Activated human monocytes inhibit the intracellular multiplication of legionnaires’ disease bacteria

We have examined the interaction between virulent egg yolk-grown L. pneumophila, Philadelphia 1 strain, and in vitro-activated human monocytes, under antibiotic-free conditions. Freshly explanted human monocytes activated by incubation with concanavalin A (Con A) and human lymphocytes inhibited the intracellular multiplication of L. pneumophila. Both Con A and lymphocytes were required for activation. Con A was consistently maximally effective at greater than or equal to 4 μg/ml. Monocytes activated by incubation with cell-free filtered supernatant from Con A-sensitized mononuclear cell cultures also inhibited the intracellular multiplication of L. pneumophil a. The most potent supernatant was obtained from mononuclear cell cultures incubated with greater than or equal to 15 μg/ml Con A for 48 h. The degree of monocyte inhibition of L. pneumophila multiplication was proportional to the length of time monocytes were preincubated with supernatant (48 {greater than} 24 {greater than} 12 h) and to the concentration of supernatant added (40 percent {greater than} 20 percent {greater than} 10 percent {greater than} 5 percent). Monocytes treated with supernatant daily were more inhibitory than monocytes treated initially only. With time in culture, monocytes progressively lost a limited degree of spontaneous inhibitory capacity and also lost their capacity to respond to supernatant with inhibition of L. pneumophila multiplication. Supernatant-activated monocytes inhibited L. pneumophila multiplication in two ways. They phagocytosed fewer bacteria, and they slowed the rate of intracellular multiplication of bacteria that were internalized. As was the case with nonactivated monocytes, antibody had no effect on the rate of intracellular multiplication in supernatant-activated monocytes. Neither supernatant-activated nor nonactivated monocytes killed L. pneumophila in the absence of antibody. Both killed a limited proportion of these bacteria in the presence of antibody and complement. We have previously reported that anti-L, pneumophila antibody and complement neither promote effective killing of L. pneumophila by human polymorphonuclear leukocytes and monocytes nor inhibit the rate of L. pneumophila multiplication in monocytes. These findings and our present report that activated monocytes do inhibit L. pneumophila multiplication indicate that cell-mediated immunity plays a major role in host defense against Legionnaires’ disease

Interaction of the legionnaires' disease bacterium (legionella pneumophila) with human phagocytes. I. L. pneumophila resists killing by polymorphonuclearleukocytes, antibody, and complement

We have previously reported that virulent egg yolk-grown Legionella pneumophila, Philadelphia 1 strain, multiplies intracellularly in human blood monocytes. We now report on the interaction between virulent L. pneumophila and human polymorphonuclear leukocytes (PMN), antibody, and complement, in vitro, under antibiotic-free conditions. L. pneumophila in concentrations ranging from 10(3) to 10(6) colony forming units (CFU)/ml are completely resistant to the bactericidal effects of 0-50 percent fresh normal human serum, even in the presence of high concentrations of rabbit or human anti-L. pneumophila antibody. L. pneumophila bacteria fix the third component of complement (C3) to their surfaces, as measured by fluorescence microscopy using rhodamine- conjugated goat anti-human C3 IgG, only when the bacteria are incubated with both specific anti-L. pneumophila antibody and complement. Similarly, L. pneumophila adhere to PMN, as measured by fluorescence microscopy, only in the presence of both specific antibody and complement. Electron microscopy revealed that these opsonized bacteria are phagocytosed by the PMN. PMN require both antibody and complement to kill L. pneumophila; even then, PMN reduced CFU of L. pneumophila by only 0.5 log under conditions in which they reduce CFU of a serum-resistant encapsulated strain of Escherichia coli by 2.5 logs. Separation of PMN-associated and nonassociated CFU of L. pneumophila revealed that the major proportion of the surviving bacteria are PMN associated. Thus, the ineffective killing of opsonized L. pneumophila is a result of a failure of PMN to kill these bacteria after they become PMN- associated. With or without antibody, PMN do not support the growth of L. pneumophila. These findings suggest that PMN, even in conjunction with the humoral immune system, do not play a decisive role in defense against the Legionnaires' disease bacterium

The Atp-dependent Rna Helicase Hrpb Plays An Important Role In Motility And Biofilm Formation In Xanthomonas Citri Subsp Citri

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)RNA helicases are enzymes that catalyze the separation of double-stranded RNA (dsRNA) using the free energy of ATP binding and hydrolysis. DEAD/DEAH families participate in many different aspects of RNA metabolism, including RNA synthesis, RNA folding, RNA-RNA interactions, RNA localization and RNA degradation. Several important bacterial DEAD/DEAH-box RNA helicases have been extensively studied. In this study, we characterize the ATP-dependent RNA helicase encoded by the hrpB (XAC0293) gene using deletion and genetic complementation assays. We provide insights into the function of the hrpB gene in Xanthomonas citri subsp. citri by investigating the roles of hrpB in biofilm formation on abiotic surfaces and host leaves, cell motility, host virulence of the citrus canker bacterium and growth in planta. Results: The hrpB gene is highly conserved in the sequenced strains of Xanthomonas. Mutation of the hrpB gene (Delta hrpB) resulted in a significant reduction in biofilms on abiotic surfaces and host leaves. Delta hrpB also exhibited increased cell dispersion on solid medium plates. Delta hrpB showed reduced adhesion on biotic and abiotic surfaces and delayed development in disease symptoms when sprayed on susceptible citrus leaves. Quantitative reverse transcription-PCR assays indicated that deletion of hrpB reduced the expression of four type IV pili genes. The transcriptional start site of fimA (XAC3241) was determined using rapid amplification of 5'-cDNA Ends (5' RACE). Based on the results of fimA mRNA structure predictions, the fimA 5' UTR may contain three different loops. HrpB may be involved in alterations to the structure of fimA mRNA that promote the stability of fimA RNA. Conclusions: Our data show that hrpB is involved in adherence of Xanthomonas citri subsp. citri to different surfaces. In addition, to the best of our knowledge, this is the first time that a DEAH RNA helicase has been implicated in the regulation of type IV pili in Xanthomonas.16INCT CitrusCAPES/PSDE fellowship [99999.002657/2014-07]CNPqCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Geomorfologia e a biodiversidade no Pantanal

The present study deals with the inter-relations in the relief which forms the Bacia do Alto Rio Paraguay (BAP) in mid-west Brazil. The overall aim is to discuss the relationship between relief forms and the biodiversity of the Pantanal. The BAP is a natural environmental system with contrasts in two of the compartments on which it is formed: the plateau, the most elevated compartment, highly transformed by human activities, and the plain which forms the Pantanal, which is more preserved and less transformed in relation to productive activities. The analysis was performed based on publications with a geomorphologic focus, examining the different relief units of the BAP and the dynamics of the revealing processes of landscape change which the Pantanal has undergone since the end of the Pleistocene.O presente estudo tem como objetivo abordar a inter-relação entre os compartimentos do relevo brasileiro que compõem a Bacia do Alto Paraguai - BAP, para discutir as formas de relevo e a biodiversidade do Pantanal. A abordagem fundamenta-se na compreensão das diferentes formas do terreno, associando ao fato que a BAP é um sistema ambiental natural que tem contrastes nos dois compartimentos que a compõem: o planalto, o compartimento mais elevado, fortemente transformado por atividades humanas; e a planície, que constitui o Pantanal mato-grossense, mais conservada e pouco transformada, quanto ao uso com atividades produtivas. A análise foi realizada a partir de publicações com enfoques geomorfológicos, abordando as diferentes unidades do relevo da BAP e o entendimento da dinâmica dos processos reveladores de que o Pantanal passa desde o fim do Pleistoceno, com processos de mudanças da paisagem.MEC - CAPE

Relationship between performance competence and cardiorespiratory fitness in contemporary dance

This is an accepted manuscript of an article published by Science & Medicine, Inc. in Medical Problems of Performing Artists on 01/06/2019, available online: https://doi.org/10.21091/mppa.2019.2014 The accepted version of the publication may differ from the final published version.© 2019 Science & Medicine. Aims: While a foundation of basic cardiorespiratory fitness is beneficial for coping with the physiological demands of dance training and performance, the extent to which cardiorespiratory fitness levels are related to performance ability is not all-together clear. This study aimed to directly compare aerobic capacity (VO2peak) and anaerobic threshold (AT) to an aesthetic competence measure (ACM) in student contemporary dancers. Methods: Participants were 18 contemporary dance students and all undertook a one-off treadmill test to volitional exhaustion in the week leading up to a performance to determine VO2peak and AT. In the same week, a final rehearsal for the performance was filmed to allow retrospective analysis of specific performance competence. Results: Mean VO2peak values of 47.67 ± 5.84 ml/kg/min and AT values of 43.18 ± 7.72 ml/kg/min (90.68 ± 11.87 %VO2peak) were recorded, and the mean total ACM score was 52.67 ± 8.74. No significant correlations were found between cardiorespiratory fitness variables and ACM scores. Regression analyses revealed experience level to be the only significant predictor of total ACM score (p<0.05, R2=0.12, SEE=11.91). Conclusions: The range of choreography used for assessment may limit the present study; nevertheless, as level of experience did significantly predict ACM total score, it is suggested that vocational dance training may be developing the performance and technical skills of students but not sufficiently developing their physical conditioning.Published versio