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1.
Proc Natl Acad Sci U S A ; 120(33): e2218869120, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37549251

RESUMO

In this paper, we introduce an efficient method for computing curves minimizing a variant of the Euler-Mumford elastica energy, with fixed endpoints and tangents at these endpoints, where the bending energy is enhanced with a user-defined and data-driven scalar-valued term referred to as the curvature prior. In order to guarantee that the globally optimal curve is extracted, the proposed method involves the numerical computation of the viscosity solution to a specific static Hamilton-Jacobi-Bellman (HJB) partial differential equation (PDE). For that purpose, we derive the explicit Hamiltonian associated with this variant model equipped with a curvature prior, discretize the resulting HJB PDE using an adaptive finite difference scheme, and solve it in a single pass using a generalized fast-marching method. In addition, we also present a practical method for estimating the curvature prior values from image data, designed for the task of accurately tracking curvilinear structure centerlines. Numerical experiments on synthetic and real-image data illustrate the advantages of the considered variant of the elastica model with a prior curvature enhancement in complex scenarios where challenging geometric structures appear.

2.
Plant J ; 2024 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-39073886

RESUMO

Genetic screens are powerful tools for biological research and are one of the reasons for the success of the thale cress Arabidopsis thaliana as a research model. Here, we describe the whole-genome sequencing of 871 Arabidopsis lines from the Homozygous EMS Mutant (HEM) collection as a novel resource for forward and reverse genetics. With an average 576 high-confidence mutations per HEM line, over three independent mutations altering protein sequences are found on average per gene in the collection. Pilot reverse genetics experiments on reproductive, developmental, immune and physiological traits confirmed the efficacy of the tool for identifying both null, knockdown and gain-of-function alleles. The possibility of conducting subtle repeated phenotyping and the immediate availability of the mutations will empower forward genetic approaches. The sequence resource is searchable with the ATHEM web interface (https://lipm-browsers.toulouse.inra.fr/pub/ATHEM/), and the biological material is distributed by the Versailles Arabidopsis Stock Center.

3.
Brain ; 2024 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-39049445

RESUMO

Perivascular macrophages (PVMs) and, to a lesser degree, microglia are targets and reservoirs of HIV and simian immunodeficiency virus (SIV) in the brain. Previously, we demonstrated that colony-stimulating factor 1 receptor (CSF1R) in PVMs was upregulated and activated in chronically SIV-infected rhesus macaques with encephalitis, correlating with SIV infection of PVMs. Herein, we investigated the role of CSF1R in the brain during acute SIV infection using BLZ945, a brain-penetrant CSF1R kinase inhibitor. Apart from three uninfected historic controls, nine Indian rhesus macaques were infected acutely with SIVmac251 and divided into three groups (n = 3 each): an untreated control and two groups treated for 20-30 days with low- (10 mg/kg/day) or high- (30 mg/kg/day) dose BLZ945. With the high-dose BLZ945 treatment, there was a significant reduction in cells expressing CD163 and CD206 across all four brain areas examined, compared with the low-dose treatment and control groups. In 9 of 11 tested regions, tissue viral DNA (vDNA) loads were reduced by 95%-99% following at least one of the two doses, and even to undetectable levels in some instances. Decreased numbers of CD163+ and CD206+ cells correlated significantly with lower levels of vDNA in all four corresponding brain areas. In contrast, BLZ945 treatment did not significantly affect the number of microglia. Our results indicate that doses as low as 10 mg/kg/day of BLZ945 are sufficient to reduce the tissue vDNA loads in the brain with no apparent adverse effect. This study provides evidence that infected PVMs are highly sensitive to CSF1R inhibition, opening new possibilities to achieve viral clearance.

4.
BMC Genomics ; 25(1): 777, 2024 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-39123115

RESUMO

BACKGROUND: Bacteria of the genus Xanthomonas cause economically significant diseases in various crops. Their virulence is dependent on the translocation of type III effectors (T3Es) into plant cells by the type III secretion system (T3SS), a process regulated by the master response regulator HrpG. Although HrpG has been studied for over two decades, its regulon across diverse Xanthomonas species, particularly beyond type III secretion, remains understudied. RESULTS: In this study, we conducted transcriptome sequencing to explore the HrpG regulons of 17 Xanthomonas strains, encompassing six species and nine pathovars, each exhibiting distinct host and tissue specificities. We employed constitutive expression of plasmid-borne hrpG*, which encodes a constitutively active form of HrpG, to induce the regulon. Our findings reveal substantial inter- and intra-specific diversity in the HrpG* regulons across the strains. Besides 21 genes directly involved in the biosynthesis of the T3SS, the core HrpG* regulon is limited to only five additional genes encoding the transcriptional activator HrpX, the two T3E proteins XopR and XopL, a major facility superfamily (MFS) transporter, and the phosphatase PhoC. Interestingly, genes involved in chemotaxis and genes encoding enzymes with carbohydrate-active and proteolytic activities are variably regulated by HrpG*. CONCLUSIONS: The diversity in the HrpG* regulon suggests that HrpG-dependent virulence in Xanthomonas might be achieved through several distinct strain-specific strategies, potentially reflecting adaptation to diverse ecological niches. These findings enhance our understanding of the complex role of HrpG in regulating various virulence and adaptive pathways, extending beyond T3Es and the T3SS.


Assuntos
Proteínas de Bactérias , Regulação Bacteriana da Expressão Gênica , Regulon , Xanthomonas , Xanthomonas/patogenicidade , Xanthomonas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Virulência/genética , Transcriptoma , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo , Perfilação da Expressão Gênica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
5.
Mol Plant Microbe Interact ; 36(11): 737-748, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37470457

RESUMO

Pseudomonas simiae WCS417 is a plant growth-promoting rhizobacterium that improves plant health and development. In this study, we investigate the early leaf responses of Arabidopsis thaliana to WCS417 exposure and the possible involvement of formate dehydrogenase (FDH) in such responses. In vitro-grown A. thaliana seedlings expressing an FDH::GUS reporter show a significant increase in FDH promoter activity in their roots and shoots after 7 days of indirect exposure (without contact) to WCS417. After root exposure to WCS417, the leaves of FDH::GUS plants grown in the soil also show an increased FDH promoter activity in hydathodes. To elucidate early foliar responses to WCS417 as well as FDH involvement, the roots of A. thaliana wild-type Col and atfdh1-5 knock-out mutant plants grown in soil were exposed to WCS417, and proteins from rosette leaves were subjected to proteomic analysis. The results reveal that chloroplasts, in particular several components of the photosystems PSI and PSII, as well as members of the glutathione S-transferase family, are among the early targets of the metabolic changes induced by WCS417. Taken together, the alterations in the foliar proteome, as observed in the atfdh1-5 mutant, especially after exposure to WCS417 and involving stress-responsive genes, suggest that FDH is a node in the early events triggered by the interactions between A. thaliana and the rhizobacterium WCS417. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteoma/metabolismo , Proteômica , Raízes de Plantas/microbiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Solo , Regulação da Expressão Gênica de Plantas
6.
New Phytol ; 238(4): 1593-1604, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36764921

RESUMO

Functional analysis of large gene families in plant pathogens can be cumbersome using classical insertional mutagenesis. Additionally, Cas9 toxicity has limited the application of CRISPR-Cas9 for directed mutagenesis in bacteria. Here, we successfully applied a CRISPR interference strategy to investigate the cryptic role of the transcription activator-like effector (tale) multigene family in several plant-pathogenic Xanthomonas bacterial species, owing to their contribution to pathogen virulence. Single guide RNAs (sgRNAs) designed against Xanthomonas phaseoli pv manihotis tale conserved gene sequences efficiently silenced expression of all tales, with concomitant decrease in virulence and TALE-induced host gene expression. The system is readily translatable to other Xanthomonas species infecting rice, citrus, Brassica, and cassava, silencing up to 16 tales in a given strain using a single sgRNA. Complementation with plasmid-borne designer tales lacking the sgRNA-targeted sequence restored molecular and virulence phenotypes in all pathosystems. Our results evidenced that X. campestris pv campestris CN08 tales are relevant for symptom development in cauliflower. They also show that the MeSWEET10a sugar transporter is surprisingly targeted by the nonvascular cassava pathogen X. cassavae, highlighting a new example of TALE functional convergence between phylogenetically distant Xanthomonas. Overall, this novel technology provides a platform for discovery and rapid functional understanding of highly conserved gene families.


Assuntos
Oryza , Xanthomonas , Efetores Semelhantes a Ativadores de Transcrição/genética , Xanthomonas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Virulência/genética , Transporte Biológico , Doenças das Plantas/microbiologia , Oryza/genética
7.
J Arthroplasty ; 38(6S): S246-S252, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36931358

RESUMO

BACKGROUND: This study evaluated the ability to achieve the targeted soft-tissue balance in terms of medio-lateral (ML) laxity and gap values when using a computer-assisted orthopedic surgery (CAOS) system featuring an intra-articular force-controlled distractor and assessed learning curves associated with the adoption of this technology. METHODS: The first 273 cases using this technology were reported without exclusions comparing 1) final ML laxity and 2) final average gap to their predefined targets. For both parameters, the signed and unsigned differentials were reported. The linear mixed model was used to evaluate laxity curve differences between surgeons. A cumulative sum control chart (CUSUM) was applied to assess surgeon learning curves regarding surgical time. RESULTS: Both the average signed ML laxity and gap differentials were neutral throughout the full arc of motion. Both the average unsigned ML laxity and gap differentials were linear. Signature of ML laxity and gap differential curves tended to be surgeon-specific. The CUSUM analyses of surgical times demonstrated either a short learning curve or the absence of a discernible learning pattern for surgeons. CONCLUSION: Data from all users involved with the pilot release of the balancing device were considered to capture variability in familiarity with the technique and learning curve cases were included. A high ability to achieve targeted gap balance throughout the arc of motion using the proposed method was observed.


Assuntos
Artroplastia do Joelho , Osteoartrite do Joelho , Cirurgia Assistida por Computador , Humanos , Articulação do Joelho/cirurgia , Artroplastia do Joelho/métodos , Amplitude de Movimento Articular , Movimento (Física) , Osteoartrite do Joelho/cirurgia
8.
Mol Plant Microbe Interact ; 35(9): 791-802, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35536128

RESUMO

Pathovars of Xanthomonas campestris cause distinct diseases on different brassicaceous hosts. The genomic relationships among pathovars as well as the genetic determinants of host range and tissue specificity remain poorly understood despite decades of research. Here, leveraging advances in multiplexed long-read technology, we fully sequenced the genomes of a collection of X. campestris strains isolated from cruciferous crops and weeds in New York and California as well as strains from global collections, to investigate pathovar relationships and candidate genes for host- and tissue-specificity. Pathogenicity assays and genomic comparisons across this collection and publicly available X. campestris genomes revealed a correlation between pathovar and genomic relatedness and provide support for X. campestris pv. barbareae, the validity of which had been questioned. Linking strain host range with type III effector repertoires identified AvrAC (also 'XopAC') as a candidate host-range determinant, preventing infection of Matthiola incana, and this was confirmed experimentally. Furthermore, the presence of a copy of the cellobiosidase gene cbsA with coding sequence for a signal peptide was found to correlate with the ability to infect vascular tissues, in agreement with a previous study of diverse Xanthomonas species; however, heterologous expression in strains lacking the gene gave mixed results, indicating that factors in addition to cbsA influence tissue specificity of X. campestris pathovars. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.


Assuntos
Xanthomonas campestris , Xanthomonas , Genômica , Especificidade de Órgãos , Sinais Direcionadores de Proteínas , Xanthomonas/genética , Xanthomonas campestris/genética
9.
New Phytol ; 236(1): 235-248, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35706385

RESUMO

Plant diseases are an important threat to food production. While major pathogenicity determinants required for disease have been extensively studied, less is known on how pathogens thrive during host colonization, especially at early infection stages. Here, we used randomly barcoded-transposon insertion site sequencing (RB-TnSeq) to perform a genome-wide screen and identify key bacterial fitness determinants of the vascular pathogen Xanthomonas campestris pv campestris (Xcc) during infection of the cauliflower host plant (Brassica oleracea). This high-throughput analysis was conducted in hydathodes, the natural entry site of Xcc, in xylem sap and in synthetic media. Xcc did not face a strong bottleneck during hydathode infection. In total, 181 genes important for fitness were identified in plant-associated environments with functional enrichment in genes involved in metabolism but only few genes previously known to be involved in virulence. The biological relevance of 12 genes was independently confirmed by phenotyping single mutants. Notably, we show that XC_3388, a protein with no known function (DUF1631), plays a key role in the adaptation and virulence of Xcc possibly through c-di-GMP-mediated regulation. This study revealed yet unsuspected social behaviors adopted by Xcc individuals when confined inside hydathodes at early infection stages.


Assuntos
Brassica , Xanthomonas campestris , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brassica/microbiologia , Doenças das Plantas/microbiologia , Virulência/genética , Xilema/metabolismo
10.
New Phytol ; 219(1): 391-407, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29677397

RESUMO

Xanthomonas transcription activator-like effectors (TALEs) are injected inside plant cells to promote host susceptibility by enhancing transcription of host susceptibility genes. TALE-encoding (tal) genes were thought to be absent from Brassicaceae-infecting Xanthomonas campestris (Xc) genomes based on four reference genomic sequences. We discovered tal genes in 26 of 49 Xc strains isolated worldwide and used a combination of single molecule real time (SMRT) and tal amplicon sequencing to yield a near-complete description of the TALEs found in Xc (Xc TALome). The 53 sequenced tal genes encode 21 distinct DNA binding domains that sort into seven major DNA binding specificities. In silico analysis of the Brassica rapa promoterome identified a repertoire of predicted TALE targets, five of which were experimentally validated using quantitative reverse transcription polymerase chain reaction. The Xc TALome shows multiple signs of DNA rearrangements that probably drove its evolution from two ancestral tal genes. We discovered that Tal12a and Tal15a of Xcc strain Xca5 contribute together in the development of disease symptoms on susceptible B. oleracea var. botrytis cv Clovis. This large and polymorphic repertoire of TALEs opens novel perspectives for elucidating TALE-mediated susceptibility of Brassicaceae to black rot disease and for understanding the molecular processes underlying TALE evolution.


Assuntos
Interações Hospedeiro-Patógeno/genética , Efetores Semelhantes a Ativadores de Transcrição/genética , Xanthomonas campestris/genética , Xanthomonas campestris/patogenicidade , Brassica/microbiologia , Genoma Bacteriano , Filogenia , Doenças das Plantas/microbiologia
11.
Plant Physiol ; 174(2): 700-716, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28184011

RESUMO

Hydathodes are water pores found on leaves of a wide range of vascular plants and are the sites of guttation. We report here on the detailed anatomy of cauliflower (Brassicaoleracea) and Arabidopsis (Arabidopsis thaliana) hydathodes. Hydathode surface presents pores resembling stomata giving access to large cavities. Beneath, the epithem is composed of a lacunar and highly vascularized parenchyma offering a direct connection between leaf surface and xylem vessels. Arabidopsis hydathode pores were responsive to ABA and light similar to stomata. The flg22 flagellin peptide, a well-characterized elicitor of plant basal immunity, did not induce closure of hydathode pores in contrast to stomata. Because hydathodes are natural infection routes for several pathogens, we investigated hydathode infection by the adapted vascular phytopathogenic bacterium Xanthomonas campestris pv campestris (Xcc), the causal agent of black rot disease of Brassicaceae. Microscopic observations of hydathodes six days postinoculation indicated a digestion of the epithem cells and a high bacterial multiplication. Postinvasive immunity was shown to limit pathogen growth in the epithem and is actively suppressed by the type III secretion system and its effector proteins. Altogether, these results give a detailed anatomic description of Brassicaceae hydathodes and highlight the efficient use of this tissue as an initial niche for subsequent vascular systemic dissemination of Xcc in distant plant tissues.


Assuntos
Brassica/anatomia & histologia , Brassica/imunologia , Doenças das Plantas/imunologia , Xanthomonas campestris/patogenicidade , Ácido Abscísico/farmacologia , Arabidopsis/anatomia & histologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/imunologia , Arabidopsis/microbiologia , Brassica/microbiologia , Interações Hospedeiro-Patógeno , Folhas de Planta/microbiologia , Estômatos de Plantas/anatomia & histologia , Plantas Geneticamente Modificadas , Xanthomonas campestris/genética
12.
J Sports Sci ; 36(11): 1269-1276, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28873044

RESUMO

This study aimed to investigate the acute effects of two barbell hip thrust-based (BHT) post-activation potentiation (PAP) protocols on subsequent sprint performance. Using a crossover design, eighteen handball athletes performed maximal 15-m sprints before and 15s, 4min and 8min after two experimental protocols consisting of BHT loaded with either 50% or 85% 1RM (50PAP and 85PAP, respectively), in order to profile the transient PAP effects. The resulting sprint performances were significantly impaired at 15s only after the 85PAP protocol, which induced likely and very likely greater decreases compared to the 50PAP. At 4min and 8min, significant improvements and very likely beneficial effects were observed in the 10m and 15m performances following both protocols. Significant differences were found when comparing the two PAPs over time; the results suggested very likely greater performance improvements in 10m following the 85PAP after 4min and 8min, and possible greater performance improvements in 15m after 4min. Positive correlations between BHT 1RMs values and the greatest individual PAP responses on sprint performance were found. This investigation showed that both moderate and intensive BHT exercises can induce a PAP response, but the effects may differ according to the recovery following the potentiating stimulus and the individual`s strength level.


Assuntos
Desempenho Atlético/fisiologia , Treinamento Resistido/métodos , Corrida/fisiologia , Esportes/fisiologia , Aceleração , Estudos Cross-Over , Quadril/fisiologia , Humanos , Força Muscular/fisiologia , Adulto Jovem
13.
Knee Surg Sports Traumatol Arthrosc ; 26(11): 3325-3332, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29476198

RESUMO

PURPOSE: Posterior tibial slope (PTS) for cruciate-retaining (CR) total knee arthroplasty (TKA) is usually pre-determined by the surgeon. Limited information is available comparing different choices of PTS on the kinematics of the CR TKA, independent of the balancing of the extension gap. This study hypothesized that with the same balanced extension gap, the choice of PTS significantly impacts the intraoperatively measured kinematics of CR TKA. METHODS: Navigated CR TKAs were performed on seven fresh-frozen cadavers with healthy knees and intact posterior cruciate ligament (PCL). A custom designed tibial baseplate was implanted to allow in situ modification of the PTS, which altered the flexion gap but maintained the extension gap. Knee kinematics were measured by performing passive range of motion (ROM) tests from full extension to 120° of flexion on the intact knee and CR TKAs with four different PTSs (1°, 4°, 7°, and 10°). The measured kinematics were compared across test conditions to assess the impact of PTS. RESULTS: With a consistent extension gap, the change of PTS had significant impact on the anteroposterior (AP) kinematics of the CR TKA knees in mid-flexion range (45°-90°), but not so much for the high-flexion range (90°-120°). No considerable impacts were found on internal/external (I/E) rotation and hip-knee-ankle (HKA) angle. However, the findings on the individual basis suggested the impact of PTS on I/E rotation and HKA angle may be patient-specific. CONCLUSIONS: The data suggested that the choice of PTS had the greatest impact on the mid-flexion AP translation among the intraoperatively measured kinematics. This impact may be considered while making surgical decisions in the context of AP kinematics. When using a tibial component designed with "center" pivoting PTS, a surgeon may be able to fine tune the PTS to achieve proper mid-flexion AP stability.


Assuntos
Artroplastia do Joelho/métodos , Articulação do Joelho/fisiologia , Tíbia/anatomia & histologia , Idoso , Fenômenos Biomecânicos/fisiologia , Cadáver , Feminino , Humanos , Prótese do Joelho , Masculino , Ligamento Cruzado Posterior , Amplitude de Movimento Articular/fisiologia
15.
Diabetes Obes Metab ; 18(1): 6-15, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26228188

RESUMO

Diabetes mellitus is a growing worldwide epidemic disease, currently affecting 1 in 12 adults. Treatment of disease complications typically consumes ∼10% of healthcare budgets in developed societies. Whilst immune-mediated destruction of insulin-secreting pancreatic ß cells is responsible for Type 1 diabetes, both the loss and dysfunction of these cells underly the more prevalent Type 2 diabetes. The establishment of robust drug development programmes aimed at ß-cell restoration is still hampered by the absence of means to measure ß-cell mass prospectively in vivo, an approach which would provide new opportunities for understanding disease mechanisms and ultimately assigning personalized treatments. In the present review, we describe the progress towards this goal achieved by the Innovative Medicines Initiative in Diabetes, a collaborative public-private consortium supported by the European Commission and by dedicated resources of pharmaceutical companies. We compare several of the available imaging methods and molecular targets and provide suggestions as to the likeliest to lead to tractable approaches. Furthermore, we discuss the simultaneous development of animal models that can be used to measure subtle changes in ß-cell mass, a prerequisite for validating the clinical potential of the different imaging tracers.


Assuntos
Diabetes Mellitus/patologia , Células Secretoras de Insulina/patologia , Imagem Molecular/métodos , Adulto , Animais , Adesão Celular , Receptor do Peptídeo Semelhante ao Glucagon 1/metabolismo , Humanos , Células Secretoras de Insulina/metabolismo , Medições Luminescentes , Manganês , Glicoproteínas de Membrana/metabolismo , Camundongos , Ratos , Receptores de Sulfonilureias/metabolismo , Proteínas Vesiculares de Transporte de Monoamina/metabolismo , Zinco
16.
Acta Anaesthesiol Scand ; 60(7): 995-1002, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26860837

RESUMO

BACKGROUND: Ultrasound-guided regional anesthesia (UGRA) requires acquisition of new skills. Learning requires one-on-one teaching, and can be limited by time and mentor availability. We investigate whether the skills required for UGRA can be developed and subsequently assessed remotely using a novel online teaching platform. This platform was developed at the University of Toronto to teach laparoscopic surgery remotely and has been termed Telesimulation. METHODS: Anesthesia Site Chiefs at 10 hospitals across Ontario were sent a letter inviting their anesthesia teams to participate in an UGRA remote training program. Four to five anesthetists from each site were recruited from the first four hospitals expressing interest. Simulation models and ultrasound machines were set up at each location and connected via Skype(™) and web cameras with the Telesimulation center at our hospital. Training consisted of four online sessions and one offline lecture in order to teach an ultrasound-guided supraclavicular block. Participants were evaluated before and after training by on-site and off-site assessors using a validated Checklist and Global Rating Scale (GRS). RESULTS: Nineteen staff anesthetists were recruited. Post-training scores were significantly higher across both assessment tools, on-site (P < 0.001) and off-site training locations (P = 0.003). The inter-rater reliability between on-site and remote training site ratings was good for the Checklist (ICC = 0.672, 95% CI: 0.369-0.830) and excellent for the GRS (ICC = 0.847, 95% CI: 0.706-0.921). CONCLUSION: This study demonstrates that UGRA can be taught remotely. Future research will focus on comparing this method to on-site teaching and its application in resource-restricted countries.


Assuntos
Anestesia por Condução/métodos , Anestesiologia/educação , Telecomunicações , Ultrassonografia de Intervenção/métodos , Anestesiologia/métodos , Canadá , Competência Clínica , Estudos de Viabilidade , Humanos , Reprodutibilidade dos Testes
18.
Anaesthesia ; 71(6): 648-56, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27062151

RESUMO

Limited data exist on platelet transfusion during postpartum haemorrhage. We retrospectively analysed a consecutive cohort from a single centre of 347 women with moderate or severe postpartum haemorrhage, transfused according to national guidelines. Twelve (3%) women required a platelet transfusion. There were no differences between women who did and did not receive platelets with respect to age, mode of initiation of labour or mode of delivery. Women receiving a platelet transfusion had a lower median (IQR [range]) platelet count at study entry than women who did not receive platelets before haemorrhage (135 (97-175 [26-259])×10(9) .l(-1) vs 224 (186-274 [91-1006])×10(9) .l(-1) ), respectively), and at diagnosis of postpartum haemorrhage (median 114 (78-153 [58-238])×10(9) .l(-1) vs 193 (155-243 [78-762])×10(9) .l(-1) respectively). Six women were thrombocytopenic pre-delivery. The cause of haemorrhage that was associated with the highest rate of platelet transfusion was placental abruption, with three of 14 women being transfused. If antenatal thrombocytopenia or consumptive coagulopathy were not present, platelets were only required for haemorrhage > 5000 ml. Early formulaic platelet transfusion would have resulted in many women receiving platelets unnecessarily. Using current guidelines, the need for platelet transfusion is uncommon without antenatal thrombocytopenia, consumptive coagulopathy or haemorrhage > 5000 ml. We found no evidence to support early fixed-ratio platelet transfusion.


Assuntos
Contagem de Plaquetas , Transfusão de Plaquetas , Hemorragia Pós-Parto/terapia , Adolescente , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Hemorragia Pós-Parto/sangue , Gravidez , Estudos Retrospectivos , Adulto Jovem
19.
BMC Genomics ; 16: 975, 2015 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-26581393

RESUMO

BACKGROUND: The bacterial species Xanthomonas campestris infects a wide range of Brassicaceae. Specific pathovars of this species cause black rot (pv. campestris), bacterial blight of stock (pv. incanae) or bacterial leaf spot (pv. raphani). RESULTS: In this study, we extended the genomic coverage of the species by sequencing and annotating the genomes of strains from pathovar incanae (CFBP 1606R and CFBP 2527R), pathovar raphani (CFBP 5828R) and a pathovar formerly named barbareae (CFBP 5825R). While comparative analyses identified a large core ORFeome at the species level, the core type III effectome was limited to only three putative type III effectors (XopP, XopF1 and XopAL1). In Xanthomonas, these effector proteins are injected inside the plant cells by the type III secretion system and contribute collectively to virulence. A deep and strand-specific RNA sequencing strategy was adopted in order to experimentally refine genome annotation for strain CFBP 5828R. This approach also allowed the experimental definition of novel ORFs and non-coding RNA transcripts. Using a constitutively active allele of hrpG, a master regulator of the type III secretion system, a HrpG-dependent regulon of 141 genes co-regulated with the type III secretion system was identified. Importantly, all these genes but seven are positively regulated by HrpG and 56 of those encode components of the Hrp type III secretion system and putative effector proteins. CONCLUSIONS: This dataset is an important resource to mine for novel type III effector proteins as well as for bacterial genes which could contribute to pathogenicity of X. campestris.


Assuntos
Perfilação da Expressão Gênica , Genômica , Xanthomonas campestris/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Anotação de Sequência Molecular , Fases de Leitura Aberta , Regulon/genética , Xanthomonas campestris/imunologia
20.
Epidemiol Infect ; 143(5): 910-21, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25050615

RESUMO

SUMMARY We analysed Mycobacterium tuberculosis strains from children, hospitalized from January 2004 to July 2008 in the largest paediatric hospital complex in Cambodia. Specimens were tested for drug susceptibility and genotypes. From the 260 children, 161 strains were available. The East African-Indian genotype family was the most common (59.0%), increasing in frequency with distance from the Phnom Penh area, while the frequency of the Beijing genotype family strains decreased. The drug resistance pattern showed a similar geographical gradient: lowest in the northwest (4.6%), intermediate in the central (17.1%), and highest in the southeastern (30.8%) parts of the country. Three children (1.9%) had multidrug-resistant tuberculosis. The Beijing genotype and streptomycin resistance were significantly associated (P < 0.001). As tuberculosis in children reflects recent transmission patterns in the community, multidrug resistance levels inform about the current quality of the tuberculosis programme.


Assuntos
DNA Bacteriano/análise , Mycobacterium tuberculosis/genética , Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose/epidemiologia , Adolescente , Antituberculosos/farmacologia , Camboja/epidemiologia , Criança , Pré-Escolar , Farmacorresistência Bacteriana Múltipla/genética , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Isoniazida/farmacologia , Masculino , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/efeitos dos fármacos , Rifampina/farmacologia , Estreptomicina/farmacologia , Tuberculose/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia
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