A novel pathogenic pathway of immune activation detectable before clinical onset in Huntington's disease

Huntington's disease (HD) is an inherited neurodegenerative disorder characterized by both neurological and systemic abnormalities. We examined the peripheral immune system and found widespread evidence of innate immune activation detectable in plasma throughout the course of HD. Interleukin 6 levels were increased in HD gene carriers with a mean of 16 years before the predicted onset of clinical symptoms. To our knowledge, this is the earliest plasma abnormality identified in HD. Monocytes from HD subjects expressed mutant huntingtin and were pathologically hyperactive in response to stimulation, suggesting that the mutant protein triggers a cell-autonomous immune activation. A similar pattern was seen in macrophages and microglia from HD mouse models, and the cerebrospinal fluid and striatum of HD patients exhibited abnormal immune activation, suggesting that immune dysfunction plays a role in brain pathology. Collectively, our data suggest parallel central nervous system and peripheral pathogenic pathways of immune activation in HD

Cytoprotective Effect of Lactobacillus crispatus CTV-05 against Uropathogenic E. coli

The vaginal flora consists of a subset of different lactic acid producing bacteria, typically creating a hostile environment for infecting pathogens. However, the flora can easily be disrupted, creating a favorable milieu for uropathogenic Escherichia coli (UPEC), making it possible to further infect the urinary system via the urethra. Probiotic use of different lactobacilli to restore the normal flora of the vagina has been proposed as a potential prophylactic treatment against urinary tract infections. This project evaluated the protective- and anti-inflammatory roles of the probiotic Lactobacillus crispatus strain CTV-05 in an in vitro system. The inflammatory response and the cytotoxic effect were studied by Enzyme-linked immunosorbent assays and by trypan blue exclusion of cells inoculated with L. crispatus CTV-05 and comparing it to non-infected controls and UPEC infected cells. L. crispatus CTV-05 showed no cytotoxicity to vaginal epithelial cells compared to non-infected controls and provided significant protection against UPEC infection (p < 0.05). Further more, L. crispatus CTV-05 did not create a pro-inflammatory response in vitro, with no significant increase of IL-1β or IL-6. These results demonstrate the protective effect of using L. crispatus CTV-05 as a probiotic treatment to reduce the risk of recurrent urinary tract infections

Evaluation of the Genotoxicity of Aqueous Extracts of Ilex paraguariensis St. Hil. (Aquifoliaceae) Using the Allium Test

Infusions and concoctions of Ilex paraguariensis are used as medicinal, nutritional and stimulant beverages in southern South America. Crop production is about 300,000 tons/year in Argentina, where the consumption rate reaches 5–9 kg/capita/year. In this study, we assessed the cytotoxicity of aqueous extracts of I. paraguariensis in the Allium test and Artemia salina microwell test. The extracts were prepared from commercial and “home processed” (laboratory) and were tested at concentrations of 5–40 g/l. Both extracts significantly decreased root growth and the mitotic index (MI). These effects were greater for the commercial material for which concentrations ≥10 g/l virtually abolished mitosis. The disturbance of mitotic behaviour was significant at 5–10 g/l of the “home-processed” product and included c-mitotic phenomena (over-condensed and disorganized metaphases, sticky metaphases, arrested anaphases, binucleated interphases) which could contribute to the increase in metaphase and anaphase indexes. None of the extracts were cytotoxic in the Artemia salina test. It is concluded that the Allium test is adequate for a preliminary screening of genotoxicity of medicinal plants and that genotoxic effects can be increased by the commercial manipulation of the raw product.Fil: Bidau, Claudio Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Amat, Anibal Gumersindo. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Yajia, Marta Esther. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Marti, Dardo Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Riglos, Ana G.. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Silvestroni, Aurelio. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentin

Characterization of the melA Locus for α-Galactosidase in Lactobacillus plantarum

Alpha-galactosides are abundant sugars in legumes such as soy. Because of the lack of α-galactosidase (α-Gal) in the digestive tract, humans are unable to digest these sugars, which consequently induce flatulence. To develop the consumption of the otherwise highly nutritional soy products, the use of exogenous α-Gal is promising. In this framework, we characterized the melA gene for α-Gal in Lactobacillus plantarum. The melA gene encodes a cytoplasmic 84-kDa protein whose enzymatically active form occurs as oligomers. The melA gene was cloned and expressed in Escherichia coli, yielding an active α-Gal. We show that melA is transcribed from its own promoter, yielding a monocistronic mRNA, and that it is regulated at the transcriptional level, i.e., it is induced by melibiose but is not totally repressed by glucose. Posttranscriptional regulation by the carbon source could also occur. Upstream of melA, a putative galactoside transporter, designated RafP, was identified that shows high homology to LacS, the unique transporter for both α- and β-galactosides in Streptococcus thermophilus. rafP is also expressed as a monocistronic mRNA. Downstream of melA, the lacL and lacM genes were identified that encode a heterodimeric β-galactosidase. A putative galM gene identified in the same cluster suggests the presence of a galactose operon. These results indicate that the genes involved in galactoside catabolism are clustered in L. plantarum ATCC 8014. This first genetic characterization of melA and of its putative associated transporter, rafP, in a lactobacillus opens doors to various applications both in the manufacture of soy-derived products and in probiotic and nutraceutical issues

126. Robust and Persistent Vaginal Colonization with LACTIN-V Vaginal Lactobacillus crispatus Probiotic in a Double-Blind, Placebo-Controlled (DBPC) Phase 2b Trial to Prevent Recurrent UTI (rUTI)

Abstract Background We investigated vaginal colonization using repetitive sequence PCR (repPCR) and 16S rRNA sequencing in a Phase 2b DBPC trial of a L. crispatus intravaginal suppository probiotic for prevention of rUTI in premenopausal women. Methods Twenty-four young women with a history of rUTI and current culture-confirmed symptomatic UTI were enrolled and treated (Visit 0), then randomized (Visit 1) to receive an intravaginal suppository containing L. crispatus CTV-05 (LACTIN-V®, Osel, Inc.) or placebo daily for 5 days, then once weekly for 2 months. Participants were followed up during the 2-month probiotic/placebo intervention (Visits 2 to 4; active intervention) and during 2 months following the intervention (Visits 5 and 6; post-intervention). At each visit, vaginal swabs were collected for repPCR to determine the presence or absence of the probiotic strain and the duration of its presence in the vagina and for 16S rRNA-based sequence analysis to determine relative abundance of any L. crispatus. Results LACTIN-V vaginal suppository induced selective and sustained colonization in the probiotic but not the placebo recipients, as follows. Pre-intervention: Probiotic lactobacillus strain, not found in vaginal specimens obtained from participants in either arm of study. Active intervention: (1) Probiotic lactobacillus strain, (a) Probiotic arm: 100% of participants positive at one or more visits and (b) Placebo arm: 0% of participants positive at any time. (2) L. crispatus relative abundance, (a) Probiotic arm: above 90%, all specimens, all visits and (b) Placebo arm: below 15%, all specimens, all visits. Post-intervention: (1) Probiotic lactobacillus strain, (a) Probiotic arm: 75% of participants positive at Visit 5, 58% at Visit 6 and (b) Placebo arm: 0% of participants positive at Visits 5 and 6. (2) L. crispatus relative abundance, (a) Probiotic arm: 70% to 100% and (b) Placebo arm: below 15%. Conclusion LACTIN-V L. crispatus vaginal probiotic achieved robust and persistent colonization throughout 2 months of weekly dosing and for 2 months after the last dose in most participants. Disclosures All authors: No reported disclosures

Reduction of α-galactooligosaccharides in soyamilk by Lactobacillus fermentum CRL 722: In vitro and in vivo evaluation of fermented soyamilk

Aims: Consumption of soya-derived products has been hampered by the presence of α-galactooligosaccharides (α-GOS) because mammals lack pancreatic α-galactosidase (α-Gal) which is necessary for their hydrolysis. These sugars thus reach the large intestine causing gastrointestinal disorders in sensitive individuals. The use of lactic acid bacteria (LAB) expressing α-Gal is a promising solution for the degradation of α-GOS in soyamilk. Methods and Results: The capacity of the LAB Lactobacillus fermentum CRL 722 to properly degrade α-GOS was studied in vitro using controlled fermentation conditions and in vivo using a rat model. Lactobacillus fermentum CRL 722 was able to grow on commercial soyamilk and completely eliminated stachyose and raffinose during fermentation because of its high α-Gal activity. Rats fed soyamilk fermented by this LAB had smaller caecums compared with rats fed unfermented soyamilk. Conclusions: Soyamilk fermentation by Lact. fermentum CRL 722 results in the reduction of α-GOS concentrations in soyamilk, thus eliminating possible undesirable physiological effects normally associated with its consumption. Significance and Impact of the Study: Fermentation with Lact. fermentum CRL 722 could prevent gastrointestinal disorders in sensitive individuals normally associated with the consumption of soya-based products. This LAB could thus be used in the elaboration of novel fermented vegetable products which better suit the digestive capacities of consumers.Fil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Garro, Marisa Selva. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Silvestroni, Aurelio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Connes, Cristelle. Institut National de la Recherche Agronomique; FranciaFil: Piard, Jean Christophe. Institut National de la Recherche Agronomique; FranciaFil: Sesma, Fernando Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Evaluation of cytotoxic and mitodepressive activity of aqueous extracts from thirteen Argentine medicinal plants

Trece especies de plantas vasculares usadas en la medicina popular argentina, fueron estudiadas para evaluar las actividades citotóxicas y mitodepresivas de sus extractos acuosos a diferentes concentraciones usando los tests de Artemia salina y Allium. No se observó actividad citotóxica de los extractos analizados con el test de Artemia salina y, en relación a los parámetros macroscópicos de toxicidad general, todas las especies mostraron correlaciones negativas aunque no significativas entre la longitud media de las raíces y la concentración del extracto y el test de Allium. La frecuencia de las anormalidades macroscópicas de las raíces mostró correlaciones negativas en todas las especies analizadas siendo la mayoría, estadísticamente significativas, indicando un efecto de la concentración. En casi todas las muestras ensayadas, se observaron efectos de tipo C-mitótico ya que el Indice Mitótico disminuyó en tanto que las anormalidades mitóticas se incrementaron con las concentraciones crecientes de los extractos. En ningún caso se observó actividad clastogénica significativa.Thirteen vascular plant species used in argentine folk medicine were studied in order to evaluate the cytotoxic and mitodepressive activity of their aqueous extracts at different concentrations, using both the Artemia salina Test and the Allium Test. No cytotoxic activity was observed for analyzed extracts with the Artemia salina Test, and in relation to macroscopic parameters of general toxicity, all species exhibited negative albeit non-significant correlations between average root length and extract concentration in the Allium Test. Frequency of root macroscopic abnormalities and average root length showed significant or highly significant negative correlations in all but two of the analyzed species which nevertheless exhibited comparable negative trends, indicating a concentration effect. In almost all assayed samples a Cmitotic effects of the extracts was observed since the mitotic index decreased and mitotic abnormalities increased with increasing extract concentrations. Clastogenic activity was not found in the analyzed extracts.Colegio de Farmacéuticos de la Provincia de Buenos Aire