Genome-wide association and Meta-analysis of age at onset in Parkinson Disease

Background and Objectives Considerable heterogeneity exists in the literature concerning genetic determinants of the age at onset (AAO) of Parkinson disease (PD), which could be attributed to a lack of well-powered replication cohorts. The previous largest genome-wide association studies (GWAS) identified SNCA and TMEM175 loci on chromosome (Chr) 4 with a significant influence on the AAO of PD; these have not been independently replicated. This study aims to conduct a meta-analysis of GWAS of PD AAO and validate previously observed findings in worldwide populations. Methods A meta-analysis was performed on PD AAO GWAS of 30 populations of predominantly European ancestry from the Comprehensive Unbiased Risk Factor Assessment for Genetics and Environment in Parkinson's Disease (COURAGE-PD) Consortium. This was followed by combining our study with the largest publicly available European ancestry dataset compiled by the International Parkinson Disease Genomics Consortium (IPDGC). Results The COURAGE-PD Consortium included a cohort of 8,535 patients with PD (91.9%: Europeans and 9.1%: East Asians). The average AAO in the COURAGE-PD dataset was 58.9 years (SD = 11.6), with an underrepresentation of females (40.2%). The heritability estimate for AAO in COURAGE-PD was 0.083 (SE = 0.057). None of the loci reached genome-wide significance (p < 5 × 10−8). Nevertheless, the COURAGE-PD dataset confirmed the role of the previously published TMEM175 variant as a genetic determinant of the AAO of PD with Bonferroni-corrected nominal levels of significance (p < 0.025): (rs34311866: β(SE)COURAGE = 0.477(0.203), pCOURAGE = 0.0185). The subsequent meta-analysis of COURAGE-PD and IPDGC datasets (Ntotal = 25,950) led to the identification of 2 genome-wide significant association signals on Chr 4, including the previously reported SNCA locus (rs983361: β(SE)COURAGE+IPDGC = 0.720(0.122), pCOURAGE+IPDGC = 3.13 × 10−9) and a novel BST1 locus (rs4698412: β(SE)COURAGE+IPDGC = −0.526(0.096), pCOURAGE+IPDGC = 4.41 × 10−8). Discussion Our study further refines the genetic architecture of Chr 4 underlying the AAO of the PD phenotype through the identification of BST1 as a novel AAO PD locus. These findings open a new direction for the development of treatments to delay the onset of PD

Streptococcus mutans GlnK protein: an unusual PII family member

Streptococcus mutans is a Gram-positive bacterium present in the oral cavity, and is considered to be one of the leading causes of dental caries. S. mutans has a glnK gene, which codes for a PII-like protein that is possibly involved in the integration of carbon, nitrogen and energy metabolism in several organisms. To characterize the GlnK protein of S. mutans, the glnK gene was amplified by PCR, and cloned into the expression vectors pET29a(+) and pET28b(+). The native GlnK-Sm was purified by anion exchange (Q-Sepharose) and affinity (Hi-Trap Heparin) chromatography. The GlnK-His-Sm protein was purified using a Hi-Trap Chelating-Ni2+ column. The molecular mass of the GlnK-His-Sm proteins was 85 kDa as determined by gel filtration, indicating that this protein is a hexamer in solution. The GlnK-His-Sm protein is not uridylylated by the Escherichia coli GlnD protein. The activities of the GlnK-Sm and GlnK-His-Sm proteins were assayed in E. coli constitutively expressing the Klebsiella pneumoniae nifLA operon. In K. pneumoniae, NifL inhibits NifA activity in the presence of high ammonium levels and the GlnK protein is required to reduce the inhibition of NifL in the presence of low ammonium levels. The GlnK-Sm protein was unable to reduce NifL inhibition of NifA protein. Surprisingly, the GlnK-His-Sm protein was able to partially reduce NifL inhibition of the NifA protein under nitrogen-limiting conditions, in a manner similar to the GlnK protein of E. coli. These results suggested that S. mutans GlnK is functionally different from E. coli PII proteins

The Streptococcus mutans GlnR protein exhibits an increased affinity for the glnRA operon promoter when bound to GlnK

The control of nitrogen metabolism in pathogenic Gram-positive bacteria has been studied in a variety of species and is involved with the expression of virulence factors. To date, no data have been reported regarding nitrogen metabolism in the odontopathogenic species Streptococcus mutans. GlnR, which controls nitrogen assimilation in the related bacterial species, Bacillus subtilis, was assessed in S. mutans for its DNA and protein binding activity. Electrophoretic mobility shift assay of the S. mutans GlnR protein indicated that GlnR binds to promoter regions of the glnRA and amtB-glnK operons. Cross-linking and pull-down assays demonstrated that GlnR interacts with GlnK, a signal transduction protein that coordinates the regulation of nitrogen metabolism. Upon formation of this stable complex, GlnK enhances the affinity of GlnR for the glnRA operon promoter. These results support an involvement of GlnR in transcriptional regulation of nitrogen metabolism-related genes and indicate that GlnK relays information regarding ammonium availability to GlnR

Use Of A Lactic Acid Plus Lactoserum Intimate Liquid Soap For External Hygiene In The Prevention Of Bacterial Vaginosis Recurrence After Metronidazole Oral Treatment [uso Do ácido Láctico Com Lactoserum Em Sabonete Líquido íntimo Para Higiene Externa Na Prevenção Da Recorrência De Vaginose Bacteriana Após Tratamento Oral Com Metronidazol]

Objective: To determine the recurrence of bacterial vaginosis (BV) after the use of a lactic acid plus lactoserum liquid soap starting immediately after the treatment with oral metronidazole and the quality of life of the participants. Methods: A total of 123 women with diagnosis of BV with at least three of the following criteria: 1) homogeneous vaginal discharge without inflammation of the vagina or vulva; 2) vaginal pH > 4.5; 3) positive Whiff test; and 4) clue cells in more than 20% of the epithelial cells in the vagina. A Nugent score > 4 in the vaginal bacterioscopy was also used. After BV diagnosis, metronidazole 500 mg was administered orally bid during 7 days. Patients cured of BV were then instructed to use 7.5 to 10 mL of a lactic acid plus lactoserum liquid soap once-a-day for hygiene of the external genital region. Three subsequent control visits after starting the hygiene treatment (30, 60, and 90 days; ± 5 days) were scheduled. A questionnaire was applied in the form of visual analogue scale (VAS) in all the visits regarding: 1) level of comfort at the genital region; 2) malodorous external genitalia; 3) comfort in sexual intercourse; 4) satisfaction with intimate hygiene; and 5) self-esteem. Results: Ninety two (74.8%) women initiated the use of a lactic acid plus lactoserum liquid soap at visit 1. At visit 2, 3, and 4 there were 84, 62 and 42 women available for evaluation, respectively. The rate of recurrence of BV was 19.0%, 24.2% and 7.1%, respectively in the three visits and vaginal candidiasis was observed in five treated women. Quality of life was evaluated in the 42 women who completed the four visits schedule and there were significant improvement in the five domains assessed. Conclusion: A lactic acid plus lactoserum liquid soap for external intimate hygiene may be an option for the prevention of BV recurrence after treatment and cure with oral metronidazole. © 2011 Elsevier Editora Ltda.574415420Hillier, S., Holmes, K.K., Bacterial vaginosis (1999) Sexually Transmitted Diseases, pp. 563-586. , In: Holmes KK, Mardh PA, Sparling PF, Lemon SM, Stamm WE, Piot P et al., editors, 3 rd ed. New York: McGraw-HillSpiegel, C.A., Gardnerella vaginalis and Mobiluncus Species (2000) Principles and Practice of Infectious Diseases, 2, pp. 2383-2386. , In: Mandell GL, Bennett JE, Dolin R, editors, 5 th Philadelphia: Churchill LivingstoneMartin, H.L., Richardson, B.A., Nyange, P.M., Lavreys, L., Hiller, S.L., Chohan, B., Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition (1999) J Infect Dis, 180, pp. 1863-1868Guidelines for treatment of sexually transmitted diseases (1998) MMWR Morb Mortal Wkly Rep, 47 (RR-1), pp. 70-79. , Centers for Disease Control and Prevention, 1998Livengood, C.H., Soper, D.E., Sheehan, K.L., Fenner, D.E., Martens, M.G., Nelson, A.L., Comparison of once-daily and twice-daily dosing of 0.75% metronidazole gel in the treatment of bacterial vaginosis (1999) Sex Transm Dis, 26, pp. 137-142Sobel, J.D., Schmitt, C., Meriwether, C., Long-term follow-up of patients with bacterial vaginosis treated with oral metronidazole and topical clindamycin (1993) J Infect Dis, 167, pp. 783-784Boris, J., Pahlson, C., Larsson, P.G., Six years observation after successful treatment of bacterial vaginosis (1997) Infect Dis Obstet Gynecol, 5, pp. 297-302Ferris, D.G., Francis, S.L., Dickman, E.D., Miler-Miles, K., Waller, J.L., McClendon, N., Variability of vaginal pH determination by patients and clinicians (2006) J Am Board Fam Med, 19, pp. 368-373Soper, D.E., Gynecologic complications of bacterial vaginosis: Fact or fiction? (1999) Curr Infect Dis Rep, 1, pp. 393-397Andersch, B., Forssman, L., Lincoln, K., Torstensson, P., Treatment of bacterial vaginosis with an acid cream: A comparison between the effect of lactate-gel and metronidazole (1986) Gynecol Obstet Invest, 21, pp. 19-25Simoes, J.A., Discacciati, M.G., Brolazo, E.M., Portugal, P.M., Dini, D.V., Dantas, M.C., Clinical diagnosis of bacterial vaginosis (2006) Int J Gynaecol Obstet, 94, pp. 28-32Amsel, R., Totten, P.A., Spiegel, C.A., Chen, K.C.S., Eschenbach, D., Holmes, K.K., Non-specific vaginitis. Diagnostic criteria and microbial and epidemiological associations (1983) Am J Med, 74, pp. 14-22Nugent, R.P., Krohn, M.A., Hillier, S.L., Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation (1991) J Clin Microbiol, 29, pp. 297-301Bradshaw, C.S., Morton, A.N., Hocking, J., Garland, S.M., Morris, M.B., Moss, L.M., High recurrence rates of bacterial vaginosis over the course of 12 months after oral metronidazole therapy and factors associated with recurrence (2006) J Infect Dis, 193, pp. 1478-1486Bradshaw, C.S., Tabrizi, S.N., Fairley, C.K., Morton, A.N., Rudland, E., Garland, S.M., The association of Atopobium vaginae and Gardnerella vaginalis with bacterial vaginosis and recurrence after oral metronidazole therapy (2006) J Infect Dis, 194, pp. 828-836Gray, R.H., Wabwire-Mangen, F., Kigozi, G., Serwadda, D., Moulton, L.H., Quinn, T.C., Randomized trial of presumptive sexually transmitted disease therapy during pregnancy in Rakai, Uganda (2001) Am J Obstet Gynecol, 185, pp. 1209-1217Camargo, R.P., Simões, J.A., Cecatti, J.G., Alves, V.M., Faro, S., Impact of treatment for bacterial vaginosis on prematurity among Brazilian pregnant women: A retrospective cohort study (2005) São Paulo Med J, 123, pp. 108-112Hay, P., Recurrent bacterial vaginosis (2009) Curr Opin Infect Dis, 22, pp. 82-86Alfonsi, G.A., Shlay, J.C., Parker, S., Neher, J.O., What is the best approach for managing recurrent bacterial vaginosis? (2004) J Fam Pract, 53, pp. 650-652CDC Sexually Transmitted Disease Treatment Guidelines 2006 (2006) MMWR Morb Mortal Wkly Rep, 55, pp. 50-52. , 2006(2006) National Guideline For the Management of Bacterial Vaginosis, , http://www.guide-line.gov, Clinical Effectiveness Group British Association for Sexual Health and HIV, cited 2009 oct. 22. Available fromSobel, J.D., Ferris, D., Schwebke, J., Nyirjesy, P., Wiesenfeld, H.C., Peipert, J., Suppressive antibacterial therapy with 0.75% metronidazole vaginal gel to prevent recurrent bacterial vaginosis (2006) Am J Obstet Gynecol, 194, pp. 1283-1289Holley, R.L., Richter, H.E., Varner, R.E., Pair, L., Schwebke, J.R., A randomized, double-blind clinical trial of vaginal acidification versus placebo for the treatment of symptomatic bacterial vaginosis (2004) Sex Transm Dis, 31, pp. 236-238Boeke, A.J., Dekker, J.H., van Eijk, J.T., Kostense, P.J., Bezemer, P.D., Effect of lactic acid suppositories compared with oral metronidazole and placebo in bacterial vaginosis: A randomized clinical trial (1993) Genitourin Med, 69, pp. 388-392Decena, D.C.D., Co, J.T., Manalastas, R.M., Palaypayon, E.P., Padolina, C.S., Sison, J.M., Metronidazole with Lactacyd vaginal gel in bacterial vaginosis (2006) J Obstet Gynaecol Res, 32, pp. 243-251Ness, R.B., Kip, K.E., Soper, D.E., Stamm, C.A., Rice, P., Richter, H.E., Variability of bacterial vaginosis over 6 to 12-month intervals (2006) Sex Transm Dis, 33, pp. 381-385Ferraz do Lago, R., Simões, J.A., Bahamondes, L., Camargo, R.P., Perrotti, M., Monteiro, I., Follow-up of users of intrauterine device with and without bacterial vaginosis and other cervicovaginal infections (2003) Contraception, 68, pp. 105-109Guaschino, S., Benvenuti, C., SOPHY project: An observational study of vaginal pH, lifestyle and correct intimate hygiene in women of different ages and in different physiopathological conditions (2008) Minerva Ginecol, 60, pp. 353-362. , SOPHY Study Grou

Mendelian randomisation study of smoking, alcohol, and coffee drinking in relation to Parkinson’s Disease

Background:Previous studies showed that lifestyle behaviors (cigarette smoking, alcohol, coffee) are inversely associated with Parkinson’s disease (PD). The prodromal phase of PD raises the possibility that these associations may be explained by reverse causation. Objective:To examine associations of lifestyle behaviors with PD using two-sample Mendelian randomisation (MR) and the potential for survival and incidence-prevalence biases. Methods:We used summary statistics from publicly available studies to estimate the association of genetic polymorphisms with lifestyle behaviors, and from Courage-PD (7,369 cases, 7,018 controls; European ancestry) to estimate the association of these variants with PD. We used the inverse-variance weighted method to compute odds ratios (ORIVW) of PD and 95%confidence intervals (CI). Significance was determined using a Bonferroni-corrected significance threshold (p = 0.017). Results:We found a significant inverse association between smoking initiation and PD (ORIVW per 1-SD increase in the prevalence of ever smoking = 0.74, 95%CI = 0.60–0.93, p = 0.009) without significant directional pleiotropy. Associations in participants ≤67 years old and cases with disease duration ≤7 years were of a similar size. No significant associations were observed for alcohol and coffee drinking. In reverse MR, genetic liability toward PD was not associated with smoking or coffee drinking but was positively associated with alcohol drinking. Conclusion:Our findings are in favor of an inverse association between smoking and PD that is not explained by reverse causation, confounding, and survival or incidence-prevalence biases. Genetic liability toward PD was positively associated with alcohol drinking. Conclusions on the association of alcohol and coffee drinking with PD are hampered by insufficient statistical power