RNA-Sensing Pattern Recognition Receptors and Their Effects on T-Cell Immune Responses: A Dissertation

Virus infection is sensed by the innate immune system through germline encoded pattern recognition receptors (PRRs). Toll-like receptors (TLRs), retinoic acid-inducible gene-I-like receptors (RLRs) and nucleotide-binding oligomerization domain-like receptors (NLRs) serve as PRRs that recognize different viral components. Microbial nucleic acids such as Ribonucleic acid (RNA) are important virus-derived pathogen-associated molecular patterns (PAMPs) to be recognized by PRRs. Virus recognition may occur at multiple stages of the viral life cycle. Replication intermediates such as single-stranded RNA (ssRNA) and double-stranded RNA (dsRNA) are detected by the RNA-sensing PRRs that initiate innate and adaptive immune responses. Triggering of the innate immune system is a critical event that can shape the adaptive immune response to virus infection. Better vaccination strategies that lead to improved T-cell and antibody responses are needed for protection against pathogens. We sought to delineate the RNA-sensing PRR pathways that are activated during infection with an RNA virus, the signaling mediators involved and the influence on subsequent virus-specific adaptive immune responses. To analyze the role of RNA-sensing PRRs in T-cell immune responses in vitro, we performed direct co-stimulation experiments on CD4+ T-cells of high purity. We utilized synthetic RNA-like immune response modifiers (IRMs) R-848 (MyD88-dependent) and poly I:C (MyD88-independent) as RNA PAMPs to determine the direct effects of RNA-sensing PRR activation on CD4+ T-cells. RNA PAMPs can act directly on CD4+ T-cells and modulate their function and phenotype. Maximal direct co-stimulatory effects were observed in CD4+ T-cells cultured with poly I:C compared to R-848. The cytoplasmic dsRNA-dependent protein kinase R (PKR) was also involved in poly I:C-mediated signaling in CD4+ T-cells. We found differences in the RNA-sensing PRRs activated by R-848 between mouse and human CD4+ T-cells. We observed minimal direct co-stimulatory effects by R-848 in mouse CD4+ T-cells. In contrast, augmentation of Th1 responses by R-848 was observed in human CD4+ T-cells. TLR8 activation in human CD4+ T-cells may explain the observed differences. We next explored the signaling pathways activated by RNA PAMPs in conventional dendritic cells (cDCs) and CD4+ T-cells that drive Th1 CD4 T-cell responses in isolated cDC/CD4 T-cell interactions. Allogeneic cDCs and CD4+ T-cells of high purity were cultured together with R-848 and poly I:C in MHC congenic mixed leukocyte reactions (MLRs). R-848 and poly I:C stimulation of type I IFN production and signaling was essential but not sufficient for driving CD4+ Th1 responses. The early production of IL-1α and IL-1β was equally critical. To analyze the role of RNA-sensing PRRs in T-cell immune responses in vivo, we utilized a mouse model of heterosubtypic influenza A virus (IAV) infections. Using MyD88-/-, TLR7-/- and IL-1-deficient mice, we explored the role of MyD88-signaling in the generation of heterosubtypic memory CD4+ T-cell, CD8+ T-cell and antibody responses. We found that MyD88 signaling played an important role in anti-IAV spleen and lung CD4+ T-cell, spleen CD8+ T-cell and Th1 antibody immune responses. Anti-IAV lung heterosubtypic CD8+ T-cell responses were not dependent on MyD88 signaling. Our in vitro and in vivo results show the pivotal role of RNA-sensing PRR pathway activation in T-cell immune responses. Understanding the complexity of the PRR pathways involved during viral infections and defining the subsequent immune response would have important implications for the generation of more effective vaccine strategies

EVOLUTION OF ANTHROPOMETRICAL AND KINEMATIC PARAMETERS IN YOUNG SWIMMERS: A LONGITUDINAL STUDY

Swimming kinematics parameters have been widely applied to measure the swimming skill in training and competitive situations. The descriptive study aim was to analyse the evolution of the swimming kinematics parameters related to anthropometrical data in the age-group. The data was obtained during the Winter National Age-Group (1994) and Junior (1996) Championships organized by the Royal Spanish Swimming Federation. Only those participanting in both championships were analysed in this study. The obtained results allow to conclude that the evolution observed in the kinematics variables and antropometrical variables in young swimmers shows the improvement in the speed depends SL resulting in part from the increase in anthropometrical variables

The combination of early and rapid type I IFN, IL-1alpha, and IL-1beta production are essential mediators of RNA-like adjuvant driven CD4+ Th1 responses

There is a growing need for novel vaccine adjuvants that can provide safe and potent T-helper type 1 (Th1) activity. RNA-like immune response modifiers (IRMs) are candidate T-cell adjuvants that skew acquired immune responses towards a Th1 phenotype. We set out to delineate the essential signaling pathways by which the RNA-like IRMs, resiquimod (R-848) and polyinosinic:polycytidylic acid (poly I:C), augment CD4+ T-helper 1 (Th1) responses. Highly purified murine conventional dendritic cells (cDCs) and conventional CD4+ T-cells were co-cultured in allogeneic and MHC congenic mixed leukocyte reactions. The activation of CD4+ Th1 cells was examined utilizing cells from mice deficient in specific RNA-sensing pattern recognition receptors and signaling mediators. R-848 and poly I:C stimulation of Type I interferon production and signaling in cDCs was essential but not sufficient for driving CD4+ Th1 responses. The early and rapid production of IL-1alpha and IL-1beta was equally critical for the optimal activation of Th1 CD4+ T-cells. R-848 activation of Toll-like receptor 7/MyD88-dependent signaling in cDCs led to a rapid upregulation of pro-IL-1alpha and pro-IL-1beta production compared to poly I:C activation of MyD88-independent signaling pathways. The in vitro data show that CD4+ T-cell adjuvant activity of RNA-like IRMs is mediated by a critical combination of early and rapid Type I interferon, IL-1alpha and IL-1beta production. These results provide important insights into the key signaling pathways responsible for RNA-like IRM CD4+ Th1 activation. A better understanding of the critical signaling pathways by which RNA-like IRMs stimulate CD4+ Th1 responses is relevant to the rational design of improved vaccine adjuvants

Google Web Toolkit application solution for mild aphasia therapy

Aphasia is a disorder that results from damage to the parts of the brain that contain language. It typically occurs after a stroke, brain tumors, head injuries, infections or degenerative diseases leading to damage in areas of the brain cells. Aphasia causes problems with any or all of the following: speaking, listening, reading, and writing. Thousands of people of all ages and gender suffer from this disease and the trend is growing with the longevity of population. This article describes the design and development of specific software that can be useful for treatment of mild aphasia. This software is based on a Google Web Toolkit (GWT) application. GWT is an open source framework that allows developing a user friendly interface. The web application can be accessible to speech and language therapists (SLT) professionals and patients at different contexts and learning situations.Sociedad Argentina de Informática e Investigación Operativ

Google Web Toolkit application solution for mild aphasia therapy

Aphasia is a disorder that results from damage to the parts of the brain that contain language. It typically occurs after a stroke, brain tumors, head injuries, infections or degenerative diseases leading to damage in areas of the brain cells. Aphasia causes problems with any or all of the following: speaking, listening, reading, and writing. Thousands of people of all ages and gender suffer from this disease and the trend is growing with the longevity of population. This article describes the design and development of specific software that can be useful for treatment of mild aphasia. This software is based on a Google Web Toolkit (GWT) application. GWT is an open source framework that allows developing a user friendly interface. The web application can be accessible to speech and language therapists (SLT) professionals and patients at different contexts and learning situations.Sociedad Argentina de Informática e Investigación Operativ

Google Web Toolkit application solution for mild aphasia therapy

Aphasia is a disorder that results from damage to the parts of the brain that contain language. It typically occurs after a stroke, brain tumors, head injuries, infections or degenerative diseases leading to damage in areas of the brain cells. Aphasia causes problems with any or all of the following: speaking, listening, reading, and writing. Thousands of people of all ages and gender suffer from this disease and the trend is growing with the longevity of population. This article describes the design and development of specific software that can be useful for treatment of mild aphasia. This software is based on a Google Web Toolkit (GWT) application. GWT is an open source framework that allows developing a user friendly interface. The web application can be accessible to speech and language therapists (SLT) professionals and patients at different contexts and learning situations.Sociedad Argentina de Informática e Investigación Operativ

Plasmacytoid dendritic cell interferon-alpha production to R-848 stimulation is decreased in male infants

BACKGROUND: Sex differences in response to microbial infections, especially viral ones, may be associated with Toll-like receptor (TLR)-mediated responses by plasmacytoid dendritic cells (pDCs). RESULTS: In this study, we identified sex differences in human infant pDC interferon-alpha production following challenge with the TLR7/8 agonist R-848. Male pDC responses were significantly lower than those of females during early infancy. This difference may be attributed to the androgen surge experienced by males during the early infancy period. Pretreatment of human pDCs with dihydrotestosterone produced a significant reduction in interferon-alpha production following R-848 challenge. CONCLUSIONS: Androgen-mediated regulation of pDC TLR7-driven innate immune responses may contribute to the observed sex differences in response to infections during early infancy

Reúso de aguas residuales: impacto en los atributos químicos y macronutricionales en un suelo inceptisol irrigado con aguas residuales domésticas tratadas

(Eng) This study aimed to estimate the potential impact of wastewater reuse on the chemical and macronutritional attributes of an inceptisol cropped with sugarcane. The experimental design was in three randomized blocks, in split-plot design (three replicates per treatment), with three different irrigation treatments: treated wastewater (TWW) from Cañaveralejo wastewater treatment plant (WWTP-C); groundwater without any fertilizer (GW); and groundwater with chemical fertilizer (GW+CF). At the end of the study, plots with treated wastewater irrigation improved slightly Ca/Mg, Mg/K, and Ca+Mg/K ratios, increased OM, Na, P, and K contents and decreased pronouncedly inorganic nitrogen (N-NH4, N-NO3) of an inceptisol. Besides, we found that the type of treatment did not influence the variation of chemical attributes in soil, since statistically differences were no found in comparison among treatments. Therefore, our results suggest no adverse impact on chemical soil attributes due to wastewater reuse on sugarcane irrigation.(Spa) Este estudio tuvo como objetivo estimar el potencial impacto en los atributos químicos y macronutricionales de un suelo inceptisol sembrado con caña de azúcar y sometido a tres diferentes tratamientos de riego: Efluente de la PTAR-C (T1), Agua de Pozo (T2) y Agua de Pozo más fertilización química (T3). Se instaló un experimento en bloques completos al alzar con parcelas divididas con tres repeticiones. Al final del estudio se encontró que las relaciones de bases químicas (Ca/Mg, Mg/K, Ca+Mg/K) mejoraron en todas las parcelas, en todos los tratamientos, situación que se ratifica al no encontrarse diferencias estadísticas significativas entre los tratamientos. Así mismo, se encontró que el tipo de tratamiento no influyó en la variación en los contenidos de los macronutrientes (N, P, K) del suelo. Los contenidos de materia orgánica, P, Na y K aumentaron; y disminuyeron los de nitrógeno inorgánico (N-NH4, N-NO3) independientemente del tratamiento, lo que permite inferir la no asociación de impactos adversos en el suelo por el reúso de agua residual tratada en riego de caña de azúcar

Risk factors for otitis media and carriage of multiple strains of Haemophilus influenzae and Streptococcus pneumoniae.

We studied genetic diversity in Streptococcus pneumoniae and Haemophilus influenzae in throat culture isolates from 38 children attending two day-care centers in Michigan. Culture specimens were collected weekly; 184 S. pneumoniae and 418 H. influenzae were isolated from the cultures. Pulsed-field gel electrophoresis identified 29 patterns among the S. pneumoniae isolates and 87 among the H. influenzae isolates. Of the cultures, 5% contained multiple genetic types of S. pneumoniae, and 43% contained multiple types of H. influenzae. Carriage of multiple H. influenzae isolates, which was associated with exposure to smoking, history of allergies, and age 36 to 47 months, may increase risk for otitis media in children

Seed viability and potential germination rate of nine endemic Boswellia taxa (Burseraceae) from Socotra Island (Yemen)

The endemic Boswellia species (Burseraceae) on Socotra Island (Yemen) are of great local significance due to their various local ethnobotanical uses. However, despite the fact that these trees are endangered, little is known about their biology. We tested seed germination rates in controlled experiments (trials of 21 days) for two subsequent years and for nine endemic taxa of Boswellia occurring on Socotra Island. For this, seeds were collected island-wide from a wide range of localities and for several populations per species. We observed differences in germination among Boswellia species, among species and localities and among both years, which indicates that the development of seeds is strongly affected by external ecological factors. Although we noted a large variation in seed germination (relatively high in Boswellia socotrana), and half of the species showed relatively low mean daily germination, our study indicated that all endangered endemic Frankincense Tree taxa of Socotra harbor the potential for in situ conservation through recruitment, given that known impacts can be reduced in local replantation areas (e.g., grazing)