than in localized (stages I, II, and III tumors (48% (P=.035, and normal controls (38% (P=.024. In addition, the codon 201Gly type was significantly more common in tumors found clinically (65% than in those found by mass screening (35% (P=.002. The results suggested that the codon 201Gly type of the DCC gene might be associated with a higher risk of disseminating neuroblastoma.

  17. A novel syndrome of autosomal-dominant hyperinsulinemic hypoglycemia linked to a mutation in the human insulin receptor gene

    Højlund, Kurt; Hansen, Torben; Lajer, Maria;

    2004-01-01

    revealed a missense mutation (Arg1174Gln) in the tyrosine kinase domain of the insulin receptor gene that cosegregated with the disease phenotype (logarithm of odds [LOD] score 3.21). In conclusion, we report a novel syndrome of autosomal-dominant hyperinsulinemic hypoglycemia. The findings demonstrate the...

  18. Relationship between β3-AR Gene and Obesity, Type 2 Diabetes, Insulin Resistance in Chinese Han Population

    HEWei; MAXiang-hua; SHENJie

    2004-01-01

    Objective: To explore the relationship between the β3-adrenergic receptor(β3-AR)gene and obesity, T2DM. insulin resistance in Chinese Han population. Methods: Fifty-three healthy subjects, 105 subjects with simple obesity, 63 type 2 diabetic patients without obesity, and 114 type 2 diabetic patients with obesity were studied with the technique of PCR-RFLP in codon 64 of the exon region of β3-AR gene representing the variation Trp/Arg. Results:Compared with the subjects of Trp homozygous group, the individuals with Arg allele were more elevated in WHR,MBP,SBP,DBP,FBS,PBS, FINS,PINS, FCP,PCP and lower in ISI. Frequency of Arg allele was higher in HINS sub-group without T2DM. Cnclusion: The results indicate that the Trp/Arg variation might lead to insulin resistance, obesity and T2DM.β3-AR gene is supposed to be the candidate gene of insulin resistance, obesity and T2DM in ChineseHan population.

  19. Utilización de otros láseres en odontología: Argón, Nd:YAP y Ho:YAG

    Guinot Moya, Rosa; España Tost, A. J. (Antonio Jesús); Berini Aytés, Leonardo; Gay Escoda, Cosme

    2004-01-01

    Las ventajas que la incorporación del láser ha proporcionado a la Odontología abarcan todos los ámbitos. Así, los láseres de Argón, de Nd:YAP y de Ho:YAG, tienen aplicaciones muy concretas y de gran interés tanto en el campo de la terapéutica dental como en la especialida de cirugía bucal. Las aplicaciones principales del láser de Argón se centran en la polimerización de los materiales de restauración, en la endodoncia y dentro del ámbito de la cirugía bucal, en el corte de tejidos blandos, p...

  20. The MECP2 variant c.925C>T (p.Arg309Trp) causes intellectual disability in both males and females without classic features of Rett syndrome.

    Schönewolf-Greulich, B; Tejada, M-I; Stephens, K; Hadzsiev, K; Gauthier, J; Brøndum-Nielsen, K; Pfundt, R; Ravn, K; Maortua, H; Gener, B; Martínez-Bouzas, C; Piton, A; Rouleau, G; Clayton-Smith, J; Kleefstra, T; Bisgaard, A-M; Tümer, Z

    2016-06-01

    Missense MECP2 variants can have various phenotypic effects ranging from a normal phenotype to typical Rett syndrome (RTT). In females, the phenotype can also be influenced by the X-inactivation pattern. In this study, we present detailed clinical descriptions of six patients with a rare base-pair substitution affecting Arg309 at the C-terminal end of the transcriptional repression domain (TRD). All patients have intellectual disability and present with some RTT features, but they do not fulfill the clinical criteria for typical or atypical RTT. Most of the patients also have mild facial dysmorphism. Intriguingly, the mother of an affected male patient is an asymptomatic carrier of this variant. It is therefore likely that the p.(Arg309Trp) variation does not necessarily lead to male lethality, and it results in a wide range of clinical features in females, probably influenced by different X-inactivation patterns in target tissues. PMID:26936630

  1. High affinity binding of the heat-stable protein kinase inhibitor to the catalytic subunit of cAMP-dependent protein kinase is selectively abolished by mutation of Arg133.

    Wen, W; Taylor, S S

    1994-03-18

    The two classes of physiological inhibitors of the catalytic subunit of cAMP-dependent protein kinase are the regulatory subunits and the heat-stable protein kinase inhibitors (PKIs), and both share a common mechanism of inhibition. Each has a similar inhibitor site that resembles a peptide substrate, and this occupies the P-3 to P+1 portion of the peptide recognition site. However, in addition to this consensus site, each inhibitor requires a peripheral binding site to achieve high affinity binding. Arg134 and Arg133 lie on the surface of the catalytic subunit with Arg133 coming close to the amphipathic helix of PKI(5-24) (Knighton, D. R., Zheng, J., Ten Eyck, L. F., Xuong, N.-h., Taylor, S. S., and Sowadski, J. M. (1991) Science 253, 414-420). Replacement of Arg134 and Arg133 with Ala selectively abolishes the high affinity binding of PKI. Replacement of Arg133 alone is sufficient to give the same phenotype. In the presence of MgATP, the Kd,app, is increased from < 0.2 to 105 nM and, in the absence of ATP, the Kd is too large to be reliably measured. Based on the crystal structure, Arg133 hydrogen bonds to the P-7 backbone carbonyl of PKI(5-24). However, more importantly, it also contributes to the hydrophobicity of the P-11 binding site in the C.PKI(5-24) complex. We predict that it is the perturbation of this hydrophobic pocket that accounts for the effects of this mutation. In the absence of peptide, Arg133 may help to stabilize Glu230, a buried carboxylate that binds to the P-2 Arg in the crystal structure of C.PKI(5-24). Replacement of Arg133 and Arg134 with Ala has little effect on catalysis using a heptapeptide substrate and has no effect on the inhibition of the catalytic subunit by the regulatory subunit. The results thus demonstrate that these two inhibitor proteins that both bind to the catalytic subunit with a high affinity utilize different sites on the enzyme to achieve tight binding. The gamma isoform of the catalytic subunit is insensitive to

  2. High incidence of Noonan syndrome features including short stature and pulmonic stenosis in patients carrying NF1 missense mutations affecting p.Arg1809: genotype-phenotype correlation

    Rojnueangnit, Kitiwan; Xie, Jing; Gomes, Alicia; Sharp, Angela; Callens, Tom; Chen, Yunjia; Liu, Ying; Cochran, Meagan; Abbott, Mary-Alice; Atkin, Joan; Babovic-Vuksanovic, Dusica; Barnett, Christopher P; Crenshaw, Melissa; Bartholomew, Dennis W; Basel, Lina

    2015-01-01

    Neurofibromatosis type 1 (NF1) is one of the most frequent genetic disorders, affecting 1:3,000 worldwide. Identification of genotype-phenotype correlations is challenging because of the wide range clinical variability, the progressive nature of the disorder, and extreme diversity of the mutational spectrum. We report 136 individuals with a distinct phenotype carrying one of five different NF1 missense mutations affecting p.Arg1809. Patients presented with multiple cafe-au-lait macules (CALM)...

  3. Expression of FOXP3, CD14, and ARG1 in Neuroblastoma Tumor Tissue from High-Risk Patients Predicts Event-Free and Overall Survival

    Sara Stigliani

    2015-01-01

    Full Text Available The prognosis of children with metastatic neuroblastoma (NB > 18 months at diagnosis is dismal. Since the immune status of the tumor microenvironment could play a role in the history of disease, we evaluated the expression of CD45, CD14, ARG1, CD163, CD4, FOXP3, Perforin-1 (PRF1, Granzyme B (GRMB, and IL-10 mRNAs in primary tumors at diagnosis from children with metastatic NB and tested whether the transcript levels are significantly associated to event-free and overall survival (EFS and OS, resp.. Children with high expression of CD14, ARG1 and FOXP3 mRNA in their primary tumors had significantly better EFS. Elevated expression of CD14, and FOXP3 mRNA was significantly associated to better OS. CD14 mRNA expression levels significantly correlated to all markers, with the exception of CD4. Strong positive correlations were found between PRF1 and CD163, as well as between PFR1 and FOXP3. It is worth noting that the combination of high levels of CD14, FOXP3, and ARG1 mRNAs identified a small group of patients with excellent EFS and OS, whereas low levels of CD14 were sufficient to identify patients with dismal survival. Thus, the immune status of the primary tumors of high-risk NB patients may influence the natural history of this pediatric cancer.

  4. Synthesis, purification, and characterization of an Arg152 → Glu site-directed mutant of recombinant human blood clotting factor VII

    Coagulation factor VII circulates in blood as a single-chain zymogen of a serine protease and is converted to its activated two-chain form, factor VIIa, by cleavage of an internal peptide bond located at Arg152-Ile153. Previous studies using serine protease active-site inhibitors suggest that zymogen factor VII may possess sufficient proteolytic activity to initiate the extrinsic pathway of blood coagulation. In order to assess the putative intrinsic proteolytic activity of single-chain factor VII, the authors have constructed a site-specific mutant of recombinant human factor VII in which arginine-152 has been replaced with a glutamic acid residue. Mutant factor VII was purified in a single step from culture supernatants of baby hamster kidney cells transfected with a plasmid containing the sequence for Arg152 → Glu factor VII using a calcium-dependent, murine anti-factor VII monoclonal antibody column. The clotting activity of mutant factor VII was completely inhibited following incubation with dansyl-Glu-Gly-Arg chloromethyl ketone, suggesting that the apparent clotting activity of mutant factor VII was due to a contaminating serine protease. Immunoblots of mutant factor VII with human factor IXa revealed no cleavage, whereas incubation of mutant factor VII with human factor Xa resulted in cleavage of mutant factor VII and the formation of a lower molecular weight degradation product migrating at Mr∼40 000. The results are consistent with the proposal that zymogen factor VII possesses no intrinsic proteolytic activity toward factor X or factor IX

  5. The structure of salt bridges between Arg(+) and Glu(-) in peptides investigated with 2D-IR spectroscopy: Evidence for two distinct hydrogen-bond geometries.

    Huerta-Viga, Adriana; Amirjalayer, Saeed; Domingos, Sérgio R; Meuzelaar, Heleen; Rupenyan, Alisa; Woutersen, Sander

    2015-06-01

    Salt bridges play an important role in protein folding and in supramolecular chemistry, but they are difficult to detect and characterize in solution. Here, we investigate salt bridges between glutamate (Glu(-)) and arginine (Arg(+)) using two-dimensional infrared (2D-IR) spectroscopy. The 2D-IR spectrum of a salt-bridged dimer shows cross peaks between the vibrational modes of Glu(-) and Arg(+), which provide a sensitive structural probe of Glu(-)⋯Arg(+) salt bridges. We use this probe to investigate a β-turn locked by a salt bridge, an α-helical peptide whose structure is stabilized by salt bridges, and a coiled coil that is stabilized by intra- and intermolecular salt bridges. We detect a bidentate salt bridge in the β-turn, a monodentate one in the α-helical peptide, and both salt-bridge geometries in the coiled coil. To our knowledge, this is the first time 2D-IR has been used to probe tertiary side chain interactions in peptides, and our results show that 2D-IR spectroscopy is a powerful method for investigating salt bridges in solution. PMID:26049464

  6. The structure of salt bridges between Arg+ and Glu- in peptides investigated with 2D-IR spectroscopy: Evidence for two distinct hydrogen-bond geometries

    Huerta-Viga, Adriana; Amirjalayer, Saeed; Domingos, Sérgio R.; Meuzelaar, Heleen; Rupenyan, Alisa; Woutersen, Sander

    2015-06-01

    Salt bridges play an important role in protein folding and in supramolecular chemistry, but they are difficult to detect and characterize in solution. Here, we investigate salt bridges between glutamate (Glu-) and arginine (Arg+) using two-dimensional infrared (2D-IR) spectroscopy. The 2D-IR spectrum of a salt-bridged dimer shows cross peaks between the vibrational modes of Glu- and Arg+, which provide a sensitive structural probe of Glu-⋯Arg+ salt bridges. We use this probe to investigate a β-turn locked by a salt bridge, an α-helical peptide whose structure is stabilized by salt bridges, and a coiled coil that is stabilized by intra- and intermolecular salt bridges. We detect a bidentate salt bridge in the β-turn, a monodentate one in the α-helical peptide, and both salt-bridge geometries in the coiled coil. To our knowledge, this is the first time 2D-IR has been used to probe tertiary side chain interactions in peptides, and our results show that 2D-IR spectroscopy is a powerful method for investigating salt bridges in solution.

  7. [The association between beta-adrenergic receptor gene polymorphisms and personality traits].

    Numajiri, Maki; Aoki, Jun; Nishizawa, Daisuke; Kasai, Shinya; Ogai, Yasukazu; Ikeda, Kazutaka; Iwahashi, Kazuhiko

    2012-08-01

    The relationship between the polymorphisms (SNPs) of the beta-adrenergic receptor (beta-AR) gene and personality assessed by TCI (Temperament and Character Inventory), was studied among 192 healthy Japanese subjects (121 male subjects and 71 female subjects). In this study, the statistical analyses were performed overall and separately for each sex. As a result, it was shown that there were significant relationships between SD (self-directedness) and 49Ser/Gly (rs1801252) in ADRB1, P (persistence) and 389Arg/Gly (rs1801253) in ADRB1, and ST (self-transcendence) and 27Gln/Glu (rs1042714) in ADRB2 overall. Among the male subjects, there were further significant relationships between ST and 49Ser/Gly in ADRB1, NS (novelty-seeking), HA (harm avoidance) and P and 389Arg/Gly in ADRB1, and P and 64Arg/Trp(rsrs4994) in ADRB3. Among the female subjects, there were also significant relationships between SD and 49Ser/Gly in ADRB1, and C (cooperativeness) and 389Arg/Gly in ADRB1. Thus it was shown that there were correlations between beta-AR gene polymorphisms and several subscales of TCI. PMID:23012891

  8. Calcined Eggshell Waste for Mitigating Soil Antibiotic-Resistant Bacteria/Antibiotic Resistance Gene Dissemination and Accumulation in Bell Pepper.

    Ye, Mao; Sun, Mingming; Feng, Yanfang; Li, Xu; Schwab, Arthur P; Wan, Jinzhong; Liu, Manqiang; Tian, Da; Liu, Kuan; Wu, Jun; Jiang, Xin

    2016-07-13

    The combined accumulation of antibiotics, heavy metals, antibiotic-resistant bacteria (ARB)/antibiotic resistance genes (ARGs) in vegetables has become a new threat to human health. This is the first study to investigate the feasibility of calcined eggshells modified by aluminum sulfate as novel agricultural wastes to impede mixed contaminants from transferring to bell pepper (Capsicum annuum L.). In this work, calcined eggshell amendment mitigated mixed pollutant accumulation in bell pepper significantly, enhanced the dissipation of soil tetracycline, sulfadiazine, roxithromycin, and chloramphenicol, decreased the water-soluble fractions of antibiotics, and declined the diversity of ARB/ARGs inside the vegetable. Moreover, quantitative polymerase chain reaction analysis detected that ARG levels in the bell pepper fruits significantly decreased to 10(-10) copies/16S copies, indicating limited risk of ARGs transferring along the food chain. Furthermore, the restoration of soil microbial biological function suggests that calcined eggshell is an environmentally friendly amendment to control the dissemination of soil ARB/ARGs in the soil-vegetable system. PMID:27333280

  9. The diversity of the HLA-E-restricted peptide repertoire explains the immunological impact of the Arg107Gly mismatch.

    Celik, Alexander A; Kraemer, Thomas; Huyton, Trevor; Blasczyk, Rainer; Bade-Döding, Christina

    2016-01-01

    Human leukocyte antigen (HLA)-E molecules are potent inhibitors of NK cell-mediated killing. Low in polymorphisms, two alleles are widely expressed among diverse populations: HLA-E*01:01 and HLA-E*01:03. Both alleles are distinguished by one SNP resulting in the substitution Arg107Gly. Both alleles present a limited set of peptides derived from class I leader sequences physiologically; however, HLA-E*01:01 presents non-canonical peptides in the absence of HLA class I molecules. To further assess the functional differences between both alleles, we analyzed the peptide repertoire of HLA-E*01:03 by applying soluble HLA technology followed by mass-spectrometric peptide sequencing. HLA-E*01:03 restricted peptides showed a length of 9-17 amino acids and differed in their biophysical properties, no overlap in the peptide repertoire of both allelic variants could be observed; however, both alleles shared marginal peptides from the same proteomic content. Artificial APCs expressing empty HLA-E*01:01 or E*01:03 molecules were generated and stabilized using cognate HLA class I-derived peptide ligands to analyze the impact of residue 107 within the HLA-E heavy chain on the NKG2/CD94 receptor engagement. Differences in peptide stabilization could be translated to the density and half-life time of peptide-HLA-E molecules on the cell surface that subsequently impacted NK cell inhibition as verified by cytotoxicity assays. Taken together, these data illustrate functional differences of HLA-E allelic variants induced by a single amino acid. Furthermore, the function of HLA-E in pathophysiologic situations when the HLA processing machinery is interrupted seems to be more emphasized than previously described, implying a crucial role for HLA-E in tumor or viral immune episodes. PMID:26552660

  10. Pharmacological evaluation of Mallotus philippinensis (Lam. Muell.-Arg. fruit hair extract for Anti-Inflammatory, Analgesic and Hypnotic Activity

    Mayank Gangwar

    2016-03-01

    Full Text Available Recently we observed wound healing activity of 50% ethanol extract of Mallotus philippinensis Muell. Arg (MP fruit hairs extract (MPE. In several intestinal infections, localized inflammation is of common occurrence and hence we evaluated the anti-inflammatory, analgesic and hypnotic activity of MPE in different rat experimental models. Anti-inflammatory activity was evaluated by carrageenan (acute and turpentine oil induced formalin (subacute induced paw oedema and while, granuloma pouch (subacute in rats. Analgesic and hypnotic activity of MPE was undertaken by tail-flick, hot and ndash;plate and acetic acid-induced writhing tests while pentobarbitone-induced hypnotic potentiation respectively in rats.\tMPE at a dose of 200 mg/kg at 3 hr after their administration showed inhibition of formalin-induced paw oedema by 41.60% (P<0.001 and carrageenan-induced paw oedema by 55.30% (P<0.001. After 7 days of treatments, MPE showed 38.0% (P<0.001 inhibition against formalin-induced paw oedema and reduced weight of turpentine-induced granuloma pouch by 29.6% (P<0.01 and volume of exudates by 26.1% (P<0.01 respectively. MPE (200 mg/kg showed dose-dependent elevation in pain threshold and peak analgesic effect at 120 min as evidenced by increased latency period in tail flick method and increased reaction time in hot-plate test while reduction in the number of acetic acid-induced writhes by 45.7% (P<0.001. The pentobarbitone-induced hypnosis model showed potentiation, as defined by increased duration of sleep in treated group rats as compared to control. Thus, the study revealed MPE is effective in reducing acute and sub-acute inflammation and showed effective and similar analgesic activity. This seemed to be safe in the treatment of pain and inflammation. [J Intercult Ethnopharmacol 2016; 5(1.000: 14-21

  11. 124I labeled Arg-Gly-Asp (RGD) peptide: PET/MR fusion imaging of tumor expressing integrin v3

    Tumor-induced angiogenesis and metastasis express high level of integrin v3 which has become a promising diagnostic biomarker and therapeutic target for various tumors. Radiolabeled RGD (Arg-Gly-Asp) peptides are well known to specifically bind to integrin and can be used not only for noninvasive imaging of integrin over-expression but also for integrin-targeted radionuclide therapy. We used three RGDyK derivatives, monomeric-, dimeric- and tetrameric-RGD for radio labeling. They were labeled with [124I]NaI using iodo-bead. Labeled RGD peptide was purified by FPLC. Xenografts were induced by subcutaneous injection of 1X107 U87MG cells into the right thigh of female Balb/c nude mice. After injection of 124I-labeled monomeric-RGD via the tail vein, the microPET imaging was obtained during 2 hours. Right after microPET imaging, T2-weighted MR imaging was performed with fast gradient spin echo sequence in 0.8 mm of a slice thickness. PET/MR fusion images were constructed by AMIDE program. For immobilization and reproducible positioning of animal, animal-specific positioning molds were fabricated. Monomeric-RGD peptide was labeled with 124I in labeling yield of 50%. After FPLC purification, the radiochemical purity was above 90%. U87MG tumor was clearly seen in both microPET and MR imaging of tumor-bearing nude mouse. The fusion imaging was well constructed by using AMIDE with little discrepancy thanks to animal-specific molds. Three RGD derivatives were also labeled with 131I in labeling yield of 20-55%. All three RGD peptides were successfully radioiodinated with 124I or 131I. U87MG glioblastoma tumor was clearly visualized in PET/MR fusion imaging. MicroPET/MR fusion imaging of dimeric- and tetrameric-RGD peptides in tumor bearing mice will also be presented during conference

  12. Additive effect of polymorphisms in the β2 -adrenoceptor and NADPH oxidase p22 phox genes contributes to the loss of estimated glomerular filtration rate in Chinese.

    Wang, Tao; Zhang, Yan; Ma, JingTao; Feng, Zhen; Niu, Kai; Liu, Bing

    2014-09-01

    Because increased oxidative stress may mediate the detrimental actions of enhanced sympathetic nervous activity on renal function and vice versa, we investigated the effect of the polymorphic Arg16Gly in the β2 -adrenoceptor (ADRB2) gene, Trp64Arg in the β3 -adrenoceptor (ADRB3) gene and C242T in the NADPH oxidase p22phox (CYBA) gene on estimated glomerular filtration rate (eGFR) in a Chinese population. Initially recruited from different outpatient services of HeBei General Hospital in northern China, 668 individuals were finally included in the study, with complete demographic information. Laboratory tests were performed and estimated glomerular filtration rate (eGFR) was derived from the Modification of Diet in Renal Disease (MDRD) equation for the Chinese population. Plasma noradrenaline levels and genotype were determined by HPLC and the TaqMan method, respectively. Only across the Arg16Gly polymorphism did eGFR show significant difference: it was lower in individuals with the Gly16Gly variation, who also had the highest plasma noradrenaline levels. This polymorphism remained a significant determinant of eGFR after multivariate analysis. Of importance, the multifactor dimensionality reduction method further detected a significant synergism between the Arg16Gly and C242T polymorphisms in reducing eGFR. These observations clarify the effects of the studied polymorphisms on eGFR and exemplify gene-gene interactions influencing renal function. PMID:24890187

  13. Haplotype analysis of TP53 polymorphisms, Arg72Pro and Ins16, in BRCA1 and BRCA2 mutation carriers of French Canadian descent

    The TP53 polymorphisms Arg72Pro (Ex4+199 G>C) and Ins16 (IVS3+24 ins16) have been proposed to modify risk of breast cancer associated with germline BRCA1 and BRCA2 mutations. Allele frequencies of these polymorphisms were investigated to determine if they modify risk in BRCA mutation carriers in breast cancer cases drawn from French Canadian cancer families, a population shown to exhibit strong founder effects. The frequencies of the TP53 alleles, genotypes and haplotypes of 157 index breast cancer cases comprised of 42 BRCA1 mutation carriers, 57 BRCA2 mutation carriers, and 58 BRCA mutation-negative cases, where each case was drawn from independently ascertained families were compared. The effect of TP53 variants on the age of diagnosis was also investigated for these groups. The TP53 polymorphisms were also investigated in 112 women of French Canadian descent with no personal history of cancer. The BRCA mutation-positive groups had the highest frequency of homozygous carriers of the 72Pro allele compared with mutation-negative group. The TP53 polymorphisms exhibited linkage disequilibrium (p < 0.001), where the 72Arg and Ins16minus alleles occurred in strong disequilibrium. The highest frequency of carriers of Ins16minus-72Arg haplotype occurred in the BRCA mutation-negative groups. The BRCA1 mutation carriers homozygous for the 72Pro allele had the youngest ages of diagnosis of breast cancer. However none of these observations were statistically significant. In contrast, the BRCA2 mutation carriers homozygous for the 72Pro allele had a significantly older age of diagnosis of breast cancer (p = 0.018). Moreover, in this group, the mean age of diagnosis of breast cancer in carriers of the Ins16minus-72Arg haplotype was significantly younger than that of the individuals who did not this carry this haplotype (p = 0.009). We observed no significant association of breast cancer risk with TP53 genetic variants based on BRCA1/2 mutation carrier status. Although the

  14. Impact of GNB3-C825T, ADRB3-Trp64Arg, UCP2-3'UTR 45 bp del/ins, and PPARγ-Pro12Ala polymorphisms on Bofutsushosan response in obese subjects: a randomized, double-blind, placebo-controlled trial.

    Park, Junghyun; Bose, Shambhunath; Hong, Sun-Woo; Lee, Dong-Ki; Yoo, Jae-Wook; Lim, Chi-Yeon; Lee, Myeongjong; Kim, Hojun

    2014-05-01

    Obesity is known to be influenced by a number of genes, including the β3 subunit of G protein (GNB3), β3-adrenergic receptor (ADRB3), uncoupling protein 2 (UCP2), and peroxisome proliferator activated receptor gamma (PPARγ). The single nucleotide polymorphisms (SNPs) of the above genes, such as GNB3-C825T, ADRB3-Trp64Arg, UCP2-3'UTR 45 bp del/ins, and PPARγ-Pro12Ala, are associated with obesity and body mass index. The present study evaluates the impact of Bofutsushosan, a traditional Eastern Asian herbal medicine with known anti-obesity properties, on obese subjects according to the presence of the above-mentioned SNPs. Upon randomization, the volunteers were allocated to receive Bofutsushosan (n=55) or placebo (n=56) treatments for 8 weeks. Following the treatment schedule, significant reductions in total cholesterol and significant improvement in the Korean version of obesity-related quality of life scale were seen in the Bofutsushosan-treated group, but not in placebo. Bofutsushosan exerted significant anti-obesity effects on a number of parameters in the carriers of the GNB3-825T allele, but only on waist circumference in the GNB3-C/C homozygote. Significant anti-obesity impact of Bofutsushosan was also seen on a number of obesity-indices in both ADRB3-Arg64 carriers and ADRB3-Trp64 homozygotes, as well as in UCP2-D/D carriers, but not in UCP2-D/I+I/I variants. The effect of Bofutsushosan was more pronounced in PPARγ-Pro/Pro genotype compared to PPARγ-Pro/Ala variants. Thus, the results revealed differential responses of the subjects to the anti-obesity effects of Bofutsushosan treatment according to the polymorphism of the vital obesity-related genes. Our study provides new insight into individualized clinical applications of Bofutsushosan for obesity. PMID:24827746

  15. β3-adrenergic receptor gene, body mass index, bone mineral density and fracture risk in elderly men and women: the Dubbo Osteoporosis Epidemiology Study (DOES)

    Center Jacqueline R; Eisman John A; Morrison Nigel A; Nguyen Nguyen D; Wang Claire Y; Nguyen Tuan V

    2006-01-01

    Abstract Background Recent studies have suggested that the Arg allele of β3-adrenergic receptor (ADRB3) gene is associated with body mass index (BMI), which is an important predictor of bone mineral density (BMD) and fracture risk. However, whether the ADRB3 gene polymorphism is associated with fracture risk has not been investigated. The aim of study was to examine the inter-relationships between ADRB3 gene polymorphisms, BMI, BMD and fracture risk in elderly Caucasians. Methods Genotypes of...

  16. Performance of a constructed wetland in Grand Marais, Manitoba, Canada: Removal of nutrients, pharmaceuticals, and antibiotic resistance genes from municipal wastewater

    Anderson, Julie C; Carlson, Jules C; Low, Jennifer E; Challis, Jonathan K; Wong, Charles S.; Charles W Knapp; Hanson, Mark L

    2013-01-01

    Background The discharge of complex mixtures of nutrients, organic micropollutants, and antibiotic resistance genes from treated municipal wastewater into freshwater systems are global concerns for human health and aquatic organisms. Antibiotic resistance genes (ARGs) are genes that have the ability to impart resistance to antibiotics and reduce the efficacy of antibiotics in the systems in which they are found. In the rural community of Grand Marais, Manitoba, Canada, wastewater is treated p...

  17. Bioinformatic analysis of an unusual gene-enzyme relationship in the arginine biosynthetic pathway among marine gamma proteobacteria: implications concerning the formation of N-acetylated intermediates in prokaryotes

    Labedan Bernard

    2006-01-01

    Full Text Available Abstract Background The N-acetylation of L-glutamate is regarded as a universal metabolic strategy to commit glutamate towards arginine biosynthesis. Until recently, this reaction was thought to be catalyzed by either of two enzymes: (i the classical N-acetylglutamate synthase (NAGS, gene argA first characterized in Escherichia coli and Pseudomonas aeruginosa several decades ago and also present in vertebrates, or (ii the bifunctional version of ornithine acetyltransferase (OAT, gene argJ present in Bacteria, Archaea and many Eukaryotes. This paper focuses on a new and surprising aspect of glutamate acetylation. We recently showed that in Moritella abyssi and M. profunda, two marine gamma proteobacteria, the gene for the last enzyme in arginine biosynthesis (argH is fused to a short sequence that corresponds to the C-terminal, N-acetyltransferase-encoding domain of NAGS and is able to complement an argA mutant of E. coli. Very recently, other authors identified in Mycobacterium tuberculosis an independent gene corresponding to this short C-terminal domain and coding for a new type of NAGS. We have investigated the two prokaryotic Domains for patterns of gene-enzyme relationships in the first committed step of arginine biosynthesis. Results The argH-A fusion, designated argH(A, and discovered in Moritella was found to be present in (and confined to marine gamma proteobacteria of the Alteromonas- and Vibrio-like group. Most of them have a classical NAGS with the exception of Idiomarina loihiensis and Pseudoalteromonas haloplanktis which nevertheless can grow in the absence of arginine and therefore appear to rely on the arg(A sequence for arginine biosynthesis. Screening prokaryotic genomes for virtual argH-X 'fusions' where X stands for a homologue of arg(A, we retrieved a large number of Bacteria and several Archaea, all of them devoid of a classical NAGS. In the case of Thermus thermophilus and Deinococcus radiodurans, the arg(A-like sequence

  18. Removal of antibiotics and antibiotic resistance genes from domestic sewage by constructed wetlands: Optimization of wetland substrates and hydraulic loading.

    Chen, Jun; Wei, Xiao-Dong; Liu, You-Sheng; Ying, Guang-Guo; Liu, Shuang-Shuang; He, Liang-Ying; Su, Hao-Chang; Hu, Li-Xin; Chen, Fan-Rong; Yang, Yong-Qiang

    2016-09-15

    This study aimed to assess removal potential of antibiotics and antibiotic resistance genes (ARGs) in raw domestic wastewater by various mesocosm-scale horizontal subsurface-flow constructed wetlands (CWs) planted Cyperus alternifolius L. with different design parameters. Twelve CWs with three hydraulic loading rates (HLR 10, 20 and 30cm/day) and four substrates (oyster shell, zeolite, medical stone and ceramic) were set up in order to select the best optimized wetland. The result showed that 7 target antibiotics compounds including erythromycin-H2O, lincomycin, monensin, ofloxacin, sulfamerazine, sulfamethazine and novobiocin were detected, and all selected 18 genes (three sulfonamide resistance genes (sul1, sul2 and sul3), four tetracycline resistance genes (tetG, tetM, tetO and tetX), two macrolide resistance genes (ermB and ermC), three quinolone resistance genes (qnrB, qnrD and qnrS) and four chloramphenicol resistance genes (cmlA, fexA, fexB and floR)) and two integrase genes (int1 and int2) were positively detected in the domestic wastewaters. The aqueous removal rates of the total antibiotics ranged from17.9 to 98.5%, while those for the total ARGs varied between 50.0 and 85.8% by the mesocosm-scale CWs. After considering their aqueous removal rates in combination with their mass removals, the CW with zeolite as the substrate and HLR of 20cm/day was selected as the best choice. Combined chemical and biological analyses indicate that both microbial degradation and physical sorption processes were responsible for the fate of antibiotics and ARGs in the wetlands. The findings from this study suggest constructed wetlands could be a promising technology for the removal of emerging contaminants such as antibiotics and ARGs in domestic wastewater. PMID:27173842

  19. Sulfonamide and tetracycline resistance genes in total- and culturable-bacterial assemblages in South African aquatic environments

    Satoru eSuzuki; Mitsuko eOgo; Tatsuya eKoike; Hideshige eTakada; Brent eNewman

    2015-01-01

    Antibiotic resistant bacteria (ARB) are ubiquitous in the natural environment. The introduction of effluent derived antibiotic resistance genes (ARGs) into aquatic environments is of concern in the spreading of genetic risk. This study showed the prevalence of sulfonamide and tetracycline resistance genes, sul1, sul2, sul3 and tet(M), in the total bacterial assemblage and colony forming bacterial assemblage in river and estuarine water and sewage treatment plants (STP) in South Africa. There ...

  20. Sulfonamide and tetracycline resistance genes in total- and culturable-bacterial assemblages in South African aquatic environments

    Suzuki, Satoru; Ogo, Mitsuko; Koike, Tatsuya; Takada, Hideshige; Newman, Brent

    2015-01-01

    Antibiotic resistant bacteria are ubiquitous in the natural environment. The introduction of effluent derived antibiotic resistance genes (ARGs) into aquatic environments is of concern in the spreading of genetic risk. This study showed the prevalence of sulfonamide and tetracycline resistance genes, sul1, sul2, sul3, and tet(M), in the total bacterial assemblage and colony forming bacterial assemblage in river and estuarine water and sewage treatment plants (STP) in South Africa. There was no ...

  1. A Comprehensive Insight into Tetracycline Resistant Bacteria and Antibiotic Resistance Genes in Activated Sludge Using Next-Generation Sequencing

    Kailong Huang; Junying Tang; Xu-Xiang Zhang; Ke Xu; Hongqiang Ren

    2014-01-01

    In order to comprehensively investigate tetracycline resistance in activated sludge of sewage treatment plants, 454 pyrosequencing and Illumina high-throughput sequencing were used to detect potential tetracycline resistant bacteria (TRB) and antibiotic resistance genes (ARGs) in sludge cultured with different concentrations of tetracycline. Pyrosequencing of 16S rRNA gene revealed that tetracycline treatment greatly affected the bacterial community structure of the sludge. Nine genera cons...

  2. Diversity, Distribution and Quantification of Antibiotic Resistance Genes in Goat and Lamb Slaughterhouse Surfaces and Meat Products

    Lavilla Lerma, Leyre; Benomar, Nabil; Knapp, Charles W.; Correa Galeote, David; Gálvez, Antonio; Abriouel, Hikmate

    2014-01-01

    The distribution and quantification of tetracycline, sulfonamide and beta-lactam resistance genes were assessed in slaughterhouse zones throughout meat chain production and the meat products; this study represents the first to report quantitatively monitor antibiotic resistance genes (ARG) in goat and lamb slaughterhouse using a culture independent approach, since most studies focused on individual bacterial species and their specific resistance types. Quantitative PCR (qPCR) revealed a high ...

  3. Diversity, distribution and quantification of antibiotic resistance genes in goat and lamb slaughterhouse surfaces and meat products.

    Lavilla Lerma, Leyre; Benomar, Nabil; Knapp, Charles W; Correa Galeote, David; Gálvez, Antonio; Abriouel, Hikmate

    2014-01-01

    The distribution and quantification of tetracycline, sulfonamide and beta-lactam resistance genes were assessed in slaughterhouse zones throughout meat chain production and the meat products; this study represents the first to report quantitatively monitor antibiotic resistance genes (ARG) in goat and lamb slaughterhouse using a culture independent approach, since most studies focused on individual bacterial species and their specific resistance types. Quantitative PCR (qPCR) revealed a high prevalence of tetracycline resistance genes tetA and tetB in almost all slaughterhouse zones. Sulfonamide resistance genes were largely distributed, while beta-lactam resistance genes were less predominant. Statistical analysis revealed that resistant bacteria, in most cases, were spread by the same route in almost all slaughterhouse zones, except for tetB, blaCTX and blaTEM genes, which occurred in few zones as isolated 'hot spots.' The sum of all analyzed ARG indicated that slaughterhouse surfaces and end products act as reservoirs of ARG, mainly tet genes, which were more prevalent in slaughtering room (SR), cutting room (CR) and commercial meat products (MP). Resistance gene patterns suggest they were disseminated throughout slaughterhouse zones being also detected in commercial meat products, with significant correlations between different sampling zones/end products and total resistance in SR, CR and white room (WR) zones, and also refrigerator 4 (F4) and MP were observed. Strategically controlling key zones in slaughterhouse (SR, CR and WR) by adequate disinfection methods could strategically reduce the risks of ARG transmission and minimize the issues of food safety and environment contamination. PMID:25479100

  4. 164Ile allele in the beta2-Adrenergic receptor gene is associated with risk of elevated blood pressure in women. The Copenhagen City Heart Study

    Sethi, Amar A; Tybjaerg-Hansen, Anne; Jensen, Gorm B;

    2005-01-01

    Since beta2-adrenergic receptors are important regulators of blood pressure, genetic variation in this receptor could explain risk of elevated blood pressure in selected individuals. We tested the hypothesis that Gly16Arg, Gln27Glu, and Thr164Ile in the beta2-adrenergic receptor gene associated w...

  5. A polymorphism in the glucocorticoid receptor gene, which decreases sensitivity to glucocorticoids in vivo, is associated with low insulin and cholesterol levels

    J.W. Koper (Jan); N.A.T.M. Huizenga (Nannette); A.G. Uitterlinden (André); J.A.M.J.L. Janssen (Joop); A.O. Brinkmann (Albert); D.E. Grobbee (Diederick); C.M. van Duijn (Cock); F.H. de Jong (Frank); E.F.C. van Rossum (Liesbeth)

    2002-01-01

    textabstractWe investigated whether a polymorphism in codons 22 and 23 of the glucocorticoid (GC) receptor gene [GAGAGG(GluArg) 3GAAAAG(GluLys)] is associated with altered GC sensitivity, anthropometric parameters, cardiovascular risk factors, and sex steroid hormones. In a subgroup of 202 healthy e

  6. A polymorphism in the glucocorticoid receptor gene, which decreases sensitivity to glucocorticoids in vivo, is associated with low insulin and cholesterol levels

    S.W.J. Lamberts (Steven); J.W. Koper (Jan); N.A.T.M. Huizenga (Nannette); A.G. Uitterlinden (André); A.O. Brinkmann (Albert); D.E. Grobbee (Diederick); F.H. de Jong (Frank); C.M. van Duijn (Cock); H.A.P. Pols (Huib); J.A.M.J.L. Janssen (Joop); E.F.C. van Rossum (Liesbeth)

    2002-01-01

    textabstractWe investigated whether a polymorphism in codons 22 and 23 of the glucocorticoid (GC) receptor gene [GAGAGG(GluArg) --> GAAAAG(GluLys)] is associated with altered GC sensitivity, anthropometric parameters, cardiovascular risk factors, and sex steroid hormones. In a subg

  7. [Cloning, sequence analysis and expression of N-acetylglutamate kinase gene in Corynebacterium crenatum].

    Hao, Ning; Zhao, Zhi; Wang, Yu; Zhang, Ying-zi; Ding, Jiu-yuan

    2006-02-01

    N-Acetylglutamate kinase (EC 2.7.2.8;NAGK) genes from wild-type Corynebacterium crenatum AS 1.542 and a L-arginine-producing mutant C. crenatum 971.1 were cloned and sequenced. Analysis of argB sequences revealed that only one ORF existed, which used ATG as the initiation codon and coded a peptide of 317 amino acids with a calculated molecular weight of 33.6kDa. Only one nucleotide difference was found in the structure gene and the difference did not cause a change of amino acid by comparison of the gene sequences between the wild type C. crenatum AS 1.542 and the mutant 971.1. The ORF sequence of argB from C. crenatum AS 1.542 showed homologies of 99.89%, 76.62%, 37.94% to those from Corynebacterium glutamicum ATCC 13032, Corynebacterium efficient YS-314 and Escherichia coli k12. And the amino acid sequence deduced from ORF displayed homologies of 100%, 78.55%, 25.25% to those from microorganisms above, respectively. An internal promoter was found in the upstream of the argB gene from C. crenatum. The argB gene from C. crenatum AS 1.542 was expressed both in C. crenatum AS 1.542 and 971.1. The NAGK activity of transformed C. crenatum AS 1.542 was greatly increased by the induction of IPTG. The NAGK activity of transformed C. crenatum 971.1 was almost twice as much as that of C. crenatum 971.1 under the same induction. The amplification of the NAGK activity yielded 25% increase of L-arginine production in C. crenatum 971.1. PMID:16579472

  8. Acromesomelic dysplasia, type maroteaux caused by novel loss-of-function mutations of the NPR2 gene: Three case reports.

    Wang, Wei; Song, Mi Hyun; Miura, Kohji; Fujiwara, Makoto; Nawa, Nobutoshi; Ohata, Yasuhisa; Kitaoka, Taichi; Kubota, Takuo; Namba, Noriyuki; Jin, Dong Kyu; Kim, Ok Hwa; Ozono, Keiichi; Cho, Tae-Joon

    2016-02-01

    The C-type natriuretic peptide (CNP)-natriuretic peptide receptor 2 (NPR2) signaling pathway plays an important role in chondrocyte development. Homozygous loss-of-function mutations of the NPR2 gene cause acromesomelic dysplasia, type Maroteaux (AMDM). The aim of this study was to identify and characterize NPR2 loss-of-function mutations in patients with AMDM. The NPR2 gene was sequenced in three Korean patients with AMDM and functional analysis of the mutated proteins was performed in vitro. Five novel NPR2 mutations were found in the three patients: two compound heterozygous mutations [c.1231T>C (Tyr411His) and c.2761C>T (Arg921X) in Patient 1 and c.1663A>T (Lys555X) and c.1711-1G>C (M571VfsX12) in Patient 3] and a homozygous mutation [c.2762G>A (Arg921Gln) in Patient 2]. Serum NT-proCNP concentration was significantly increased in each patient compared to control subjects. Cells transfected with the expression vector of each mutant except those found in Patient 3 showed a negligible or a markedly low cGMP response after treatment with CNP. HA-tagged wild-type (wt) and HA-mutant NPR2 were expressed at comparable levels: there were two bands of ∼130 and ∼120 kDa in wt and Arg921Gln, a single ∼120 kDa band in Tyr411His, and a single ∼110 kDa in the nonsense mutant. With respect to subcellular localization, Arg921Gln as well as wt-NPR2 reached the cell surface, whereas Tyr411His and Arg921X mutants did not. The Tyr411His and Arg921X NPR2 proteins were co-localized with an endoplasmic reticulum (ER) marker and failed to traffic from the ER to the Golgi apparatus. These results are consistent with deglycosylation experiments. Tyr411His and Arg921X NPR2 are complete loss-of-function mutations, whereas Arg921Gln behaves as a receptor for CNP with limited function. PMID:26567084

  9. The KMO allele encoding Arg452 is associated with psychotic features in bipolar disorder type 1, and with increased CSF KYNA level and reduced KMO expression.

    Lavebratt, C; Olsson, S; Backlund, L; Frisén, L; Sellgren, C; Priebe, L; Nikamo, P; Träskman-Bendz, L; Cichon, S; Vawter, M P; Osby, U; Engberg, G; Landén, M; Erhardt, S; Schalling, M

    2014-03-01

    The kynurenine pathway metabolite kynurenic acid (KYNA), modulating glutamatergic and cholinergic neurotransmission, is increased in cerebrospinal fluid (CSF) of patients with schizophrenia or bipolar disorder type 1 with psychotic features. KYNA production is critically dependent on kynurenine 3-monooxygenase (KMO). KMO mRNA levels and activity in prefrontal cortex (PFC) are reduced in schizophrenia. We hypothesized that KMO expression in PFC would be reduced in bipolar disorder with psychotic features and that a functional genetic variant of KMO would associate with this disease, CSF KYNA level and KMO expression. KMO mRNA levels were reduced in PFC of bipolar disorder patients with lifetime psychotic features (P=0.005, n=19) or schizophrenia (P=0.02, n=36) compared with nonpsychotic patients and controls. KMO genetic association to psychotic features in bipolar disorder type 1 was studied in 493 patients and 1044 controls from Sweden. The KMO Arg(452) allele was associated with psychotic features during manic episodes (P=0.003). KMO Arg(452) was studied for association to CSF KYNA levels in an independent sample of 55 Swedish patients, and to KMO expression in 717 lymphoblastoid cell lines and 138 hippocampal biopsies. KMO Arg(452) associated with increased levels of CSF KYNA (P=0.03) and reduced lymphoblastoid and hippocampal KMO expression (P≤0.05). Thus, findings from five independent cohorts suggest that genetic variation in KMO influences the risk for psychotic features in mania of bipolar disorder patients. This provides a possible mechanism for the previous findings of elevated CSF KYNA levels in those bipolar patients with lifetime psychotic features and positive association between KYNA levels and number of manic episodes. PMID:23459468

  10. Virome-associated antibiotic-resistance genes in an experimental aquaculture facility.

    Colombo, Stefano; Arioli, Stefania; Guglielmetti, Simone; Lunelli, Fernando; Mora, Diego

    2016-03-01

    We report the comprehensive characterization of viral and microbial communities within an aquaculture wastewater sample, by a shotgun sequencing and 16S rRNA gene profiling metagenomic approach. Caudovirales had the largest representation within the sample, with over 50% of the total taxonomic abundance, whereas approximately 30% of the total open reading frames (ORFs) identified were from eukaryotic viruses (Mimiviridae and Phycodnaviridae). Antibiotic resistance genes (ARGs) within the virome accounted for 0.85% of the total viral ORFs and showed a similar distribution both in virome and in microbiome. Among the ARGs, those encoding proteins involved in the modulation of antibiotic efflux pumps were the most abundant. Interestingly, the taxonomy of the bacterial ORFs identified in the viral metagenome did not reflect the microbial taxonomy as deduced by 16S rRNA gene profiling and shotgun metagenomic analysis. A limited number of ARGs appeared to be mobilized from bacteria to phages or vice versa, together with other bacterial genes encoding products involved in general metabolic functions, even in the absence of any antibiotic treatment within the aquaculture plant. Thus, these results confirm the presence of a complex phage-bacterial network in the aquaculture environment. PMID:26738553

  11. Cellular localization of kinin B1 receptor in the spinal cord of streptozotocin-diabetic rats with a fluorescent [Nα-Bodipy]-des-Arg9-bradykinin

    Gaudreau Pierrette

    2009-03-01

    Full Text Available Abstract Background The kinin B1 receptor (B1R is upregulated by pro-inflammatory cytokines, bacterial endotoxins and hyperglycaemia-induced oxidative stress. In animal models of diabetes, it contributes to pain polyneuropathy. This study aims at defining the cellular localization of B1R in thoracic spinal cord of type 1 diabetic rats by confocal microscopy with the use of a fluorescent agonist, [Nα-Bodipy]-des-Arg9-BK (BdABK and selective antibodies. Methods Diabetes was induced by streptozotocin (STZ; 65 mg/kg, i.p.. Four days post-STZ treatment, B1R expression was confirmed by quantitative real-time PCR and autoradiography. The B1R selectivity of BdABK was determined by assessing its ability to displace B1R [125I]-HPP-desArg10-Hoe140 and B2R [125I]-HPP-Hoe 140 radioligands. The in vivo activity of BdABK was also evaluated on thermal hyperalgesia. Results B1R was increased by 18-fold (mRNA and 2.7-fold (binding sites in the thoracic spinal cord of STZ-treated rats when compared to control. BdABK failed to displace the B2R radioligand but displaced the B1R radioligand (IC50 = 5.3 nM. In comparison, IC50 values of B1R selective antagonist R-715 and B1R agonist des-Arg9-BK were 4.3 nM and 19 nM, respectively. Intraperitoneal BdABK and des-Arg9-BK elicited dose-dependent thermal hyperalgesia in STZ-treated rats but not in control rats. The B1R fluorescent agonist was co-localized with immunomarkers of microglia, astrocytes and sensory C fibers in the spinal cord of STZ-treated rats. Conclusion The induction and up-regulation of B1R in glial and sensory cells of the spinal cord in STZ-diabetic rats reinforce the idea that kinin B1R is an important target for drug development in pain processes.

  12. Argümantasyon ve Matematiksel Kanıt Süreçleri Arasındaki İlişkiler

    Ali BÜLBÜL

    2016-06-01

    Full Text Available Bu çalışmanın amacı, lise son sınıf öğrencilerinin argümantasyon ve matematiksel kanıt yapma süreçlerini Toulmin modeline göre analiz etmek ve karşılaştırmaktır. Çalışmaya Ankara’da bir özel okulda lise son sınıfa devam eden dört öğrenci katılmıştır. Nitel araştırma olarak tasarlanan bu çalışmada veri toplamak için literatürden alınan bir problem kullanılmış ve çiftler halinde çalışması sağlanan öğrencilerden problemin çözümü ile ilgili üretecekleri hipotezin kanıtını yapmaları istenmiştir. Analiz sonuçlarına göre, öğrencilerin abdüktif argümantasyon ile dedüktif kanıt arasındaki yapısal boşluğu tamamlayarak dedüktif kanıta geçebildikleri durumda, kanıt sürecini başarıyla tamamladıkları; yapısal boşluğu tamamlayamadıkları durumda ise argümantasyondaki abdüktif yapıyı devam ettirdikleri ve dedüktif kanıt yapamadıkları görülmüştür. Matematik eğitiminde argümantasyon ve matematiksel kanıt süreçlerinin karşılaştırmalı olarak analiz edilmesi ve aralarındaki ilişkilerin belirlenmesi, öğrenciler için kanıtlama sürecini kolaylaştırmak adına gerekli ve önemlidir.

  13. Chapter 20: In vitro and in vivo evaluation of ['99mTc(CO)3-cyclo[Arg-Gly-Asp-D-Tyr-Lys(PZ1)

    As part of a coordinated research project, Development of 99mTc Based Small Biomolecules Using Novel 99mTc Cores, with the International Atomic Energy Agency, we report the design and development of a new cyclic Rd analogue cyclo[Arg-Gly- Asp-D-Tyr-Lys(P Z1)] (P Z1 3,5Me2-Pz(CH2)2N((CH2)3COOH)(CH2)2NH2) that can be radiolabelled with the 99mTc(CO)3 metal fragment. Radiochemical evaluation, including synthesis, in vitro uptake analyses and in vivo pharmacokinetics in normal mice, is discussed. (author)

  14. Enhanced horizontal transfer of antibiotic resistance genes in freshwater microcosms induced by an ionic liquid.

    Qing Wang

    Full Text Available The spread and propagation of antibiotic resistance genes (ARGs is a worldwide public health concern. Ionic liquids (ILs, considered as "environmentally friendly" replacements for industrial organic solvents, have been widely applied in modern industry. However, few data have been collected regarding the potential ecological and environmental risks of ILs, which are important for preparing for their potential discharge into the environment. In this paper, the IL 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIm][PF6] (0.001-5.0 g/L was tested for its effects on facilitating ARGs horizontal transfer mediated by plasmid RP4 in freshwater microcosms. In the horizontal transfer microcosms, the transfer frequency of plasmid RP4 was significantly enhanced (60-fold higher than untreated groups by the IL [BMIm][PF6] (1.0 g/L. Meanwhile, two strains of opportunistic pathogen Acinetobacter spp. and Salmonella spp. were isolated among the transconjugants, illustrating plasmid RP4 mediated horizontal transfer of ARGs occurred in pathogen. This could increase the risk of ARGs dissemination to human pathogens and pose great threat to public health. The cause that [BMIm[PF6] enhanced the transfer frequency of plasmid RP4 was proposed by suppressed cell membrane barrier and enhanced cell membrane permeability, which was evidenced by flow cytometry (FCM. This is the first report that some ILs facilitate horizontal transfer of plasmid RP4 which is widely distributed in the environment and thus add the adverse effects of the environmental risk of ILs.

  15. [Investigation of pollution characteristics of erythromycin resistance genes in a sewage treatment plant and the relevant selective factors].

    Li, Kan-Zhu; Wu, Li-Le; Huang, Sheng-Lin; He, Shi; Liu, Zhen-Hong; Xue, Gang; Gao, Pin

    2014-12-01

    Occurrence and distribution of twelve pharmaceutical and personal care products (PPCPs) were investigated in a sewage treatment plant in Shanghai using solid-phase extraction combined with high-performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS). Quantitative PCR (qPCR) was used to determine the distribution and removal of seven erythromycin resistance genes (ERY-ARGs). The results showed that five PPCPs including sulfamethoxazole, erythromycin, tetracycline, carbamazepine and triclosan were detected in the collected wastewater samples with concentrations in the ranges of 24.5- 38.7, 47.5-49.2, 43.1-85.4, 2.5-3.9 and 423.2-8 973.3 ng x L(-1), respectively. During the wastewater treatment process, a significant reduction of triclosan was observed, but the removal efficiencies for the other detected PPCPs were relatively low. Additionally, all target ERY-ARGs were detected in the wastewater samples ranging from 9.28 x 10(3) (ermA) to 1.83 x 10(8) (ereA) copies x L(-1) in raw influent. Though significant reductions (1.19 log-3.97 log) of ERY-ARGs were obtained, their concentrations found in the final effluent were still high. Moreover, the concentration of ERY-ARGs exhibited significant positive correlation with the concentration of erythromycin and triclosan (P < 0.05), respectively, elucidating that erythromycin played an important role in the occurrence and spread of ERY-ARGs, while triclosan may confer cross-selection for ERY-ARGs. PMID:25826929

  16. Occurrence and distribution of antibiotics, antibiotic resistance genes in the urban rivers in Beijing, China.

    Xu, Yan; Guo, Changsheng; Luo, Yi; Lv, Jiapei; Zhang, Yuan; Lin, Haixia; Wang, Li; Xu, Jian

    2016-06-01

    The occurrence and distribution of sulfonamide and tetracycline, corresponding bacterial resistant rate and resistance genes (ARGs) and two integrase genes were investigated in seven urban rivers in Beijing, China. The total concentration of sulfonamide and tetracycline ranged from 1.3 × 10(1)-1.5 × 10(3) ng/L and 3.9 × 10(1)-5.4 × 10(4) ng/L for water, and 1.0 × 10(0)-2.7 × 10(2) and 3.1 × 10(1)-1.6 × 10(2) ng/g for sediment, respectively. The sul resistant rate was 2-3 times higher than tet resistant rate in both surface water and sediment. The average rate of sul resistance and tet resistance were up to 81.3% and 38.6% in surface water, 89.1% and 69.4% in the sediment, respectively. The sul1, tetA and tetE genes were predominant in term of the absolute abundance. The absolute abundance of ARGs in Wenyu River and Qinghe River, which were close to the direct discharging sites, were 5-50 times higher than those in the other investigated urban rivers, suggesting that the source release played an important role in the distribution of ARGs. The sul1 and sul2 genes had positive correlation (p sources of pollution. Considering principal component analysis, sampling sites (QH5, QH6, B1, B2, B3 and BX2) intimated that a complex interplay of processes govern fate and transport of ARGs in the junction of rivers. These results are significant to understand the fate, and the contribution of ARGs from the source release. In view of the large-scale investigation of urban rivers system in Beijing, it reflected the bacterial resistance in sewage drainage system. Such investigation highlights the management on controlling the pollutant release which was seemed as a major driving force for the maintenance and propagation of many ARGs during the development of urbanization in the future. PMID:27038570

  17. Preliminary Study on the Single Nucleotide Polymorphism (SNP of XRCC1 Gene Identificationto Improve the Outcomes of Radiotherapy for Cervical Cancer

    Devita Tetriana

    2015-09-01

    Full Text Available Cervical cancer is the most fatal disease among Indonesian women. In recognition of the substantial variation in the intrinsic response of individuals to radiation, an effort had been done to identify the genetic markers, primarily Single Nucleotide polymorphisms (SNPs, which are associated with responsiveness of cancer cells to radiation therapy. One of these SNPs is X-ray repair cross-complementing protein 1 (XRCC1 that is one of the most important genes in deoxyribonucleic acid (DNA repair pathways. Meta-analysis in the determination of the association of XRCC1 polymorphisms with cervical cancer revealed the potential role of XRCC1 polymorphisms in predicting cell response to radiotherapy.Our preliminary study with real-time polymerase chain reaction (RT-PCR showed that radiotherapy affected the XRCC1 gene analyzed in blood of cervical cancer patient. Other published study found three SNPs of XRCC1 (Arg194Trp, Arg280His, and Arg399Gln that cause amino acid substitutions. Arg194Trp is only SNPs that associated with high risk of cervical cancer but not others. Additionally, structure and function of this protein can be altered by functional SNPs, which may lead to the susceptibility of individuals to cancers. Anotherstudy found G399A polymorphisms. We concluded that SNP of this DNA repair genes have been found to be good predictors of efficacy of radiotherapy.Kanker serviks adalah penyakit yang paling fatal pada perempuan di Indonesia. Untuk memahami variasi substansial respon intrinsik individual terhadap radiasi, suatu usaha telah dilakukan untuk mengidentifikasi petanda genetik, terutama Single Nucleotide polymorphism (SNP, yang berkaitan dengan responsel kanker terhadap terapi radiasi. Satu dari SNP tersebut adalah X-ray repair cross-complementing protein 1 (XRCC1 yang merupakan satu dari gen paling penting dalam lajur perbaikan asam deoksiribonukleat (DNA. Meta-analysis dalam penentuan hubungan polimorfisme XRCC1 dengan kanker serviks

  18. ARGE Weser (Working Group for Water Pollution Abatement in the Weser River). Weser water quality report 1996; Arbeitsgemeinschaft zur Reinhaltung der Weser. Weserguetebericht 1996

    NONE

    1997-08-01

    The ARGE Weser (Arbeitsgemeinschaft zur Reinhaltung der Weser), which comprises representatives of the German states of Bremen, Hessen, Niedersachsen, Nordrhein-Westfalen and Thueringen, publishes a yearly water quality report based on the measuring programmes for the Weser river and for the Werra/Ulster rivers (the latter is a joint project of the states of Hessen and Thrueringen). The Weser river today has a water quality of II-III after the successful implementation of the 1989 action programme. Chloride concentrations were reduced as well, but efforts will still be made by communal authorities, agriculture, industry and administrative bodies if the goals set for 2000 are to be reached. (orig/AJ) [Deutsch] Die Arbeitsgemeinschaft zur Reinhaltung der Weser (ARGE Weser), der die Bundeslaender Bremen, Hessen, Niedersachsen, Nordrhein-Westfalen und Thueringen angehoeren, dokumentiert jaehrlich im Weserguetebericht die Ergebnisse des Weser- und des von Hessen und Thueringen gemeinsam gefuehrten Messprogrammes Werra/Ulster. Die Weser weist heute ueberwiegend die Gewaesserguete II-III auf. Dies ist ein Ergebnis des 1989 beschlossenen Aktionsprogrammes. Deutliche Erfolge konnten auch im Hinblick auf die Reduzierung der Chloridbelastung erzielt werden. Die volle Verwirklichung des Aktionsprogrammes Weser bis zum Jahr 2000 wird allerdings noch erhebliche Anstrengungen auf Seiten der Kommunen, der Landwirtschaft, der Industrie und der Verwaltungen erfordern. (orig./AJ)

  19. Un arma no solo de prestigio: la espada argárica de Peñalosa (Baños de la Encina, Jaén

    Moreno Onorato, Auxilio

    2015-12-01

    Full Text Available The sword found in a house at the Bronze Age settlement of Peñalosa (Baños de la Encina, province of Jaén is the only example from a domestic context in the El Argar culture. We analize the circumstances of the discovery, and discuss the results of the various analyses that have been performed. Finally, we compare the Peñalosa find to other contemporaneous swords from the Iberian Peninsula and discuss whether the role of Argaric weapons was symbolic or functional.Se estudia una espada singular de la cultura argárica tanto por su tipología, estado de conservación (hoja, piezas de la empuñadura y metal (bronce y plata como por proceder de un contexto doméstico del poblado de la Edad del Bronce de Peñalosa (Baños de la Encina, Jaén. Se analizan las circunstancias del hallazgo, presentándose para su discusión los resultados de los análisis FRX y ICP-SFMS realizados. Se compara con otras espadas contemporáneas aparecidas en la Península Ibérica y se debate el papel simbólico o funcional de las armas argáricas.

  20. Interaction pattern of Arg 62 in the A-pocket of differentially disease-associated HLA-B27 subtypes suggests distinct TCR binding modes.

    Elisa Nurzia

    Full Text Available The single amino acid replacement Asp116His distinguishes the two subtypes HLA-B*2705 and HLA-B*2709 which are, respectively, associated and non-associated with Ankylosing Spondylitis, an autoimmune chronic inflammatory disease. The reason for this differential association is so far poorly understood and might be related to subtype-specific HLA:peptide conformations as well as to subtype/peptide-dependent dynamical properties on the nanoscale. Here, we combine functional experiments with extensive molecular dynamics simulations to investigate the molecular dynamics and function of the conserved Arg62 of the α1-helix for both B27 subtypes in complex with the self-peptides pVIPR (RRKWRRWHL and TIS (RRLPIFSRL, and the viral peptides pLMP2 (RRRWRRLTV and NPflu (SRYWAIRTR. Simulations of HLA:peptide systems suggest that peptide-stabilizing interactions of the Arg62 residue observed in crystal structures are metastable for both B27 subtypes under physiological conditions, rendering this arginine solvent-exposed and, probably, a key residue for TCR interaction more than peptide-binding. This view is supported by functional experiments with conservative (R62K and non-conservative (R62A B*2705 and B*2709 mutants that showed an overall reduction in their capability to present peptides to CD8+ T cells. Moreover, major subtype-dependent differences in the peptide recognition suggest distinct TCR binding modes for the B*2705 versus the B*2709 subtype.

  1. A canine orthologue of the human GFAP c.716G>A (p.Arg239His) variant causes Alexander disease in a Labrador retriever.

    Van Poucke, Mario; Martlé, Valentine; Van Brantegem, Leen; Ducatelle, Richard; Van Ham, Luc; Bhatti, Sofie; Peelman, Luc J

    2016-06-01

    Alexander disease (AxD) is a fatal neurodegenerative disorder of astrocyte dysfunction in man, for which already a number of causal variants are described, mostly de novo dominant missense variants in the glial fibrillary acidic protein (GFAP). A similar disorder was already phenotypically described in animals but without the identification of causal variants. We diagnosed a Labrador retriever with a juvenile form of AxD based on clinical (tetraparesis with spastic front limbs mimicking 'swimming puppy syndrome') and pathological (the detection of GFAP containing Rosenthal fibers in astrocytes) features. In order to identify a causal variant, the coding sequences of the four detected GFAP transcript variants (orthologues from human transcript variants α, γ, δ/ɛ and κ) were sequenced. From the five detected variants, a heterozygous c.719G>A nucleotide substitution resulting in a p.Arg240His substitution was considered to be causal, because it is orthologous to the heterozygous de novo dominant c.716G>A (p.Arg239His) hotspot variant in man, proven to cause a severe phenotype. In addition, the variant was not found in 50 unrelated healthy Labrador retrievers. Because the condition in dogs is morphologically similar to man, it could be a promising animal model for further elucidating the genotype/phenotype correlation in order to treat or prevent this disease. PMID:26486469

  2. Population Structure and Gene Flow of the Yellow Anaconda (Eunectes notaeus) in Northern Argentina

    McCartney-Melstad, Evan; Waller, Tomás; Micucci, Patricio A.; Barros, Mariano; Draque, Juan; Amato, George; Mendez, Martin

    2012-01-01

    Yellow anacondas (Eunectes notaeus) are large, semiaquatic boid snakes found in wetland systems in South America. These snakes are commercially harvested under a sustainable management plan in Argentina, so information regarding population structuring can be helpful for determination of management units. We evaluated genetic structure and migration using partial sequences from the mitochondrial control region and mitochondrial genes cyt-b and ND4 for 183 samples collected within northern Arge...

  3. Aerobic digestion reduces the quantity of antibiotic resistance genes in residual municipal wastewater solids

    Burch, Tucker R.; Sadowsky, Michael J.; LaPara, Timothy M.

    2013-01-01

    Numerous initiatives have been undertaken to circumvent the problem of antibiotic resistance, including the development of new antibiotics, the use of narrow spectrum antibiotics, and the reduction of inappropriate antibiotic use. We propose an alternative but complimentary approach to reduce antibiotic resistant bacteria (ARB) by implementing more stringent technologies for treating municipal wastewater, which is known to contain large quantities of ARB and antibiotic resistance genes (ARGs)...

  4. Reduction of antibiotic resistance genes in municipal wastewater effluent by advanced oxidation processes.

    Zhang, Yingying; Zhuang, Yao; Geng, Jinju; Ren, Hongqiang; Xu, Ke; Ding, Lili

    2016-04-15

    This study investigated the reduction of antibiotic resistance genes (ARGs), intI1 and 16S rRNA genes, by advanced oxidation processes (AOPs), namely Fenton oxidation (Fe(2+)/H2O2) and UV/H2O2 process. The ARGs include sul1, tetX, and tetG from municipal wastewater effluent. The results indicated that the Fenton oxidation and UV/H2O2 process could reduce selected ARGs effectively. Oxidation by the Fenton process was slightly better than that of the UV/H2O2 method. Particularly, for the Fenton oxidation, under the optimal condition wherein Fe(2+)/H2O2 had a molar ratio of 0.1 and a H2O2 concentration of 0.01molL(-1) with a pH of 3.0 and reaction time of 2h, 2.58-3.79 logs of target genes were removed. Under the initial effluent pH condition (pH=7.0), the removal was 2.26-3.35 logs. For the UV/H2O2 process, when the pH was 3.5 with a H2O2 concentration of 0.01molL(-1) accompanied by 30min of UV irradiation, all ARGs could achieve a reduction of 2.8-3.5 logs, and 1.55-2.32 logs at a pH of 7.0. The Fenton oxidation and UV/H2O2 process followed the first-order reaction kinetic model. The removal of target genes was affected by many parameters, including initial Fe(2+)/H2O2 molar ratios, H2O2 concentration, solution pH, and reaction time. Among these factors, reagent concentrations and pH values are the most important factors during AOPs. PMID:26815295

  5. Convulsant doses of a dopamine D1 receptor agonist result in Erk-dependent increases in Zif268 and Arc/Arg3.1 expression in mouse dentate gyrus.

    Gangarossa, Giuseppe; Di Benedetto, Manuela; O'Sullivan, Gerard J; Dunleavy, Mark; Alcacer, Cristina; Bonito-Oliva, Alessandra; Henshall, David C; Waddington, John L; Valjent, Emmanuel; Fisone, Gilberto

    2011-01-01

    Activation of dopamine D1 receptors (D1Rs) has been shown to induce epileptiform activity. We studied the molecular changes occurring in the hippocampus in response to the administration of the D1-type receptor agonist, SKF 81297. SKF 81297 at 2.5 and 5.0 mg/kg induced behavioural seizures. Electrophysiological recordings in the dentate gyrus revealed the presence of epileptiform discharges peaking at 30-45 min post-injection and declining by 60 min. Seizures were prevented by the D1-type receptor antagonist, SCH 23390, or the cannabinoid CB1 receptor agonist, CP 55,940. The effect of SKF 81297 was accompanied by increased phosphorylation of the extracellular signal-regulated protein kinases 1 and 2 (ERK), in the granule cells of the dentate gyrus. This effect was also observed in response to administration of other D1-type receptor agonists, such as SKF83822 and SKF83959. In addition, SKF 81297 increased the phosphorylation of the ribosomal protein S6 and histone H3, two downstream targets of ERK. These effects were prevented by genetic inactivation of D1Rs, or by pharmacological inhibition of ERK. SKF 81297 was also able to enhance the levels of Zif268 and Arc/Arg3.1, two immediate early genes involved in transcriptional regulation and synaptic plasticity. These changes may be involved in forms of activity-dependent plasticity linked to the manifestation of seizures and to the ability of dopamine to affect learning and memory. PMID:21559295

  6. Association of the ADRA1A gene and the severity of metabolic abnormalities in patients with schizophrenia.

    Cheng, Chin; Chiu, Hsien-Jane; Loh, El-Wui; Chan, Chin-Hong; Hwu, Tzong-Ming; Liu, Yun-Ru; Lan, Tsuo-Hung

    2012-01-10

    Patients with schizophrenia have a higher risk of developing metabolic abnormalities and their associated diseases. Some studies found that the accumulative number of metabolic syndrome components was associated with the severity of metabolic abnormalities. The purpose of this study was to examine the roles of the ADRA1A, ADRA2A, ADRB3, and 5HT2A genes in the risk of having more severe metabolic abnormalities among patients with schizophrenia. We studied a sample of 232 chronic inpatients with schizophrenia (120 males and 112 females) to explore the associations between the four candidate genes and the severity of metabolic syndrome by accumulative number of the components. Four single nucleotide polymorphisms in the candidate genes were genotyped, including the Arg347Cys in ADRA1A, the C1291G in ADRA2A, the Try64Arg in ADRB3, and the T102C in 5HT2A. An association between the accumulative number of metabolic syndrome components and the ADRA1A gene was found after adjusting age, sex, and other related variables (p-value=0.036). Presence of the Arg347 allele in the ADRA1A gene is a risk factor for having more severe metabolic abnormalities. These findings suggest a medical attention of closely monitoring metabolic risks for schizophrenia patients with high-risk genotypes. PMID:22037178

  7. Abundances of tetracycline, sulphonamide and beta-lactam antibiotic resistance genes in conventional wastewater treatment plants (WWTPs) with different waste load

    Laht, Mailis; Karkman, Antti; Voolaid, Veiko;

    2014-01-01

    Antibiotics and antibiotic resistant bacteria enter wastewater treatment plants (WWTPs), an environment where resistance genes can potentially spread and exchange between microbes. Several antibiotic resistance genes (ARGs) were quantified using qPCR in three WWTPs of decreasing capacity located in...... abundances with 16S rRNA gene abundances while assessing if the respective genes increased or decreased during treatment. ARGs were detected in most samples; sul1, sul2, and tetM were detected in all samples. Statistically significant differences (adjusted p<0.01) between the inflow and effluent were...... detected in only four cases. Effluent values for blaoxa-58 and tetC decreased in the two larger plants while tetM decreased in the medium-sized plant. Only blashv-34 increased in the effluent from the medium-sized plant. In all other cases the purification process caused no significant change in the...

  8. Co-occurrence of integrase 1, antibiotic and heavy metal resistance genes in municipal wastewater treatment plants.

    Di Cesare, Andrea; Eckert, Ester M; D'Urso, Silvia; Bertoni, Roberto; Gillan, David C; Wattiez, Ruddy; Corno, Gianluca

    2016-05-01

    The impact of human activities on the spread and on the persistence of antibiotic resistances in the environment is still far from being understood. The natural background of resistances is influenced by human activities, and the wastewater treatment plants (WWTPs) are among the main sources of the release of antibiotic resistance into the environment. The various treatments of WWTPs provide a number of different environmental conditions potentially favoring the selection of antibiotic resistance genes (ARGs) and thereby their well-documented spread in the environment. Although the distribution of different ARGs in WWTPs has been deeply investigated, very little is known on the ecology and on the molecular mechanisms underlying the selection of specific ARGs. This study investigates the fate of diverse ARGs, heavy metal resistance genes (HMRGs) and of a mobile element (the class I integron) in three WWTPs. Abundances of the different genetic markers were correlated to each other and their relation to biotic and abiotic factors (total organic carbon, total nitrogen, prokaryotic cell abundance and its relative distribution in single cells and aggregates) influencing the microbial communities in the different treatment phases in three WWTPs, were investigated. Water samples were analyzed for the abundance of six ARGs (tetA, sulII, blaTEM, blaCTXM,ermB, and qnrS), two HMRGs (czcA and arsB), and of the class I integron (int1). The measured variables clustered in two well-defined groups, the first including tetA, ermB, qnrS and the different biotic and abiotic factors, and a second group around the genes sulII, czcA, arsB and int1. Moreover, the dynamics of sulII, HMRGs, and int1 correlated strongly. Our results suggest a potentially crucial role of HMRGs in the spread, mediated by mobile elements, of some ARGs, i.e. sulII. The possibility of a relation between heavy metal contamination and the spread of ARGs in WWTPs calls for further research to clarify the mechanisms

  9. The Gln27Glu Polymorphism in β2-Adrenergic receptor gene is linked to hypertriglyceridemia, hyperinsulinemia and hyperleptinemia in Saudis

    Daghestani Maha H; Warsy Arjumand; Daghestani Mazin H; Al-odaib Ali N; Eldali Abdelmoneim; Al-Eisa Nadia A; Al-zhrani Sabah

    2010-01-01

    Abstract Background β2-adrenoceptor (β2AR) gene polymorphism glutamine 27 glutamic acid (Gln27Glu) and Arg16Gly were reported to have an association with obesity and obesity related disorders in some population. We evaluated Gln27Glu polymorphism in the β2AR gene in obese Saudi populations to investigate the association of β2AR gene with obesity and other related metabolic parameters. Design We studied possible association of Gln27Glu in β2AR gene with body mass index (BMI), anthropometric me...

  10. Molecular Characterization of TP53 Gene in Human Populations Exposed to Low-Dose Ionizing Radiation

    Igor Brasil-Costa

    2013-01-01

    Full Text Available Ionizing radiation, such as that emitted by uranium, may cause mutations and consequently lead to neoplasia in human cells. The TP53 gene acts to maintain genomic integrity and constitutes an important biomarker of susceptibility. The present study investigated the main alterations observed in exons 4, 5, 6, 7, and 8 of the TP53 gene and adjacent introns in Amazonian populations exposed to radioactivity. Samples were collected from 163 individuals. Occurrence of the following alterations was observed: (i a missense exchange in exon 4 (Arg72Pro; (ii 2 synonymous exchanges, 1 in exon 5 (His179His, and another in exon 6 (Arg213Arg; (iii 4 intronic exchanges, 3 in intron 7 (C → T at position 13.436; C → T at position 13.491; T → G at position 13.511 and 1 in intron 8 (T → G at position 13.958. Alteration of codon 72 was found to be an important risk factor for cancer development (P=0.024; OR=6.48; CI: 1.29–32.64 when adjusted for age and smoking. Thus, TP53 gene may be an important biomarker for carcinogenesis susceptibility in human populations exposed to ionizing radiation.

  11. Hypokalemic periodic paralysis; two different genes responsible for similar clinical manifestations

    Hunmin Kim

    2011-11-01

    Full Text Available Primary hypokalemic periodic paralysis (HOKPP is an autosomal dominant disorder manifesting as recurrent periodic flaccid paralysis and concomitant hypokalemia. HOKPP is divided into type 1 and type 2 based on the causative gene. Although 2 different ion channels have been identified as the molecular genetic cause of HOKPP, the clinical manifestations between the 2 groups are similar. We report the cases of 2 patients with HOKPP who both presented with typical clinical manifestations, but with mutations in 2 different genes (CACNA1S p.Arg528His and SCN4A p.Arg672His. Despite the similar clinical manifestations, there were differences in the response to acetazolamide treatment between certain genotypes of SCN4A mutations and CACNA1S mutations. We identified p.Arg672His in the SCN4A gene of patient 2 immediately after the first attack through a molecular genetic testing strategy. Molecular genetic diagnosis is important for genetic counseling and selecting preventive treatment.

  12. Hypokalemic periodic paralysis; two different genes responsible for similar clinical manifestations.

    Kim, Hunmin; Hwang, Hee; Cheong, Hae Il; Park, Hye Won

    2011-11-01

    Primary hypokalemic periodic paralysis (HOKPP) is an autosomal dominant disorder manifesting as recurrent periodic flaccid paralysis and concomitant hypokalemia. HOKPP is divided into type 1 and type 2 based on the causative gene. Although 2 different ion channels have been identified as the molecular genetic cause of HOKPP, the clinical manifestations between the 2 groups are similar. We report the cases of 2 patients with HOKPP who both presented with typical clinical manifestations, but with mutations in 2 different genes (CACNA1Sp.Arg528His and SCN4A p.Arg672His). Despite the similar clinical manifestations, there were differences in the response to acetazolamide treatment between certain genotypes of SCN4A mutations and CACNA1S mutations. We identified p.Arg672His in the SCN4A gene of patient 2 immediately after the first attack through a molecular genetic testing strategy. Molecular genetic diagnosis is important for genetic counseling and selecting preventive treatment. PMID:22253645

  13. Development of antibiotic resistance genes in microbial communities during long-term operation of anaerobic reactors in the treatment of pharmaceutical wastewater.

    Aydin, Sevcan; Ince, Bahar; Ince, Orhan

    2015-10-15

    Biological treatment processes offer the ideal conditions in which a high diversity of microorganisms can grow and develop. The wastewater produced during these processes is contaminated with antibiotics and, as such, they provide the ideal setting for the acquisition and proliferation of antibiotic resistance genes (ARGs). This research investigated the occurrence and variation in the ARGs found during the one-year operation of the anaerobic sequencing batch reactors (SBRs) used to treat pharmaceutical wastewater that contained combinations of sulfamethoxazole-tetracycline-erythromycin (STE) and sulfamethoxazole-tetracycline (ST). The existence of eighteen ARGs encoding resistance to sulfamethoxazole (sul1, sul2, sul3), erythromycin (ermA, ermF, ermB, msrA, ereA), tetracycline (tetA, tetB, tetC, tetD, tetE, tetM, tetS, tetQ, tetW, tetX) and class Ι integron gene (intΙ 1) in the STE and ST reactors was investigated by quantitative real-time PCR. Due to the limited availability of primers to detect ARGs, Illumina sequencing was also performed on the sludge and effluent of the STE and ST reactors. Although there was good reactor performance in the SBRs, which corresponds to min 80% COD removal efficiency, tetA, tetB, sul1, sul2 and ermB genes were among those ARGs detected in the effluent from STE and ST reactors. A comparison of the ARGs acquired from the STE and ST reactors revealed that the effluent from the STE reactor had a higher number of ARGs than that from the ST reactor; this could be due to the synergistic effects of erythromycin. According to the expression of genes results, microorganisms achieve tetracycline and erythromycin resistance through a combination of three mechanisms: efflux pumping protein, modification of the antibiotic target and modifying enzymes. There was also a significant association between the presence of the class 1 integron and sulfamethoxazole resistance genes. PMID:26188597

  14. Polymorphisms of the XRCC1, XRCC3 and XPD genes and risk of colorectal adenoma and carcinoma, in a Norwegian cohort: a case control study

    Genetic polymorphisms in DNA repair genes may influence individual variation in DNA repair capacity, which may be associated with risk of developing cancer. For colorectal cancer the importance of mutations in mismatch repair genes has been extensively documented. Less is known about other DNA repair pathways in colorectal carcinogenesis. In this study we have focused on the XRCC1, XRCC3 and XPD genes, involved in base excision repair, homologous recombinational repair and nucleotide excision repair, respectively. We used a case-control study design (157 carcinomas, 983 adenomas and 399 controls) to test the association between five polymorphisms in these DNA repair genes (XRCC1 Arg194Trp, Arg280His, Arg399Gln, XRCC3 Thr241Met and XPD Lys751Gln), and risk of colorectal adenomas and carcinomas in a Norwegian cohort. Odds ratio (OR) and 95% confidence interval (95% CI) were estimated by binary logistic regression model adjusting for age, gender, cigarette smoking and alcohol consumption. The XRCC1 280His allele was associated with an increased risk of adenomas (OR 2.30, 95% CI 1.19–4.46). The XRCC1 399Gln allele was associated with a reduction of risk of high-risk adenomas (OR 0.62, 95% CI 0.41–0.96). Carriers of the variant XPD 751Gln allele had an increased risk of low-risk adenomas (OR 1.40, 95% CI 1.03–1.89), while no association was found with risk of carcinomas. Our results suggest an increased risk for advanced colorectal neoplasia in individuals with the XRCC1 Arg280His polymorphism and a reduced risk associated with the XRCC1 Arg399Gln polymorphism. Interestingly, individuals with the XPD Lys751Gln polymorphism had an increased risk of low-risk adenomas. This may suggest a role in regression of adenomas

  15. β3-adrenergic receptor gene, body mass index, bone mineral density and fracture risk in elderly men and women: the Dubbo Osteoporosis Epidemiology Study (DOES

    Center Jacqueline R

    2006-07-01

    Full Text Available Abstract Background Recent studies have suggested that the Arg allele of β3-adrenergic receptor (ADRB3 gene is associated with body mass index (BMI, which is an important predictor of bone mineral density (BMD and fracture risk. However, whether the ADRB3 gene polymorphism is associated with fracture risk has not been investigated. The aim of study was to examine the inter-relationships between ADRB3 gene polymorphisms, BMI, BMD and fracture risk in elderly Caucasians. Methods Genotypes of the ADRB3 gene were determined in 265 men and 446 women aged 60+ in 1989 at entry into the study, whose BMD were measured by DXA (GE Lunar, WI USA at baseline. During the follow-up period (between 1989 and 2004, fractures were ascertained by reviewing radiography reports and personal interviews. Results The allelic frequencies of the Trp and the Arg alleles were 0.925 and 0.075 respectively, and the relative frequencies of genotypes Trp/Trp, Trp/Arg and Arg/Arg 0.857, 0.138 and 0.006 respectively. There was no significant association between BMI and ADRB3 genotypes (p = 0.10 in women and p = 0.68 in men. There was also no significant association between ADRB3 genotypes and lumbar spine or femoral neck BMD in either men and women. Furthermore, there were no significant association between ADRB3 genotypes and fracture risk in both women and men, either before or after adjusting for and, BMD and BMI. Conclusion The present data suggested that in Caucasian population the contribution of ADRB3 genotypes to the prediction of BMI, BMD and fracture risk is limited.

  16. A three-step programmed method for the identification of causative gene mutations of maturity onset diabetes of the young (MODY).

    Li, Qian; Cao, Xi; Qiu, Hai-Yan; Lu, Jing; Gao, Rui; Liu, Chao; Yuan, Ming-Xia; Yang, Guang-Ran; Yang, Jin-Kui

    2016-08-22

    To establish a three-step programmed method to find gene mutations related to maturity onset diabetes of the young (MODY). Target region capture and next-generation sequencing (NGS) were performed using customized oligonucleotide probes designed to capture suspected genes for MODY in 11 probands with clinically diagnosed MODY. The suspected associations of certain genes with MODY were then confirmed by Sanger sequencing in the probands and their family members. Finally, to validate variants of one of the genes of interest (glucokinase, GCK) as pathogenic mutations, protein function editing by the variant genes was assessed. In the target region capture and NGS phase, a total of nine variants of seven genes (GCK, WFS1, SLC19A2, SH2B1, SERPINB4, RFX6, and GATA6) were identified in eight probands. Two heterozygous GCK mutations located on the same allele (p.Leu77Arg and p.Val101Met) were identified in a MODY family. Sanger sequencing was used to confirm the variants identified by NGS to be present in probands and their diabetic family members, but not in non-diabetic family members. Finally, enzyme kinetic and thermal stability analyses revealed that the p.Leu77Arg mutation or the p.Leu77Arg mutation in combination with the p.Val101Met mutation inactivates GCK function and stability, while mutation of p.Val101Met alone does not. The p.Leu77Arg but not p.Val101Met GCK mutation is therefore considered a pathogenic mutation associated with MODY. Genetic screening coupled with gene-editing protein function testing is an effective and reliable method by which causative gene mutations of MODY can be identified. PMID:27185633

  17. Association between Β3-Adrenergic receptor (ADRB3) gene polymorphism with body mass index and bone mineral density in Turkish postmenopausal women

    Turgay İşbir2, Ayşe Can1, Özlem Kurt-Şirin1, Hülya Yılmaz-Aydoğan2 Mehmet Uyar3, Mehmet Fatih Seyhan2,

    2016-01-01

    Abstract: Previous studies have suggested that β3-adrenergic receptor (ADRB3) gene is associated with body mass index (BMI), which is an important predictor of bone mineral density (BMD). However, little is known concerning the effect of the ADRB3 gene on BMD. The present study investigated the relationship between ADRB3 Trp64Arg polymorphism, BMI and BMD in Turkish postmenopausal women. 133 postmenopausal women (81 osteoporotic and 52 healthy control) were recruited. For the detection of ADR...

  18. Partial rescue of in vivo insulin signalling in skeletal muscle by impaired insulin clearance in heterozygous carriers of a mutation in the insulin receptor gene

    Højlund, K.; Wojtaszewski, Jørgen; Birk, Jesper Bratz;

    2006-01-01

    AIMS/HYPOTHESIS: Recently we reported the coexistence of postprandial hypoglycaemia and moderate insulin resistance in heterozygous carriers of the Arg1174Gln mutation in the insulin receptor gene (INSR). Controlled studies of in vivo insulin signalling in humans with mutant INSR are unavailable,...

  19. Dose and polymorphic genes xrcc1, xrcc3, gst play a role in the risk of articledeveloping erythema in breast cancer patients following single shot partial breast irradiation after conservative surgery

    To evaluate the association between polymorphisms involved in DNA repair and oxidative stress genes and mean dose to whole breast on acute skin reactions (erythema) in breast cancer (BC) patients following single shot partial breast irradiation (SSPBI) after breast conservative surgery. Acute toxicity was assessed using vers.3 criteria. single nucleotides polymorphisms(SNPs) in genes: XRCC1(Arg399Gln/Arg194Trp), XRCC3 (A4541G-5'UTR/Thr241Met), GSTP1(Ile105Val), GSTA1 and RAD51(untranslated region). SNPs were determined in 57 BC patients by the Pyrosequencing analysis. Univariate(ORs and 95% CI) and logistic multivariate analyses (MVA) were performed to correlate polymorphic genes with the risk of developing acute skin reactions to radiotherapy. After SSPBI on the tumour bed following conservative surgery, grade 1 or 2 acute erythema was observed in 19 pts(33%). Univariate analysis indicated a higher significant risk of developing erythema in patients with polymorphic variant wt XRCC1Arg194Trp, mut/het XRCC3Thr241Met, wt/het XRCC3A4541G-5'UTR. Similarly a higher erythema rate was also found in the presence of mut/het of XRCC1Arg194Trp or wt of GSTA1. Whereas, a lower erythema rate was observed in patients with mut/het of XRCC1Arg194Trp or wt of XRCC1Arg399Gln. The mean dose to whole breast(p = 0.002), the presence of either mut/het XRCC1Arg194Trp or wt XRCC3Thr241Met (p = 0.006) and the presence of either mut/het XRCC1Arg194Trp or wt GSTA1(p = 0.031) were confirmed as predictors of radiotherapy-induced erythema by MVA. The Whole breast mean dose together with the presence of some polymorphic genes involved in DNA repair or oxidative stress could explain the erythema observed after SSPBI, but further studies are needed to confirm these results in a larger cohort. ClinicalTrials.gov Identifier: http://www.clinicaltrials.gov/ct2/show/NCT01316328

  20. 长链人胰岛素样生长因子-1的分离纯化%Isolation and purification of long chain Arg3 human insulin-like growth factor-1

    张文明; 洪静; 林殿海; 孙天玮; 梁军; 黄国团

    2012-01-01

    Objective To develop a method for isolation and purification of long chain Arg3 human insulin-like growth factor-1 (LR3IGF-1). Methods Recombinant Pichia pastoris with LR3IGF-1 gene was inoculated to a 30 L fermenter for high density fermentation. The fermentation liquid was concentrated by centrifugation and ultrafiltration,purified by SP Sepharose FF cation exchange chromatography,Phenyl Sepharose FF hydrophobic chromatDgraphy,S-100 gel filtration chromatDgraphy and DEAE Sepharose FF chromatography,then determined for protein concentration,based on which the recovery rale of protein was calculated,and the concentration of purified sample was analyzed. The activities of samples at various steps of purification in promoting the proliferation of HIN-3T3 cells were determined by MTT method. Results The total recovery rate and RP-HPLC purity of purified LR3IGF-1 was 28% and more than 95% respectively. The activity of LR3IGF-1 increased with the increasing purity during purification,which was basically uninfluenced by the purification steps. Conclusion A simple and effective procedure for isolation and purification of LR3IGF-1 was preliminarily developed,which laid a foundation of large-scale production of LR3iGF-1.%目的 建立长链人胰岛素样生长因子-1(Long chain Arg3 human insulin-like growth factor-1,LR3IGF-1)的分离纯化方法.方法 将LR31GF-1毕赤酵母工程菌接种至30 L发酵罐进行高密度发酵,发酵液经离心、超滤浓缩后,采用SPSepharose FF阳离子交换层析、Phenyl SepharoseFF疏水层析、S-100凝胶过滤层析和DEAE Sepharose FF层析对发酵上清液中的表达产物进行分离纯化,测定蛋白浓度,计算蛋白收率,并分析纯化样品的浓度.采用MTT法检测各步纯化样品促NIH-3T3细胞增殖的活性.结果 纯化LR3IGF-1的蛋白总收率为28%,RP-HPLC纯度可达95%以上.随着纯化过程中样品纯度的增加,LR3IGF-1的活性也相应增强,各纯化步骤基本不影响LR3IGF-1

  1. Monitoring and Comparison of Antibiotic Resistant Bacteria and Their Resistance Genes in Municipal and Hospital Wastewaters

    Rahim Aali

    2014-01-01

    Full Text Available Background: Human exposure to antibiotic resistant bacteria (ARB is a public health concern which could occur in a number of ways. Wastewaters seem to play an important role in the dissemination of bacteria and antibiotic resistant genes (ARGs in our environment. The aim of this study was to evaluate the occurrence of three groups of ARB and their resistance genes in hospital and municipal wastewaters (MWs as possible sources. Methods: A total of 66 samples were collected from raw MWs and hospital wastewaters (HWs and final effluents of related wastewater treatment plants (WWTPs. Samples were analyzed for the detection of three groups of ARB including gentamicin (GM, chloramphenicol (CHL and ceftazidime resistant bacteria and their ARGs (aac (3-1, cmlA1 and ctx-m-32, respectively. Results: The mean concentration of GM, CHL and ceftazidime resistant bacteria in raw wastewater samples was 1.24 × 10 7 , 3.29 × 10 7 and 5.54 × 10 7 colony forming unit/100 ml, respectively. There is a variation in prevalence of different groups of ARB in MWs and HWs. All WWTPs decreased the concentration of ARB. However, high concentration of ARB was found in the final effluent of WWTPs. Similar to ARB, different groups of ARGs were found frequently in both MWs and HWs. All genes also detected with a relative high frequency in effluent samples of MWs WWTPs. Conclusions: Discharge of final effluent from conventional WWTPs is a potential route for dissemination of ARB and ARGs into the natural environment and poses a hazard to environmental and public health.

  2. Occurrence and persistence of antibiotic resistance genes in river biofilms after wastewater inputs in small rivers.

    Proia, Lorenzo; von Schiller, Daniel; Sànchez-Melsió, Alexandre; Sabater, Sergi; Borrego, Carles M; Rodríguez-Mozaz, Sara; Balcázar, José Luis

    2016-03-01

    The extensive use of antibiotics in human and veterinary medicine and their subsequent release into the environment may have direct consequences for autochthonous bacterial communities, especially in freshwater ecosystems. In small streams and rivers, local inputs of wastewater treatment plants (WWTPs) may become important sources of organic matter, nutrients and emerging pollutants, such as antibiotic resistance genes (ARGs). In this study, we evaluated the effect of WWTP effluents as a source of ARGs in river biofilms. The prevalence of genes conferring resistance to main antibiotic families, such as beta-lactams (blaCTX-M), fluoroquinolones (qnrS), sulfonamides (sul I), and macrolides (ermB), was determined using quantitative PCR (qPCR) in biofilm samples collected upstream and downstream WWTPs discharge points in four low-order streams. Our results showed that the WWTP effluents strongly modified the hydrology, physico-chemistry and biological characteristics of the receiving streams and favoured the persistence and spread of antibiotic resistance in microbial benthic communities. It was also shown that the magnitude of effects depended on the relative contribution of each WWTP to the receiving system. Specifically, low concentrations of ARGs were detected at sites located upstream of the WWTPs, while a significant increase of their concentrations was observed in biofilms collected downstream of the WWTP discharge points (particularly ermB and sul I genes). These findings suggest that WWTP discharges may favour the increase and spread of antibiotic resistance among streambed biofilms. The present study also showed that the presence of ARGs in biofilms was noticeable far downstream of the WWTP discharge (up to 1 km). It is therefore reasonable to assume that biofilms may represent an ideal setting for the acquisition and spread of antibiotic resistance determinants and thus be considered suitable biological indicators of anthropogenic pollution by active

  3. Clinical variability and novel mutations in the NHEJ1 gene in patients with a Nijmegen breakage syndrome-like phenotype.

    Dutrannoy, Véronique; Demuth, Ilja; Baumann, Ulrich; Schindler, Detlev; Konrat, Kateryna; Neitzel, Heidemarie; Gillessen-Kaesbach, Gabriele; Radszewski, Janina; Rothe, Susanne; Schellenberger, Mario T; Nürnberg, Gudrun; Nürnberg, Peter; Teik, Keng Wee; Nallusamy, Revathy; Reis, André; Sperling, Karl; Digweed, Martin; Varon, Raymonda

    2010-09-01

    We have previously shown that mutations in the genes encoding DNA Ligase IV (LIGIV) and RAD50, involved in DNA repair by nonhomologous-end joining (NHEJ) and homologous recombination, respectively, lead to clinical and cellular features similar to those of Nijmegen Breakage Syndrome (NBS). Very recently, a new member of the NHEJ repair pathway, NHEJ1, was discovered, and mutations in patients with features resembling NBS were described. Here we report on five patients from four families of different ethnic origin with the NBS-like phenotype. Sequence analysis of the NHEJ1 gene in a patient of Spanish and in a patient of Turkish origin identified homozygous, previously reported mutations, c.168C>G (p.Arg57Gly) and c.532C>T (p.Arg178Ter), respectively. Two novel, paternally inherited truncating mutations, c.495dupA (p.Asp166ArgfsTer20) and c.526C>T (p.Arg176Ter) and two novel, maternal genomic deletions of 1.9 and 6.9 kb of the NHEJ1 gene, were found in a compound heterozygous state in two siblings of German origin and in one Malaysian patient, respectively. Our findings confirm that patients with NBS-like phenotypes may have mutations in the NHEJ1 gene including multiexon deletions, and show that considerable clinical variability could be observed even within the same family. PMID:20597108

  4. Co-Circulation of the Rare CPV-2c with Unique Gln370Arg Substitution, New CPV-2b with Unique Thr440Ala Substitution, and New CPV-2a with High Prevalence and Variation in Heilongjiang Province, Northeast China.

    Geng, Yufei; Guo, Donghua; Li, Chunqiu; Wang, Enyu; Wei, Shan; Wang, Zhihui; Yao, Shuang; Zhao, Xiwen; Su, Mingjun; Wang, Xinyu; Wang, Jianfa; Wu, Rui; Feng, Li; Sun, Dongbo

    2015-01-01

    To trace evolution of canine parvovirus-2 (CPV-2), a total of 201 stool samples were collected from dogs with diarrhea in Heilongjiang province of northeast China from May 2014 to April 2015. The presence of CPV-2 in the samples was determined by PCR amplification of the VP2 gene (568 bp) of CPV-2. The results revealed that 95 samples (47.26%) were positive for CPV-2, and they showed 98.8%-100% nucleotide identity and 97.6%-100% amino acid identity. Of 95 CPV-2-positive samples, types new2a (Ser297Ala), new2b (Ser297Ala), and 2c accounted for 64.21%, 21.05%, and 14.74%, respectively. The positive rate of CPV-2 and the distribution of the new2a, new2b and 2c types exhibited differences among regions, seasons, and ages. Immunized dogs accounted for 48.42% of 95 CPV-2-positive samples. Coinfections with canine coronavirus, canine kobuvirus, and canine bocavirus were identified. Phylogenetic analysis revealed that the identified new2a, new2b, and CPV-2c strains in our study exhibited a close relationship with most of the CPV-2 strains from China; type new2a strains exhibited high variability, forming three subgroups; type new2b and CPV-2c strains formed one group with reference strains from China. Of 95 CPV-2 strains, Tyr324Ile and Thr440Ala substitutions accounted for 100% and 64.21%, respectively; all type new2b strains exhibited the Thr440Ala substitution, while the unique Gln370Arg substitution was found in all type 2c strains. Recombination analysis using entire VP2 gene indicated possible recombination events between the identified CPV-2 strains and reference strains from China. Our data revealed the co-circulation of new CPV-2a, new CPV-2b, and rare CPV-2c, as well as potential recombination events among Chinese CPV-2 strains. PMID:26348721

  5. Intervención educativa sobre higiene bucal en escolares del seminternado “Jesús Argüelles Hidalgo”

    Nivia M. Aguilera Trotman; Risquet Joel Menéndez Cabrera; Neiva de la Caridad Nápoles Rodríguez

    2015-01-01

    Se realizó una intervención educativa en la rama de las Ciencias Médicas, específicamente en la estomatología, con el objetivo de elevar el nivel de conocimiento sobre higiene bucal en escolares de ocho a doce años de edad del seminternado “Jesús Argüelles Hidalgo”, perteneciente a la clínica estomatológica “Guillermo Tejas” del municipio de Las Tunas, en el período comprendido de mayo de 2010 a mayo de 2012. El universo estuvo constituido por 392 escolares, seleccionándose una muestra de 150...

  6. Effects of antibiotic resistance genes on the performance and stability of different microbial aggregates in a granular sequencing batch reactor.

    Zou, Wenci; Xue, Bin; Zhi, Weijia; Zhao, Tianyu; Yang, Dong; Qiu, Zhigang; Shen, Zhiqiang; Li, Junwen; Zhang, Bin; Wang, Jingfeng

    2016-03-01

    Antibiotic resistance genes (ARGs) have emerged as key factors in wastewater environmental contaminants and continue to pose a challenge for wastewater treatment processes. With the aim of investigating the performance of granular sludge system when treating wastewater containing a considerable amount of ARGs, a lab-scale granular sequencing batch reactor (GSBR) where flocculent and granular sludge coexisted was designed. The results showed that after inoculation of donor strain NH4(+)-N purification efficiency diminished from 94.7% to 32.8% and recovered to 95.2% after 10 days. Meanwhile, RP4 plasmid had varying effects on different forms of microbial aggregates. As the size of aggregates increased, the abundance of RP4 in sludge decreased. The residence time of RP4 in granules with particle size exceeding 0.9 mm (14 days) was far shorter than that in flocculent sludge (26 days). Therefore, our studies conclude that with increasing number of ARGs being detected in wastewater, the use of granular sludge system in wastewater treatment processes will allow the reduction of ARGs transmissions and lessen potential ecological threats. PMID:26590870

  7. Occurrence and removal of antibiotics and the corresponding resistance genes in wastewater treatment plants: effluents' influence to downstream water environment.

    Li, Jianan; Cheng, Weixiao; Xu, Like; Jiao, Yanan; Baig, Shams Ali; Chen, Hong

    2016-04-01

    In this study, the occurrence of 8 antibiotics [3 tetracyclines (TCs), 4 sulfonamides, and 1 trimethoprim (TMP)], 12 antibiotic resistance genes (ARGs) (10 tet, 2 sul), 4 types of bacteria [no antibiotics, anti-TC, anti-sulfamethoxazole (SMX), and anti-double], and intI1 in two wastewater treatment plants (WWTPs) were assessed and their influences in downstream lake were investigated. Both WWTPs' effluent demonstrated some similarities, but the abundance and removal rate varied significantly. Results revealed that biological treatment mainly removed antibiotics and ARGs, whereas physical techniques were found to eliminate antibiotic resistance bacteria (ARBs) abundance (about 1 log for each one). UV disinfection did not significantly enhance the removal efficiency, and the release of the abundantly available target contaminants from the excess sludge may pose threats to human and the environment. Different antibiotics showed diverse influences on the downstream lake, and the concentrations of sulfamethazine (SM2) and SMX were observed to increase enormously. The total ARG abundance ascended about 0.1 log and some ARGs (e.g., tetC, intI1, tetA) increased due to the high input of the effluent. In addition, the abundance of ARB variation in the lake also changed, but the abundance of four types of bacteria remained stable in the downstream sampling sites. PMID:26658782

  8. Feasibility of lettuce cultivation in sophoroliplid-enhanced washed soil originally polluted with Cd, antibiotics, and antibiotic-resistant genes.

    Ye, Mao; Sun, Mingming; Wan, Jinzhong; Feng, Yanfang; Zhao, Yu; Tian, Da; Hu, Feng; Jiang, Xin

    2016-02-01

    Vegetable cultivation in soils polluted with heavy metals, antibiotics and a high abundance of antibiotic-resistance genes (ARGs) can seriously threaten human health through the food chain. Therefore, novel techniques that not only remediate soil, but also ensure food security are urgently required. In the present study, two successive washings with 20gL(-1) of sophoroliplid solution plus ultrasonication (35kHz) were effective in extracting 71.2% Cd, 88.2% tetracycline, 96.6% sulfadiazine, and 100% roxithromycin. Simultaneously, relative abundance of ARGs (tetM, tetX, sulI, and sulII) was decreased to 10(-7)-10(-8) (ARG copies/16S copies). Further, lettuce cultivation in the 2nd washed soil showed significant improvement in vegetable growth indices (fresh/dry weight, root surface area, chlorophyll content and soluble protein content) and a decrease in isolate counts for antibiotic-resistant bacterial endophytes and ARG abundance in lettuce tissues. This combined cleanup strategy provides an environmentally friendly technology for ensuring vegetable security in washed soils. PMID:26590696

  9. Effect of red mud addition on tetracycline and copper resistance genes and microbial community during the full scale swine manure composting.

    Wang, Rui; Zhang, Junya; Sui, Qianwen; Wan, Hefeng; Tong, Juan; Chen, Meixue; Wei, Yuansong; Wei, Dongbin

    2016-09-01

    Swine manure has been considered as the reservoir of antibiotic resistance genes (ARGs). Composting is one of the most suitable technologies for treating livestock manures, and red mud was proved to have a positive effect on nitrogen conservation during composting. This study investigated the abundance of eight tetracycline and three copper resistance genes, the bacterial community during the full scale swine manure composting with or without addition of red mud. The results showed that ARGs in swine manure could be effectively removed through composting (reduced by 2.4log copies/g TS), especially during the thermophilic phase (reduced by 1.5log copies/g TS), which the main contributor might be temperature. Additionally, evolution of bacterial community could also have a great influence on ARGs. Although addition of red mud could enhance nitrogen conservation, it obviously hindered removal of ARGs (reduced by 1.7log copies/g TS) and affected shaping of bacterial community during composting. PMID:27367291

  10. Lack of Association between Toll Like Receptor-2 and Toll Like Receptor-4 Gene Polymorphisms and Other Feature in Iranian Asthmatics Patients.

    Hamid Bahrami

    2015-02-01

    Full Text Available Asthma as a chronic inflammatory airway disease is considered to be the most common chronic disease that is involving genetic and environmental factors. Toll like receptors (TLRs and other inflammatory mediators are important in modulation of inflammation. In this study, we evaluated the role of TLR2 Arg753Gln and TLR4 Asp299Gly polymorphisms in the asthma susceptibility, progress, control levels and lung functions in Iranian patients. On 99 asthmatic patients and 120 normal subjects, TLR2 Arg753Gln and TLR4 Asp299Gly polymorphisms were evaluated by PCR-RFLP method recruiting Msp1 and Nco1 restriction enzymes, respectively. IgE serum levels by ELISA technique were determined and asthma diagnosis, treatment and control levels were considered using standard schemes and criteria. Our results indicated that the genotype and allele frequencies of the TLR2 Arg753Gln and TLR4 Asp299Gly polymorphisms were not significantly different between control subjects and asthmatics and were not related to in asthma features such as IgE levels, asthma history and pulmonary factors. Wherease some previous studies indicated TLRs and their polymorphisms might have some role in asthma incidence and features, our data demonstrated that TLR2 Arg753Gln and TLR4 Asp299Gly gene variants were not risk factors for asthma or its features in Iranian patients. Genetic complexity, ethnicity, influence of other genes or polymorphisms may overcome these polymorphisms in our asthmatics.

  11. Covalent attachment of cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) to poly(etheretherketone) surface by tailored silanization layers technique

    Highlights: • The carbonyl groups on PEEK surface were effectively reduced to hydroxyl groups using sodium borohydride. • Silanization layers technique was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on hydroxylation-pretreated PEEK sheet surface by covalent chemical attachment. • XPS, surface profiler and water contact angle measurements proved the presence of GRGD on PEEK surface. • Osteoblast-like cells (MC3T3-E1) attachment and proliferation were improved effectively on GRGD-modified PEEK surface. - Abstract: Poly(etheretherketone) (PEEK) is a rigid semicrystalline polymer that combines excellent mechanical properties, broad chemical resistance and bone-like stiffness and is widely used in biomedical fields. However, PEEK is naturally bioinert, leading to limited biomedical applications, especially when a direct bone-implant osteointegration is desired. In this study, a three-step reaction procedure was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on the surface of PEEK sheet by covalent chemical attachment to favor cell adhesion and proliferation. First, hydroxylation-pretreated PEEK surfaces were silanized with 7-Oct-1-enyltrichlorosilane (OETS) in dry cyclohexane, resulting in a silanization layer with terminal ethenyl. Second, the terminal ethylenic double bonds of the silanization layer on PEEK surface were converted to carboxyl groups through acidic potassium manganate oxidation. Finally, GRGD was covalently attached by carbodiimide mediated condensation between the carboxyl on PEEK surface and amine presents in GRGD. X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, surface profiler and water contact angle measurements were applied to characterize the modified surfaces. The effect of cells attachment and proliferation on each specimen was investigated. Pre-osteoblast cells (MC3T3-E1) attachment, spreading and proliferation

  12. Covalent attachment of cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) to poly(etheretherketone) surface by tailored silanization layers technique

    Zheng, Yanyan [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Xiong, Chengdong [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China); Li, Xiaoyu [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Zhang, Lifang, E-mail: zhanglfcioc@163.com [Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041 (China)

    2014-11-30

    Highlights: • The carbonyl groups on PEEK surface were effectively reduced to hydroxyl groups using sodium borohydride. • Silanization layers technique was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on hydroxylation-pretreated PEEK sheet surface by covalent chemical attachment. • XPS, surface profiler and water contact angle measurements proved the presence of GRGD on PEEK surface. • Osteoblast-like cells (MC3T3-E1) attachment and proliferation were improved effectively on GRGD-modified PEEK surface. - Abstract: Poly(etheretherketone) (PEEK) is a rigid semicrystalline polymer that combines excellent mechanical properties, broad chemical resistance and bone-like stiffness and is widely used in biomedical fields. However, PEEK is naturally bioinert, leading to limited biomedical applications, especially when a direct bone-implant osteointegration is desired. In this study, a three-step reaction procedure was employed to immobilize the cell-adhesive peptide Gly-Arg-Gly-Asp (GRGD) on the surface of PEEK sheet by covalent chemical attachment to favor cell adhesion and proliferation. First, hydroxylation-pretreated PEEK surfaces were silanized with 7-Oct-1-enyltrichlorosilane (OETS) in dry cyclohexane, resulting in a silanization layer with terminal ethenyl. Second, the terminal ethylenic double bonds of the silanization layer on PEEK surface were converted to carboxyl groups through acidic potassium manganate oxidation. Finally, GRGD was covalently attached by carbodiimide mediated condensation between the carboxyl on PEEK surface and amine presents in GRGD. X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, surface profiler and water contact angle measurements were applied to characterize the modified surfaces. The effect of cells attachment and proliferation on each specimen was investigated. Pre-osteoblast cells (MC3T3-E1) attachment, spreading and proliferation

  13. The conserved residue Arg46 in the N-terminal heptad repeat domain of HIV-1 gp41 is critical for viral fusion and entry.

    Xiaoyi Wang

    Full Text Available During the process of HIV-1 fusion with the target cell, the N-terminal heptad repeat (NHR of gp41 interacts with the C-terminal heptad repeat (CHR to form fusogenic six-helix bundle (6-HB core. We previously identified a crucial residue for 6-HB formation and virus entry--Lys63 (K63 in the C-terminal region of NHR (aa 54-70, which forms a hydrophobic cavity. It can form an important salt bridge with Asp121 (D121 in gp41 CHR. Here, we found another important conserved residue for virus fusion and entry, Arg46 (R46, in the N-terminal region of NHR (aa 35-53, which forms a hydrogen bond with a polar residue, Asn43 (N43, in NHR, as a part of the hydrogen-bond network. R46 can also form a salt bridge with a negatively charged residue, Glu137 (E137, in gp41 CHR. Substitution of R46 with the hydrophobic residue Ala (R46A or the negatively charged residue Glu (R46E resulted in disruption of the hydrogen bond network, breakage of the salt bridge and reduction of 6-HB's stability, leading to impairment of viral fusion and decreased inhibition of N36, an NHR peptide. Similarly, CHR peptide C34 with substitution of E137 for Ala (E137A or Arg (E137R also exhibited reduced inhibitory activity against HIV-1 infection and HIV-1-mediated cell-to-cell fusion. These results suggest that the positively charged residue R46 and its hydrogen bond network, together with the salt bridge between R46 and E137, are important for viral fusion and entry and may therefore serve as a target for designing novel HIV fusion/entry inhibitors.

  14. Himatanthus lancifolius (Müll. Arg.) Woodson, Apocynaceae: estudo farmacobotânico de uma planta medicinal da Farmacopeia brasileira 1ª edição Himatanthus lancifolius (Müll. Arg.) Woodson, Apocynaceae: morpho-anatomical study of a medicinal plant described in the Brazilian Pharmacopoeia 1st edition

    Leopoldo C. Baratto; Sandra V. A. Hohlemwerger; Maria Lenise S. Guedes; Márcia R. Duarte; Cid A. M. Santos

    2010-01-01

    Himatanthus lancifolius (Müll. Arg.) Woodson é um arbusto nativo do Brasil, latescente, popularmente conhecido como agoniada e utilizado principalmente para distúrbios menstruais. O presente trabalho teve como objetivo caracterizar morfoanatomicamente a folha, o caule e a casca caulinar dessa planta medicinal, a fim de contribuir para o controle de qualidade e a autenticidade dessa espécie. O material vegetal foi fixado e submetido às microtécnicas usuais. A folha é simples, glabra e obovado-...

  15. Contribuição ao estudo anatômico do caule de Himatanthus sucuuba (Spruce ex Müll. Arg.) Woodson, Apocynaceae Contribution to the anatomical study of the stem of Himatanthus sucuuba (Spruce ex Müll. Arg.) Woodson, Apocynaceae

    C.R.R. Larrosa; M. R. Duarte

    2005-01-01

    Himatanthus sucuuba (Spruce ex Müll. Arg.) Woodson, Apocynaceae, denominada comumente de sucuuba ou janaguba, é uma espécie arbórea, latescente, da região amazônica, cujo tronco é ereto e a casca rugosa. As atividades antiinflamatória e analgésica e a baixa toxicidade foram estabelecidas em diferentes pesquisas. Com o objetivo de estabelecer caracteres anatômicos aplicáveis à identificação da planta e da droga vegetal, analisou-se o caule em estrutura secundária, que consiste da casca e da re...

  16. La contabilidad pública en las Cortes de Cádiz: la "Memoria sobre la Cuenta y Razón de España" (1811) de José Canga Argüelles. (Public Sector Accounting at the Parliament in Cadiz: the "Report on Spain's Accounting" (1811) by José Canga Argüelles).

    López Castellano, Fernando

    2009-01-01

    En este trabajo se narra un intento relevante, siquiera desde el punto de vista doctrinal, de la historia institucional de la Hacienda: el ambicioso proyecto de reforma de la Contabilidad Pública llevado a cabo por las Cortes Generales y Extraordinarias instaladas en Cádiz durante la invasión napoleónica. En este capítulo poco conocido de la historia institucional de la Hacienda, destacan las ideas sobre la Cuenta y Razón expuestas por el Secretario del Despacho de Hacienda, José Canga Argüel...

  17. Behavior of antibiotics and antibiotic resistance genes in eco-agricultural system: A case study.

    Cheng, Weixiao; Li, Jianan; Wu, Ying; Xu, Like; Su, Chao; Qian, Yanyun; Zhu, Yong-Guan; Chen, Hong

    2016-03-01

    This study aims to determine abundance and persistence of antibiotics and antibiotic resistance genes (ARGs) in eco-agricultural system (EAS), which starts from swine feces to anaerobic digestion products, then application of anaerobic digestion solid residue (ADSR) and anaerobic digestion liquid residue (ADLR) to the soil to grow ryegrass, one of swine feed. Oxytetracycline had the highest concentration in manure reaching up to 138.7 mg/kg. Most of antibiotics could be effectively eliminated by anaerobic digestion and removal rates ranged from 11% to 86%. ARGs abundance fluctuated within EAS. TetQ had the highest relative abundance and the relative abundance of tetG had the least variation within the system, which indicates that tetG is persistent in the agricultural environment and requires more attention. Compared to the relative abundance in manure, tetC and tetM increased in biogas residue while three ribosomal protection proteins genes (tetO, tetQ, tetW) decreased (p0.05). Most ARGs in downstream components (soils and fishpond) of EAS showed significantly higher relative abundance than the control agricultural system (p<0.05), except for tetG and sulI. PMID:26546700

  18. Prevalence of tetracycline resistance genes among multi-drug resistant bacteria from selected water distribution systems in southwestern Nigeria

    Adesoji, Ayodele. T.; Ogunjobi, Adeniyi. A.; Olatoye, Isaac. O.; Douglas, Douglas. R.

    2015-01-01

    Background Antibiotic resistance genes [ARGs] in aquatic systems have drawn increasing attention they could be transferred horizontally to pathogenic bacteria. Water treatment plants (WTPs) are intended to provide quality and widely available water to the local populace they serve. However, WTPs in developing countries may not be dependable for clean water and they could serve as points of dissemination for antibiotic resistant bacteria. Only a few studies have investigated the occurrence of ...

  19. Development of pre-implantation porcine embryos cultured within a three-dimensional alginate hydrogel system either conjugated with Arg-Gly-Asp (RGD) peptide or supplemented with secreted phosphoprotein 1 (SPP1)

    Many uterine specific factors have been shown to be increased within the uterine milieu as the porcine embryo initiates elongation. Secreted phosphoprotein 1 (SPP1) is increased during this time and contains an Arg-Gly-Asp (RGD) peptide sequence that has been shown to bind to cell surface integrins ...

  20. Mutational analysis of the human MAOA gene

    Tivol, E.A.; Shalish, C.; Schuback, D.E.; Breakefield, X.O. [Harvard Medical School, Boston, MA (United States); Hsu, Yun-Pung [VA Medical Center, West Roxbury, MA (United States)

    1996-02-16

    The monoamine oxidases (MAO-A and MAO-B) are the enzymes primarily responsible for the degradation of amine neurotransmitters, such as dopamine, norepinephrine, and serotonin. Wide variations in activity of these isozymes have been reported in control humans. The MAOA and MAOB genes are located next to each other in the p11.3-11.4 region of the human X chromosome. Our recent documentation of an MAO-A-deficiency state, apparently associated with impulsive aggressive behavior in males, has focused attention on genetic variations in the MAOA gene. In the present study, variations in the coding sequence of the MAOA gene were evaluated by RT-PCR, SSCP, and sequencing of mRNA or genomic DNA in 40 control males with >100-fold variations in MAOA activity, as measured in cultured skin fibroblasts. Remarkable conservation of the coding sequence was found, with only 5 polymorphisms observed. All but one of these were in the third codon position and thus did not alter the deduced amino acid sequence. The one amino acid alteration observed, lys{r_arrow}arg, was neutral and should not affect the structure of the protein. This study demonstrates high conservation of coding sequence in the human MAOA gene in control males, and provides primer sets which can be used to search genomic DNA for mutations in this gene in males with neuropsychiatric conditions. 47 refs., 1 fig., 2 tabs.

  1. Splicing mutation of a gene within the Duchenne muscular dystrophy family.

    Zhu, Y B; Gan, J H; Luo, J W; Zheng, X Y; Wei, S C; Hu, D

    2016-01-01

    The aim of this study was to identify the mutation site and phenotype of the Duchenne muscular dystrophy (DMD) gene in a DMD family. The DMD gene is by far the largest known gene in humans. Up to 34% of the point mutations reported to date affect splice sites of the DMD gene. However, no hotspot mutation has been reported. Capture sequencing of second-generation exons was used to investigate the DMD gene in a proband. Sanger sequencing was performed for mutation scanning in eight family members. Scale-invariant feature transform and PolyPhen were applied to predict the functional impact of protein mutations. A hemizygous splicing mutation IVS44ds +1G-A (c.6438 +1G>A) that induces abnormal splicing variants during late transcription and produces abnormal proteins was located in intron 44. Four missense mutations (p.Arg2937Gln, p.Asp882Gly, p.Lys2366Gln, and p.Arg1745His) that are known multiple-polymorphic sites were found in the coding region of the DMD gene. A heterozygous c.6438+1G>A mutation was detected on the X chromosome of the proband's mother and maternal grandmother. PMID:27421007

  2. Shift in antibiotic resistance gene profiles associated with nanosilver during wastewater treatment.

    Ma, Yanjun; Metch, Jacob W; Yang, Ying; Pruden, Amy; Zhang, Tong

    2016-03-01

    This study investigated the response of antibiotic resistance genes (ARGs) to nanosilver (Ag) in lab-scale nitrifying sequencing batch reactors (SBRs), compared to Ag(+)-dosed and undosed controls. Quantitative polymerase chain reaction (q-PCR) targeting sul1, tet(O), ermB and the class I integron gene intI1 and corresponding RNA expression did not indicate measureable effects of nanoAg or Ag(+) on abundance or expression of these genes. However, high-throughput sequencing based metagenomic analysis provided a much broader profile of gene responses and revealed a greater abundance of aminoglycoside resistance genes (mainly strA) in reactors dosed with nanoAg. In contrast, bacitracin and macrolide-lincosamide-streptogramin (MLS) resistance genes were more abundant in the SBRs dosed with Ag(+). The distinct ARG profiles associated with nanoAg and Ag(+) were correlated with the taxonomic composition of the microbial communities. This study indicates that nanoAg may interact with bacteria differently from Ag(+) during biological wastewater treatment. Therefore, it cannot necessarily be assumed that nanosilver behaves identically as Ag(+) when conducting a risk assessment for release into the environment. PMID:26850160

  3. Effect of biochar amendment on the control of soil sulfonamides, antibiotic-resistant bacteria, and gene enrichment in lettuce tissues.

    Ye, Mao; Sun, Mingming; Feng, Yanfang; Wan, Jinzhong; Xie, Shanni; Tian, Da; Zhao, Yu; Wu, Jun; Hu, Feng; Li, Huixin; Jiang, Xin

    2016-05-15

    Considering the potential threat of vegetables growing in antibiotic-polluted soil with high abundance of antibiotic-resistant genes (ARGs) against human health through the food chain, it is thus urgent to develop novel control technology to ensure vegetable safety. In the present work, pot experiments were conducted in lettuce cultivation to assess the impedance effect of biochar amendment on soil sulfonamides (SAs), antibiotic-resistant bacteria (ARB), and ARG enrichment in lettuce tissues. After 100 days of cultivation, lettuce cultivation with biochar amendment exhibited the greatest soil SA dissipation as well as the significant improvement of lettuce growth indices, with residual soil SAs mainly existing as the tightly bound fraction. Moreover, the SA contents in roots and new/old leaves were reduced by one to two orders of magnitude compared to those without biochar amendment. In addition, isolate counts for SA-resistant bacterial endophytes in old leaves and sul gene abundances in roots and old leaves also decreased significantly after biochar application. However, neither SA resistant bacteria nor sul genes were detected in new leaves. It was the first study to demonstrate that biochar amendment can be a practical strategy to protect lettuce safety growing in SA-polluted soil with rich ARB and ARGs. PMID:26896719

  4. Genetic polymorphisms of MDM2 and TP53 genes are associated with risk of nasopharyngeal carcinoma in a Chinese population

    The tumor suppressor TP53 and its negative regulator MDM2 play crucial roles in carcinogenesis. Previous case-control studies also revealed TP53 72Arg>Pro and MDM2 309T>G polymorphisms contribute to the risk of common cancers. However, the relationship between these two functional polymorphisms and nasopharyngeal carcinoma (NPC) susceptibility has not been explored. In this study, we performed a case-control study between 522 NPC patients and 722 healthy controls in a Chinese population by using PCR-RFLP. We found an increased NPC risk associated with the MDM2 GG (odds ratio [OR] = 2.83, 95% confidence interval [CI] = 2.08-3.96) and TG (OR = 1.49, 95% CI = 1.16-2.06) genotypes. An increased risk was also associated with the TP53 Pro/Pro genotype (OR = 2.22, 95% CI = 1.58-3.10) compared to the Arg/Arg genotype. The gene-gene interaction of MDM2 and TP53 polymorphisms increased adult NPC risk in a more than multiplicative manner (OR for the presence of both MDM2 GG and TP53 Pro/Pro genotypes = 7.75, 95% CI = 3.53-17.58). The findings suggest that polymorphisms of MDM2 and TP53 genes may be genetic modifier for developing NPC

  5. Probing the interaction of human serum albumin with vitamin B2 (riboflavin) and L-Arginine (L-Arg) using multi-spectroscopic, molecular modeling and zeta potential techniques

    Memarpoor-Yazdi, Mina [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of); Mahaki, Hanie, E-mail: hanieh.mahaki@gmail.com [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of)

    2013-04-15

    This study was designed to examine the interaction of Riboflavin (RB) and L-Arginine (L-Arg) with human serum albumin (HSA) using different spectroscopic, zeta potential and molecular modeling techniques under imitated physiological conditions. The resonance light scattering (RLS) method determined the critical aggregation concentration of RB on HSA in the presence and absence of L-Arg which confirmed the zeta potential results. The binding constants (K{sub a}) of HSA–RB were 2.5×10{sup 4} and 9.7×10{sup 3} M{sup −1}, respectively in binary and ternary system at the excitation wavelength of 280 nm, also were 7.5×10{sup 3} and 7.3×10{sup 3}, respectively in binary and ternary system at the excitation wavelength of 295 nm. Fluorescence spectroscopy demonstrated that in the presence of L-Arg, the binding constant of HSA–RB was increased. Static quenching was confirmed to results in the fluorescence quenching and FRET. The binding distances between HSA and RB in two- and three-component systems were estimated by the Forster theory which revealed that nonradiative energy transfer from HSA to RB occurred with a high probability. The effect of RB on the conformation of HSA was analyzed using synchronous fluorescence spectroscopy and circular dichroism (CD) in both systems. Docking studies demonstrated a reduction in the binding affinity between RB and HSA in the presence of L-Arg. -- Highlights: ► We studied the interaction of riboflavin with HSA in presence and absence of L-Arg. ► Molecular modeling and zeta-potential used to describe competitive interaction. ► We compared the binding mechanism of riboflavin (RB) to HSA in both systems. ► We determined critical aggregation concentration of RB on HSA in both systems. ► The binding site of RB on HSA in both systems has been determined.

  6. Clustering of genes for L-fucose dissimilation by Escherichia coli.

    Chakrabarti, T.; Y.M. Chen; Lin, E C

    1984-01-01

    Aerobic and anaerobic L-fucose utilization by Escherichia coli involves an inducible trunk pathway mediated by a permease, an isomerase, a kinase, and an aldolase. Tn5 insertion mutants of a parental strain expressing this pathway constitutively were used to map the positions of the structural genes by transduction. Results from this and previous studies show that all of the structural genes of the L-fucose trunk pathway map between eno and argA at minute 60.2 of the chromosome.

  7. Screening of bacterial biocontrols against sapstain fungus (Lasiodiplodia theobromae Pat.) of rubberwood (Hevea brasiliensis Muell.Arg.).

    Sajitha, K L; Maria Florence, E J; Dev, Suma Arun

    2014-09-01

    Diverse bacterial biocontrol agents from various sources of aerobic composts against the sapstain fungus Lasiodiplodia theobromae in rubberwood (Hevea brasiliensis) were isolated, screened and identified by various morphological, biochemical and molecular techniques. The inhibitory effect of seventeen bacterial isolates was examined and seven exhibited inhibition towards the sapstain fungus. Among the seven antagonists, six were conclusively identified as Bacillus subtilis and one as Paenibacillus polymyxa using 16S rRNA-encoding gene sequencing. This is the first report on the occurrence of P. polymyxa, a potent biofertilizer and antagonist in vermicompost. HiCrome Bacillus agar was identified as an effective medium for differentiation of B. subtilis from other Bacillus species. The present work demonstrates the efficacy of the antagonistic property of B. subtilis strains against rubberwood sapstain fungus. Culture-based antagonistic inhibition displayed by B. subtilis can be extended to cater to the biocontrol requirements of wood-based industries against the stain fungus. The study showed the utility of an integrated approach, employing morphological, biochemical and molecular tools for conclusive identification of several bacterial isolates present in aerobic composts from diverse sources. PMID:25049165

  8. Phenotypic expressions of a Gly154Arg mutation in type II collagen in two unrelated patients with spondyloepimetaphyseal dysplasia (SEMD)

    Kaitila, I.; Marttinen, E. [Helsinki Univ. Hospital (Finland); Koerkkoe, J.; Ala-Kokko, L. [Thomas Jefferson Univ., Philadelphia, PA (United States)

    1996-05-03

    Type II collagenopathies consist of chondrodysplasia ranging from lethal to mild in severity. A large number of mutations has been found in the COL2A1 gene. Glycine substitutions have been the most common types of mutation. Genotype-phenotype correlations in type II collagenopathies have not been established, partly because of insufficient clinical and radiographic description of the patients. We found a glycine-to-arginine substitution at position 154 in type II collagen in two unrelated isolated propositi with spondyloepimetaphyseal dysplasia and provide a comparative clinical and radiographic analysis from birth to young adulthood for this condition. The clinical phenotype was disproportionate short stature with varus/valgus deformities of the lower limbs requiring corrective osteotomies, and lumbar lordosis. The skeletal radiographs showed an evolution from short tubular bones, delayed epiphyseal development, and mild vertebral involvement to severe metaphyseal dysplasia with dappling irregularities, and hip {open_quotes}dysplasia.{close_quotes} The metaphyseal abnormalities disappeared by adulthood. 27 refs., 11 figs., 1 tab.

  9. O uso do laser de argônio no tratamento da triquíase Argon laser in the treatment of trichiasis

    Nilson Lopes da Fonseca Junior

    2004-04-01

    Full Text Available OBJETIVO: Avaliar a eficácia da técnica de termoablação dos folículos pilosos dos cílios em triquíase com laser de argônio e observar a preferência dos pacientes submetidos a este tratamento, pela anestesia tópica com colírio anestésico ou pela anestesia local injetável. MÉTODOS: Estudo prospectivo de 55 pálpebras de 39 pacientes com triquíase, tratados com fotocoagulação dos folículos pilosos com "Argon green laser" (Alcon® - EUA. Neste estudo avaliou-se a idade e o sexo dos pacientes, o número de sessões realizadas, a evolução após as aplicações e a preferência pelo tipo de anestesia. Os parâmetros utilizados foram: (1 Mira - 150 µm; (2 Potência- 750 mW; (3 Tempo de exposição- 0,2 s. Cada cílio recebeu, no máximo, 9 disparos por sessão. RESULTADOS: Dentre os 39 participantes do estudo, 58,9% eram do sexo feminino e 41% do sexo masculino. A idade média foi de 71 anos. Houve cura em 69% (38 pálpebras, sendo 29% (16 pálpebras com apenas uma sessão de laser. Houve preferência estatisticamente significativa pelo procedimento realizado sob anestesia local injetável. CONCLUSÃO: Os resultados sugerem que a anestesia local no tratamento da triquíase com laser de argônio é a preferência da maioria dos pacientes e que a termoablação dos folículos pilosos com laser de argônio é tratamento efetivo para a triquíase.PURPOSE: To evaluate the effectiveness of thermoablation of eyelashes in trichiasis with Argon laser and to observe the patients' choice of topic or local anesthesia. METHODS: Prospective study of 55 eyelids of 39 patients with trichiasis treated with photocoagulation of the eyelash follicle with Argon green laser (Alcon® - USA. Age, sex, number of applications in each session, evolution after the application and preference for the anesthesic method were evaluated. The laser settings were: 150 µm aim 750 mW potency; 0.2 seconds exhibition. Each eyelash received no more than 9 applications per

  10. Profile of milk fatty acids from moxotó goats fed with different levels of manicoba (Manihot Glaziovii Muel Arg. silage

    Ítala Viviane Ubaldo Mesquita

    2008-12-01

    Full Text Available The objective of this work was to evaluate the effects of the manicoba silage use (Manihot glaziovii Muel Arg. in different roughage:concentrate ratios (30:70; 40:60; 50:50 and 60:40 % on the fatty acids profile of the milk from Moxotó goats. Eight multiparous goats with approximately 60 post-birth days and weigh 44 kg on average were used in a Double Latin Square with four treatments, four periods and four animals. Each period lasted 15 days with 10 days of adaptation to experimental diets and 5 days of milk collection. The results of the fatty acids analyses were submitted to the analysis of variance (ANOVA and regression. An increasing linear effect for the miristic acid (C14:0 and decreasing for the linoleic acid (C18:3 was observed in function of the silage levels in diet. The milk fat presented high contents of desirable fatty acids (C18:0 + unsaturated acids, considered as nutritionally important due to their benefits to the human health. It could be concluded that the manicoba silage could be included in the diet with the participation of up to 60%, presenting a nutrient supply of high nutritional value.O objetivo deste trabalho foi avaliar os efeitos da utilização de silagem de maniçoba (Manihot glaziovii Muel Arg. em diferentes relações volumoso:concentrado (30:70; 40:60; 50:50 e 60:40 % no perfil de ácidos graxos do leite de cabras da raça Moxotó. Foram utilizadas oito cabras multíparas com aproximadamente 60 dias pós-parto, pesando em média 44 kg, em um Quadrado Latino duplo 4 x 4, com quatro tratamentos, quatro períodos e quatro animais. Cada período teve duração de 15 dias, com 10 de adaptação às dietas experimentais e cinco dias de colheita de leite. Foi verificado efeito linear crescente do ácido mirístico (C14:0 e decrescente para o ácido linolênico (C18:3, em função dos níveis de silagem da dieta. A gordura do leite apresentou elevado teor de ácidos graxos desejáveis (C18:0 + insaturados

  11. Influences of XDH genotype by gene-gene interactions with SUCLA2 for thiopurine-induced leukopenia in Korean patients with Crohn's disease.

    Park, Soo-Kyung; Hong, Myunghee; Ye, Byong Duk; Kim, Kyung-Jo; Park, Sang Hyoung; Yang, Dong-Hoon; Hwang, Sung-Wook; Kwak, Min Seob; Lee, Ho-Su; Song, Kyuyoung; Yang, Suk-Kyun

    2016-06-01

    Background The impact of genetic variation in the thiopurine S-methyltransferase (TPMT) gene on thiopurine-induced leukopenia has been well demonstrated. Although xanthine dehydrogenase (XDH) is the second major contributor to azathioprine breakdown, polymorphisms in XDH have rarely been studied in IBD patients. We aim to access association between XDH variants and thiopurine-induced leukopenia by gene-gene interaction in a Crohn's disease (CD) population. Study A total of 964 CD patients treated with thiopurines were recruited from a tertiary referral center. The association between four XDH variants (p.Gly172Arg, p.Asn1109Thr, p.Arg149Cys, and p.Thr910Lys) and thiopurine-induced leukopenia was analyzed in cases with early leukopenia (n = 66), late leukopenia (n = 264), and in controls without leukopenia (n = 632). Three non-synonymous SNPs, which we previously reported association with thiopurine-induced leukopenia, NUDT15 (p.Arg139Cys), SUCLA2 (p.Ser199Thr), and TPMT *3C were selected for epistasis analysis with the XDH variants. Results There was no significant association for two variants of XDH and thiopurine-induced leukopenia. In the epistasis analysis, only XDH (p.Asn1109Thr) * SUCLA2 (p.Ser199Thr) showed a statistically significant association with early leukopenia [odds ratio (OR) = 0.16; p = 0.03]. After genotype stratification, a positive association on the background of SUCLA2 wild-type (199Ser) between the XDH (p.Asn1109Thr) and early leukopenia (OR = 4.39; p = 0.01) was detected. Conclusion Genes associated with thiopurine-induced leukopenia can act in a complex interactive manner. Further studies are warranted to explore the mechanisms underlying the effects of the combination of XDH (p.Asn1109Thr) and SUCLA2 (199Ser) on thiopurine-induced leukopenia. PMID:26863601

  12. Clinical study of DMD gene point mutation causing Becker muscular dystrophy

    Ji-qing CAO

    2015-07-01

    Full Text Available Background  DMD gene point mutation, mainly nonsense mutation, always cause the most severe Duchenne muscular dystrophy (DMD. However, we also observed some cases of Becker muscular dystrophy (BMD carrying DMD point mutation. This paper aims to explore the mechanism of DMD point mutation causing BMD, in order to enhance the understanding of mutation types of BMD.  Methods  Sequence analysis was performed in 11 cases of BMD confirmed by typical clinical manifestations and muscle biopsy. The exon of DMD gene was detected non-deletion or duplication by multiplex ligation-dependent probe amplification (MLPA.  Results  Eleven patients carried 10 mutation types without mutational hotspot. Six patients carried nonsense mutations [c.5002G>T, p.(Glu1668X; c.1615C > T, p.(Arg539X; c.7105G > T, p.(Glu2369X; c.5287C > T, p.(Arg1763X; c.9284T > G, p.(Leu3095X]. One patient carried missense mutation [c.5234G > A, p.(Arg1745His]. Two patients carried frameshift mutations (c.10231dupT, c.10491delC. Two patients carried splicing site mutations (c.4518 + 3A > T, c.649 + 2T > C.  Conclusions  DMD gene point mutation may result in BMD with mild clinical symptoms. When clinical manifestations suggest the possibility of BMD and MLPA reveals non?deletion or duplication mutation of DMD gene, BMD should be considered. Study on the mechanism of DMD point mutation causing BMD is very important for gene therapy of DMD. DOI: 10.3969/j.issn.1672-6731.2015.06.005

  13. Single nucleotide polymorphisms in caprine calpastatin gene.

    Sharma, R; Maitra, A; Pandey, A K; Singh, L V; Mishra, B P

    2013-04-01

    The calpains and calpastatin (CAST) make up a major cytosolic proteolytic system, the calpain-calpastatin system, found in mammalian tissues. The relative levels of the components of the calpain-calpastatin system determine the extent of meat tenderization during postmortem storage. Calpastatin (CAST) is a protein inhibitor of the ubiquitous calcium-dependent proteases-micro-calpain and m-calpain. Polymorphisms in the bovine, ovine and pig CAST gene have been associated with meat tenderness but little is known about how caprine CAST gene may affect goat meat quality traits. In this study we selected different parts of the CAST gene: 1) that have been previously reported to be polymorphic, intron 5 and 12 and 3'UTR; 2) first time explored (exon 3, 7 and 8 and part of intron 7 and 8) to investigate polymorphic status of caprine CAST gene. Using comparative sequencing ten novel SN Ps located in exon 3 and intron 5, 7 and 8 were identified. Previously reported SNPs in intron 5, 3'UTR and intron 12 were absent. Sequence analysis revealed a non synonymous amino acid variation in exon 3, which would result in Lys/Arg substitution in the corresponding protein sequence. Considerable variation was detected in intronic regions. Twenty-four InDel were also recognized in intronic regions (15) and 3'UTR (9). All the sequences shared high homology with published bovine and ovine sequences. Three PCR-RFLP loci have been established for further analyzing genetic polymorphism in indigenous goats. PMID:23866627

  14. Metagenomic analysis reveals that bacteriophages are reservoirs of antibiotic resistance genes.

    Subirats, Jéssica; Sànchez-Melsió, Alexandre; Borrego, Carles M; Balcázar, José Luis; Simonet, Pascal

    2016-08-01

    A metagenomics approach was applied to explore the presence of antibiotic resistance genes (ARGs) in bacteriophages from hospital wastewater. Metagenomic analysis showed that most phage sequences affiliated to the order Caudovirales, comprising the tailed phage families Podoviridae, Siphoviridae and Myoviridae. Moreover, the relative abundance of ARGs in the phage DNA fraction (0.26%) was higher than in the bacterial DNA fraction (0.18%). These differences were particularly evident for genes encoding ATP-binding cassette (ABC) and resistance-nodulation-cell division (RND) proteins, phosphotransferases, β-lactamases and plasmid-mediated quinolone resistance. Analysis of assembled contigs also revealed that blaOXA-10, blaOXA-58 and blaOXA-24 genes belonging to class D β-lactamases as well as a novel blaTEM (98.9% sequence similarity to the blaTEM-1 gene) belonging to class A β-lactamases were detected in a higher proportion in phage DNA. Although preliminary, these findings corroborate the role of bacteriophages as reservoirs of resistance genes and thus highlight the necessity to include them in future studies on the emergence and spread of antibiotic resistance in the environment. PMID:27312355

  15. Allelic variants of XRCC1 and XRCC3 repair genes and susceptibility of oral cancer in Brazilian patients

    Dos Reis, Mariana Bisarro; Losi-Guembarovski, Roberta; de Souza Fonseca Ribeiro, Enilze Maria; Cavalli, Iglenir João; Morita, Maria Celeste; Ramos, Gyl Henrique Albrecht; de Oliveira, Benedito Valdecir; Mizuno, Lauro Toyoshi; Rogatto, Silvia Regina; de Syllos Cólus, Ilce Mara

    2013-01-01

    genes have been found to be associated with oral cancer. The aim of this study was to investigate the relationship between the presence of allelic variants Arg194Trp (rs:1799782) and Arg399Gln (rs: 25487) of XRCC1 gene and Thr241Met (rs: 861539) of XRCC3 gene and susceptibility to oral cancer. We also...... variants of the XRCC1 gene within codon 194 (OR 0.82, 95% CI: 0.44-1.51) and codon 399 (OR 0.94, 95% CI: 0.59-1.50) and within the XRCC3 gene (OR 0.72; 95% CI: 0.45-1.16) were not associated with an increased risk of oral cancer. A combinational analysis of SNPs in both genes indicated no association. The......BACKGROUND: The capacity for DNA repair is essential in maintaining cellular functions and homeostasis; however, this capacity can be altered based on DNA sequence variations in DNA repair genes, which may contribute to the onset of cancer. Many single-nucleotide polymorphisms (SNPs) in repair...

  16. Use of Arbuscular Mycorrhiza and Organic Amendments to Enhance Growth of Macaranga peltata (Roxb.) Müll. Arg. in Iron Ore Mine Wastelands.

    Rodrigues, Cassie R; Rodrigues, Bernard F

    2015-01-01

    Macaranga peltata (Roxb.) Mull. Arg. is a disturbance tolerant plant species with potential in mine wasteland reclamation. Our study aims at studying the phyto-extraction potential of M. peltata and determining plant-soil interaction factors effecting plant growth in iron ore mine spoils. Plants were grown in pure mine spoil and spoil amended with Farm Yard Manure (FYM) and Vermicompost (VC) along with arbuscular mycorrhizal (AM) species Rhizophagus irregularis. Pure and amended mine spoils were evaluated for nutrient status. Plant growth parameters and foliar nutrient contents were determined at the end of one year. FYM amendment in spoil significantly increased plant biomass compared to pure mine spoil and VC amended spoil. Foliar Fe accumulation was recorded highest (594.67 μg/g) in pure spoil with no mortality but considerably affecting plant growth, thus proving to exhibit phyto-extraction potential. FYM and VC amendments reduced AM colonization (30.4% and 37% resp.) and plants showed a negative mycorrhizal dependency (-30.35 and -39.83 resp.). Soil pH and P levels and, foliar Fe accumulation are major factors determining plant growth in spoil. FYM amendment was found to be superior to VC as a spoil amendment for hastening plant growth and establishment in iron ore mine spoil. PMID:25495939

  17. A triad of lys12, lys41, arg78 spatial domain, a novel identified heparin binding site on tat protein, facilitates tat-driven cell adhesion.

    Jing Ai

    Full Text Available Tat protein, released by HIV-infected cells, has a battery of important biological effects leading to distinct AIDS-associated pathologies. Cell surface heparan sulfate protoglycans (HSPGs have been accepted as endogenous Tat receptors, and the Tat basic domain has been identified as the heparin binding site. However, findings that deletion or substitution of the basic domain inhibits but does not completely eliminate Tat-heparin interactions suggest that the basic domain is not the sole Tat heparin binding site. In the current study, an approach integrating computational modeling, mutagenesis, biophysical and cell-based assays was used to elucidate a novel, high affinity heparin-binding site: a Lys12, Lys41, Arg78 (KKR spatial domain. This domain was also found to facilitate Tat-driven β1 integrin activation, producing subsequent SLK cell adhesion in an HSPG-dependent manner, but was not involved in Tat internalization. The identification of this new heparin binding site may foster further insight into the nature of Tat-heparin interactions and subsequent biological functions, facilitating the rational design of new therapeutics against Tat-mediated pathological events.

  18. Production of diketopiperazine derivative cyclo (l-leu-l-arg) by streptomyces sp. tn262 after exposure to heat-killed fungus fusarium sp

    In a screening program for new active secondary metabolites producers, a strain of Streptomyces called TN262 was isolated from Tunisian soil and selected for its ability to produce eleven active compounds in pure culture conditions. In this work, the effect of different concentrations of heat-killed fungus Fusarium sp. on the production of active compounds by TN262 strain was studied. The ethyl acetate extract from the culture of Streptomyces sp. TN262 combined with heat-killed Fusarium sp. at 50 micro g/ml inhibited the growth of the three used indicator microorganisms. In fact, an increase of 36%, 21% and 20% in inhibitory activity was obtained against Micrococcus luteus LB 14110, Escherichia coli ATCC 8739 and Fusarium sp. respectively. The HPLC chromatographic profiles of the ethyl acetate extracts from both culture conditions were different and an additional active compound was produced only under induced conditions. This active component was isolated and identified as Cyclo (L-Leu-L-Arg) (1), a diketopiperazine derivative, possessing antibacterial and antifungal activity. Consequently, this study showed that the addition of heat-killed fungus is a useful method for inducing the production of bioactive compounds. (author)

  19. IL-4RaArg55、IgE与API阳性婴幼儿喘息的相关性研究%Correlation of IL-4RaArg55 and IgE Levels with API Positive Infantile Wheezing

    王建荣; 齐英征; 吴英杰

    2015-01-01

    目的 探讨白介素4受体基因Arg55(IL-4RaArg55)、IgE在哮喘预测指数(asthma predicting index,API)阳性患儿中的应用价值.方法 选取同期的年龄在1月至3岁喘息婴幼儿356例,分为API阳性组167例,API阴性组189例,同时设立对照组203例,首先应用PCR聚合酶链反应和DNA测序法对两组基因进行分型,并用ELISA法检测IgE水平,进一步根据年龄、性别进行组内分层,进行IgE水平统计分析.结果 ①IL-4RaArg55位点各基因型频率分布在API(+)组、API(-)组、对照组3组中差异无统计学意义(AA基因:x2 =2.16,P=0.35;x2=0.77,P=0.68;x2 =4.11,P=0.13;A等位基因:x2=0.48,P =0.49;x2=0.71,P=0.40;x2 =2.58,P=0.11).②API(+)、API(-)及对照组间IgE水平分别为93.18±40.79、54.16±22.66、48.82±21.42 U/mL,3组差异有统计学意义(H=377.419,P=0.000),API(+)组远高于API(-)及对照组.③API(+)组内<2岁与≥2岁IgE水平相互比较,差异有统计学意义(t'=9.281,P<0.001);API(-)组内<2岁与≥2岁IgE水平相互比较,差异无统计学意义(t=0.693,P=0.489).④API(+)组及API(-)组内男性与女性IgE水平相互比较,差异无统计学意义[API(+) t=1.598,P=0.112,API(-)t=0.330,P=0.742].结论 ①IL-4RaArg55多态性与API结果无相关性;②血清IgE水平与API阳性有相关性,对≥2岁的婴幼儿喘息的预后判断有一定的意义.

  20. Various pAQU plasmids possibly contribute to disseminate tetracycline resistance gene tet(M among marine bacterial community

    LisaNonaka

    2014-05-01

    Full Text Available Emergence of antibiotic-resistant bacteria in the aquaculture environment is a significant problem for disease control of cultured fish in as well as in human public health. Conjugative mobile genetic elements (MGEs are involved in dissemination of antibiotic resistance genes (ARGs among marine bacteria. In the present study, we first designed a PCR targeting traI gene encoding essential relaxase for conjugation. By this new PCR, we demonstrated that five of 83 strains isolated from a coastal aquaculture site had traI-positive MGEs. While one of the five strains that belonged to Shewanella sp. was shown to have an integrative conjugative element of the SXT/R391 family (ICEVchMex, the MGEs of the other four strains of Vibrio spp. were shown to have the backbone structure similar to that of previously described in pAQU1. The backbone structure shared by the pAQU1-like MGEs in the four strains corresponded to a ~100-kbp highly conserved region required for replication, partition and conjugative transfer, suggesting that these MGEs are plasmids that constitute “pAQU group”. The pAQU group plasmids were shown to be capable of conjugative transfer of tet(M and other ARGs from the Vibrio strains to E. coli. The pAQU group plasmid in one of the examined strains was designated as pAQU2, and its complete nucleotide sequence was determined and compared with that of pAQU1. The results revealed that pAQU2 contained fewer ARGs than pAQU1 did, and most of the ARGs in both of these plasmids were located in the similar region where multiple transposases were found, suggesting that the ARGs were introduced by several events of DNA transposition into an ancestral plasmid followed by drug selection in the aquaculture site. The results of the present study indicate that the “pAQU group” plasmids may play an important role in dissemination of ARGs in the marine environment.

  1. Polymorphism of XRCC1, XRCC3, and XPD genes and risk of chronic myeloid leukemia.

    Bănescu, Claudia; Trifa, Adrian P; Demian, Smaranda; Benedek Lazar, Erzsebeth; Dima, Delia; Duicu, Carmen; Dobreanu, Minodora

    2014-01-01

    The genetic polymorphisms of X-ray repair cross complementing group 1 (XRCC1), X-ray repair cross complementing group 3 (XRCC3), and xeroderma pigmentosum complementation group D (XPD) repair genes may lead to genetic instability and leukemogenesis. The purpose of the study was to evaluate the association between XRCC1 Arg399Gln, Arg280His and Arg194Trp, XRCC3 Thr241Met, and XPD Lys751Gln polymorphisms and the risk of developing CML in Romanian patients. A total of 156 patients diagnosed with CML and 180 healthy controls were included in this study. We found no association between CML and XRCC1 or XRCC3 variant genotypes in any of the investigated cases. A significant difference was observed in the variant genotype frequencies of the XPD Lys751Gln polymorphism between the patients with CML and control group (for variant homozygous genotypes, OR = 2.37; 95% CI = 1.20-4.67; P value = 0.016 and for combined heterozygous and variant homozygous genotypes, OR = 1.72; 95% CI = 1.10-2.69; P value = 0.019). This was also observed when analyzing the variant 751Gln allele (OR = 1.54; 95% CI = 1.13-2.11; P value = 0.008). Our results suggest that the XPD Lys751Gln variant genotype increases the risk of CML. PMID:24955348

  2. Polymorphism of XRCC1, XRCC3, and XPD Genes and Risk of Chronic Myeloid Leukemia

    Claudia Bănescu

    2014-01-01

    Full Text Available The genetic polymorphisms of X-ray repair cross complementing group 1 (XRCC1, X-ray repair cross complementing group 3 (XRCC3, and xeroderma pigmentosum complementation group D (XPD repair genes may lead to genetic instability and leukemogenesis. The purpose of the study was to evaluate the association between XRCC1 Arg399Gln, Arg280His and Arg194Trp, XRCC3 Thr241Met, and XPD Lys751Gln polymorphisms and the risk of developing CML in Romanian patients. A total of 156 patients diagnosed with CML and 180 healthy controls were included in this study. We found no association between CML and XRCC1 or XRCC3 variant genotypes in any of the investigated cases. A significant difference was observed in the variant genotype frequencies of the XPD Lys751Gln polymorphism between the patients with CML and control group (for variant homozygous genotypes, OR=2.37; 95% CI=1.20–4.67; P value = 0.016 and for combined heterozygous and variant homozygous genotypes, OR=1.72; 95% CI=1.10–2.69; P value = 0.019. This was also observed when analyzing the variant 751Gln allele (OR=1.54; 95% CI=1.13–2.11; P value = 0.008. Our results suggest that the XPD Lys751Gln variant genotype increases the risk of CML.

  3. Fungal treatment for the removal of antibiotics and antibiotic resistance genes in veterinary hospital wastewater.

    Lucas, D; Badia-Fabregat, M; Vicent, T; Caminal, G; Rodríguez-Mozaz, S; Balcázar, J L; Barceló, D

    2016-06-01

    The emergence and spread of antibiotic resistance represents one of the most important public health concerns and has been linked to the widespread use of antibiotics in veterinary and human medicine. The overall elimination of antibiotics in conventional wastewater treatment plants is quite low; therefore, residual amounts of these compounds are continuously discharged to receiving surface waters, which may promote the emergence of antibiotic resistance. In this study, the ability of a fungal treatment as an alternative wastewater treatment for the elimination of forty-seven antibiotics belonging to seven different groups (β-lactams, fluoroquinolones, macrolides, metronidazoles, sulfonamides, tetracyclines, and trimethoprim) was evaluated. 77% of antibiotics were removed after the fungal treatment, which is higher than removal obtained in conventional treatment plants. Moreover, the effect of fungal treatment on the removal of some antibiotic resistance genes (ARGs) was evaluated. The fungal treatment was also efficient in removing ARGs, such as ermB (resistance to macrolides), tetW (resistance to tetracyclines), blaTEM (resistance to β-lactams), sulI (resistance to sulfonamides) and qnrS (reduced susceptibility to fluoroquinolones). However, it was not possible to establish a clear link between concentrations of antibiotics and corresponding ARGs in wastewater, which leads to the conclusion that there are other factors that should be taken into consideration besides the antibiotic concentrations that reach aquatic ecosystems in order to explain the emergence and spread of antibiotic resistance. PMID:26991378

  4. Molecular characterization of the PK-LR gene in pyruvate kinase deficient Spanish patients. Red Cell Pathology Group of the Spanish Society of Haematology (AEHH).

    Zarza, R; Alvarez, R; Pujades, A; Nomdedeu, B; Carrera, A; Estella, J; Remacha, A; Sánchez, J M; Morey, M; Cortes, T; Pérez Lungmus, G; Bureo, E; Vives Corrons, J L

    1998-11-01

    The PK-LR gene has been studied in 12 unrelated patients with red cell pyruvate kinase deficiency and hereditary nonspherocytic haemolytic anaemia (CNSHA). The entire codifying region of the R-type PK gene and the flanking intronic regions were analysed by single-stranded conformation polymorphism (SSCP) followed by direct sequencing of abnormal DNA. 10 different mutations were identified in 22/24 alleles at risk. Eight of these were missense mutations that caused the following single amino acid changes: G514C (172Glu-Gln), G1010A (337Arg-Gln), G1015C (339Asp-Gln), T1070C (357Ile-Thr), C1223T (408Thr-Ile), G1291A (431Ala-Thr), C1456T (486Arg-Trp) and G1595A (532Arg-Gln). Two were nonsense mutations: G721T (241Glu-Stop) and C1675T (559Arg-Stop). 7/22 alleles demonstrated the same C1456 --> T mutation. The study of the polymorphic site at nucleotide (nt) 1705 performed in all cases disclosed a 1705 C/C mutation in 10 and a 1705 A/C mutation in three. This is the first report on the presence of several different L-type PK gene mutations within Spanish population. Furthermore, from the PK gene mutations found, six were unique and not previously described (1015C, 1070C, 1223T, 1291A, 1595A and 1675T) and one (C1456T) seems to be predominant in Spain. Interestingly, no case with the 1529A mutation commonly found in Northern European populations was present here. PMID:9827908

  5. The alpha7 nicotinic receptor agonist SSR180711 increases activity regulated cytoskeleton protein (Arc) gene expression in the prefrontal cortex of the rat

    Kristensen, Søren; Thomsen, Morten Skøtt; Hansen, Henrik H;

    2007-01-01

    Nicotinic alpha7 acetylcholine receptors (alpha7 nAChR) have been shown to enhance attentional function and aspects of memory function in experimental models and in man. The protein Arc encoded by the effector immediate early gene arc or arg3.1 has been shown to be strongly implicated in long-ter...... a subset of neurons in the rat prefrontal cortex and this activation likely is important for the attentional effects of this new class of drugs....

  6. MDM2 SNP309, gene-gene interaction, and tumor susceptibility: an updated meta-analysis

    Wu Wei

    2011-05-01

    Full Text Available Abstract Background The tumor suppressor gene p53 is involved in multiple cellular pathways including apoptosis, transcriptional control, and cell cycle regulation. In the last decade it has been demonstrated that the single nucleotide polymorphism (SNP at codon 72 of the p53 gene is associated with the risk for development of various neoplasms. MDM2 SNP309 is a single nucleotide T to G polymorphism located in the MDM2 gene promoter. From the time that this well-characterized functional polymorphism was identified, a variety of case-control studies have been published that investigate the possible association between MDM2 SNP309 and cancer risk. However, the results of the published studies, as well as the subsequent meta-analyses, remain contradictory. Methods To investigate whether currently published epidemiological studies can clarify the potential interaction between MDM2 SNP309 and the functional genetic variant in p53 codon72 (Arg72Pro and p53 mutation status, we performed a meta-analysis of the risk estimate on 27,813 cases with various tumor types and 30,295 controls. Results The data we reviewed indicated that variant homozygote 309GG and heterozygote 309TG were associated with a significant increased risk of all tumor types (homozygote comparison: odds ratio (OR = 1.25, 95% confidence interval (CI = 1.13-1.37; heterozygote comparison: OR = 1.10, 95% CI = 1.03-1.17. We also found that the combination of GG and Pro/Pro, TG and Pro/Pro, GG and Arg/Arg significantly increased the risk of cancer (OR = 3.38, 95% CI = 1.77-6.47; OR = 1.88, 95% CI = 1.26-2.81; OR = 1.96, 95% CI = 1.01-3.78, respectively. In a stratified analysis by tumor location, we also found a significant increased risk in brain, liver, stomach and uterus cancer (OR = 1.47, 95% CI = 1.06-2.03; OR = 2.24, 95%CI = 1.57-3.18; OR = 1.54, 95%CI = 1.04-2.29; OR = 1.34, 95%CI = 1.07-1.29, respectively. However, no association was seen between MDM2 SNP309 and tumor susceptibility

  7. MDM2 SNP309, gene-gene interaction, and tumor susceptibility: an updated meta-analysis

    The tumor suppressor gene p53 is involved in multiple cellular pathways including apoptosis, transcriptional control, and cell cycle regulation. In the last decade it has been demonstrated that the single nucleotide polymorphism (SNP) at codon 72 of the p53 gene is associated with the risk for development of various neoplasms. MDM2 SNP309 is a single nucleotide T to G polymorphism located in the MDM2 gene promoter. From the time that this well-characterized functional polymorphism was identified, a variety of case-control studies have been published that investigate the possible association between MDM2 SNP309 and cancer risk. However, the results of the published studies, as well as the subsequent meta-analyses, remain contradictory. To investigate whether currently published epidemiological studies can clarify the potential interaction between MDM2 SNP309 and the functional genetic variant in p53 codon72 (Arg72Pro) and p53 mutation status, we performed a meta-analysis of the risk estimate on 27,813 cases with various tumor types and 30,295 controls. The data we reviewed indicated that variant homozygote 309GG and heterozygote 309TG were associated with a significant increased risk of all tumor types (homozygote comparison: odds ratio (OR) = 1.25, 95% confidence interval (CI) = 1.13-1.37; heterozygote comparison: OR = 1.10, 95% CI = 1.03-1.17). We also found that the combination of GG and Pro/Pro, TG and Pro/Pro, GG and Arg/Arg significantly increased the risk of cancer (OR = 3.38, 95% CI = 1.77-6.47; OR = 1.88, 95% CI = 1.26-2.81; OR = 1.96, 95% CI = 1.01-3.78, respectively). In a stratified analysis by tumor location, we also found a significant increased risk in brain, liver, stomach and uterus cancer (OR = 1.47, 95% CI = 1.06-2.03; OR = 2.24, 95%CI = 1.57-3.18; OR = 1.54, 95%CI = 1.04-2.29; OR = 1.34, 95%CI = 1.07-1.29, respectively). However, no association was seen between MDM2 SNP309 and tumor susceptibility in the stratified analysis by p53 mutation status

  8. Enhanced uptake of antibiotic resistance genes in the presence of nanoalumina.

    Ding, Chengshi; Pan, Jie; Jin, Min; Yang, Dong; Shen, Zhiqiang; Wang, Jingfeng; Zhang, Bin; Liu, Weili; Fu, Jialun; Guo, Xuan; Wang, Daning; Chen, Zhaoli; Yin, Jing; Qiu, Zhigang; Li, Junwen

    2016-10-01

    Nanomaterial pollution and the spread of antibiotic resistance genes (ARGs) are global public health and environmental concerns. Whether nanomaterials could aid the transfer of ARGs released from dead bacteria into live bacteria to cause spread of ARGs is still unknown. Here, we demonstrated that nano-Al2O3 could significantly promote plasmid-mediated ARGs transformation into Gram-negative Escherichia coli strains and into Gram-positive Staphylococcus aureus; however, bulk Al2O3 did not have this effect. Under suitable conditions, 7.4 × 10(6) transformants of E. coli and 2.9 × 10(5) transformants of S. aureus were obtained from 100 ng of a pBR322-based plasmid when bacteria were treated with nano-Al2O3. Nanoparticles concentrations, plasmid concentrations, bacterial concentrations, interaction time between the nanomaterial and bacterial cells and the vortexing time affected the transformation efficiency. We also explored the mechanisms underlying this phenomenon. Using fluorescence in situ hybridization and scanning electron microscopy, we found that nano-Al2O3 damaged the cell membrane to produce pores, through which plasmid could enter bacterial cells. Results from reactive oxygen species (ROS) assays, genome-wide expression microarray profiling and quantitative real-time polymerase chain reactions suggested that intracellular ROS damaged the cell membrane, and that an SOS response promoted plasmid transformation. Our results indicated the environmental and health risk resulting from nanomaterials helping sensitive bacteria to obtain antibiotic resistance. PMID:26946995

  9. The biologic activity of melittin-88ArgIL-2 chimeric protein in vitro%蜂毒素与基因变构IL-2嵌合蛋白的体外生物活性研究

    宋旭霞; 刘明军; 王斌; 钱冬萌; 闫志勇; 丁守怡

    2009-01-01

    目的:IL-2具有多种与增强免疫功能相关的生物学活性,蜂毒素对多种肿瘤细胞有杀伤作用.文中研究蜂毒素与基因变构IL-2(melittin-88ArgIL-2)嵌合蛋白在体外的生物学活性. 方法:用甲基噻唑基四唑(MTT)法研究纯化制备的melittin-88ArgIL-2嵌合蛋白在体外对T细胞增殖和NK细胞杀伤活性的影响及对人卵巢癌细胞SKOV3生长抑制的作用. 结果:melittin-88ArgIL-2嵌合蛋白在体外可促进T细胞的增值,增强NK细胞的杀伤活性,抑制人卵巢癌细胞SKOV3的生长增殖. 结论:melittin-88ArgIL-2嵌合蛋白在体外具有一定的增强免疫功能和抗肿瘤活性.值得进一步对其体内的生物学活性进行研究,为大规模制备的中试放大研究和临床前期研究提供了资料.

  10. Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells.

    Bai, M.; Harfe, B; Freimuth, P

    1993-01-01

    The adenovirus penton base protein has a cell rounding activity and may lyse endosomes during virus entry into the cytoplasm. We found that penton base that was expressed in Escherichia coli also caused cell rounding and that cells adhered to polystyrene wells that were coated with the protein. Mutant analysis showed that both properties required an Arg-Gly-Asp (RGD) sequence at residues 340 to 342 of penton base. In flat adherent cells, virus mutants with amino acid substitutions in the RGD ...

  11. Morphology and anatomy of the developing fruit and seed of Dalechampia stipulacea Müll. Arg. (Euphorbiaceae) - DOI: 10.4025/actascibiolsci.v31i4.4834

    Luiz Antonio de Souza; Aparecido Caetano da Silva

    2009-01-01

    In forest remnants of Maringá, Paraná, Brazil, there is abundance of the liana species, with little studied reproductive structures, morphologically and anatomically. Among these species occurs Dalechampia stipulacea Müll. Arg., which is the object of the present study. It is a Euphorbiaceae with pseudanthium and trichomes that cause irritation of the skin. The fruits and seeds were fixed and cut freehand and in a rotation microtome, according to the usual techniques. The fruit is a schizocar...

  12. Expression of cold-adapted β-1,3-xylanase as a fusion protein with a ProS2 tag and purification using immobilized metal affinity chromatography with a high concentration of ArgHCl.

    Kudou, Motonori; Okazaki, Fumiyoshi; Asai-Nakashima, Nanami; Ogino, Chiaki; Kondo, Akihiko

    2015-01-01

    Cold-adapted β-1,3-xylanase (P.t.Xyn26A) from the psychrotrophic bacterium, Psychroflexus torquis, was expressed as a fusion protein with tandem repeats of the N-terminal domain of Protein S from Myxocuccus xanthus (ProS2) in Escherichia coli. After cell lysis in phosphate buffer, most of the ProS2-P.t.Xyn26A was located in the insoluble fraction and aggregated during purification. Arginine hydrochloride (ArgHCl) efficiently solubilized the ProS2-P.t.Xyn26A. The solubilized ProS2-P.t.Xyn26A was purified using immobilized metal affinity chromatography (IMAC) with 500 mM ArgHCl. After cleavage of ProS2-P.t.Xyn26A by human rhinovirus 3C protease, we confirmed that recombinant P.t.Xyn26A maintained its native fold. This is the first report of the expression of a cold-adapted enzyme fused with a ProS2 tag under IMAC purification using a high concentration of ArgHCl. These insights into the expression and purification should be useful during the handling of cold-adapted enzymes. PMID:25214227

  13. A seringueira (Hevea brasiliensisMuell.-Arg. no Vale do Rio Paraíba The rubber plant in the Valley of the Paraiba river

    João Ferreira da Cunha

    1966-01-01

    Full Text Available Foi feito um estudo sôbre o cultivo de seringueira (Hevea brasiliensisMuell. - Arg. no vale do rio Paraíba, no Estado de São Paulo, onde se fêz pela primeira vez o plantio com a finalidade de pesquisar a adaptação dessa espécie vegetal ao ambiente e as possibilidades da exploração comercial na região. O seringal foi plantado em terreno terciário, da série Guatemala, próximo de baixada, na Estação Experimental de Pindamonhangaba, localizada mais ou menos no centro do referido vale, a 22°55' de latitude sul, 45°25' de longitude e à altitude de 570 metros. O desenvolvimento das plantas foi normal. Ao completar sete anos de idade, havia 70% das plantas com as dimensões convencionais para início de exploração do seringal. A produção obtida de borracha foi de cêrca de 460 quilogramas por hectare no primeiro ano de colheita, 620 no segundo e 835 no terceiro. O aparecimento da moléstia das fôlhas, causada por Dothidella ulei,nos dois últimos anos de colheita, parece não ter tido influência apreciável na produção, que continuou aumentando. Os exames tecnológicos efetuados na borracha obtida revelaram características de fina qualidade.A study was made about the culture of the rubber plant (Hevea brasiliensisMuell.-Arg. in the Paraíba Valley of the State of São Paulo, in a region where this planting was done for the first time, with a view to investigate about the adaptability of its culture to this habitat and the possibility of its exploitation on this land. The rubber culure was established in the lowlands of the valley on tertiary soil that belongs to the Guatemala series, at the Pindamonhangaba Experiment station, located practically in the center of the valley, at 22°55' S. and 45°22' W., 570 m above sea level. The development of the plants was normal. After 7 years, when these plants had been settled, 70% were found to have the conventional dimensions for starting their exploitation. Rubber production was

  14. Influence of two-phase anaerobic digestion on fate of selected antibiotic resistance genes and class I integrons in municipal wastewater sludge.

    Wu, Ying; Cui, Erping; Zuo, Yiru; Cheng, Weixiao; Rensing, Christopher; Chen, Hong

    2016-07-01

    The response of representative antibiotic resistance genes (ARGs) to lab-scale two-phase (acidogenic/methanogenic phase) anaerobic digestion processes under thermophilic and mesophilic conditions was explored. The associated microbial communities and bacterial pathogens were characterized by 16S rRNA gene sequencing. A two-phase thermophilic digestion reduced the presence of tetA, tetG, tetX, sul1, ermB, dfrA1, dfrA12 and intI1 exhibiting 0.1-0.72 log unit removal; in contrast, tetO, tetW, sul3, ermF and blaTEM even increased relative to the feed, and sul2 showed no significant decrease. The acidogenic phase of thermophilic digestion was primarily responsible for reducing the quantity of these genes, while the subsequent methanogenic phase caused a rebound in their quantity. In contrast, a two-phase mesophilic digestion process did not result in reducing the quantity of all ARGs and intI1 except for ermB and blaTEM. ARGs patterns were correlated with Proteobacteria and Actinobacteria during the two-phase anaerobic digestion. PMID:27035472

  15. Trp homozygotes at codon 64 of ADRB3 gene are protected against the risk of type 2 diabetes in the Kashmiri population.

    Hameed, Iqra; Masoodi, Shariq R; Afroze, Dil; Naykoo, Niyaz A; Bhat, Riyaz A; Ganai, Bashir A

    2013-10-01

    The prevalence of type 2 diabetes mellitus has reached epidemic proportions worldwide. Type 2 diabetes is a consequence of complex interactions among multiple genetic variants and environmental risk factors. Polymorphisms in various candidate genes confer susceptibility to diabetes. This study was undertaken to analyse a single nucleotide polymorphism Trp64Arg (C↔T) in the ADRB3 gene and elucidate its effects on type 2 diabetes and its associated risk factors. The study included 200 type 2 diabetes patients and 300 age and gender matched healthy controls belonging to the ethnic Kashmiri population. Polymerase chain reaction-restriction fragment length polymorphism technique was used for genotyping and the results were validated by direct sequencing assay. Genotypes for Trp64Arg polymorphism were in Hardy-Weinberg equilibrium (χ(2)=0.48, p=NS). Frequency of the Arg64 allele was 40% and 10.2% in cases and controls, respectively (pADRB3 gene is a genotypic risk factor and confers susceptibility to type 2 diabetes, whereas the homozygous Trp64 genotype exerted a protective effect in our population. PMID:23968135

  16. Analysis of rare variants in the C3 gene in patients with age-related macular degeneration.

    Maheswara R Duvvari

    Full Text Available Age-related macular degeneration (AMD is a progressive retinal disorder affecting over 33 million people worldwide. Genome-wide association studies (GWASs for AMD identified common variants at 19 loci accounting for 15-65% of the heritability and it has been hypothesized that the missing heritability may be attributed to rare variants with large effect sizes. Common variants in the complement component 3 (C3 gene have been associated with AMD and recently a rare C3 variant (Lys155Gln was identified which exerts a large effect on AMD susceptibility independent of the common variants. To explore whether additional rare variants in the C3 gene are associated with AMD, we sequenced all coding exons in 84 unrelated AMD cases. Subsequently, we genotyped all identified variants in 1474 AMD cases and 2258 controls. Additionally, because of the known genetic overlap between AMD and atypical hemolytic uremic syndrome (aHUS, we genotyped two recurrent aHUS-associated C3 mutations in the entire cohort. Overall, we identified three rare variants (Lys65Gln (P=0.04, Arg735Trp (OR=17.4, 95% CI=2.2-136; P=0.0003, and Ser1619Arg (OR=5.2, 95% CI=1.0-25; P=0.05 at the C3 locus that are associated with AMD in our EUGENDA cohort. However, the Arg735Trp and Ser1619Arg variants were not found to be associated with AMD in the Rotterdam Study. The Lys65Gln variant was only identified in patients from Nijmegen, the Netherlands, and thus may represent a region-specific AMD risk variant.

  17. Identification of polymorphisms of XRCC1 gene in patients with cancer in a city of northern Brazil

    Artemis Socorro N. Rodrigues

    2015-06-01

    Full Text Available ABSTRACT Introduction: Cancer is considered a genetic disease. For this reason, identification and characterization of the genes involved in its origin and progression are of fundamental importance in understanding its molecular basis. Objective: Our objective was to determine whether people from Macapá with a diagnosis of cancer have genetic polymorphisms related to the XRCC1 gene. Materials and methods: We analyzed 30 samples of deoxyribonucleic acid (DNA of cases with cancer and 30 control samples. All samples were amplified and analyzed by the polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP method, with the use of restriction enzyme MspI. Results: Regarding the 194T polymorphism, we found that all samples of the cases presented the polymorphic allele Trp (Arg/Trp. In control samples, 96.6% also identify the polymorphic allele Trp and, among these, one was homozygous for the same allele (Trp/Trp. Regarding the 399A polymorphism, 83.3% of the cases and 23.3% of the controls had the Arg/ Gln genotype, respectively. We found that 73.3% of controls and 16.6% of cases had the Arg/Arg genotype. Among the controls, we found only a sample that was homozygous for the polymorphic allele Trp/Trp. Conclusion: Our results demonstrated the allele frequency of 194Trp polymorphism in both sample groups analyzed. We also found a significant number of polymorphic allele 399A in people with cancer. Thus, we can highlight 399Gln polymorphism as a genetic marker of cancer risk in this population.

  18. Effects of Condensed Tannins in Mao (Antidesma thwaitesianum Muell. Arg.) Seed Meal on Rumen Fermentation Characteristics and Nitrogen Utilization in Goats.

    Gunun, P; Wanapat, M; Gunun, N; Cherdthong, A; Sirilaophaisan, S; Kaewwongsa, W

    2016-08-01

    Mao seed is a by-product of the wine and juice industry, which could be used in animal nutrition. The current study was designed to determine the effect of supplementation of mao (Antidesma thwaitesianum Muell. Arg.) seed meal (MOSM) containing condensed tannins (CT) on rumen fermentation, nitrogen (N) utilization and microbial protein synthesis in goats. Four crossbred (Thai Native×Anglo Nubian) goats with initial body weight (BW) 20±2 kg were randomly assigned to a 4×4 Latin square design. The four dietary treatments were MOSM supplementation at 0%, 0.8%, 1.6%, and 2.4% of total dry matter (DM) intake, respectively. During the experimental periods, all goats were fed a diet containing roughage to concentrate ratio of 60:40 at 3.0% BW/d and pangola grass hay was used as a roughage source. Results showed that supplementation with MOSM did not affect feed intake, nutrient intakes and apparent nutrient digestibility (p>0.05). In addition, ruminal pH and ammonia nitrogen (NH3-N) were not influenced by MOSM supplementation, whilst blood urea nitrogen was decreased quadraticly (p0.05). There were linear decreases in urinary N (p0.05). From the current study, it can be concluded that supplementation of MOSM at 1.6% to 2.4% of total DM intake can be used to modify ruminal fermentation, especially propionate and N utilization in goats, without affecting the nutrient digestibility, microbial populations and microbial protein synthesis. PMID:26954153

  19. Photodynamic therapy of a 2-methoxyestradiol tumor-targeting drug delivery system mediated by Asn-Gly-Arg in breast cancer

    Shi J

    2013-04-01

    Full Text Available Jinjin Shi, Zhenzhen Wang, Lei Wang, Honghong Wang, Lulu Li, Xiaoyuan Yu, Jing Zhang, Rou Ma, Zhenzhong ZhangSchool of Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, People's Republic of ChinaAbstract: Fullerene (C60 has shown great potential in drug delivery. In this study we exploited modified fullerene (diadduct malonic acid-fullerene-Asn-Gly-Arg peptide [DMA-C60-NGR] as an antitumor drug carrier in order to build a new tumor-targeting drug delivery system. We also investigated the synergistic enhancement of cancer therapy using photodynamic therapy (PDT induced by DMA-C60-NGR and 2-methoxyestradiol (2ME. Cytotoxicity tests indicated that DMA-C60-NGR had no obvious toxicity, while our drug delivery system (DMA-C60-2ME-NGR had a high inhibition effect on MCF-7 cells compared to free 2ME. The tumor-targeting drug delivery system could efficiently cross cell membranes, and illumination induced the generation of intracellular reactive oxygen species and DNA damage. Furthermore, DMA-C60-2ME-NGR with irradiation had the highest inhibition effect on MCF-7 cells compared to the other groups. DMA-C60-NGR combined with 2ME showed a good synergistic photosensitization effect for inhibiting the growth of MCF-7 cells, demonstrating that DMA-C60-2ME-NGR may be promising for high treatment efficacy with minimal side effects in future therapy.Keywords: fullerene, drug delivery system, photodynamic therapy, tumor targeting

  20. Study of root distribution of Macaranga tanarius (L. Müll. Arg. (Parasol leaf tree on East-West highway slope, Malaysia

    Nazi Avani

    2013-01-01

    Full Text Available Vegetation influences the stability of slope by both hydrological events and mechanical reinforcement of the soil. The magnitude of such effects is subject to the root system growth, which in turn is a purpose of the genetic properties of the species and site appearances. In this study, we investigated the root distribution of Macaranga tanarius (L. Müll. Arg. as a native species in Malaysia, with an aim to rise our knowledge on root area ratio distribution inside the soil. Concerning the estimation of root distribution within the soil, we assessed the root area ratio (the proportion between the area occupied by the roots in a slice area of soil according to its depth for four samples in the rainforest. Results show that the root area ratio (RAR declined with depth, and the higher RAR values were calculated in the higher layers. RAR values were between 0.950% and 18.477%. There is a plentiful diversity of root density in depth classes. Also, the result showed that about 50% of the roots are located in the first 10 cm layer, and about 87% of roots were in the fine root diameter (>2 mm. There is not a significant difference between RAR and soil depth. Spearman correlation showed no significant and negative correlation between RAR and depth. The maximum RAR percentage was in the first layer 0-10 cm (44.59%. Also, the results showed that the amount of RAR decreased with depth in fine roots (d<2mm, but RAR in thin roots (2

  1. Intervención educativa sobre higiene bucal en escolares del seminternado “Jesús Argüelles Hidalgo”

    Nivia M. Aguilera Trotman

    2015-11-01

    Full Text Available Se realizó una intervención educativa en la rama de las Ciencias Médicas, específicamente en la estomatología, con el objetivo de elevar el nivel de conocimiento sobre higiene bucal en escolares de ocho a doce años de edad del seminternado “Jesús Argüelles Hidalgo”, perteneciente a la clínica estomatológica “Guillermo Tejas” del municipio de Las Tunas, en el período comprendido de mayo de 2010 a mayo de 2012. El universo estuvo constituido por 392 escolares, seleccionándose una muestra de 150 que cumplió con los criterios establecidos. La intervención educativa se desarrolló con previa organización de los temas en ocho sesiones, y con el empleo de técnicas educativas y diferentes medios de enseñanza. A los escolares se les aplicó un cuestionario, determinándose así el nivel de conocimiento sobre salud bucal y para determinar el comportamiento de la higiene bucal se utilizó el índice de higiene bucal de Love, al inicio y final del estudio; además, se ejecutó una lluvia de ideas para conocer las necesidades sentidas al inicio de la intervención. Se aplicó el modelo de David Leyva, determinándose las necesidades de aprendizaje. Prevaleció el grupo de ocho a diez años de edad, así como el sexo masculino. Se compararon los resultados iniciales con los obtenidos al finalizar la intervención, concluyendo que se logró elevar el nivel de conocimientos sobre salud bucal de los escolares y se mejoró la higiene bucal de los mismos; la intervención educativa fue satisfactoria.

  2. Angiotensin I-converting enzyme Gln1069Arg mutation impairs trafficking to the cell surface resulting in selective denaturation of the C-domain.

    Sergei M Danilov

    Full Text Available BACKGROUND: Angiotensin-converting enzyme (ACE; Kininase II; CD143 hydrolyzes small peptides such as angiotensin I, bradykinin, substance P, LH-RH and several others and thus plays a key role in blood pressure regulation and vascular remodeling. Complete absence of ACE in humans leads to renal tubular dysgenesis (RTD, a severe disorder of renal tubule development characterized by persistent fetal anuria and perinatal death. METHODOLOGY/PRINCIPAL FINDINGS: Patient with RTD in Lisbon, Portugal, maintained by peritoneal dialysis since birth, was found to have a homozygous substitution of Arg for Glu at position 1069 in the C-terminal domain of ACE (Q1069R resulting in absence of plasma ACE activity; both parents and a brother who are heterozygous carriers of this mutation had exactly half-normal plasma ACE activity compared to healthy individuals. We hypothesized that the Q1069R substitution impaired ACE trafficking to the cell surface and led to accumulation of catalytically inactive ACE in the cell cytoplasm. CHO cells expressing wild-type (WT vs. Q1069R-ACE demonstrated the mutant accumulates intracellularly and also that it is significantly degraded by intracellular proteases. Q1069R-ACE retained catalytic and immunological characteristics of WT-ACE N domain whereas it had 10-20% of the nativity of the WT-ACE C domain. A combination of chemical (sodium butyrate or pharmacological (ACE inhibitor chaperones with proteasome inhibitors (MG 132 or bortezomib significantly restored trafficking of Q1069R-ACE to the cell surface and increased ACE activity in the cell culture media 4-fold. CONCLUSIONS/SIGNIFICANCE: Homozygous Q1069R substitution results in an ACE trafficking and processing defect which can be rescued, at least in cell culture, by a combination of chaperones and proteasome inhibitors. Further studies are required to determine whether similar treatment of individuals with this ACE mutation would provide therapeutic benefits such as

  3. Premilinary studies for optimiziing a protocol for obtaining embryogenic calluses in two rubber (Hevea brasiliensis Mull. Arg clones from different geographical origins

    Santiago Cadavid Ruiz

    2007-02-01

    Full Text Available The influence of growth regulators on obtaining friable rubber (Hevea brasiliensis Müll. Arg. calluses with no plant regeneration as investigated. Two clones having different geographical origin were used in all trails carried out in this study: FX 3864 (South-American and PB 254 (Asian. Young leaves and eight- to ten-week-old seed integument from both clones were used as explants in several experiments; they were initially cultured in MH medium (Carron, et ál., 1989, modified MH medium (Montoro, et ál., 1993, 2000 and modified MS medium (Carron, et at,. 1992, no positive response being obtained by days 25 or 50. However, other trials were carried out with the integument in modified MS medium (1962, 0.67 mg/L BAP and 0.66 mg/L 2-4 D being added as medium for initiating embryogenesis, the formation of white, friable calluses being observed by day 25 in the two selected clones. These calluses were sub-cultured in MS supplemented with 0.35 mg/L BAP and 0.2 mg/L 2-4 D as callogenesis expression medium, embryogenic expression being observed in both clones by day 50. Equally friable white calluses were obtained from young leaves in the two clones in MS medium supplemented with 1.0 mg/L BAP, 1.0 mg/L ANA but without IBA and kinetin by day 25. Calluses sub-cultured in the same medium supplemented with 0.5 mg/L BAP and 0.5 mg/L ANA began to show increased friability after 50 days. culture. This work is a partial report of a macro-project for optimising a protocol for rubber (Hevea brasiliensis  multiplication by somatic embryogenesis.

  4. Assessment of MC1R and α-MSH gene sequences in Iranian vitiligo patients

    Eskandani M

    2010-01-01

    Full Text Available Background: Vitiligo is an acquired pigmentary disorder of the skin that is caused by unknown factors and is characterized by white and depigmented patches that enlarge and become more numerous with time. Genetic factors, oxidative stress, autoimmunity, and neurochemical agents, such as catecholamines might also contribute to vitiligo. Cutaneous pigmentation is determined by the amounts of eumelanin and pheomelanin synthesized by the epidermal melanocytes and interference of melanocortin-1 receptor (MC1R, a G-protein coupled receptor, its normal agonist, alpha-melanocyte stimulating hormone (α-MSH, and key enzymes, such as tyrosinase, to protect against sun-induced DNA damage. The MC1R, a 7 pass trans-membrane G-protein coupled receptor, is a key control point in melanogenesis. Loss-of-function mutations at the MC1R are associated with a switch from eumelanin to pheomelanin production, resulting in a red or yellow coat color. Aim: In this research, we aim to examine the genetic variety of MC1R and α-MSH gene in 20 Iranian vitiligo patients and 20 healthy controls. Materials and Methods: Analysis of the MC1R coding gene was performed with direct sequencing. Results: We found the following 9 MC1R coding region variants: Arg163Gl (G488A, Arg227Leu (G680A, Val 97Phe (G289T, Asp184Asn (G550A, Arg227Lys (G680A, Arg142His (G425A, Val60Leu (G178T, Val247Met (C739A, and Val174Ile (G520A. We also found 2 frameshift changes: one of them was the Insertion of C (frameshift in Pro136, stop at Trp148 and the other, Insertion of G (frameshift in Pro256, stop at Trp 333. Of all the changes, the most common was Val60Leu at 5% in patients vs 20% in controls, Val247Met at 15% in patients vs 0% in controls and Val174Ile at 15% in controls and 0% in patients. The other variants showed a frequency <5% in both patients and controls. Also in this study, we have examined the frequency of single nucleotide polymorphisms within the α-MSH genes with direct sequencing in

  5. Antibiotic-Resistant Gonorrhea (ARG)

    ... 1993, ciprofloxacin, a fluoroquinolone, and cephalosporins ceftriaxone and cefixime were the recommended treatments for gonorrhea. However, in ... people in the United States. The cephalosporins, either cefixime or ceftriaxone, were the only remaining recommended treatments. ...

  6. Who possesses drug resistance genes in the aquatic environment?: sulfamethoxazole (SMX) resistance genes among the bacterial community in water environment of Metro-Manila, Philippines

    Suzuki, Satoru; Ogo, Mitsuko; Miller, Todd W.; Shimizu, Akiko; Takada, Hideshige; Siringan, Maria Auxilia T.

    2013-01-01

    Recent evidence has shown that antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are ubiquitous in natural environments, including sites considered pristine. To understand the origin of ARGs and their dynamics, we must first define their actual presence in the natural bacterial assemblage. Here we found varying distribution profiles of sul genes in “colony forming bacterial assemblages” and “natural bacterial assemblages.” Our monitoring for antibiotic contamination revealed that sulfamethoxazole (SMX) is a major contaminant in aquatic environments of Metro-Manila, which would have been derived from human and animal use, and subsequently decreased through the process of outflow from source to the sea. The SMX-resistant bacterial rate evaluated by the colony forming unit showed 10 to 86% of the total colony numbers showed higher rates from freshwater sites compared to marine sites. When sul genes were quantified by qPCR, colony-forming bacteria conveyed sul1 and sul2 genes in freshwater and seawater (10−5–10−2 copy/16S) but not sul3. Among the natural bacterial assemblage, all sul1, sul2, and sul3 were detected (10−5–10−3 copy/16S), whereas all sul genes were at an almost non-detectable level in the freshwater assemblage. This study suggests that sul1 and sul2 are main sul genes in culturable bacteria, whereas sul3 is conveyed by non-culturable bacteria in the sea. As a result marine bacteria possess sul1, sul2 and sul3 genes in the marine environment. PMID:23641240

  7. Human-specific SNP in obesity genes, adrenergic receptor beta2 (ADRB2, Beta3 (ADRB3, and PPAR γ2 (PPARG, during primate evolution.

    Akiko Takenaka

    Full Text Available UNLABELLED: Adrenergic-receptor beta2 (ADRB2 and beta3 (ADRB3 are obesity genes that play a key role in the regulation of energy balance by increasing lipolysis and thermogenesis. The Glu27 allele in ADRB2 and the Arg64 allele in ADRB3 are associated with abdominal obesity and early onset of non-insulin-dependent diabetes mellitus (NIDDM in many ethnic groups. Peroxisome proliferator-activated receptor γ (PPARG is required for adipocyte differentiation. Pro12Ala mutation decreases PPARG activity and resistance to NIDDM. In humans, energy-expense alleles, Gln27 in ADRB2 and Trp64 in ADRB3, are at higher frequencies than Glu27 and Arg64, respectively, but Ala12 in PPARG is at lower frequency than Pro12. Adaptation of humans for lipolysis, thermogenesis, and reduction of fat accumulation could be considered by examining which alleles in these genes are dominant in non-human primates (NHP. All NHP (P. troglodytes, G. gorilla, P. pygmaeus, H. agilis and macaques had energy-thrifty alleles, Gly16 and Glu27 in ADRB2, and Arg64 in ADRB3, but did not have energy-expense alleles, Arg16, Gln27 and Trp64 alleles. In PPARG gene, all NHP had large adipocyte accumulating type, the Pro12 allele. CONCLUSIONS: These results indicate that a tendency to produce much more heat through the energy-expense alleles developed only in humans, who left tropical rainforests for savanna and developed new features in their heat-regulation systems, such as reduction of body hair and increased evaporation of water, and might have helped the protection of entrails from cold at night, especially in glacial periods.

  8. Performance of vertical up-flow constructed wetlands on swine wastewater containing tetracyclines and tet genes.

    Huang, Xu; Liu, Chaoxiang; Li, Ke; Su, Jianqiang; Zhu, Gefu; Liu, Lin

    2015-03-01

    Antibiotics and antibiotic resistance genes (ARGs) pollution in animal feeding farms received more public attention recently. Livestock wastewater contains large quantities of antibiotics and ARGs even after traditional lagoon treatment. In this study, the performance of vertical up-flow constructed wetlands (VUF-CWs) on swine wastewater containing tetracycline compounds (TCs) and tet genes was evaluated based on three aspects, TCs and tet genes removal efficiencies, residual TCs and tet genes in soils and plants, and the effect of TCs accumulation on nutrients removal and tet genes development. High removal efficiencies (69.0-99.9%) were achieved for oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC) with or without OTC spiked in the influent additionally. TCs concentrations in surface soils increased at first two sampling periods and then decreased after plants were harvested. Satisfactory nutrients removal efficiencies were also obtained, but TN and NH4-N removal efficiencies were significantly negative correlated with total concentration of TCs (∑TCs) in the soils (p < 0.01). The absolute abundances of all the target genes (tetO, tetM, tetW, tetA, tetX and intI1) were greatly reduced with their log units ranging from 0.26 to 3.3. However, the relative abundances of tetO, tetM and tetX in some effluent samples were significantly higher than those in the influent (p < 0.05). The relative abundances of tet genes except for tetO were significantly correlated with ∑TCs in the soils (p < 0.05). In summary, the proposed VUF-CWs are effective alternative for the removal of TCs and tet genes. But it is of great importance to prevent large accumulation of TCs in the soils. PMID:25528541

  9. Inorganic and organic fertilizers impact the abundance and proportion of antibiotic resistance and integron-integrase genes in agricultural grassland soil.

    Nõlvak, Hiie; Truu, Marika; Kanger, Kärt; Tampere, Mailiis; Espenberg, Mikk; Loit, Evelin; Raave, Henn; Truu, Jaak

    2016-08-15

    Soil fertilization with animal manure or its digestate may facilitate an important antibiotic resistance dissemination route from anthropogenic sources to the environment. This study examines the effect of mineral fertilizer (NH4NO3), cattle slurry and cattle slurry digestate amendment on the abundance and proportion dynamics of five antibiotic resistance genes (ARGs) and two classes of integron-integrase genes (intI1 and intI2) in agricultural grassland soil. Fertilization was performed thrice throughout one vegetation period. The targeted ARGs (sul1, tetA, blaCTX-M, blaOXA2 and qnrS) encode resistance to several major antibiotic classes used in veterinary medicine such as sulfonamides, tetracycline, cephalosporins, penicillin and fluoroquinolones, respectively. The non-fertilized grassland soil contained a stable background of tetA, blaCTX-M and sul1 genes. The type of applied fertilizer significantly affected ARGs and integron-integrase genes abundances and proportions in the bacterial community (pfertilizer's application event, but this increase was followed by a stage of decrease, suggesting that microbes possessing these genes were predominantly entrained into soil via cattle slurry or its digestate application and had somewhat limited survival potential in a soil environment. However, the abundance of these three target genes did not decrease to a background level by the end of the study period. TetA was most abundant in mineral fertilizer treated soil and blaCTX-M in cattle slurry digestate amended soil. Despite significantly different abundances, the abundance dynamics of bacteria possessing these genes were similar (p<0.05 in all cases) in different treatments and resembled the dynamics of the whole bacterial community abundance in each soil treatment. PMID:27115621

  10. Contribuição ao estudo anatômico do caule de Himatanthus sucuuba (Spruce ex Müll. Arg. Woodson, Apocynaceae Contribution to the anatomical study of the stem of Himatanthus sucuuba (Spruce ex Müll. Arg. Woodson, Apocynaceae

    C.R.R. Larrosa

    2005-06-01

    Full Text Available Himatanthus sucuuba (Spruce ex Müll. Arg. Woodson, Apocynaceae, denominada comumente de sucuuba ou janaguba, é uma espécie arbórea, latescente, da região amazônica, cujo tronco é ereto e a casca rugosa. As atividades antiinflamatória e analgésica e a baixa toxicidade foram estabelecidas em diferentes pesquisas. Com o objetivo de estabelecer caracteres anatômicos aplicáveis à identificação da planta e da droga vegetal, analisou-se o caule em estrutura secundária, que consiste da casca e da região caulinar mais interna. A primeira periderme é observada nas camadas subepidérmicas e o córtex constitui-se de numerosos laticíferos ramificados, idioblastos com cristais de oxalato de cálcio e células parenquimáticas contendo amiloplastos. Uma bainha esclerenquimática descontínua de fibras com paredes espessadas circunda o sistema vascular, cuja organização é bicolateral. As células do parênquima medular apresentam as mesmas características do córtex. Em níveis mais inferiores, a região cortical se reduz e numerosas fibras e células pétreas localizam-se no floema externo.Himatanthus sucuuba (Spruce ex Müll. Arg. Woodson, Apocynaceae, popularly known as sucuuba and janaguba, is a laticiferous woody species from the Amazonian region. The stem is upright and the bark is rugous. Different investigations have confirmed its anti-inflammatory and analgesic activities as well as low toxicity. Aiming to establish anatomical characters for the plant and drug identification, the stem in secondary growth, consisting of the bark and the most internal region, has been analysed. The first periderm is observed in the sub-epidermal layers and the cortex is composed of several branched laticifers, idioblasts containing calcium oxalate crystals and parenchymatic cells having amiloplasts. A non-continuous sclerenchymatic sheath with thick cell-walled fibres encircles the vascular system, whose organization is bicollateral. The pith cells

  11. Himatanthus lancifolius (Müll. Arg. Woodson, Apocynaceae: estudo farmacobotânico de uma planta medicinal da Farmacopeia brasileira 1ª edição Himatanthus lancifolius (Müll. Arg. Woodson, Apocynaceae: morpho-anatomical study of a medicinal plant described in the Brazilian Pharmacopoeia 1st edition

    Leopoldo C. Baratto

    2010-11-01

    Full Text Available Himatanthus lancifolius (Müll. Arg. Woodson é um arbusto nativo do Brasil, latescente, popularmente conhecido como agoniada e utilizado principalmente para distúrbios menstruais. O presente trabalho teve como objetivo caracterizar morfoanatomicamente a folha, o caule e a casca caulinar dessa planta medicinal, a fim de contribuir para o controle de qualidade e a autenticidade dessa espécie. O material vegetal foi fixado e submetido às microtécnicas usuais. A folha é simples, glabra e obovado-lanceolada. A epiderme é uniestratificada, revestida por cutícula estriada e possui estômatos anisocíticos na face abaxial. O mesofilo é dorsiventral. A nervura central é biconvexa e o pecíolo é circular, ambos apresentando feixes vasculares bicolaterais. Laticíferos, amiloplastos e idioblastos fenólicos estão presentes no parênquima fundamental da nervura central e do pecíolo. O sistema vascular do caule é tipicamente bicolateral. Laticíferos e idioblastos fenólicos ocorrem no córtex, no floema e na medula. Esses caracteres morfoanatômicos, em conjunto, podem ser utilizados como parâmetros para o controle de qualidade dessa espécie.Himatanthus lancifolius (Müll. Arg. Woodson is a Brazilian native shrub, laticiferous, popularly known as "agoniada" and it is mainly used for uterine disorders. The present work aimed to study the leaf, stem and stem bark morpho-anatomy of this medicinal plant, in order to contribute to its quality control and identification. The plant material was fixed and submitted to standard microtechniques. The leaf is simple, glabrous and obovate-lanceolate. The epidermis is uniseriate, coated with striated cuticle and it has anysocitic stomata on the abaxial surface. The mesophyll is dorsiventral. The midrib is biconvex and the petiole is circular, both presenting bicollateral vascular bundles. Laticiferous ducts, amyloplasts and phenolic idioblasts are found in ground parenchyma of the midrib and petiole. The

  12. Synthesis and In Vitro Testing of New Potent Polyacridine-Melittin Gene Delivery Peptides

    Baumhover, Nicholas J.; Anderson, Kevin; Fernandez, Christian A.; Rice, Kevin G.

    2010-01-01

    The combination of a polyacridine peptide modified with a melittin fusogenic peptide results in a potent gene transfer agent. Polyacridine peptides of the general formula (Acr-X)n-Cys were prepared by solid phase peptide synthesis, where Acr is Lys modified on its ε-amine with acridine, X is Arg, Leu or Lys and n is 2, 3 or 4 repeats. The Cys residue was modified by either a maleimide-melittin or a thiolpyridine-Cys-melittin fusogenic peptide resulting in reducible or non-reducible polyacridi...

  13. Research on carrier detection and prenatal gene diagnosis of hemophilia B%血友病B携带者及产前基因诊断的研究

    王莉; 侯巧芳; 廖世秀; 王应太; 雷平冲

    2011-01-01

    目的 探讨血友病B家系分子致病机制及基因诊断携带者和高危胎儿.方法 采集5个典型血友病B家系成员的标本,提取基因组DNA后PCR扩增和序列比对凝血因子Ⅸ的8个外显子及其侧翼序列;利用基因外6个STR位点进行连锁分析.结果 序列分析表明5个家系患者分别存在6485C→T(Arg37stop)、6364C→T( Arg-4Trp)、6409G→A (Gly12Arg)、6364C→T (Arg-4Trp)、30383G→C (Val135Leu)突变;家系1的Ⅲ2和Ⅳ1、家系2的Ⅰ1、家系4的Ⅱ8和Ⅲ6均携带家系中患者的杂合突变;发现1例女性胎儿携带致病基因,其余2例胎儿未见突变;连锁分析结果与序列分析一致.结论 替换突变可能是这5个家系致病的分子机制;在严格的质量控制下,序列突变分析结合连锁分析是血友病B携带者和产前基因诊断准确而快速的方法.%[Objective] To explore the molecular mechanism of collected five hemophilia B families, and perform gene diagnosis for carriers and high-risk fetus. [Methods] Specimens from five typical Hemophilia B families were collected. After extraction of genomic DNA, all of the 8 exons and their flanks of FIX gene were amplified by polymerase chain reaction (PCR) and direct sequencing. The linkage relationships were analyzed by extra-genic six STR loci of FIX gene. [Results] Sequence analysis indicated that the probands of 5 families have 6485C→T (Arg 37 stop), 6364C→T (Arg-4 Trp), 6409 G→A (Gly 12 Arg), 6364C→T (Arg-4Trp), 30383G→C (Val 135 Leu) mutation, respectively. Member III2 and IV1 from family 1, 11 from family 2, 118 and III6 from family 4 carried the corresponding heterozygous mutation. STR linkage analysis results were consistent with sequence analysis. A female fetus was found to carry the pathogenesis gene, and other two fetuses carried normal X chromosome. [Conclusion] Replacement mutations are likely to be the main mechanisms of the 5 hemophilia B families. To strict quality control, the DNA

  14. Molecular evolution of B6 enzymes: Binding of pyridoxal-5'-phosphate and Lys41Arg substitution turn ribonuclease A into a model B6 protoenzyme

    Marra Ersilia

    2008-06-01

    Full Text Available Abstract Background The pyridoxal-5'-phosphate (PLP-dependent or vitamin B6-dependent enzymes that catalyze manifold reactions in the metabolism of amino acids belong to no fewer than four evolutionarily independent protein families. The multiple evolutionary origin and the essential mechanistic role of PLP in these enzymes argue for the cofactor having arrived on the evolutionary scene before the emergence of the respective apoenzymes and having played a dominant role in the molecular evolution of the B6 enzyme families. Here we report on an attempt to re-enact the emergence of a PLP-dependent protoenzyme. The starting protein was pancreatic ribonuclease A (RNase, in which active-site Lys41 or Lys7 readily form a covalent adduct with PLP. Results We screened the PLP adduct of wild-type RNase and two variant RNases (K7R and K41R for catalytic effects toward L- and D-amino acids. RNase(K41R-PLP, in which the cofactor is bound through an imine linkage to Lys7, qualifies for a model proto-B6 enzyme by the following criteria: (1 covalent linkage of PLP (internal aldimine; (2 catalytic activity toward amino acids that depends on formation of an imine linkage with the substrate (external aldimine; (3 adjoining binding sites for the cofactor and amino acid moiety that facilitate the transimination reaction of the internal to the external aldimine and stabilize the resulting noncovalent complex of the coenzyme-substrate adduct with the protein; (4 reaction specificity, the only detectable reactions being racemization of diverse amino acids and β-decarboxylation of L-aspartate; (5 acceleration factors for racemization and β-decarboxylation of >103 over and above that of PLP alone; (6 ribonuclease activity that is 103-fold lower than that of wild-type RNase, attenuation of a pre-existing biological activity being indispensable for the further evolution as a PLP-dependent protoenzyme. Conclusion A single amino acid substitution (Lys41Arg and covalent

  15. A gênese social do homo-economicus: a Argélia e a sociologia da economia em Pierre Bourdieu

    Marie-France Garcia-Parpet

    2006-10-01

    Full Text Available O artigo se propõe a mostrar que o interesse de P. Bourdieu pelas questões econômicas já estava presente desde seus primeiros trabalhos - sobre o processo de adaptação ao capitalismo das populações autóctones na Argélia - nos quais já se encontram as formas mais elaboradas de uma sociologia da economia. Nesses trabalhos, fortemente ancorados em uma pesquisa realizada na mesma época sobre o celibato e a crise da reprodução camponesa no Béarn, sua região de origem, P. Bourdieu opõe-se radicalmente à concepção da teoria neoclássica, e demonstra que as disposições econômicas mais fundamentais - necessidades, preferências, propensões ao trabalho, à poupança e ao investimento - não são exógenas, colocando em evidência as condições sócio-históricas do comportamento econômico racional. Tais trabalhos revelaram-se essenciais para todas as pesquisas posteriores como fontes de questões a serem examinadas, como matrizes de inovações conceituais e como posturas teóricas posteriormente afirmadas.The present article analyze P. Bourdieu's interest in economic questions, showing how this interest was present since the author's initial work - regarding the autochthonous population of Algeria's adaptation to capitalism - in which more elaborate forms of a sociology of the economy can already be found. In these works, which where anchored on simultaneous research conducted regarding celibacy and the crisis of peasant reproduction in Bearn, P. Bourdieu's birth region, the author radically opposed neo-classical theoretical concepts. Bourdieu demonstrated that the most fundamental economic dispositions (needs, preferences, propensity towards work, savings or investment where not exogenous, exposing the socio-historical conditions of rational economic behavior. These works are essential elements in the forging of an understanding of all of Bourdieu's later research. They are sources of the questions which will later be examined

  16. {sup 124}I labeled Arg-Gly-Asp (RGD) peptide: PET/MR fusion imaging of tumor expressing integrin v3

    Yun, Seon Young; Park, Jeong Chan [Kyungpook National University School of Medicine, Daegu (Korea, Republic of); Chen, Xiaoyuan [Standford Univ. School of Medicine, Palo Alto (United States)] (and others)

    2007-07-01

    Tumor-induced angiogenesis and metastasis express high level of integrin {sup v3} which has become a promising diagnostic biomarker and therapeutic target for various tumors. Radiolabeled RGD (Arg-Gly-Asp) peptides are well known to specifically bind to integrin and can be used not only for noninvasive imaging of integrin over-expression but also for integrin-targeted radionuclide therapy. We used three RGDyK derivatives, monomeric-, dimeric- and tetrameric-RGD for radio labeling. They were labeled with [{sup 124}I]NaI using iodo-bead. Labeled RGD peptide was purified by FPLC. Xenografts were induced by subcutaneous injection of 1X10{sup 7} U87MG cells into the right thigh of female Balb/c nude mice. After injection of {sup 124}I-labeled monomeric-RGD via the tail vein, the microPET imaging was obtained during 2 hours. Right after microPET imaging, T{sub 2}-weighted MR imaging was performed with fast gradient spin echo sequence in 0.8 mm of a slice thickness. PET/MR fusion images were constructed by AMIDE program. For immobilization and reproducible positioning of animal, animal-specific positioning molds were fabricated. Monomeric-RGD peptide was labeled with {sup 124}I in labeling yield of 50%. After FPLC purification, the radiochemical purity was above 90%. U87MG tumor was clearly seen in both microPET and MR imaging of tumor-bearing nude mouse. The fusion imaging was well constructed by using AMIDE with little discrepancy thanks to animal-specific molds. Three RGD derivatives were also labeled with {sup 131}I in labeling yield of 20-55%. All three RGD peptides were successfully radioiodinated with {sup 124}I or {sup 131}I. U87MG glioblastoma tumor was clearly visualized in PET/MR fusion imaging. MicroPET/MR fusion imaging of dimeric- and tetrameric-RGD peptides in tumor bearing mice will also be presented during conference.

  17. Gene expression

    We prepared probes for isolating functional pieces of the metallothionein locus. The probes enabled a variety of experiments, eventually revealing two mechanisms for metallothionein gene expression, the order of the DNA coding units at the locus, and the location of the gene site in its chromosome. Once the switch regulating metallothionein synthesis was located, it could be joined by recombinant DNA methods to other, unrelated genes, then reintroduced into cells by gene-transfer techniques. The expression of these recombinant genes could then be induced by exposing the cells to Zn2+ or Cd2+. We would thus take advantage of the clearly defined switching properties of the metallothionein gene to manipulate the expression of other, perhaps normally constitutive, genes. Already, despite an incomplete understanding of how the regulatory switch of the metallothionein locus operates, such experiments have been performed successfully

  18. Analysis of similarity of the SI gene in infectious bronchitis virus (IBV isolates in Shanghai, China Análisis de similaridad del gen S1 de aislados del virus de la bronquitis infecciosa (IBV en Shanghai, China

    J G Zhu

    2007-01-01

    Full Text Available The aim of the study was to analyze the similarity of the SI gene in 5 novel viral strains of infectious bronchitis virus (IBV isolated in Shanghai, China, using a pair of specific primers which were designed based on the published sequences of the S gene of IBV. The 5 full-length cDNA segments of the SI gene were amplified from the 5 isolated strains with RT-PCR. It was confirmed that the nucleotide sequences of the SI gene in the 5 strains isolated from chickens and pigeon have a length of 1626 bp and encode 541 amino acid residues. Compared to the sequence of selected reference IBV strains published in GenBank, the 4 strains from chickens exhibited 77.4%-82.9% identity in the SI gene sequence, with the deduced similarity of the protein sequence being 74.7%-82.6%, while the strain from pigeon exhibited 79.3%-99.6% similarity in the SI gene sequence, with a deduced identity of the protein sequence of 81.6%-99.6%. The SI protein cleavage site sequence of these isolates contains 5 consecutive basic amino acids, named, Arg-Arg-Phe-Arg-Arg (RRFRR similar to most of IBV strains. The results indicated that the variability range of the SI gene structure is high, corresponding with the biological character of IBV, and that molecularly the five isolated strains from Shanghai have a close relationship with the IBV reference strains. It was concluded that the five isolated strains belong to IBV, even though one of them was isolated from pigeonsEl objetivo del estudio fue analizar la similaridad del gen SI de 5 nuevas cepas virales del virus de la bronquitis infecciosa (IBV aislado en Shangai, China, para lo cual se usaron un par de partidores específicos, los cuales fueron designados en base a las secuencias publicadas del gen S del IBV. Los 5 segmentos completos de cDNA del gen SI fueron amplificados de las 5 cepas aisladas con RT-PCR. Se pudo confirmar que las secuencias de nucleótidos de genes SI en las 5 cepas aisladas de gallinas y palomas tienen

  19. Leptin receptor gene polymorphisms in severely pre-eclamptic women.

    Rigó, János; Szendei, György; Rosta, Klára; Fekete, Andrea; Bögi, Krisztina; Molvarec, Attila; Rónai, Zsolt; Vér, Agota

    2006-09-01

    Variants of the leptin receptor gene (LEPR) may modulate the effect of elevated serum leptin levels in pre-eclampsia. The aim of our study was to evaluate the LEPR gene polymorphisms Lys109Arg (A109G) and Gln223Arg (A223G) in severely pre-eclamptic women. In a case-control study, we analyzed blood samples from 124 severely pre-eclamptic patients and 107 healthy control women by the polymerase chain reaction-restriction fragment length polymorphism method. The Pearson chi2 test was used to estimate odds ratios (OR) and 95% confidence intervals (CI). The association was adjusted for maternal age, pre-pregnancy body mass index and primiparity with logistic regression analysis. Pregnant women with the LEPR 223G allele (223A/G or 223G/G genotype) had almost double the risk of developing severe pre-eclampsia compared with patients with the 223A/A genotype (adjusted OR = 1.92, 95% CI: 1.07-3.41). Genotype variants of LEPR A109G alone did not affect the risk of severe pre-eclampsia. Haplotype estimation of A109G and A223G polymorphisms of the LEPR gene revealed that the G-A haplotype versus other pooled haplotypes was significantly less common in the pre-eclamptic group (p < 0.01), while the G-G haplotype versus others was overrepresented among severely pre-eclamptic patients (p < 0.01), compared with controls. In conclusion, our data indicate that LEPR A223G polymorphism may individually modify the risk of severe pre-eclampsia. PMID:17071538

  20. Risk factors and gene polymorphisms of inflammatory bowel disease in population of Zhejiang,China

    Zi-Wei Wang; Feng Ji; Wei-Jun Teng; Xiao-Gang Yuan; Xiao-Ming Ye

    2011-01-01

    AIM:To identify the risk factors and three single nucleotide polymorphisms(SNPs)of NOD2 /CARD15 gene in inflammatory bowel disease(IBD)of the population in Zhejiang,China.METHODS:A case-control study was conducted using recall questionnaire to collect data on demographic,socioeconomic,lifestyle characteristics and dietary behaviors from 136 determined IBD patients and 136 paired healthy controls.COX regression method was used to screen the statistically significant risk factors for IBD.The polymorphisms of NOD2 /CARD15 gene Arg702Trp ,Gly908Arg and Leu1007fsinsC were genotyped and further compared between 60 patients with IBD and 60 healthy controls by polymerase chain reaction and restriction fragment length polymorphism.RESULTS:IBD occurred primarily in young and middle- aged people.The mean age for IBD patients was 42.6 years.The ratio of males to females was 1.23:1.COX regression indicated a higher statistical significance in milk,fried food and stress compared with the other postulated risk factors for IBD.None of the patients with IBD and healthy controls had heterozygous or homozygous SNPs variants.CONCLUSION:Milk,fried food and stress are associated with increased risk of IBD.The common variants in NOD2 /CARD15 gene are not associated with IBD in China's Zhejiang population.

  1. Arginine 16 Glycine Polymorphism in β2-Adrenergic Receptor Gene Is Associated with Obesity, Hyperlipidemia, Hyperleptinemia, and Insulin Resistance in Saudis

    Maha H. Daghestani

    2012-01-01

    Full Text Available Background. Several studies have shown an association between codon 16 polymorphism of the β2AR gene and obesity. Methods. We studied the association between Arg16Gly polymorphism and obesity and its influence on anthropometric parameters, lipids, insulin resistance and leptin in Saudi individuals. The study group included 329 individuals (males: 109 and females: 220. Metabolic parameters, including glucose, lipids, insulin, and leptin were analyzed and anthropometric parameters including waist and hip circumference, waist/hip (W/H ratio, and body mass index (BMI were measured and HOMA-IR was calculated. Genotyping was conducted by DNA sequencing of 353 bp fragments, carrying the Arg16Gly polymorphic site. Results and Conclusion. Overweight and obese subjects had a significantly higher frequency of Gly16 (0.375 and 0.38, resp. compared with normal-weight subjects (0.200. In addition, subjects carrying Gly16 allele regardless of their BMI had greater waist and hip circumference, W/H ratio, plasma lipids, leptin, glucose level, and insulin resistance as judged from the HOMA-IR, compared to those with the wild-type allele. The findings of this study show a significant association between the Arg16Gly polymorphism in β2AR gene and the development of insulin resistance, overweight, and obesity in Saudi populations with an influence on the levels of lipid and leptin.

  2. Beta 2-adrenergic receptor gene association with overweight and asthma in children and adolescents and its relationship with physical fitness

    Neiva Leite

    2015-12-01

    Full Text Available Objective: To investigate the association of Arg16Gly and Gln27Glu polymorphisms of β2-adrenergic receptor gene (ADRB2 with the occurrence of asthma and overweight and the gene's influence on anthropometric, clinic, biochemical and physical fitness variables in children and adolescents. Methods: Subjects were evaluated for allelic frequencies of the β2-adrenergic receptor gene, height, weight, body mass index (BMI, BMI Z-score, waist circumference (WC, pubertal stage, resting heart rate (HRres, blood pressure (BP, total cholesterol (TC, glucose, insulin, high density lipoprotein (HDL-C, low density lipoprotein (LDL-C, triglyceride (TG, Homeostasis Metabolic Assessment (HOMA2-IR, Quantitative Insulin Sensitivity Check Index (QUICKI and maximal oxygen uptake (VO2max. The participants were divided in four groups: overweight asthmatic (n=39, overweight non-asthmatic (n=115, normal weight asthmatic (n=12, and normal weight non-asthmatic (n=40. Results: Regarding the Gln27Glu polymorphism, higher total cholesterol was observed in usual genotype individuals than in genetic variant carriers (p=0.04. No evidence was found that the evaluated polymorphisms are influencing the physical fitness. The Arg16 allele was found more frequently among the normal weight asthmatic group when compared to the normal weight non-asthmatic group (p=0.02, and the Glu27 allele was more frequently found in the overweight asthmatics group when compared to the normal weight non-asthmatic group (p=0.03. Conclusions: The association of Arg16 allele with the occurrence of asthma and of the Glu27 allele with overweight asthmatic adolescents evidenced the contribution of the β2-adrenergic receptor gene to the development of obesity and asthma.

  3. Lactotransferrin gene functional polymorphisms do not influence susceptibility to human immunodeficiency virus-1 mother-to-child transmission in different ethnic groups

    Luisa Zupin

    2015-04-01

    Full Text Available Lactotransferrin, also known as lactoferrin, is an iron binding glycoprotein that displays antiviral activity against many different infectious agents, including human immunodeficiency virus (HIV-1. Lactotransferrin is present in the breast milk and in the female genitourinary mucosa and it has been hypothesised as a possible candidate to prevent mother-to-child HIV-1 transmission. To verify if two functional polymorphisms, Thr29Ala and Arg47Lys, in the lactotransferrin encoding gene (LTF could affect HIV-1 infection and vertical transmission, a preliminary association study was performed in 238 HIV-1 positive and 99 HIV-1 negative children from Brazil, Italy, Africa and India. No statistically significant association for the Thr29Ala and Arg47Lys LTF polymorphisms and HIV-1 susceptibility in the studied populations was found. Additionally LTF polymorphisms frequencies were compared between the four different ethnic groups.

  4. Superior analgesic effect of H-Dmt-D-Arg-Phe-Lys-NH2 ([Dmt1]DALDA), a multifunctional opioid peptide, compared to morphine in a rat model of neuropathic pain

    Shimoyama, Megumi; Schiller, Peter W.; Shimoyama, Naohito; Toyama, Satoshi; Szeto, Hazel H.

    2012-01-01

    H-Dmt-D-Arg-Phe-Lys-NH2 ([Dmt1]DALDA), is a synthetic tetrapeptide with extraordinary selectivity for the mu-opioid receptor and is an extremely potent analgesic. [Dmt1]DALDA is unusual in the way that the greater part of its analgesic potency appears to be produced by its actions in the spinal cord. Furthermore, [Dmt1]DALDA inhibits norepinephrine re-uptake and is a mitochondria-targeted antioxidant. Such characteristics may make [Dmt1]DALDA particularly effective against neuropathic pain. T...

  5. La contabilidad pública en las Cortes de Cádiz: la “Memoria sobre la Cuenta y Razón de España” (1811) de José Canga Argüelles.

    Fernando López Castellano

    2009-01-01

    En este trabajo se narra un intento relevante, siquiera desde el punto de vista doctrinal, de la historia institucional de la Hacienda: el ambicioso proyecto de reforma de la Contabilidad Pública llevado a cabo por las Cortes Generales y Extraordinarias instaladas en Cádiz durante la invasión napoleónica. En este capítulo poco conocido de la historia institucional de la Hacienda, destacan las ideas sobre la Cuenta y Razón expuestas por el Secretario del Despacho de Hacienda, José Canga Argüel...

  6. [Arg6,D-Trp7,9,NmePhe8]-substance P (6–11) activates JNK and induces apoptosis in small cell lung cancer cells via an oxidant-dependent mechanism

    MacKinnon, A. C.; Armstrong, R. A.; Waters, C. M.; Cummings, J.; Smyth, J. F.; Haslett, C.; Sethi, T.

    1999-01-01

    [Arg6,D-Trp7,9,NmePhe8]-substance P (6–11) (antagonist G) is a novel class of anti-cancer agent that inhibits small-cell lung cancer (SCLC) cell growth in vitro and in vivo and is entering phase II clinical investigation for the treatment of SCLC. Although antagonist G blocks SCLC cell growth (IC50 = 24.5 ± 1.5 and 38.5 ± 1.5 μM for the H69 and H510 cell lines respectively), its exact mechanism of action is unclear. This study shows that antagonist G stimulates apoptosis as assessed by morpho...

  7. Gene therapy

    2005-01-01

    2005147 CNHK200-hA-a gene-viral therapeutic system and its antitumor effect on lung cancer. WANG Wei-guo(王伟国),et al. Viral & Gene Ther Center, Eastern Hepatobilli Surg Instit 2nd Milit Univ, Shanghai 200438. Chin J Oncol,2005:27(2):69-72. Objective: To develop a novel vector system, which combines the advantages of the gene therapy,

  8. Aerobic digestion reduces the quantity of antibiotic resistance genes in residual municipal wastewater solids

    TimothyMLaPara

    2013-02-01

    Full Text Available Numerous initiatives have been undertaken to circumvent the problem of antibiotic resistance, including the development of new antibiotics, the use of narrow spectrum antibiotics, and the reduction of inappropriate antibiotic use. We propose an alternative but complimentary approach to reduce antibiotic resistant bacteria by implementing more stringent technologies for treating municipal wastewater, which is known to contain large quantities of antibiotic resistant bacteria and antibiotic resistance genes (ARGs. In this study, we investigated the ability of conventional aerobic digestion to reduce the quantity of ARGs in untreated wastewater solids. A bench-scale aerobic digester was fed untreated wastewater solids collected from a full-scale municipal wastewater treatment facility. The reactor was operated under semi-continuous flow conditions for more than 200 days at a residence time of approximately 40 days. During this time, the quantities of tet(A, tet(W, and erm(B decreased by more than 90%. In contrast, intI1 did not decrease, and tet(X increased in quantity by 5-fold. Following operation in semi-continuous flow mode, the aerobic digester was converted to batch mode to determine the first-order decay coefficients, with half-lives ranging from as short as 2.8 days for tet(W to as long as 6.3 days for intI1. These results demonstrated that aerobic digestion can be used to reduce the quantity of antibiotic resistance genes in untreated wastewater solids, but that rates can vary substantially depending on the reactor design (i.e., batch versus continuous-flow and the specific ARG.

  9. Adrenergic gene polymorphisms and cardiovascular risk in the NHLBI-sponsored Women's Ischemia Syndrome Evaluation

    Sharaf Barry L

    2008-03-01

    Full Text Available Abstract Background Adrenergic gene polymorphisms are associated with cardiovascular and metabolic phenotypes. We investigated the influence of adrenergic gene polymorphisms on cardiovascular risk in women with suspected myocardial ischemia. Methods We genotyped 628 women referred for coronary angiography for eight polymorphisms in the α1A-, β1-, β2- and β3-adrenergic receptors (ADRA1A, ADRB1, ADRB2, ADRB3, respectively, and their signaling proteins, G-protein β 3 subunit (GNB3 and G-protein α subunit (GNAS. We compared the incidence of death, myocardial infarction, stroke, or heart failure between genotype groups in all women and women without obstructive coronary stenoses. Results After a median of 5.8 years of follow-up, 115 women had an event. Patients with the ADRB1 Gly389 polymorphism were at higher risk for the composite outcome due to higher rates of myocardial infarction (adjusted hazard ratio [HR] 3.63, 95% confidence interval [95%CI] 1.17–11.28; Gly/Gly vs. Arg/Arg HR 4.14, 95%CI 0.88–19.6. The risk associated with ADRB1 Gly389 was limited to those without obstructive CAD (n = 400, Pinteraction = 0.03, albeit marginally significant in this subset (HR 1.71, 95%CI 0.91–3.19. Additionally, women without obstructive CAD carrying the ADRB3 Arg64 variant were at higher risk for the composite endpoint (HR 2.10, 95%CI 1.05–4.24 due to subtle increases in risk for all of the individual endpoints. No genetic associations were present in women with obstructive CAD. Conclusion In this exploratory analysis, common coding polymorphisms in the β1- and β3-adrenergic receptors increased cardiovascular risk in women referred for diagnostic angiography, and could improve risk assessment, particularly for women without evidence of obstructive CAD. Trial Registration ClinicalTrials.gov NCT00000554.

  10. Autoradiography study and SPECT imaging of reporter gene HSV1-tk expression in heart

    Lan Xiaoli [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China)], E-mail: LXL730724@hotmail.com; Liu Ying; He Yong; Wu Tao; Zhang Binqing; Gao Zairong; An Rui [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China); Zhang Yongxue [Department of Nuclear Medicine, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Hubei Province Key Laboratory of Molecular Imaging, Wuhan, Hubei Province, 430022 (China)], E-mail: zhyx1229@163.com

    2010-04-15

    Aim: To demonstrate the feasibility and optimal conditions of imaging herpes simplex virus 1-thymidine kinase (HSV1-tk) gene transferred into hearts with {sup 131}I-2'-fluoro-2'-deoxy-1-{beta}-D-arabinofuranosyl-5-iodouracil ({sup 131}I-FIAU) using autoradiography (ARG) and single photon emission computed tomography (SPECT) in animal models. Methods: HSV1-tk inserted into adenovirus vector (Ad5-tk) and adenovirus (Ad5-null) was prepared. Rats or rabbits were divided into a study group receiving intramyocardial injection of Ad5-tk, and a control group receiving Ad-null injection. In the study group of rats, two sets of experiments, time-course study and dose-dependence study, were performed. In time-course experiments, rats were injected with {sup 131}I-FIAU on Days 1, 2, 3, 5 and 7, after transfection of 1x10{sup 8} pfu Ad5-tk, to study the feasibility and suitable time course for reporter gene imaging. In dose-dependence study, various titers of Ad5-tk (5x10{sup 8}, 1x10{sup 8}, 5x10{sup 7} and 1x10{sup 7} pfu) were used to determine the threshold and optimal viral titer needed for detection of gene expression. The gamma counts of hearts were measured. The rat myocardium was analyzed by ARG and reverse transcriptase-polymerase chain reaction (RT-PCR). SPECT whole-body planar imaging and cardiac tomographic imaging were performed in the rabbit models. Results: From the ARG images, rats injected with Ad5-tk showed significant {sup 131}I-FIAU activity in the anterolateral wall compared with background signals seen in the control Ad5-null rats. In time-course study, the highest radioactivity in the focal myocardium could be seen on Day 1, and then progressively declined with time. In dose-dependence study, the level of {sup 131}I-FIAU accumulation in the transfected myocardium declined with the decrease of Ad viral titers. From the ARG analysis and gamma counting, the threshold viral titer was 5x10{sup 7} pfu, and the optimal Ad titer was 1x10{sup 8} pfu

  11. Hospital Effluents Are One of Several Sources of Metal, Antibiotic Resistance Genes, and Bacterial Markers Disseminated in Sub-Saharan Urban Rivers

    Laffite, Amandine; Kilunga, Pitchouna I.; Kayembe, John M.; Devarajan, Naresh; Mulaji, Crispin K.; Giuliani, Gregory; Slaveykova, Vera I.; Poté, John

    2016-01-01

    Data concerning the occurrence of emerging biological contaminants such as antibiotic resistance genes (ARGs) and fecal indicator bacteria (FIB) in aquatic environments in Sub-Saharan African countries is limited. On the other hand, antibiotic resistance remains a worldwide problem which may pose serious potential risks to human and animal health. Consequently, there is a growing number of reports concerning the prevalence and dissemination of these contaminants into various environmental compartments. Sediments provide the opportunity to reconstruct the pollution history and evaluate impacts so this study investigates the abundance and distribution of toxic metals, FIB, and ARGs released from hospital effluent wastewaters and their presence in river sediments receiving systems. ARGs (blaTEM, blaCTX-M, blaSHV, and aadA), total bacterial load, and selected bacterial species FIB [Escherichia coli, Enterococcus (ENT)] and species (Psd) were quantified by targeting species specific genes using quantitative PCR (qPCR) in total DNA extracted from the sediments recovered from 4 hospital outlet pipes (HOP) and their river receiving systems in the City of Kinshasa in the Democratic Republic of the Congo. The results highlight the great concentration of toxic metals in HOP, reaching the values (in mg kg−1) of 47.9 (Cr), 213.6 (Cu), 1434.4 (Zn), 2.6 (Cd), 281.5 (Pb), and 13.6 (Hg). The results also highlight the highest (P effluent water is not an exclusive source of the biological contaminants entering the urban rivers. Significant correlation were observed between (i) all analyzed ARGs and total bacterial load (16S rRNA) 0.51 to 0.72 (p toxic metals (Cd, Cr, Cu, and Zn) 0.44 to 0.72, (p toxic metals and biological emerging contaminants in aquatic ecosystems. PMID:27499749

  12. Association Between Polymorphisms of DNA Repair Gene XRCC1 and DNA Damage in Asbestos-Exposed Workers

    XIAO-HONG ZHAO; GUANG JIA; YONG-QUAN LIU; SHAO-WEI LIU; LEI YAN; YU JIN; NIAN LIU

    2006-01-01

    Objective To compare the asbestos-induced DNA damage and repair capacities of DNA damage between 104 asbestos exposed workers and 101 control workers in Qingdao City of China and to investigate the possible association between polymorphisms in codon 399 of XRCC1 and susceptibility to asbestosis. Methods DNA damage levels in peripheral bloodlymphocytes were determined by comet assay, and XRCC 1 genetic polymorphisms of DNA samples from 51 asbestosis cases and 53 non-asbestosis workers with a similar asbestos exposure history were analyzed by PCR/RFLP. Results The basal comet scores (3.95±2.95) were significantly higher in asbestos-exposed workers than in control workers (0.10±0.28). After 1 h H2O2 stimulation, DNA damage of lymphocytes exhibited different increases. After a 4 h repair period, the comet scores were 50.98±19.53 in asbestos-exposed workers and 18.32±12.04 in controls. The residual DNA damage (RD) was significantly greater (P<0.01) in asbestos-exposed workers (35.62%) than in controls (27.75%). XRCC1 genetic polymorphism in 104 asbestos-exposed workers was not associated with increased risk of asbestosis. But compared with polymorphisms in the DNA repair gene XRCC1 (polymorphisms in codon 399) and the DNA damage induced by asbestos, the comet scores in asbestosis cases with Gln/Gln, Gln/Arg, and Arg/Arg were 40.26±18.94, 38.03±28.22, and 32.01±11.65, respectively, which were higher than those in non-asbestosis workers with the same genotypes (25.58±11.08, 37.08±14.74, and 29.38±10.15). There were significant differences in the comet scores between asbestosis cases and non-asbestosis workers with Gln/Gln by Student's t-test (P<0.05 or 0.01). The comet scores were higher in asbestosis workers with Gln/Gln than in those with Arg/Arg and in non-asbestosis workers exposed to asbestos, but without statistically significant difference. Conclusions Exposure to asbestos may be related to DNA damage or the capacity of cells to repair H2O2-induced

  13. Correlation between leptin receptor gene polymorphisms and atherosclerosis in patients of essential hypertension%瘦素受体基因多态性与原发性高血压患者动脉粥样硬化的相关性

    李春城; 王先梅; 周靓; 叶伟

    2012-01-01

    目的:探讨两种瘦素受体(LEPR)基因多态性Gln223Arg(rs1137101)和Lys109Arg(rs1137100)与动脉粥样硬化的相关性.方法:选取原发性高血压(EH)住院患者302例(EH组)和门诊体检健康者120例(对照组),用统计学方法分析两种多态基因型、内膜-中膜厚度(IMT)与动脉粥样硬化危险因素的相关性.结果:EH组双侧颈总动脉(CCA)、颈动脉分叉处(BIF)及平均颈动脉(CBI) IMT值均高于对照组(均P<0.05);Gln223Arg AA基因型携带者有更高的颈内动脉(ICA)及CBI(均P<0.05);Lys109Arg GG基因型携带者有更低的BIF及CBI(均P<0.05).结论:LEPR基因多态性与动脉粥样硬化发生发展密切相关.%Objective: To explore the correlation between two polymorphisms of LEPR and atherosclerosis. Method:Three hundred and two inpatients with essential hypertension (EH group) and 120 health people (control group) were selected. Then we detected two kinds of genotypes, Gln223Arg and Lysl09Arg, intima-media thickness (IMT) and atherosclerosis risk factors by statistical method. Result:Compared to control group, the IMT of CCA, BIF and CBI in EH group increased (both P<0. 05). Gln223Arg polymorphismsn AA+AG homozygotes had higher ICA and CBI, while LyslO9Arg polymorphismsn GG homozygotes had lower BIF and CBI (all Pgene polymorphismare associated with atherosclerosis.

  14. Evaluation of the antinociceptive activities of the aqueous root extract of Alchornea cordifolia (Schumach and Thonn Müll. Arg. (Euphorbiaceace

    Ismaila O Ishola

    2012-08-01

    Full Text Available Summary. Alchornea cordifolia (Schumach and Thonn Müll. Arg. (Euphorbiaceace is used in traditional African medicine for the treatment of painful and inflammatory disorders. This study was carried out to investigate the antinociceptive effect of aqueous root extract of Alchornea cordifolia in mice. The antinociceptive activity of the oral doses of 100 – 400 mg/kg of Alchornea cordifolia was assessed using the acetic acid-induced mouse writhing reflex, formalin-induced paw licking, hot plate and tail clip models of pain.The extract (100–400 mg/kg produced significant (P< 0.05 dose dependent inhibition of writhing reflex as well as significant dose-dependent inhibition of the neurogenic and inflammatory pains associated with the formalin test. The formalin test result showed that the extract (100-400 mg/kg, p.o increased pain threshold in a dose dependent manner in both phases but the effect is more prominent against the inflammatory phase (second phase.This findings showed that A.cordifolia possesses both central and peripheral analgesic activity, as shown by its ability to significantly (p<0.05 inhibit nociceptive response associated with both phases. The ability of the oral doses of the extract (100-400 mg/kg to prolonged the reaction time of the animals to noxious heat in hot plate and tail clip models of pain, in a dose dependent manner suggests a central analgesic effect. Acute toxicity studies of oral doses of aqueous root extract of Alchornea cordifolia in mice revealed that it has a wide margin of safety as it well tolerated by the animals.The results of the study suggest an analgesic property demonstrated by the aqueous root extract of Alchornea cordifolia possibly mediated by central and peripheral mechanisms and thus confirm the folkloric uses of the plant in painful disorders. Industrial relevance. In the recent past there has been a growing interest in Traditional medicine/Complementary and Alternative Medicine (TCAM and their

  15. Familial dilated cardiomyopathy associated with congenital defects in the setting of a novel VCL mutation (Lys815Arg in conjunction with a known MYPBC3 variant

    Quinn S. Wells

    2011-08-01

    Full Text Available Idiopathic dilated cardiomyopathy (DCM is a primary myocardial disorder characterized by ventricular chamber enlargement and systolic dysfunction. Twenty to fifty percent of idiopathic DCM cases are thought to have a genetic cause. Of more than 30 genes known to be associated with DCM, rare variants in the VCL and MYBPC3 genes have been reported in several cases of DCM. In this report, we describe a family with DCM and congenital abnormalities who carry a novel missense mutation in the VCL gene. More severely affected family members also possess a second missense variant in MYBPC3, raising the possibility that this variant may be a disease modifier. Intere - stingly, many of the affected individuals also have congenital defects, including two with bicuspid aortic valve with aortic regurgitation. We discuss the implications of the family history and genetic information on management of at-risk individuals with aortic regurgitation.

  16. Gene therapy.

    Mota Biosca, Anna

    1992-01-01

    Applications of gene therapy have been evaluated in virtually every oral tissue, and many of these have proved successful at least in animal models. While gene therapy will not be used routinely in the next decade, practitioners of oral medicine should be aware of the potential of this novel type of treatment that doubtless will benefit many patients with oral diseases.

  17. Trichoderma genes

    Foreman, Pamela; Goedegebuur, Frits; Van Solingen, Pieter; Ward, Michael

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  18. Horizontal Transfer of Plasmid-Mediated Cephalosporin Resistance Genes in the Intestine of Houseflies (Musca domestica).

    Fukuda, Akira; Usui, Masaru; Okubo, Torahiko; Tamura, Yutaka

    2016-06-01

    Houseflies are a mechanical vector for various types of bacteria, including antimicrobial-resistant bacteria (ARB). If the intestine of houseflies is a suitable site for the transfer of antimicrobial resistance genes (ARGs), houseflies could also serve as a biological vector for ARB. To clarify whether cephalosporin resistance genes are transferred efficiently in the housefly intestine, we compared with conjugation experiments in vivo (in the intestine) and in vitro by using Escherichia coli with eight combinations of four donor and two recipient strains harboring plasmid-mediated cephalosporin resistance genes and chromosomal-encoded rifampicin resistance genes, respectively. In the in vivo conjugation experiment, houseflies ingested donor strains for 6 hr and then recipient strains for 3 hr, and 24 hr later, the houseflies were surface sterilized and analyzed. In vitro conjugation experiments were conducted using the broth-mating method. In 3/8 combinations, the in vitro transfer frequency (Transconjugants/Donor) was ≥1.3 × 10(-4); the in vivo transfer rates of cephalosporin resistance genes ranged from 2.0 × 10(-4) to 5.7 × 10(-5). Moreover, cephalosporin resistance genes were transferred to other species of enteric bacteria of houseflies such as Achromobacter sp. and Pseudomonas fluorescens. These results suggest that houseflies are not only a mechanical vector for ARB but also a biological vector for the occurrence of new ARB through the horizontal transfer of ARGs in their intestine. PMID:26683492

  19. Expression of Arc/Arg3.1 in diabetic rat hippocampus and its relationship with cognitive function%Arc/Arg3.1在糖尿病性脑病大鼠海马组织中的表达及其与认知功能改变的关系

    朱奕潼; 苗雅; 何婷; 钟远

    2014-01-01

    目的:探讨糖尿病大鼠认知功能与海马Arc蛋白和β淀粉样蛋白(Aβ)表达的相关性,探讨突触可塑性在糖尿病性脑病(DE)发病机制中的作用。方法30只SPF级雄性8周龄大鼠随机分为对照组和糖尿病组,糖尿病组高脂饮食喂养4周后腹腔注射链脲佐菌素建立2型糖尿病大鼠模型,观察不同阶段动物的体质量、空腹血糖(FBG)、空腹血清胰岛素(FSI),并计算胰岛素敏感指数(ISI)。实验末行水迷宫测试评估两组大鼠认知功能;酶联免疫吸附测定法、免疫组织化学法和蛋白质印迹法检测大鼠海马组织Aβ和Arc的表达。结果糖尿病组大鼠在目标象限的探索时间较对照组缩短(P<0.01),原平台穿越次数较对照组减少(P<0.01)。糖尿病组Aβ表达较对照组升高(P<0.01),Arc/Arg3.1表达降低(P<0.01)。结论糖尿病大鼠认知功能受损可能与Arc表达降低导致突触可塑性失衡加剧Aβ的沉积和神经元损伤有关。%Objective To analyze the correlation of cognitive function with the expression of Arc protein and amyloid-beta peptide (Aβ) in the hippocampus of diabetic rats, and to assess the role of synaptic plasticity in the possible pathogenesis of diabetic encephalopathy (DE). Methods Thirty male SPF SD rats (8 weeks old) were randomly assigned to diabetes mellitus (DM) group and control group. After 4 weeks of high fat diet feeding, the rats of DM group were injected intraperitoneally with streptozotocin at a dose of 30mg/kg to establish diabetic model. Body mass, fasting blood glucose (FBG), and fasting serum insulin (FSI) were monitored at several stages, and insulin sensitivity index (ISI) was calculated. At the end of the experiment, water maze test was carried out to evaluate the cognition of all the rats. The expression of Arc and Aβin the hippocampus was detected and observed by ELISA, immunohistochemistry and Western blotting

  20. Produção de compostos fenólicos a partir de células imobilizadas do líquen Parmotrema andinum (Müll. Arg. Hale e avaliação de atividade antimicrobiana Production of phenolic compounds from immobilized cells of the lichen Pamotrema andinum (Müll. Arg. Hale and evaluation of antimicrobial activity

    Nadejda de Azevedo Nóbrega

    2012-03-01

    Full Text Available A utilização de metabólitos secundários obtidos de líquens, na indústria farmacêutica, de cosmético, têxtil e de alimentos deve ser criteriosa, visto que a extração e isolamento desses metabólitos requerem uma grande quantidade de biomassa dificilmente renovável, devido ao crescimento lento do líquen. Atualmente, é possível obter substâncias liquênicas tanto por cultivo de tecidos, como por imobilizações celulares e enzimáticas, a partir do talo in natura, utilizando pequena quantidade de material liquênico. Portanto, este trabalho objetiva investigar a produção de compostos fenólicos a partir de células imobilizadas de Parmotrema andinum (Müll. Arg. Hale utilizando acetato de sódio como precursor da biossíntese dos fenóis. Ensaios de atividade antimicrobiana com extratos orgânicos do talo in natura, eluatos celulares e do ácido lecanórico isolado de P. andinum Hale demonstraram ação contra bactérias Gram-positivas. Através de testes biocromatográficos foi possível associar a atividade antibacteriana ao ácido lecanórico e uma substância não identificada presente na espécie. As substâncias produzidas através de imobilização celular não exibiram ação inibitória frente os microrganismos testados.Hale and evaluation of antimicrobial activity. Secondary metabolites obtained from lichens should be carefully used by the pharmaceutical, cosmetic, textile and food industries because the extraction and isolation of these metabolites requires large amounts of biomass, which is not renewable due to how slow lichens grow. Nowadays, it is possible to obtain these substances either by tissue culture or cell and enzymatic immobilizations, from an in natura thallus, using a small amount of lichenous material. Therefore, this work aimed to investigate the production of phenolic compounds from Parmotrema andinum (Müll. Arg. Hale immobilized cells, using sodium acetate as a precursor in the biosynthesis of

  1. Diâmetro de estacas e substratos na propagação vegetativa de maniçoba, Manihot glaziovii Muell. Arg Diameter of cuttings and substrates in the vegetative propagation of maniçoba Manihot glaziovii Muell. Arg

    Leonardo Elias Ferreira

    2010-09-01

    Full Text Available Objetivou-se com este trabalho estudar a influência do diâmetro das estacas e dos substratos na propagação vegetativa de maniçoba, Manihot glaziovii Muell. Arg. Utilizou-se o delineamento em blocos casualizados em esquema fatorial 10 x 3, sendo 10 substratos e 3 diâmetros de estacas, totalizando 30 tratamentos, oriundos das combinações entre substratos e diâmetros de estacas, com 4 repetições e 8 estacas por parcela. Os diâmetros das estacas corresponderam a: 0,5 a 1,0 cm (D1; 1,1 a 2,0 cm (D2 e 2,1 a 3,0 cm (D3. Foram utilizados 10 substratos: terra (S1; areia (S2; terra+areia: 1:1 (S3; terra+areia: 2:1 (S4; terra+areia: 1:2 (S5; terra+esterco: 1:1 (S6; areia+esterco: 1:1 (S7; terra+esterco: 2:1 (S8; areia+esterco: 2:1 (S9 e terra+areia+esterco: 1:1:1 (S10. As estacas lenhosas foram retiradas de plantas matrizes em repouso vegetativo, com comprimento de 25 cm e base cortada em bisel. Avaliou-se: percentual de estacas enraizadas; número de raízes; número de brotações; diâmetro e comprimento das brotações; número de folhas; massa seca das brotações e massa seca das raízes, obtidos aos 70 dias após instalação do experimento. Os melhores resultados para todas as variáveis estudadas foram obtidos com estacas de diâmetros entre 1,1 a 2,0 cm e entre 2,1 a 3,0 cm, com substratos que incluíram areia + esterco na sua composição. Tais tratamentos proporcionaram percentagens de enraizamento da ordem de 70 a 80%.This study aims to evaluate substrates and cuttings diameter influence in the vegetative propagation of Manihot glaziovii Muell.Arg. A design in randomized blocks using a 10 x 3 factorial scheme was used, being 10 substrates and 3 cuttings diameter, totaling 30 treatments, from the combinations between substrates and cuttings diameter, with 4 replications and 8 cuttings per plot. Diameters corresponded: 0.5 to 1.0 cm (D1; 1.1 to 2.0 cm (D2 and 2.1 to 3.0 cm (D3. Substrates used were: soil (S1; sand (S2; soil + sand: 1

  2. Purification and Biological Activity of Recombinant Melittin-88 ArgIL-2%重组蜂毒素-基因变构IL-2的纯化与生物活性

    刘明军; 王斌; 钱冬萌; 丁守怡; 闫志勇; 宋旭霞

    2006-01-01

    目的 制备纯化重组蜂毒素-基因变构IL-2(Melittin-88 ArgIL-2)嵌合蛋白,并检测其生物活性.方法 将含表达载体的大肠杆菌DH5α,经IPTG诱导表达.表达产物经离子交换层析与亲和层析进行纯化,SDS-PAGE鉴定,MTT染色法检测嵌合蛋白对Hela细胞增殖的抑制作用.结果 所表达的可溶性蛋白,纯化后纯度达95%,蛋白浓度0.5 g/L.纯化后的嵌合蛋白在体外能抑制Hela细胞的增殖.结论 已成功地制备并纯化了重组melittin-88 ArgIL-2嵌合蛋白,该蛋白具有生物活性,为中试放大和纯化工艺研究奠定了基础.

  3. Somatic mosaicism for the COL7A1 mutation p.Gly2034Arg in the unaffected mother of a patient with dystrophic epidermolysis bullosa pruriginosa

    van den Akker, P. C.; Pasmooij, A. M. G.; Meijer, R.; Scheffer, H.; Jonkman, M. F.

    2015-01-01

    Dystrophic epidermolysis bullosa (DEB) is a heritable blistering disorder caused by mutations in the type VII collagen gene, COL7A1. Although revertant mosaicism is well known in DEB, 'forward' somatic mosaicism, in which a pathogenic mutation arises on a wild-type (WT) background, extending beyond

  4. Polyglycerol-functionalized nanodiamond as a platform for gene delivery: Derivatization, characterization, and hybridization with DNA

    Li Zhao

    2014-03-01

    Full Text Available A gene vector consisting of nanodiamond, polyglycerol, and basic polypeptide (ND-PG-BPP has been designed, synthesized, and characterized. The ND-PG-BPP was synthesized by PG functionalization of ND through ring-opening polymerization of glycidol on the ND surface, multistep organic transformations (–OH → –OTs (tosylate → –N3 in the PG layer, and click conjugation of the basic polypeptides (Arg8, Lys8 or His8 terminated with propargyl glycine. The ND-PG-BPP exhibited good dispersibility in water (>1.0 mg/mL and positive zeta potential ranging from +14.2 mV to +44.1 mV at neutral pH in Milli-Q water. It was confirmed by gel retardation assay that ND-PG-Arg8 and ND-PG-Lys8 with higher zeta potential hybridized with plasmid DNA (pDNA through electrostatic attraction, making them promising as nonviral vectors for gene delivery.

  5. ADRB2 and LEPR gene polymorphisms: synergistic effects on the risk of obesity in Japanese.

    Pereira, Tiago V; Mingroni-Netto, Regina C; Yamada, Yoshiji

    2011-07-01

    The objective of the present study was to validate a recently reported synergistic effect between variants located in the leptin receptor (LEPR) gene and in the β-2 adrenergic receptor (ADRB2) gene on the risk of overweight/obesity. We studied a middle-aged/elderly sample of 4,193 nondiabetic Japanese subjects stratified according gender (1,911 women and 2,282 men). The LEPR Gln223Arg (rs1137101) variant as well as both ADRB2 Arg16Gly (rs1042713) and Gln27Glu (rs1042714) polymorphisms were analyzed. The primary outcome was the risk of overweight/obesity defined as BMI ≥25 kg/m(2), whereas secondary outcomes included the risk of a BMI ≥27 kg/m(2) and BMI as a continuous variable. None of the studied polymorphisms showed statistically significant individual effects, regardless of the group or phenotype studied. Haplotype analysis also did not disclose any associations of ADRB2 polymorphisms with BMI. However, dimensionality reduction-based models confirmed significant interactions among the investigated variants for BMI as a continuous variable as well as for the risk of obesity defined as BMI ≥27 kg/m(2). All disclosed interactions were found in men only. Our results provide external validation for a male specific ADRB2-LEPR interaction effect on the risk of overweight/obesity, but indicate that effect sizes associated with these interactions may be smaller in the population studied. PMID:21233812

  6. Pharmacogenetic Study in Rectal Cancer Patients Treated With Preoperative Chemoradiotherapy: Polymorphisms in Thymidylate Synthase, Epidermal Growth Factor Receptor, GSTP1, and DNA Repair Genes

    Paez, David, E-mail: dpaez@santpau.cat [Department of Medical Oncology, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Salazar, Juliana; Pare, Laia [Centre for Biomedical Network Research on Rare Diseases, Barcelona (Spain); Department of Genetics, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Pertriz, Lourdes [Department of Radiotherapy, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Targarona, Eduardo [Department of Surgery, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Rio, Elisabeth del [Centre for Biomedical Network Research on Rare Diseases, Barcelona (Spain); Department of Genetics, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Barnadas, Agusti; Marcuello, Eugenio [Department of Medical Oncology, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain); Baiget, Montserrat [Centre for Biomedical Network Research on Rare Diseases, Barcelona (Spain); Department of Genetics, Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona (Spain)

    2011-12-01

    Purpose: Several studies have been performed to evaluate the usefulness of neoadjuvant treatment using oxaliplatin and fluoropyrimidines for locally advanced rectal cancer. However, preoperative biomarkers of outcome are lacking. We studied the polymorphisms in thymidylate synthase, epidermal growth factor receptor, glutathione S-transferase pi 1 (GSTP1), and several DNA repair genes to evaluate their usefulness as pharmacogenetic markers in a cohort of 128 rectal cancer patients treated with preoperative chemoradiotherapy. Methods and Materials: Blood samples were obtained from 128 patients with Stage II-III rectal cancer. DNA was extracted from the peripheral blood nucleated cells, and the genotypes were analyzed by polymerase chain reaction amplification and automated sequencing techniques or using a 48.48 dynamic array on the BioMark system. The germline polymorphisms studied were thymidylate synthase, (VNTR/5 Prime UTR, 2R G>C single nucleotide polymorphism [SNP], 3R G>C SNP), epidermal growth factor receptor (Arg497Lys), GSTP1 (Ile105val), excision repair cross-complementing 1 (Asn118Asn, 8092C>A, 19716G>C), X-ray repair cross-complementing group 1 (XRCC1) (Arg194Trp, Arg280His, Arg399Gln), and xeroderma pigmentosum group D (Lys751Gln). The pathologic response, pathologic regression, progression-free survival, and overall survival were evaluated according to each genotype. Results: The Asterisk-Operator 3/ Asterisk-Operator 3 thymidylate synthase genotype was associated with a greater response rate (pathologic complete remission and microfoci residual tumor, 59% in Asterisk-Operator 3/ Asterisk-Operator 3 vs. 35% in Asterisk-Operator 2/ Asterisk-Operator 2 and Asterisk-Operator 2/ Asterisk-Operator 3; p = .013). For the thymidylate synthase genotype, the median progression-free survival was 103 months for the Asterisk-Operator 3/ Asterisk-Operator 3 patients and 84 months for the Asterisk-Operator 2/ Asterisk-Operator 2 and Asterisk-Operator 2/ Asterisk

  7. Pharmacogenetic Study in Rectal Cancer Patients Treated With Preoperative Chemoradiotherapy: Polymorphisms in Thymidylate Synthase, Epidermal Growth Factor Receptor, GSTP1, and DNA Repair Genes

    Purpose: Several studies have been performed to evaluate the usefulness of neoadjuvant treatment using oxaliplatin and fluoropyrimidines for locally advanced rectal cancer. However, preoperative biomarkers of outcome are lacking. We studied the polymorphisms in thymidylate synthase, epidermal growth factor receptor, glutathione S-transferase pi 1 (GSTP1), and several DNA repair genes to evaluate their usefulness as pharmacogenetic markers in a cohort of 128 rectal cancer patients treated with preoperative chemoradiotherapy. Methods and Materials: Blood samples were obtained from 128 patients with Stage II-III rectal cancer. DNA was extracted from the peripheral blood nucleated cells, and the genotypes were analyzed by polymerase chain reaction amplification and automated sequencing techniques or using a 48.48 dynamic array on the BioMark system. The germline polymorphisms studied were thymidylate synthase, (VNTR/5′UTR, 2R G>C single nucleotide polymorphism [SNP], 3R G>C SNP), epidermal growth factor receptor (Arg497Lys), GSTP1 (Ile105val), excision repair cross-complementing 1 (Asn118Asn, 8092C>A, 19716G>C), X-ray repair cross-complementing group 1 (XRCC1) (Arg194Trp, Arg280His, Arg399Gln), and xeroderma pigmentosum group D (Lys751Gln). The pathologic response, pathologic regression, progression-free survival, and overall survival were evaluated according to each genotype. Results: The ∗3/∗3 thymidylate synthase genotype was associated with a greater response rate (pathologic complete remission and microfoci residual tumor, 59% in ∗3/∗3 vs. 35% in ∗2/∗2 and ∗2/∗3; p = .013). For the thymidylate synthase genotype, the median progression-free survival was 103 months for the ∗3/∗3 patients and 84 months for the ∗2/∗2 and ∗2/∗3 patients (p = .039). For XRCC1 Arg399Gln SNP, the median progression-free survival was 101 months for the G/G, 78 months for the G/A, and 31 months for the A/A patients (p = .048). Conclusions: The thymidylate

  8. Thyroglobulin Gene Mutation with Cold Nodule on Thyroid Scintigraphy

    Toshio Kahara

    2012-01-01

    Full Text Available Thyroglobulin gene mutation is a rare cause of congenital hypothyroidism, but thyroglobulin gene mutations are thought to be associated with thyroid cancer development. A 21-year-old Japanese man treated with levothyroxine for congenital hypothyroidism had an enlarged thyroid gland with undetectable serum thyroglobulin despite elevated serum TSH level. The patient was diagnosed with thyroglobulin gene mutation, with compound heterozygosity for Gly304Cys missense mutation and Arg432X nonsense mutation. Ultrasonography showed a hypovascular large tumor in the left lobe that appeared as a cold nodule on thyroid scintigraphy. He underwent total thyroidectomy, but pathological study did not reveal findings of thyroid carcinoma, but rather a hyperplastic nodule with hemorrhage. Strong cytoplasmic thyroglobulin immunostaining was observed, but sodium iodide symporter immunostaining was hardly detected in the hyperplastic nodule. The clinical characteristics of patients with thyroglobulin gene mutations are diverse, and some patients are diagnosed by chance on examination of goiter in adults. The presence of thyroid tumors that appear as cold nodules on thyroid scintigraphy should consider the potential for thyroid carcinoma, if the patient has relatively low serum thyroglobulin concentration in relation to the degree of TSH without thyroglobulin autoantibody.

  9. Association between variants in the genes for leptin, leptin receptor, and proopiomelanocortin with chronic heart failure in the Czech population.

    Bienertová-Vasků, Julie Anna; Spinarová, Lenka; Bienert, Petr; Vasků, Anna

    2009-03-01

    Patients with chronic heart failure (CHF) express enhanced catabolic metabolism finally resulting in overall weight loss, whereas adipokines might play a crucial role in signaling among tissues. The aim of this study was to investigate the possible associations of defined variability in leptin (dbSNP ID rs7799039), proopiomelanocortin (dbSNP ID rs3754860 and dbSNP ID rs1009388), and leptin receptor gene (dbSNP rs1137101) with CHF and evaluate their potential as the CHF susceptibility genes. The case-control study comprised a total of 372 patients of Caucasian origin with chronic heart failure (New York Heart Association [NYHA] functional classes II-IV, ejection fraction (EF) <40%) and 407 healthy controls. They were genotyped for the leptin (LEP) -2548 G/A, leptin receptor (LEPR) Gln223Arg, and proopiomelanocortin (POMC) RsaI (5'-untranslated region) and C1032G variants (intron 1) using PCR-based methodology. No case-control differences in genotype as well as allele frequencies were observed between CHF patients and controls. We constructed POMC RsaI/C1032G haplotypes, having found no significant association with body mass index (BMI), left ventricle ejection fraction (LVEF), left ventricle hypertrophy (LVH) and diabetes mellitus (DM). Multivariate regression analyses revealed an approximately 2-fold risk for NYHA class IV associated with the LEPR Gln223Arg (P = 0.0000001, odds ratio [OR] = 2.10, 95% confidence interval [CI] = 1.56-2.84); it also displayed an independent prediction role for LVEF in heart failure cases of all etiologies (P = 0.002, OR = 4.05, 95% CI = 1.36-10.06). In subanalyses according to CHF etiology the LEPR Gln223Arg showed an independent prediction role for NYHA IV in IHD patients (P = 0.0001, OR = 2.50, 95% CI = 1.69-3.82) and both for NYHA IV(P = 0.007, OR = 2.04, 95% CI = 1.20-3.84) and LVEF (P = 0.004, OR = 11.87, 95% CI = 2.08-55.6) in DCMP patients. The role of the polymorphic variants in the genes encoding for adipokines as potential

  10. [Language gene].

    Takahashi, Hiroshi

    2006-11-01

    The human capacity for acquiring speech and language must derive, at least in part, from the genome. Recent advance in the field of molecular genetics finally discovered 'Language Gene'. Disruption of FOXP2 gene, the firstly identified 'language gene' causes severe speech and language disorder. To elucidate the anatomical basis of language processing in the brain, we examined the expression pattern of FOXP2/Foxp2 genes in the monkey and rat brains through development. We found the preferential expression of FOXP2/Foxp2 in the striosomal compartment of the developing striatum. Thus, we suggest the striatum, particularly striosomal system may participate in neural information processing for language and speech. Our suggestion is consistent with the declarative/ procedural model of language proposed by Ullman (1997, 2001), which the procedural memory-dependent mental grammar is rooted in the basal ganglia and the frontal cortex, and the declarative memory-dependent mental lexicon is rooted in the temporal lobe. PMID:17432197

  11. Gene Silencing

    Kertbundit, Sunee; Juříček, Miloslav; Hall, T.C.

    Dordrecht : Springer, 2010 - (Jain, S.; Brar, D.), s. 631-652 ISBN 978-90-481-2966-9 Institutional research plan: CEZ:AV0Z50380511 Keywords : Gene Silencing * RISC complex Subject RIV: EB - Genetics ; Molecular Biology

  12. KOBİ'lerde teknolojik ar-ge çalışmalarının istihdam üzerine etkileri : Tekmer'lerde bir uygulama =The effects of technological research and development at smes on employment: a practice at Tekmers

    Eker, Serhan

    2011-01-01

    KOBİ'lerde Teknolojik AR-GE Çalışmalarının istihdam Üzerine Etkileri: TEKMER'lerde Bir Uygulama. isimli çalışmada AR-GE faaliyetlerinin KOBİ'lerde yarattığı değişimin görülmesi ve TEKMER'lerde desteklenen AR-GE projelerinin işletmenin büyümesi ve istihdamın geliştirilmesinde yaratılan etkilerin değerlendirilmesi amacı ile hazırlanmıştır. Burada uygulamaya katılan işletmelerin tamamı KOBİ'lerden oluşmakla birlikte uygulama yeri ülkemizdeki TEKMER'ler olarak seçilmiştir. Seçim yerinin nedeni TE...

  13. Atypical radiological findings in achondroplasia with uncommon mutation of the fibroblast growth factor receptor-3 (FGFR-3) gene (Gly to Cys transition at codon 375)

    Nishimuri, Gen [Dokkyo Univ. School of Medicine, Tochigi (Japan); Fukushima, Yoshimitsu; Ohashi, Hirofumi [Saitama Children`s Medical Center, Saitama (Japan); Ikegawa, Shiro [Tokyo Univ. (Japan)

    1995-11-20

    The recent discovery of mutations in the FGFR-3 (fibroblast growth factor receptor-3) gene (FGFR3) as the cause of achondroplasia has provided new insight into understanding genetic diseases. It was surprising from the viewpoint of molecular genetics that most patients with achondroplasia showed the same mutation at nucleotide 1138, leading to a single amino acid substitution from glycine to arginine at codon 380 (Gly380Arg). All 39 patients examined by two groups had the Gly380Arg; 38 patients and the other demonstrated a G to A and a G to C transition at nucleotide 1138, respectively. Subsequently another group disclosed a G to A transition at the same nucleotide 1138 in 21/23 patients of diverse ethnic origin, although mutations were not identified in two patients. To date, a total of 193 patients with the mutation of the G380Arg have been reported; a single patient with another mutation resulting in a substitution from glycine to cysteine at codon 375 (Gly375Cys) has been described. The presence of this common mutation is consistent with the clinical fact that achondroplastic individuals show less phenotypic variability than is unusual for autosomal dominant diseases. We encountered a Japanese boy with the Gly375Cys. His mother with achondroplasia has the same mutation. The molecular investigation of these patients was reported elsewhere. Here we report the clinical and radiological findings in this boy who demonstrated some atypical manifestations from those of typical achondroplasia. 8 refs., 1 fig.

  14. XRCC1 Arg399Gln and clinical outcome of platinum-based treatment for advanced non-small cell lung cancer:a meta-analysis in 17 studies

    Jian CHEN; Qing-wei ZHAO; Gen-ming SHI; Lin-run WANG

    2012-01-01

    Objective:XRCC1 polymorphism is a research hotpot in individual treatment for non-small cell lung cancer (NSCLC).To obtain the association between XRCC1 polymorphism and clinical outcome of platinum-based treatment for NSCLC,a meta-analysis was conducted.Methods:Databases including PubMed,Embase,Cochrane,and Chinese National Knowledge Infrastructure (CNKI) were searched for publications that met the inclusion criteria.A fixed effect model was used to estimate pooled odds ratio (OR) and hazard ratio (HR) with 95% confidence interval (CI) for the association between XRCC1 Arg399Gln and response or survival of platinum-based treatment for advanced NSCLC.A chi-squared-based Q-test was used to test the heterogeneity hypothesis.Egger's test was used to check publication bias.Results:Seventeen published case-control studies that focus on the association between XRCC1 Arg399GIn and response or survival of platinum-based treatment for advanced NSCLC in 2 256 subjects were included in this meta-analysis,of whom 522 were AA genotypes (23.2% frequency),916 AG genotypes (40.6% frequency),and 818 GG genotypes (36.2% frequency).The overall response rate (ORR) was 45.2% (110/243) for AA genotype patients,29.9% for AG genotype (73/244),and 30.7% for GG genotype (124/403).The heterogeneity test did not show any heterogeneity and the Egger's test did not reveal an obvious publication bias among the included studies.The meta-analysis indicated that AA genotype patients presented higher response rates toward platinum drug treatment compared with G model (GG+GA) patients (GG vs.AA model:OR=0.489,95% CI 0.266-0.900,P=0.021;AG vs.AA model:OR=0.608,95% CI 0.392-0.941,P=0.026; GA+AA vs.GG model:OR=1.259,95% CI 0.931-1.701,P=0.135; GG+GA vs.AA model:OR=0.455,95% CI 0.313-0.663,P=0.0001).However,no evidence validates XRCC1 associates with the survival following platinum drug therapy.Conclusions:Our meta-analysis suggested that XRCC1 Arg399GIn is related with the

  15. 壮药白楸的生药学及毒理学初步研究%Preliminary study on the pharmacognosy and toxicology of Mallotus panicutus (lam) Muell.Arg

    黄婧; 马健雄

    2014-01-01

    目的 对大戟科植物白楸进行鉴别及急性毒性试验,确保用药安全.方法 采集药材标本并对其进行鉴别;白楸醇提取物以最大浓度及最大给药体积对实验组小鼠灌胃给药,观察14 d内不良反应情况.结果 化学反应结果显示白楸含有黄酮、皂苷等成分;急性毒性试验观察中未发现任何急性毒性反应.给药后14 d,空白组和药物组小鼠体质量增长率分别为73.5%和62.9% (P >0.05).结论 该实验结果可为白楸临床用药指导和制定质量标准提供参考依据.%Objective To identify Mallotus panicutus (lam) Muell.Arg and explore the acute toxicity.Methods Pharmacognostic identification and chemical reaction methods were adopted to the identification.The maximum concentration and maximum volume of ethanol extracts of M.panicutus (lam) Muell.Arg were given via intragastric administration in mice to observe the toxic and side effects after 14 days.Results The chemical reaction showed M.panicutus(lam) Muell.Arg concluded the constituents such as flavone and saponin and there was no acute toxicity in mice.After 14 days,the growth rate of mice weight of blank group and drug group were 73.5% and 62.9% (P > 0.05).Conclusion The results could be taken as the reference for analyzing the quality of this drug and guiding the clinical medication.

  16. Antibiotic resistance gene abundances associated with waste discharges to the Almendares River near Havana, Cuba.

    Graham, David W; Olivares-Rieumont, Susana; Knapp, Charles W; Lima, Lazaro; Werner, David; Bowen, Emma

    2011-01-15

    Considerable debate exists over the primary cause of increased antibiotic resistance (AR) worldwide. Evidence suggests increasing AR results from overuse of antibiotics in medicine and therapeutic and nontherapeutic applications in agriculture. However, pollution also can influence environmental AR, particularly associated with heavy metal, pharmaceutical, and other waste releases, although the relative scale of the "pollution" contribution is poorly defined, which restric