2,566 research outputs found
Production of the Catechol Type Siderophore Bacillibactin by the Honey Bee Pathogen Paenibacillus larvae
The Gram-positive bacterium Paenibacillus larvae is the etiological agent of
American Foulbrood. This bacterial infection of honey bee brood is a
notifiable epizootic posing a serious threat to global honey bee health
because not only individual larvae but also entire colonies succumb to the
disease. In the recent past considerable progress has been made in elucidating
molecular aspects of host pathogen interactions during pathogenesis of P.
larvae infections. Especially the sequencing and annotation of the complete
genome of P. larvae was a major step forward and revealed the existence of
several giant gene clusters coding for non-ribosomal peptide synthetases which
might act as putative virulence factors. We here present the detailed analysis
of one of these clusters which we demonstrated to be responsible for the
biosynthesis of bacillibactin, a P. larvae siderophore. We first established
culture conditions allowing the growth of P. larvae under iron-limited
conditions and triggering siderophore production by P. larvae. Using a gene
disruption strategy we linked siderophore production to the expression of an
uninterrupted bacillibactin gene cluster. In silico analysis predicted the
structure of a trimeric trithreonyl lactone (DHB-Gly-Thr)3 similar to the
structure of bacillibactin produced by several Bacillus species. Mass
spectrometric analysis unambiguously confirmed that the siderophore produced
by P. larvae is identical to bacillibactin. Exposure bioassays demonstrated
that P. larvae bacillibactin is not required for full virulence of P. larvae
in laboratory exposure bioassays. This observation is consistent with results
obtained for bacillibactin in other pathogenic bacteria
Biological effects of paenilamicin, a secondary metabolite antibiotic produced by the honey bee pathogenic bacterium Paenibacillus larvae
Paenibacillus larvae is the etiological agent of American Foulbrood (AFB) a world-wide distributed devastating disease of the honey bee brood. Previous comparative genome analysis and more recently, the elucidation of the bacterial genome, provided evidence that this bacterium harbors putative functional nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) and therefore, might produce nonribosomal peptides (NRPs) and polyketides (PKs). Such biosynthesis products have been shown to display a wide-range of biological activities such as antibacterial, antifungal or cytotoxic activity. Herein we present an in silico analysis of the first NRPS/PKS hybrid of P. larvae and we show the involvement of this cluster in the production of a compound named paenilamicin (Pam). For the characterization of its in vitro and in vivo bioactivity, a knock-out mutant strain lacking the production of Pam was constructed and subsequently compared to wild-type species. This led to the identification of Pam by mass spectrometry. Purified Pam-fractions showed not only antibacterial but also antifungal and cytotoxic activities. The latter suggested a direct effect of Pam on honey bee larval death which could, however, not be corroborated in laboratory infection assays. Bee larvae infected with the non-producing Pam strain showed no decrease in larval mortality, but a delay in the onset of larval death. We propose that Pam, although not essential for larval mortality, is a virulence factor of P. larvae influencing the time course of disease. These findings are not only of significance in elucidating and understanding host-pathogen interactions but also within the context of the quest for new compounds with antibiotic activity for drug development.DFG, GRK 1121, Genetische und immunologische Determinanten von Pathogen-Wirt-InteraktionenDFG, EXC 314, Unifying Concepts in Catalysi
Paenibacillus larvae Chitin-Degrading Protein PlCBP49 Is a Key Virulence Factor in American Foulbrood of Honey Bees
Paenibacillus larvae, the etiological agent of the globally occurring
epizootic American Foulbrood (AFB) of honey bees, causes intestinal infections
in honey bee larvae which develop into systemic infections inevitably leading
to larval death. Massive brood mortality might eventually lead to collapse of
the entire colony. Molecular mechanisms of host-microbe interactions in this
system and of differences in virulence between P. larvae genotypes are poorly
understood. Recently, it was demonstrated that the degradation of the
peritrophic matrix lining the midgut epithelium is a key step in pathogenesis
of P. larvae infections. Here, we present the isolation and identification of
PlCBP49, a modular, chitin-degrading protein of P. larvae and demonstrate that
this enzyme is crucial for the degradation of the larval peritrophic matrix
during infection. PlCBP49 contains a module belonging to the auxiliary
activity 10 (AA10, formerly CBM33) family of lytic polysaccharide
monooxygenases (LPMOs) which are able to degrade recalcitrant polysaccharides.
Using chitin-affinity purified PlCBP49, we provide evidence that PlCBP49
degrades chitin via a metal ion-dependent, oxidative mechanism, as already
described for members of the AA10 family. Using P. larvae mutants lacking
PlCBP49 expression, we analyzed in vivo biological functions of PlCBP49. In
the absence of PlCBP49 expression, peritrophic matrix degradation was markedly
reduced and P. larvae virulence was nearly abolished. This indicated that
PlCBP49 is a key virulence factor for the species P. larvae. The
identification of the functional role of PlCBP49 in AFB pathogenesis broadens
our understanding of this important family of chitin-binding and -degrading
proteins, especially in those bacteria that can also act as entomopathogens
Role of bacterial community composition as a driver of the small-sized phytoplankton community structure in a productive coastal system
Financiado para publicación en acceso aberto: Universidade de Vigo/CISUGWe present here the first detailed description of the seasonal patterns in bacterial community composition (BCC) in shelf waters off the Ría de Vigo (Spain), based on monthly samplings during 2 years. Moreover, we studied the relationship between bacterial and small-sized eukaryotic community composition to identify potential biotic interactions among components of these two communities. Bacterial operational taxonomic unit (OTU) richness and diversity systematically peaked in autumn–winter, likely related to low resource availability during this period. BCC showed seasonal and vertical patterns, with Rhodobacteraceae and Flavobacteriaceae families dominating in surface waters, and SAR11 clade dominating at the base of the photic zone (30 m depth). BCC variability was significantly explained by environmental variables (e.g., temperature of water, solar radiation, or dissolved organic matter). Interestingly, a strong and significant correlation was found between BCC and small-sized eukaryotic community composition (ECC), which suggests that biotic interactions may play a major role as structuring factors of the microbial plankton in this productive area. In addition, co-occurrence network analyses revealed strong and significant, mostly positive, associations between bacteria and small-sized phytoplankton. Positive associations likely result from mutualistic relationships (e.g., between Dinophyceae and Rhodobacteraceae), while some negative correlations suggest antagonistic interactions (e.g., between Pseudo-nitzchia sp. and SAR11). These results support the key role of biotic interactions as structuring factors of the small-sized eukaryotic community, mostly driven by positive associations between small-sized phytoplankton and bacteria.Xunta de Galicia | Ref. EM2013/023Xunta de Galicia | Ref. ED481A-2019/290Xunta de Galicia | Ref. ED431I 2020/03Ministerio de Economía y Competitividad | Ref. CTM2017-83362-RMinisterio de Ciencia e Innovación | Ref. PID2019-110011RB-C3
Agreement between two photoplethysmography-based wearable devices for monitoring heart rate during different physical activity situations : a new analysis methodology
Wearables are being increasingly used to monitor heart rate (HR). However, their usefulness for analyzing continuous HR in research or at clinical level is questionable. The aim of this study is to analyze the level of agreement between different wearables in the measurement of HR based on photoplethysmography, according to different body positions and physical activity levels, and compared to a gold-standard ECG. The proposed method measures agreement among several time scales since different wearables obtain HR at different sampling rates. Eighteen university students (10 men, 8 women; 22 ± 2.45 years old) participated in a laboratory study. Participants simultaneously wore an Apple Watch and a Polar Vantage watch. ECG was measured using a BIOPAC system. HR was recorded continuously and simultaneously by the three devices, for consecutive 5-min periods in 4 different situations: lying supine, sitting, standing and walking at 4 km/h on a treadmill. HR estimations were obtained with the maximum precision offered by the software of each device and compared by averaging in several time scales, since the wearables obtained HR at different sampling rates, although results are more detailed for 5 s and 30 s epochs. Bland-Altman (B-A) plots show that there is no noticeable difference between data from the ECG and any of the smartwatches while participants were lying down. In this position, the bias is low when averaging in both 5 s and 30 s. Differently, B-A plots show that there are differences when the situation involves some level of physical activity, especially for shorter epochs. That is, the discrepancy between devices and the ECG was greater when walking on the treadmill and during short time scales. The device showing the biggest discrepancy was the Polar Watch, and the one with the best results was the Apple Watch. We conclude that photoplethysmography-based wearable devices are suitable for monitoring HR averages at regular intervals, especially at rest, but their feasibility is debatable for a continuous analysis of HR for research or clinical purposes, especially when involving some level of physical activity. An important contribution of this work is a new methodology to synchronize and measure the agreement against a gold standard of two or more devices measuring HR at different and not necessarily even paces
Genomic Potential and Virulence Mechanisms of Paenibacillus larvae
Paenibacillus larvae, a Gram positive bacterial pathogen, causes American
Foulbrood (AFB), which is the most serious infectious disease of honey bees.
In order to investigate the genomic potential of P. larvae, two strains
belonging to two different genotypes were sequenced and used for comparative
genome analysis. The complete genome sequence of P. larvae strain DSM 25430
(genotype ERIC II) consisted of 4,056,006 bp and harbored 3,928 predicted
protein-encoding genes. The draft genome sequence of P. larvae strain DSM
25719 (genotype ERIC I) comprised 4,579,589 bp and contained 4,868 protein-
encoding genes. Both strains harbored a 9.7 kb plasmid and encoded a large
number of virulence-associated proteins such as toxins and collagenases. In
addition, genes encoding large multimodular enzymes producing nonribosomally
peptides or polyketides were identified. In the genome of strain DSM 25719
seven toxin associated loci were identified and analyzed. Five of them encoded
putatively functional toxins. The genome of strain DSM 25430 harbored several
toxin loci that showed similarity to corresponding loci in the genome of
strain DSM 25719, but were non-functional due to point mutations or disruption
by transposases. Although both strains cause AFB, significant differences
between the genomes were observed including genome size, number and
composition of transposases, insertion elements, predicted phage regions, and
strain-specific island-like regions. Transposases, integrases and recombinases
are important drivers for genome plasticity. A total of 390 and 273 mobile
elements were found in strain DSM 25430 and strain DSM 25719, respectively.
Comparative genomics of both strains revealed acquisition of virulence factors
by horizontal gene transfer and provided insights into evolution and
pathogenicity
Actividad extracurricular en el ámbito de la Ingeniería para la competencia genérica de comunicación escrita
El objetivo de este trabajo es describir una actividad formativa extracurricular para la competencia de comunicación escrita, que puede ser fácilmente reproducida en otros centros y universidades. Para ello, en este trabajo se indica el marco en el que se pueden desarrollar las competencias genéricas y se analiza brevemente la actividad propuesta a través de sus resultados de aprendizaje, y cómo estos favorecen e influyen en otras competencias genéricas como la capacidad creativa y el aprendizaje para toda la vida.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech
Linking the impact of bacteria on phytoplankton growth with microbial community composition and co-occurrence patterns
The interactions between microalgae and bacteria have recently emerged as key control factors which might contribute to a better understanding on how phytoplankton communities assemble and respond to environmental disturbances. We analyzed partial 16S rRNA and 18S rRNA genes from a total of 42 antibiotic bioassays, where phytoplankton growth was assessed in the presence or absence of an active bacterial community. A significant negative impact of bacteria was observed in 18 bioassays, a significant positive impact was detected in 5 of the cases, and a non-detectable effect occurred in 19 bioassays. Thalasiossira spp., Chlorophytes, Vibrionaceae and Alteromonadales were relatively more abundant in the samples where a positive effect of bacteria was observed compared to those where a negative impact was observed. Phytoplankton diversity was lower when bacteria negatively affect their growth than when the effect was beneficial. The phytoplankton-bacteria co-occurrence subnetwork included many significant Chlorophyta-Alteromonadales and Bacillariophyceae-Alteromonadales positive associations. Phytoplankton-bacteria co-exclusions were not detected in the network, which contrasts with the negative effect of bacteria on phytoplankton growth frequently detected in the bioassays, suggesting strong competitive interactions. Overall, this study adds strong evidence supporting the key role of phytoplanktonbacteria interactions in the microbial communities.Agencia Estatal de Investigación | Ref. CTM2017-83362-RAgencia Estatal de Investigación | Ref. PID2019-110011RB-C33Xunta de Galicia | Ref. ED481A-2019/290Xunta de Galicia | Ref. ED481A-2018/288Universidade de Vigo/CISU
SOX2 Expression and Transcriptional Activity Identifies a Subpopulation of Cancer Stem Cells in Sarcoma with Prognostic Implications
Stemness in sarcomas is coordinated by the expression of pluripotency factors, like SOX2, in cancer stem cells (CSC). The role of SOX2 in tumor initiation and progression has been well characterized in osteosarcoma. However, the pro-tumorigenic features of SOX2 have been scarcely investigated in other sarcoma subtypes. Here, we show that SOX2 depletion dramatically reduced the ability of undifferentiated pleomorphic sarcoma (UPS) cells to form tumorspheres and to initiate tumor growth. Conversely, SOX2 overexpression resulted in increased in vivo tumorigenicity. Moreover, using a reporter system (SORE6) which allows to monitor viable cells expressing SOX2 and/or OCT4, we found that SORE6+ cells were significantly more tumorigenic than the SORE6- subpopulation. In agreement with this findings, SOX2 expression in sarcoma patients was associated to tumor grade, differentiation, invasive potential and lower patient survival. Finally, we studied the effect of a panel of anti-tumor drugs on the SORE6+ cells of the UPS model and patient-derived chondrosarcoma lines. We found that the mithramycin analogue EC-8042 was the most efficient in reducing SORE6+ cells in vitro and in vivo. Overall, this study demonstrates that SOX2 is a pro-tumorigenic factor with prognostic potential in sarcoma. Moreover, SORE6 transcriptional activity is a bona fide CSC marker in sarcoma and constitutes an excellent biomarker for evaluating the efficacy of anti-tumor treatments on CSC subpopulations.This work was supported by the Agencia Estatal de Investigación (AEI) [MINECO/Fondo Europeo de Desarrollo Regional (FEDER) (SAF-2016-75286-R to R.R.), ISC III/FEDER (Miguel Servet Program CPII16/00049 to R.R., Sara Borrell Program CD16/00103 to S.T.M. and PI16/00280 and PI19/00560 to J.M.G-P) and Consorcio CIBERONC CB16/12/00390)] and the Plan de Ciencia Tecnología e Innovación del Principado de Asturias/FEDER (IDI/2018/155) to J.P.R and Predoctoral Fellowship Severo Ochoa (BP-17-108) to O.E.S
Factors affecting the relationship between psychological status and quality of life in COPD patients
<p>Abstract</p> <p>Background</p> <p>This study aims to (i) evaluate the association between anxiety and depressive symptoms and health-related quality of life (HRQoL); and (ii) identify the effect modifiers of this relationship in patients with chronic obstructive pulmonary disease (COPD).</p> <p>Methods</p> <p>A total of 337 clinically stable COPD patients answered the St. George's Respiratory Questionnaire (SGRQ) (assessing HRQoL) and the Hospital Anxiety and Depression Scale (HADS). Socio-demographic information, lung function, and other clinical data were collected.</p> <p>Results</p> <p>Most patients (93%) were male; they had a mean (SD) age of 68 (9) years and mild to very severe COPD (post-bronchodilator FEV<sub>1 </sub>52 (16)% predicted). Multivariate analyses showed that anxiety, depression, or both conditions were associated with poor HRQoL (for all SGRQ domains). The association between anxiety and total HRQoL score was 6.7 points higher (indicating a worse HRQoL) in current workers than in retired individuals. Estimates for patients with "both anxiety and depression" were 5.8 points lower in stage I-II than in stage III-IV COPD, and 10.2 points higher in patients with other comorbidities than in those with only COPD.</p> <p>Conclusions</p> <p>This study shows a significant association between anxiety, depression, or both conditions and impaired HRQoL. Clinically relevant factors affecting the magnitude of this association include work status, COPD severity, and the presence of comorbidities.</p
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