Silent Phase of Johne’s Disease in Experimentally Infected Goats – A Study on New and Established Diagnostic Approaches Using Specific and Non-Specific Parameters

The current gold standard diagnostic test for Johne’s disease (JD) is detecting Mycobacterium avium subsp. paratuberculosis (MAP) from fecal samples via culture and/or PCR. Other commercially available JD diagnostic tests focus on the detection of specific antibodies within the serum or milk of infected ruminants. These tests have a high specificity but low their sensitivity and usually fail to diagnose the disease until later stages of the disease. The ideal diagnostic test should detect infected animals already during the silent phase. Here, we evaluate the use of new and established approaches to define the silent phase of JD in experimentally infected goats. None of the established diagnostic tests or new approaches for the detection of humoral and cellular immune responses were positive during the first year of infection. Only the characterization of various subsets of peripheral blood leukocytes and the weight development gave some indication for the presence of a chronic, but silent, infection. Weight differences were present throughout the first year. In addition, some of the subsets of leukocytes (WC1+ T cells, MHC class II+ leukocytes, CD1+ leukocytes, CD14+ granulocytes, and CD14+/MHC class II+ granulocytes) demonstrated significant differences, but only at certain time points

Functional drug screening reveals anticonvulsants as enhancers of mTOR-independent autophagic killing of Mycobacterium tuberculosis through inositol depletion.

Mycobacterium tuberculosis (MTB) remains a major challenge to global health made worse by the spread of multidrug resistance. We therefore examined whether stimulating intracellular killing of mycobacteria through pharmacological enhancement of macroautophagy might provide a novel therapeutic strategy. Despite the resistance of MTB to killing by basal autophagy, cell-based screening of FDA-approved drugs revealed two anticonvulsants, carbamazepine and valproic acid, that were able to stimulate autophagic killing of intracellular M. tuberculosis within primary human macrophages at concentrations achievable in humans. Using a zebrafish model, we show that carbamazepine can stimulate autophagy in vivo and enhance clearance of M. marinum, while in mice infected with a highly virulent multidrug-resistant MTB strain, carbamazepine treatment reduced bacterial burden, improved lung pathology and stimulated adaptive immunity. We show that carbamazepine induces antimicrobial autophagy through a novel, evolutionarily conserved, mTOR-independent pathway controlled by cellular depletion of myo-inositol. While strain-specific differences in susceptibility to in vivo carbamazepine treatment may exist, autophagy enhancement by repurposed drugs provides an easily implementable potential therapy for the treatment of multidrug-resistant mycobacterial infection

Increased Virulence of an Epidemic Strain of Mycobacterium massiliense in Mice

Chronic pulmonary disease and skin/soft tissue infections due to non-tuberculous mycobacteria (NTM) of the Mycobacterium chelonae-abscessus-massiliense group is an emerging health problem worldwide. Moreover, the cure rate for the infections this group causes is low despite aggressive treatment. Post-surgical outbreaks that reached epidemic proportions in Brazil recently were caused by M. massiliense isolates resistant to high-level disinfection with glutaraldehyde (GTA). Understanding the differences in the virulence and host immune responses induced by NTM differing in their sensitivity to disinfectants, and therefore their relative threat of causing outbreaks in hospitals, is an important issue.We compared the replication and survival inside macrophages of a GTA-susceptible reference Mycobacterium massiliense clinical isolate CIP 108297 and an epidemic strain from Brazil, CRM-0019, and characterized the immune responses of IFNγ knockout mice exposed to a high dose aerosol with these two isolates. CRM-0019 replicated more efficiently than CIP 108297 inside mouse bone marrow macrophages. Moreover, the animals infected with CRM-0019 showed a progressive lung infection characterized by a delayed influx of CD4+ and CD8+ T cells, culminating in extensive lung consolidation and demonstrated increased numbers of pulmonary CD4+ Foxp3+ regulatory T cells compared to those infected with the reference strain. Immunosuppressive activity of regulatory T cells may contribute to the progression and worsening of NTM disease by preventing the induction of specific protective immune responses.These results provide the first direct evidence of the increased virulence in macrophages and mice and pathogenicity in vivo of the Brazilian epidemic isolate and the first observation that NTM infections can be associated with variable levels of regulatory T cells which may impact on their virulence and ability to persist in the host

Innovación, desarrollo tecnológico y gestión : una construcción desde la investigación

Libro que compila investigaciones de carácter aplicadas y descriptivas en materia de innovación y desarrollo tecnológico, gestión organizacional y empresarial orientada a productividad, rentabilidad, competitividad y sostenibilidadBook that compiles researches of applied and descriptive character in the matter of innovation and technological development, organizational and business management oriented to productivity, profitability, competitiveness and sustainabilityCapítulo 1. Material compuesto para la construcción a partir de la celulosa del papel y cartón reciclado / Carlos Arturo Tamayo S; Nicolás Montero Camacho; Fredy Antonio Herrera -- Capítulo 2. Tecnologías de conservación para base de sopa de frijol rojo (phaseolus vulgaris) y vegetales / Yaceris Castro Escorcia; Teresa Altamar Pérez; Enedys Florez Cortés; Ángela Ortiz Ruiz. Capítulo 3. Utilización de harina compuesta de frijol caupi (vigna unguiculata) en masas para alimentos congelados / Marcela Villalba Cadavid; Matilde Rodríguez Muñoz; Beatriz Fernández; Juan Mendoza Combatt -- Capítulo 4. Elaboración de biorrecubrimiento comestible para carne de hamburguesa como alternativa de conservación y condimento natural / Camila Andrea Ubaque Beltrán. CAPÍTULO 5. Identificación de alternativas de industrialización de productos y subproductos agroindustriales en nuevos materiales bio poliméricos / Luz Henao Díaz; Deya Pérez Zúñiga; Herold Arango Gómez. Capítulo 6. SCADA inalámbrico para monitoreo de sistemas de energía solar / Nelson Giovanni Agudelo Cristancho; Juan Carlos Amezquita Tovar; Ángela María Montoya Castro. -- Capítulo 7. Evaluación de la calidad del agua para consumo humano del corregimiento de Jaraquiel, Montería, Córdoba / Carlos Burgos Galeano; Álvaro Aleán Vásquez; Paula Estrada Palencia -- Capítulo 8. Optimización del sistema de abastecimiento de agua en la comunidad de Jaraquel, Montería Colombia / Carlos Burgos Galeano; Pedro Ramos Tejada; Paula Estrada Palencia; Jhon Sánchez Correa. -- Capítulo 9. Ejecución y sostenibilidad de proyectos productivos en la microrregión cafetera del municipio de Ciénaga, Magdalena / Sugey Issa Fontalvo; Eduardo Robles Panetta; Freddy González Castillo. -- Capítulo 10. Aplicación del método cualitativo por puntos para determinar aspirante favorito a cargos directivos en IES / Zamir Martelo Ballesteros; Raúl Martelo Gómez; Luis Tovar Garrido; Natividad Villabona Gómez; David Franco Borré. -- Capítulo 11 Responsabilidad social en comunidades indígenas orientada al diseño de automatización de válvulas del gasoducto Riohacha-Maicao / Gelvis Melo Freile; César Rivera Romero; Jesús García Guiliany. -- Capítulo 12 Fortalecimiento de la gestión económica de las Mipymes a través de la consultoría / Gloria Amparo Acosta Romero; Mónica Andrade Ríos; Karen Roxana Sánchez. -- Capítulo 13. El compromiso como valor en la responsabilidad social universitaria / Maura Quintero Gutiérrez, Dubys Villarreal Torres; Jesús García Guiliany; Annherys Paz Marcano; Marieth Orcasitas Peñaloza. -- Capítulo 14. Educación financiera como alternativa de desarrollo económico y social para el distrito de Riohacha / Henitzo Martínez Pinedo; Darcy Luz Mendoza; Martha Jaramillo Acosta; Edwin Salas Solano. -- Capítulo 15. Tecnologías de información y comunicación en proceso contable y financiero en pymes del sector turístico / Martha Josefina Castrillón Rois; Edilberto Rafael Santos Moreno; Lorena Esther Gómez Bermúdez; Génesis Barros González. -- Capítulo 16. Aplicación de brainstorming y problem trees para determinar factores que inciden en enseñanza del inglés / Jesús Llerena; Raúl J. Martelo; Jhon Cuesta; Javier Pinedo; David Franco.-- Capítulo 17. Incidencia del marketing en las microempresas del sector comercio en Rionegro Antioquia: conceptualización / Santiago Álzate Carmona; María Yamile Mazo Gil; Leidy García Jaramillo. -- Capítulo 18. Turismo en el Cabo de la Vela: un acercamiento entre los imaginarios turísticos de los visitantes y la creencia de Jepirra, territorio sagrado / María Laura Aponte Aarón; Esmerlis Camargo Torres. -- Capítulo 19. Caracterización de la actividad turística en buenaventura y su integración con las comunidades locales / Víctor Cándelo Aragón; Henry Orobio García; Luis Montaño Aguilar. -- Capítulo 20. Plataforma de comercialización electrónica de un centro de abastos / Karen Ávila Suarez; Mauro Reyes Ortiz. -- Capítulo 21. El teletrabajo en la gestión administrativa / Estefanía Sandoval Cruz; René Alexander Guerrero Vergel. -- Capítulo 22. Gestión del conocimiento y alianzas estratégicas en los procesos de innovación tecnológica / Elder Rivero Gutiérrez; Fátima Bolaño Mendoza. -- Capítulo 23. Competitividad e innovación en el aprendiz SENA: perspectivas de formación / Elizabeth Tuberquia Vanegas; René Alexander Guerrero Vergel. -- Capítulo 24. Bomba de riego por goteo solar, una alternativa para aumentar la eficiencia energética en las unidades acuícolas / Sergio Gabriel Brito Brito; Daldo Ricardo Araujo Vidal; Nicolás Annicharico Jiménez. -- Capítulo 25. Herramienta digital de consultas contables y tributarias para unidades productivas creadas en el fondo emprender / Elkin Fuentes Jiménez; Alda Pérez Campuzano; Marieth Orcasitas Peñaloza; Olga Elena Guerra ArmentaPrimera ediciónna347 página

Trying to See the Forest through the Trees: Deciphering the Nature of Memory Immunity to Mycobacterium tuberculosis

The purpose of vaccination against tuberculosis and other diseases is to establish a heightened state of acquired specific resistance in which the memory immune response is capable of mediating an accelerated and magnified expression of protection to the pathogen when this is encountered at a later time. In the earliest studies in mice infected with Mycobacterium tuberculosis, memory immunity and the cells that express this were definable both in terms of kinetics of emergence, and soon thereafter by the levels of expression of markers including CD44, CD62L, and the chemokine receptor CCR7, allowing the identification of effector memory and central memory T cell subsets. Despite these initial advances in knowledge, more recent information has not revealed more clarity, but instead, has created a morass of complications—complications that, if not resolved, could harm correct vaccine design. Here, we discuss two central issues. The first is that we have always assumed that memory is induced in the same way, and consists of the same T cells, regardless of whether that immunity is generated by BCG vaccination, or by exposure to M. tuberculosis followed by effective chemotherapy. This assumption is almost certainly incorrect. Second, a myriad of additional memory subsets have now been described, such as resident, stem cell-like, tissue specific, among others, but as yet we know nothing about the relative importance of each, or whether if a new vaccine needs to induce all of these, or just some, to be fully effective

Gr1(int)CD11b+ myeloid-derived suppressor cells in Mycobacterium tuberculosis infection.

Tuberculosis is one of the world's leading killers, stealing 1.4 million lives and causing 8.7 million new and relapsed infections in 2011. The only vaccine against tuberculosis is BCG which demonstrates variable efficacy in adults worldwide. Human infection with Mycobacterium tuberculosis results in the influx of inflammatory cells to the lung in an attempt to wall off bacilli by forming a granuloma. Gr1(int)CD11b(+) cells are called myeloid-derived suppressor cells (MDSC) and play a major role in regulation of inflammation in many pathological conditions. Although MDSC have been described primarily in cancer their function in tuberculosis remains unknown. During M. tuberculosis infection it is crucial to understand the function of cells involved in the regulation of inflammation during granuloma formation. Understanding their relative impact on the bacilli and other cellular phenotypes is necessary for future vaccine and drug design.We compared the bacterial burden, lung pathology and Gr1(int)CD11b(+) myeloid-derived suppressor cell immune responses in M. tuberculosis infected NOS2-/-, RAG-/-, C3HeB/FeJ and C57/BL6 mice. Gr-1(+) cells could be found on the edges of necrotic lung lesions in NOS2-/-, RAG-/-, and C3HeB/FeJ, but were absent in wild-type mice. Both populations of Gr1(+)CD11b(+) cells expressed high levels of arginase-1, and IL-17, additional markers of myeloid derived suppressor cells. We then sorted the Gr1(hi) and Gr1(int) populations from M. tuberculosis infected NOS-/- mice and placed the sorted both Gr1(int) populations at different ratios with naïve or M. tuberculosis infected splenocytes and evaluated their ability to induce activation and proliferation of CD4+T cells. Our results showed that both Gr1(hi) and Gr1(int) cells were able to induce activation and proliferation of CD4+ T cells. However this response was reduced as the ratio of CD4(+) T to Gr1(+) cells increased. Our results illustrate a yet unrecognized interplay between Gr1(+) cells and CD4(+) T cells in tuberculosis

Stem cell analysis of fresh and frozen human breast milk

Pumping breast milk and freezing it is common practice in hospitals, neonatal intensive care units (NICU), and numerous households. Freezing milk may alter the cellular constituency of breast milk by reducing the amount of stem cells. Here, we investigate the alteration of stem cell viability and concentration in human breast milk frozen at varying temperatures and durations. This reduction may negatively impact infant development; vulnerable populations such as preterm infants have higher nutritional requirements and less volumetric capacity in their stomach. Freezing milk may undermine attempts to supply preterm infants with the nutritional requirements needed to sustain life

Gr1^int and Gr1^hi cells sorted from <i>M. tuberculosis</i> infected NOS-/- mice activate Th17 cells and induce proliferation of CD4⁺ T cells.

<p>Specific populations of Gr1^int or Gr1^hi cells were sorted from infected NOS-/- mice and placed into <i>in </i><i>vitro</i> cultures with increasing numbers of either stimulated naïve or infected splenocytes. After 2 days of incubation T cells were evaluated for activation and memory profiles (A-D). Panel A -D shows lower ratios of Gr1^high (A and C) or Gr1^int (B and D) induce increased the percentage of activation of CD4⁺CD69⁺IL-17⁺ and memory CD4⁺CD44⁺IL-17⁺, when co-incubated with naïve (A and B) or <i>M. tuberculosis</i> infected splenocytes (C and D), respectively. Panel E and F shows Gr1 ^high and Gr1^int induced of proliferating CD4 T cells. However, as the splenocyte to Gr1^high or Gr1^int ratio increased the percentage of CD4+ proliferation was reduced. Results are expressed as the mean values of mean percentage of CD4⁺CD69⁺IL-17⁺, CD4⁺CD44⁺IL-17⁺ and proliferation. ANOVA and the Tukey post-test. *p<0.05, **p<0.01, ***p<0.001.</p