A Pilot Microbiota Study in Parkinson's Disease Patients versus Control Subjects, and Effects of FTY720 and FTY720-Mitoxy Therapies in Parkinsonian and Multiple System Atrophy Mouse Models.

BACKGROUND: Parkinson's disease (PD) and multiple system atrophy (MSA) patients often suffer from gastrointestinal (GI) dysfunction and GI dysbiosis (microbial imbalance). GI dysfunction also occurs in mouse models of PD and MSA. OBJECTIVES: To assess gut dysfunction and dysbiosis in PD subjects as compared to controls, identify potential shared microbial taxa in humans and mouse models of PD and MSA, and to assess the effects of potential therapies on mouse GI microbiota. METHODS: In this human pilot study, GI function was assessed by fecal consistency/frequency measured using the Bristol Stool Form Scale and GI transit time assessed using Sitzmarks pills and abdominal radiology. Human and mouse microbiota were analyzed by extracting fecal genomic DNA followed by 16S rRNA sequencing. RESULTS: In our PD patients genera Akkermansia significantly increased while a trend toward increased Bifidobacterium and decreased Prevotella was observed. Families Bacteroidaceae and Lachnospiraceae and genera Prevotella and Bacteroides were detected in both humans and PD mice, suggesting potential shared biomarkers. In mice treated with the approved multiple sclerosis drug, FTY720, or with our FTY720-Mitoxy-derivative, we saw that FTY720 had little effect while FTY720-Mitoxy increased beneficial Ruminococcus and decreased Rickenellaceae family. CONCLUSION: Akkermansia and Prevotellaceae data reported by others were replicated in our human pilot study suggesting the use of those taxa as potential biomarkers for PD diagnosis. The effect of FTY720-Mitoxy on taxa Rikenellaceae and Ruminococcus and the relevance of S24-7 await further evaluation. It also remains to be determined if mouse microbiota have predictive power for human subjects.

Parkinsonian GM2 synthase knockout mice lacking mature gangliosides develop urinary dysfunction and neurogenic bladder.

Parkinson's disease is a neurodegenerative disorder that reduces a patients' quality of life by the relentless progression of motor and non-motor symptoms. Among the non-motor symptoms is a condition called neurogenic bladder that is associated with detrusor muscle underactivity or overactivity occurring from neurologic damage. In Parkinson's disease, Lewy-body-like protein aggregation inside neurons typically contributes to pathology. This is associated with dopaminergic neuron loss in substantia nigra pars compacta (SNc) and in ventral tegmental area (VTA), both of which play a role in micturition. GM1 gangliosides are mature glycosphingolipids that enhance normal myelination and are reduced in Parkinson's brain. To explore the role of mature gangliosides in vivo, we obtained GM2 Synthase knockout (KO) mice, which develop parkinsonian pathology including a loss of SNc dopaminergic neurons, which we reconfirmed. However, bladder function and innervation have never been assessed in this model. We compared GM2 Synthase KO and wild type (WT) littermates' urination patterns from 9 to 19 months of age by counting small and large void spots produced during 1 h tests. Because male and female mice had different patterns, we evaluated data by sex and genotype. Small void spots were significantly increased in 12-16 month GM2 Synthase KO females, consistent with overactive bladder. Similarly, at 9-12 month GM2 KO males tended to have more small void spots than WT males. As GM2 Synthase KO mice aged, both females and males had fewer small and large void spots, consistent with detrusor muscle underactivity. Ultrasounds confirmed bladder enlargement in GM2 Synthase KO mice compared to WT mice. Tyrosine hydroxylase (TH) immunohistochemistry revealed significant dopaminergic loss in GM2 Synthase KO VTA and SNc, and a trend toward TH loss in the GM2 KO periaqueductal gray (PAG) micturition centers. Levels of the nerve growth factor precursor, proNGF, were significantly increased in GM2 Synthase KO bladders and transmission electron micrographs showed atypical myelination of pelvic ganglion innervation in GM2 Synthase KO bladders. Cumulatively, our findings provide the first evidence that mature ganglioside loss affects micturition center TH neurons as well as proNGF dysregulation and abnormal innervation of the bladder. Thus, identifying therapies that will counteract these effects should be beneficial for those suffering from Parkinson's disease and related disorders.

The Candida albicans Exocyst Subunit Sec6 Contributes to Cell Wall Integrity and Is a Determinant of Hyphal Branching.

The yeast exocyst is a multiprotein complex comprised of eight subunits (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84) which orchestrates trafficking of exocytic vesicles to specific docking sites on the plasma membrane during polarized secretion. To study SEC6 function in Candida albicans, we generated a conditional mutant strain in which SEC6 was placed under the control of a tetracycline-regulated promoter. In the repressed state, the tetR-SEC6 mutant strain (denoted tSEC6) was viable for up to 27 h; thus, all phenotypic analyses were performed at 24 h or earlier. Strain tSEC6 under repressing conditions had readily apparent defects in cytokinesis and endocytosis and accumulated both post-Golgi apparatus secretory vesicles and structures suggestive of late endosomes. Strain tSEC6 was markedly defective in secretion of aspartyl proteases and lipases as well as filamentation under repressing conditions. Lack of SEC6 expression resulted in markedly reduced lateral hyphal branching, which requires the establishment of a new axis of polarized secretion. Aberrant localization of chitin at the septum and increased resistance to zymolyase activity were observed, suggesting that C. albicans Sec6 plays an important role in mediating trafficking and delivery of cell wall components. The tSEC6 mutant was also markedly defective in macrophage killing, indicating a role of SEC6 in C. albicans virulence. Taken together, these studies indicate that the late secretory protein Sec6 is required for polarized secretion, hyphal morphogenesis, and the pathogenesis of C. albicans.

Surface Enhanced Raman Scattering (SERS) Studies of Gold and Silver Nanoparticles Prepared by Laser Ablation.

Gold and silver nanoparticles (NPs) were prepared in water, acetonitrile and isopropanol by laser ablation methodologies. The average characteristic (longer) size of the NPs obtained ranged from 3 to 70 nm. 4-Aminobenzebethiol (4-ABT) was chosen as the surface enhanced Raman scattering (SERS) probe molecule to determine the optimum irradiation time and the pH of aqueous synthesis of the laser ablation-based synthesis of metallic NPs. The synthesized NPs were used to evaluate their capacity as substrates for developing more analytical applications based on SERS measurements. A highly energetic material, TNT, was used as the target compound in the SERS experiments. The Raman spectra were measured with a Raman microspectrometer. The results demonstrate that gold and silver NP substrates fabricated by the methods developed show promising results for SERS-based studies and could lead to the development of micro sensors.

Preparation of hollow magnetite microspheres and their applications as drugs carriers.

Hollow magnetite microspheres have been synthesized by a simple process through a template-free hydrothermal approach. Hollow microspheres were surface modified by coating with a silica nanolayer. Pristine and modified hollow microparticles were characterized by field-emission electron microscopy, transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, FT-IR and Raman spectroscopy, and VSM magnetometry. The potential application of the modified hollow magnetite microspheres as a drug carrier was evaluated by using Rhodamine B and methotrexate as model drugs. The loading and release kinetics of both molecules showed a clear pH and temperature dependent profile. GRAPHICAL Hollow magnetite microspheres have been synthesized. Load-release experiments with Rhodamine-B as a model drug and with Methotrexate (chemotherapy drug used in treating certain types of cancer) demonstrated the potential applications of these nanostructures in biomedical applications.

Investigación clinicoepidemiológica en envejecimiento: metodología del proyecto encuesta salud, bienestar y envejecimiento (SABE) en México / Epidemiological clinical research in aging: methodology of the project survey health, wellbeing and ageing in Mexico

La población de personas adultas mayores crece de forma paulatina y sostenida en el mundo y por ende en nuestro país. Sin embargo, la información que se tiene sobre la forma de vida, salud y bienestar de este grupo poblacional es limitada debido a que la mayor parte de las investigaciones en adultos mayores es derivada del estudio de grupos pequeños con patología bien circunscrita y características especiales. Por otro lado, la realización de estudios, en la población abierta, que permitan establecer la demografía y epidemiología de las personas mayores de 60 años requieren de una gran infraestructura y recursos tanto humanos como económicos. El Proyecto-Encuesta Salud Bienestar y Envejecimiento (SABE), mediante la realización de un estudio tipo encuesta, tiene la finalidad de obtener dicha información contando con la participación del Sector Salud, instituciones de educación superior así como organizaciones no gubernamentales lo que constituye un logro interinstitucional. Para la realización del proyecto SABE se ha seguido con rigurosidad científica el protocolo y para su puesta en marcha hubo la necesidad de la integración intersectorial de las instituciones que brindan atención a los adultos mayores mexicanos, ya sea dentro del contexto de la seguridad o asistencia social. Los alcances de SABE permitirán la creación y difusión del conocimiento derivado del mismo. En este documento, se expone y analiza de forma breve y concreta la metodología operativa de SABE México, lo que significa dar el primer paso para que otros investigadores interesados en el tema, realicen este tipo de estudios. Dada la experiencia que se ha ido generando en la realización de SABE México, este tipo de documentos sirven como base para proponer e iniciar las acciones al respecto.

The elderly population grows so gradual and sustained in the world and therefore in our country. However, information that is on the way of life, health and well-being of this population group is limited since most of the research in older adults is derived from the study of small groups with well circumscribed pathology and special features. On the other hand, open population studies to establish the demography and epidemiology of people older than 60 years require a large infrastructure and resources both human and financial. Project- Survey Health, Wellbeing and Ageing (SABE, in Spanish)), through a study type survey, has the purpose of obtaining such information with the participation of the Health Sector, institutions of higher education as well as organizations NGO constituting an interinstitutional achievement. For the implementation of the project SABE the Protocol has been followed with scientific rigor and for it simplementation underway there was the need to cross-sectoral integration of institutions providing care to Mexican older adults, either within the context of the social security or welfare. The reaches of SABE will enable the creation and dissemination of the knowledge derived from it. In this document, it exposes and analyzes operational methodology of a brief and concrete way of SABE Mexico, which means the first step so that other researchers interested in the topic, take place this type of study. Given the experience that has been generated in the implementation of SABE Mexico, this type of documents serves as the basis to propose and initiate actions in this regard.

Factores de riesgo que incrementan las infecciones intrahospitalarias en el servicio de cuneros / Risk factors that increase nosocomial infections in the nursery service / Fatores de risco que aumentam as infecções nosocomiais no serviço de berçário

Introducción: Las Unidades de Cuidados Intensivos y Neonatales; se ven afectados por la presencia de Infecciones Intrahospitalarias (IIH), se han identificado en el recién nacido una serie de factores de riesgo, que se relacionan y permiten la presencia de éstas infecciones, entre los que se encuentran los antecedentes maternos, historial del recién nacido, hacinamiento en el área neonatal y la insuficiencia en cantidad del personal de Enfermería. Objetivo: Analizar la relación entre las dimensiones: materna, del recién nacido, el hacinamiento en el área neonatal y la suficiencia en cantidad del personal de Enfermería y la presencia de las IIH, en el Servicio Cunero del Hospital General de Ticomán, pertenecen a los Servicios de Salud Pública del Distrito Federal. Material y métodos: Estudio epidemiológico, descriptivo, retrospectivo y correlacional Universo: Censo de 799 expedientes clínicos de los Recién Nacidos, que estuvieron internados en el Servicio de Cuneros del Hospital General de Ticomán, durante el periodo del 1° de enero al 30 de noviembre del 2004; fueron incluidos solo 742 casos que representaron el 92% de expedientes. La validez del instrumento, es de contenido por juicio de 8 expertos. Confiabilidad por Kunder Richarson de 0.79. Se utilizó estadística descriptiva e inferencial, obtenido los resultados utilizando los programas estadísticos SPSS, versión 121 y G-Stat 2.0. Resultados: Los factores de riesgo que determinan las IIH en el servicio de cuneros son: Peso del Recién Nacidoa 4000 gramos con una x cuadrada=7.1401, g. 1=1; p=0.0075; semanas de gestación menos a 37(SDG) presentado una X cuadrada= 8.09836, g. 1.=1 y p= 0.004; uso Catéter Umbilical con X cuadrada= 60.0588, g.1.=1 y p<0.0001; uso de Catéter Intravascular con x cuadrada=241.2030, g. 1.=1 y p<0.0001, utilización de Catéter Vesical con x cuadrada= 197559, g.1.=1 y p<0.0001; uso de Técnica Endotraqueal con x cuadrada= 89.6646, g.1.=1 y p<0.0001; los recién nacidos con los siguientes diagnósticos: Aspiración de Meconio con x cuadrada= 31.0444, g 1.=1 y p<0.0001; Prematurez con x cuadrada= 10.9662, g. 1.=1 y p=0.0009; Macrosómico con peso>a 4000 gramos presento una X cuadrada=60.0588, g.1.=1 y p<0.0001. Se corroboro que el hemocultivo con la identificación del microorganismo confirma la presencia de IIH. Discusión Los resultados obtenidos demostraron que no existe una relación entre las cuatro dimensiones, la presencia de infecciones intrahospitalarias en este Servicio en particular son debidas a los factores inherentes al recién nacido, Barroso Aguirre y cols. Mencionan que los Neonatos en Unidades especiales tendrán la presencia de IIH. En las cuatro dimensiones se observó la manifestación del riesgo relativo para la presencia IIH. Conclusiones: La presencia de IIH, en el Servicio de Cueros son consecuencia del historial del Recién Nacido, no existió relación entre las cuatro dimensiones, sin embargo el riesgo relativo se cuenta presente en todas ellas.

Introdução: as unidades de terapia intensiva e neonatal; são afectados pela presença de infecções nosocomiais (IIH), que foram identificados no recém-nascido um número de factores de risco que se relacionam e permitem que a presença destas infecções, incluindo aqueles encontrados história materna, história de recém-nascido nascido, superlotação na área neonatal e número insuficiente de pessoal de enfermagem. Objetivo: Analisar a relação entre dimensões: saúde materna, neonatal, a superlotação na área neonatal e suficiência em quantidade de pessoal de enfermagem ea presença de IIH no Hospital chaveta de Serviços Gerais Ticoman pertencem ao Serviços de Saúde Pública do Distrito Federal. Métodos: Um epidemiológico, descritivo, retrospectivo e correlacional Estudo Universe: Censo de 799 prontuários de recém-nascidos, que foram internados no Hospital serviço Cuneros Geral Ticoman durante o período de 1 janeiro - 30 novembro de 2004; Apenas 742 casos foram incluídos, o que representou 92% dos registros. A validade do instrumento é de conteúdo pelo julgamento de 8 especialistas. Confiabilidade de Kunder Richarson de 0,79. Utilizamos estatística descritiva e inferencial, obtivemos os resultados utilizando os programas estatísticos SPSS, versão 121 e G-Stat 2.0. Resultados: Os fatores de risco que determinam o HII no serviço de creches são: Peso do Recém-Nascido a 4000 gramas com um quadrado x = 7,1401, g. 1 = 1; p = 0,0075; semanas de gestação menos 37 (ODS) apresentaram um quadrado X = 8,09836, g. 1. = 1 e p = 0,004; usar cateter umbilical com X = 60,0588, g.1 = 1 e p <0,0001; Uso de cateter intravascular com o quadrado x = 241.2030, g. 1. = 1 e p <0,0001, uso de Cateter Vesical com x ao quadrado = 197559, g.1. = 1 e p <0,0001; uso de Técnica Endotraqueal com x ao quadrado = 89,6646, g.1. = 1 e p <0,0001; recém-nascidos com os seguintes diagnósticos: aspiração de mecônio com x ao quadrado = 31,0444, g1 = 1 e p <0,0001; Prematuridade com quadrado x = 10.9662, g. 1 = 1 e p = 0,0009; Macrosomic com peso> em 4000 gramas eu apresento um quadrado X = 60,0588, g.1 = 1 e p <0,0001. Foi corroborado que a hemocultura com a identificação do microrganismo confirma a presença de HII. Discussão Os resultados obtidos mostraram que não há relação entre as quatro dimensões, a presença de infecções nosocomiais neste Serviço, em particular, são devidas aos fatores inerentes ao recém-nascido, Barroso Aguirre et al. Mencionam que os Neonatos em Unidades especiais terão a presença do IIH. Nas quatro dimensões, foi observada a manifestação do risco relativo para a presença da HII. Conclusões: A presença de HII, no Serviço de Couros, é consequência da história do recém-nascido, não houve relação entre as quatro dimensões, porém o risco relativo está presente em todas elas.