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1.
Br J Radiol ; 96(1151): 20220815, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37660369

ABSTRACT

OBJECTIVES: To assess the impact of fusion imaging guidance on fluoroscopy duration and volume of contrast agent used for pulmonary artery embolization. METHODS: Thirty-four consecutive patients who underwent pulmonary artery embolization for pulmonary arterio-venous malformation (n = 28) or hemoptysis (n = 6) were retrospectively included. In the experimental group (n = 15), patients were treated using fusion imaging with 2D/3D registration. In the control group (n = 19), no fusion imaging has been used. Fluoroscopy duration and amount of contrast used were measured and intergroup comparison was performed. RESULTS: The average volume of contrast agent used for embolization in the fusion group (118.3 ml) was significantly lower than in the control group (285.3 ml) (p < 0.002). The mean fluoroscopy duration was not significantly different between both groups (19.5 min in the fusion group vs 31.4 min in the control group (p = 0.10)). No significant difference was observed regarding the average X-ray exposure (Air Kerma) (p = 0.68 in the univariate analysis). Technical success rate was 100% for both groups. CONCLUSION: Fusion imaging significantly reduces contrast medium volumes needed to perform pulmonary artery embolization. The fluoroscopy duration and the X-ray exposure did not vary significantly. ADVANCES IN KNOWLEDGE: CTA-based fusion imaging using 2D-3D registration is a valuable tool for performing pulmonary artery embolization, helpful for planning and guiding catheterization.Compared to the traditional imaging guidance, fusion imaging reduces the volume of contrast agent used.


Subject(s)
Contrast Media , Pulmonary Artery , Humans , Pulmonary Artery/diagnostic imaging , Retrospective Studies , Imaging, Three-Dimensional , Fluoroscopy/methods , Treatment Outcome
2.
Eur Radiol ; 33(12): 8521-8527, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37470824

ABSTRACT

OBJECTIVES: To assess feasibility and accuracy of aortic annulus measurements using cardiac computed tomography angiography (CTA) performed during free-breathing prior to transcatheter aortic valve replacement (TAVR). MATERIALS AND METHODS: Sixty consecutive TAVR candidates underwent free-breathing wide-detector cardiac CTA, followed by a percutaneous valve replacement. For each, a theoretical valve size was suggested through CT measurements of the annulus, then compared to the size of the actual implanted transcatheter heart valve (THV). The procedural success and the 30-day outcomes were collected. Image quality of the annulus was also studied according to subjective and objective criteria. Data of a control group of 60 patients previously evaluated on breath-holding were also evaluated. RESULTS: A total of 120 patients (mean age, 83 years ± 7, 60 men) were evaluated. All CT acquisitions provided sufficient image quality allowing precise annulus measurements. Mean attenuation (p < 0.001) and image noise (p = 0.01) were higher in the free-breathing group, while image quality was comparable (p = 0.36). The agreement rate between CT-suggested valve size and THV implanted size was comparable, estimated at 87% (κ = 0.79, 95%CI 0.566, 0.908) on free-breathing vs. 82% (κ = 0.78, 95%CI 0.634, 0.904) on breath-holding. The procedure was successful for all patients without increase in 30-day mortality or adverse events. CONCLUSIONS: Free-breathing cardiac CTA allows accurate aortic annulus measurements without compromising image quality or patients' outcome after TAVR. Elderly patients experiencing dyspnea, discomfort, or hearing loss that could prevent proper breath-holding should not be excluded from CT prior to TAVR. CLINICAL RELEVANCE STATEMENT: To decrease elderly patients' discomfort, MDCT evaluation prior to transcatheter aortic valve replacement (TAVR) may be performed on quiet breathing with no significant impact on the outcome. KEY POINTS: • Adhering to CT breathing commands can be challenging for patients with dyspnea, hearing impairment, agitation, or pulmonary diseases. • Free-breathing cardiac CT may be an alternative to breath-holding for patients unable to follow the breathing commands. • Wide-detector CT acquisition on free-breathing does not impair annulus measurements and prosthesis sizing in patients scheduled for TAVR.


Subject(s)
Aortic Valve Stenosis , Heart Valve Prosthesis , Transcatheter Aortic Valve Replacement , Male , Humans , Aged , Aged, 80 and over , Transcatheter Aortic Valve Replacement/methods , Aortic Valve/diagnostic imaging , Aortic Valve/surgery , Aortic Valve Stenosis/diagnostic imaging , Aortic Valve Stenosis/surgery , Computed Tomography Angiography , Dyspnea , Multidetector Computed Tomography/methods , Treatment Outcome , Prosthesis Design , Predictive Value of Tests
3.
J Neurosurg ; 139(4): 1070-1077, 2023 10 01.
Article in English | MEDLINE | ID: mdl-36905663

ABSTRACT

OBJECTIVE: The management of Spetzler-Martin grade (SMG) III brain arteriovenous malformations (bAVMs) may be challenging, whatever the exclusion treatment modality chosen. The purpose of this study was to evaluate the safety and effectiveness of endovascular treatment (EVT) as a first-line treatment of SMG III bAVMs. METHODS: The authors performed a retrospective, two-center, observational cohort study. Cases recorded in institutional databases between January 1998 and June 2021 were reviewed. Patients who were ≥ 18 years of age, had ruptured or unruptured SMG III bAVMs, and received EVT as first-line therapy were included. Baseline characteristics of patients and bAVMs, procedure-related complications, clinical outcome according to the modified Rankin Scale, and angiographic follow-up were assessed. The independent risk factors of procedure-related complications and poor clinical outcome were assessed using binary logistic regression. RESULTS: One hundred sixteen patients with 116 SMG III bAVMs were included. The mean age of the patients was 41.9 ± 14.0 years. The most common presentation was hemorrhage (66.4%). Forty-nine (42.2%) bAVMs were found to be completely obliterated by EVT alone at follow-up. Complications occurred in 39 patients (33.6%), including 5 (4.3%) major procedure-related complications. There was no independent predictor of procedure-related complication. Age > 40 years and poor preoperative modified Rankin Scale score were the independent predictors of poor clinical outcome. CONCLUSIONS: EVT of SMG III bAVMs provides encouraging results but needs further improvement. When the embolization procedure performed with intent to cure appears difficult and/or risky, a combined technique (with microsurgery or radiosurgery) may be a safer and more effective strategy. In terms of safety and effectiveness, the benefit of EVT (alone or included in a multimodal management strategy) for SMG III bAVMs needs to be confirmed by randomized controlled trials.


Subject(s)
Embolization, Therapeutic , Intracranial Arteriovenous Malformations , Radiosurgery , Humans , Adult , Middle Aged , Retrospective Studies , Treatment Outcome , Intracranial Arteriovenous Malformations/therapy , Intracranial Arteriovenous Malformations/surgery , Microsurgery , Brain/surgery , Embolization, Therapeutic/adverse effects , Embolization, Therapeutic/methods
4.
Front Neurol ; 13: 925763, 2022.
Article in English | MEDLINE | ID: mdl-36203983

ABSTRACT

Indications for mechanical thrombectomy in acute ischemic stroke are increasing, resulting in the continuous development of new devices and techniques. Therefore, there is a need for a realistic testing and training environment that offers the opportunity to practice different procedures and test the latest devices. Some authors have described the use of the human placenta as a model for neurointerventional surgery, with striking similarities to real-life conditions. This model has many advantages, including its relatively low cost and minimal infrastructure requirements, with fewer ethical concerns compared to animal models. So far, some preparation and set-up details were missing, and only arteries from the chorionic plate were used. This article provides the necessary clarifications and a mapping of the chorionic plate veins, so that the use of this model, which is particularly well suited for mechanical thrombectomy, can be as easy and wide as possible. A video explaining how to prepare the model is provided.

5.
Microorganisms ; 10(7)2022 Jul 01.
Article in English | MEDLINE | ID: mdl-35889056

ABSTRACT

Haptophyte microalgae are key contributors to microbial communities in many environments. It has been proposed recently that members of this group would be virtually all dependent on vitamin B12 (cobalamin), an enzymatic cofactor produced only by some bacteria and archaea. Here, we examined the processes of vitamin B12 acquisition by haptophytes. We tested whether co-cultivating the model species Tisochrysis lutea with B12-producing bacteria in vitamin-deprived conditions would allow the microalga to overcome B12 deprivation. While T. lutea can grow by scavenging vitamin B12 from bacterial extracts, co-culture experiments showed that the algae did not receive B12 from its associated bacteria, despite bacteria/algae ratios supposedly being sufficient to allow enough vitamin production. Since other studies reported mutualistic algae-bacteria interactions for cobalamin, these results question the specificity of such associations. Finally, cultivating T. lutea with a complex bacterial consortium in the absence of the vitamin partially rescued its growth, highlighting the importance of microbial interactions and diversity. This work suggests that direct sharing of vitamin B12 is specific to each species pair and that algae in complex natural communities can acquire it indirectly by other mechanisms (e.g., after bacterial lysis).

6.
Front Plant Sci ; 13: 830069, 2022.
Article in English | MEDLINE | ID: mdl-35251102

ABSTRACT

We observed differences in lhc classification in Chromista. We proposed a classification of the lhcf family with two groups specific to haptophytes, one specific to diatoms, and one specific to seaweeds. Identification and characterization of the Fucoxanthin and Chlorophyll a/c-binding Protein (FCP) of the haptophyte microalgae Tisochrysis lutea were performed by similarity analysis. The FCP family contains 52 lhc genes in T. lutea. FCP pigment binding site candidates were characterized on Lhcf protein monomers of T. lutea, which possesses at least nine chlorophylls and five fucoxanthin molecules, on average, per monomer. The expression of T. lutea lhc genes was assessed during turbidostat and chemostat experiments, one with constant light (CL) and changing nitrogen phases, the second with a 12 h:12 h sinusoidal photoperiod and changing nitrogen phases. RNA-seq analysis revealed a dynamic decrease in the expression of lhc genes with nitrogen depletion. We observed that T. lutea lhcx2 was only expressed at night, suggesting that its role is to protect \cells from return of light after prolonged darkness exposure.

7.
Insights Imaging ; 12(1): 116, 2021 Aug 12.
Article in English | MEDLINE | ID: mdl-34383170

ABSTRACT

Imaging findings after thoracic surgery can be misleading. Knowledge of the normal post-operative anatomy helps the radiologist to recognise life-threatening complications and conversely not to wrongly evoke a complication in cases of trivial post-operative abnormalities. In this educational article, we reviewed the expected patterns after thoracic surgery including sublobar resection, lobectomy, pneumonectomy and related techniques. Imaging aspects of frequent and less common complications and their typical imaging features are then presented.

9.
Mar Drugs ; 20(1)2021 Dec 24.
Article in English | MEDLINE | ID: mdl-35049877

ABSTRACT

The production of polyunsaturated fatty acids (PUFA) in Tisochrysis lutea was studied using the gradual incorporation of a 13C-enriched isotopic marker, 13CO2, for 24 h during the exponential growth of the algae. The 13C enrichment of eleven fatty acids was followed to understand the synthetic pathways the most likely to form the essential polyunsaturated fatty acids 20:5n-3 (EPA) and 22:6n-3 (DHA) in T. lutea. The fatty acids 16:0, 18:1n-9 + 18:3n-3, 18:2n-6, and 22:5n-6 were the most enriched in 13C. On the contrary, 18:4n-3 and 18:5n-3 were the least enriched in 13C after long chain polyunsaturated fatty acids such as 20:5n-3 or 22:5n-3. The algae appeared to use different routes in parallel to form its polyunsaturated fatty acids. The use of the PKS pathway was hypothesized for polyunsaturated fatty acids with n-6 configuration (such as 22:5n-6) but might also exist for n-3 PUFA (especially 20:5n-3). With regard to the conventional n-3 PUFA pathway, Δ6 desaturation of 18:3n-3 appeared to be the most limiting step for T. lutea, "stopping" at the synthesis of 18:4n-3 and 18:5n-3. These two fatty acids were hypothesized to not undergo any further reaction of elongation and desaturation after being formed and were therefore considered "end-products". To circumvent this limiting synthetic route, Tisochrysis lutea seemed to have developed an alternative route via Δ8 desaturation to produce longer chain fatty acids such as 20:5n-3 and 22:5n-3. 22:6n-3 presented a lower enrichment and appeared to be produced by a combination of different pathways: the conventional n-3 PUFA pathway by desaturation of 22:5n-3, the alternative route of ω-3 desaturase using 22:5n-6 as precursor, and possibly the PKS pathway. In this study, PKS synthesis looked particularly effective for producing long chain polyunsaturated fatty acids. The rate of enrichment of these compounds hypothetically synthesized by PKS is remarkably fast, making undetectable the 13C incorporation into their precursors. Finally, we identified a protein cluster gathering PKS sequences of proteins that are hypothesized allowing n-3 PUFA synthesis.


Subject(s)
Fatty Acids, Unsaturated/biosynthesis , Haptophyta , Animals , Aquatic Organisms , Biosynthetic Pathways , Carbon Dioxide
10.
Sci Rep ; 9(1): 8417, 2019 06 10.
Article in English | MEDLINE | ID: mdl-31182768

ABSTRACT

Vitamin B12 (cobalamin) can control phytoplankton development and community composition, with around half of microalgal species requiring this vitamin for growth. B12 dependency is determined by the absence of cobalamin-independent methionine synthase and is unrelated across lineages. Despite their important role in carbon and sulphur biogeochemistry, little is known about haptophytes utilization of vitamin B12 and their ability to cope with its limitation. Here we report the first evaluation of B12 auxotrophy among this lineage based on molecular data of 19 species from 9 families. We assume that all species encode only a B12-dependent methionine synthase, suggesting ubiquitous B12 auxotrophy in this phylum. We further address the effect of different B12 limitations on the molecular physiology of the model haptophyte Tisochrysis lutea. By coupling growth assays in batch and chemostat to cobalamin quantification and expression analyses, we propose that haptophytes use three strategies to cope with B12 limitation. Haptophytes may assimilate dissolved methionine, finely regulate genes involved in methionine cycle and B12 transport and/or limit B12 transport to the mitochondrion. Taken together, these results provide better understanding of B12 metabolism in haptophytes and represent valuable data for deciphering how B12-producing bacteria shape the structure and dynamics of this important phytoplankton community.


Subject(s)
Haptophyta/metabolism , Microalgae/metabolism , Vitamin B 12/pharmacology , 5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase/metabolism , Batch Cell Culture Techniques , Gene Expression Regulation , Haptophyta/drug effects , Haptophyta/genetics , Haptophyta/growth & development , Likelihood Functions , Microalgae/drug effects , Microalgae/genetics , Microalgae/growth & development , Phylogeny
11.
Mar Drugs ; 15(9)2017 Sep 20.
Article in English | MEDLINE | ID: mdl-28930152

ABSTRACT

The green microalgae Dunaliella genus is known for the production of high added value molecules. In this study, strain AL-1 was isolated from the Sebkha of Sidi El Hani (Sousse, Tunisia). This isolate was identified both morphologically and genetically via 18S rRNA gene sequence as a member of the genus Dunaliella. Strain AL-1 was found to be closely related to Dunaliella salina, Dunaliella quartolecta and Dunaliella polymorpha with more than 97% similarity. Response surface methodology was used to maximize carotenoid production by strain AL-1 by optimizing its growth conditions. The highest carotenoid content was obtained at salinity: 51, light intensity: 189.89 µmol photons·m-2·s-1, and nitrogen: 60 mg·L-1. Proteomic profiling, using two-dimensional gel electrophoresis, was performed from standard and optimized cultures. We detected 127 protein spots which were significantly differentially expressed between standard and optimized cultures. Among them 16 protein spots were identified with mass spectrometry and grouped into different functional categories using KEGG (Kyoto Encyclopedia of Genes and Genomes) such as photosynthetic Calvin cycle, regulation/defense, energy metabolism, glycolysis, and cellular processes. The current study could be of great interest in providing information on the effect of stressful conditions in microalgae carotenoid production.


Subject(s)
Carotenoids/biosynthesis , Microalgae/metabolism , Animals , Electrophoresis, Gel, Two-Dimensional , Mass Spectrometry , Proteome/genetics , RNA, Ribosomal, 18S/genetics
12.
Physiol Plant ; 154(4): 572-90, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25640753

ABSTRACT

Microalgae have a diversity of industrial applications such as feed, food ingredients, depuration processes and energy. However, microalgal production costs could be substantially improved by controlling nutrient intake. Accordingly, a better understanding of microalgal nitrogen metabolism is essential. Using in silico analysis from transcriptomic data concerning the microalgae Tisochrysis lutea, four genes encoding putative high-affinity nitrate/nitrite transporters (TlNrt2) were identified. Unlike most of the land plants and microalgae, cloning of genomic sequences and their alignment with complementary DNA (cDNA) sequences did not reveal the presence of introns in all TlNrt2 genes. The deduced TlNRT2 protein sequences showed similarities to NRT2 proteins of other phyla such as land plants and green algae. However, some interesting specificities only known among Haptophyta were also revealed, especially an additional sequence of 100 amino acids forming an atypical extracellular loop located between transmembrane domains 9 and 10 and the function of which remains to be elucidated. Analyses of individual TlNrt2 gene expression with different nitrogen sources and concentrations were performed. TlNrt2.1 and TlNrt2.3 were strongly induced by low NO3 (-) concentration and repressed by NH4 (+) substrate and were classified as inducible genes. TlNrt2.2 was characterized by a constitutive pattern whatever the substrate. Finally, TlNrt2.4 displayed an atypical response that was not reported earlier in literature. Interestingly, expression of TlNrt2.4 was rather related to internal nitrogen quota level than external nitrogen concentration. This first study on nitrogen metabolism of T. lutea opens avenues for future investigations on the function of these genes and their implication for industrial applications.


Subject(s)
Anion Transport Proteins/genetics , Genes, Plant , Microalgae/genetics , Nitrates/metabolism , Nitrites/metabolism , Amino Acid Sequence , Anion Transport Proteins/chemistry , Anion Transport Proteins/metabolism , Exons , Introns , Molecular Sequence Data , RNA, Messenger/genetics , Sequence Homology, Amino Acid , Transcription, Genetic
13.
PLoS One ; 9(1): e86889, 2014.
Article in English | MEDLINE | ID: mdl-24489800

ABSTRACT

The applied exploitation of microalgae cultures has to date almost exclusively involved the use of wild type strains, deposited over decades in dedicated culture collections. Concomitantly, the concept of improving algae with selection programs for particular specific purposes is slowly emerging. Studying since a decade an economically and ecologically important haptophyte Tisochrysis lutea (Tiso), we took advantage of the availability of wild type (Tiso-Wt) and selected (Tiso-S2M2) strains to conduct a molecular variations study. This endeavour presented substantial challenges: the genome assembly was not yet available, the life cycle unknown and genetic diversity of Tiso-Wt poorly documented. This study brings the first molecular data in order to set up a selection strategy for that microalgae. Following high-throughput Illumina sequencing, transcriptomes of Tiso-Wt and Tiso-S2M2 were de novo assembled and annotated. Genetic diversity between both strains was analyzed and revealed a clear conservation, while a comparison of transcriptomes allowed identification of polymorphisms resulting from the selection program. Of 34,374 transcripts, 291 were differentially expressed and 165 contained positional polymorphisms (SNP, Indel). We focused on lipid over-accumulation of the Tiso-S2M2 strain and 8 candidate genes were identified by combining analysis of positional polymorphism, differential expression levels, selection signature and by study of putative gene function. Moreover, genetic analysis also suggests the existence of a sexual cycle and genetic recombination in Tisochrysis lutea.


Subject(s)
Algal Proteins/genetics , Life Cycle Stages/genetics , Lipid Metabolism/genetics , Microalgae/genetics , Transcriptome , Algal Proteins/metabolism , Gene Expression Profiling , Genetic Variation , Genotype , High-Throughput Nucleotide Sequencing , Microalgae/growth & development , Microalgae/metabolism , Molecular Sequence Annotation , Polymorphism, Single Nucleotide , Recombination, Genetic , Selection, Genetic , Sequence Analysis, DNA
14.
Bioresour Technol ; 119: 79-87, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22728186

ABSTRACT

Association of microalgae culture and anaerobic digestion seems a promising technology for sustainable algal biomass and biogas production. The use of digestates for sustaining the growth of microalgae reduces the costs and the environmental impacts associated with the substantial algal nutrient requirements. A natural marine algae-bacteria consortium was selected by growing on a medium containing macro nutrients (ammonia, phosphate and acetate) specific of a digestate, and was submitted to a factorial experimental design with different levels of temperature, light and pH. The microalgal consortium reached a maximum C conversion efficiency (i.e. ratio between carbon content produced and carbon supplied through light photosynthetic C conversion and acetate) of 3.6%. The presence of bacteria increased this maximum C conversion efficiency up to 6.3%. The associated bacterial community was considered beneficial to the total biomass production by recycling the carbon lost during photosynthesis and assimilating organic by-products from anaerobic digestion.


Subject(s)
Bacteria, Anaerobic/physiology , Carbon/metabolism , Microalgae/physiology , Microbial Consortia/physiology , Photobioreactors/microbiology , Water Microbiology , Bacteria, Anaerobic/radiation effects , Cell Proliferation , Light , Microalgae/radiation effects , Microbial Consortia/radiation effects , Oceans and Seas
15.
Dis Aquat Organ ; 91(3): 213-21, 2010 Sep 17.
Article in English | MEDLINE | ID: mdl-21133321

ABSTRACT

Vibrio aestuarianus is frequently found in coastal areas and can infect and induce mortalities in the pacific oyster Crassostrea gigas. However, nothing is known about its distribution and seasonality in the estuarine environment, especially where oyster farming is practiced. Its occurrence was investigated in sediment and oyster haemolymph at 2 oyster farms in Brittany (France) over 2 yr during 2 periods, from June to September 2007 and from February to June 2008. Total heterotrophic bacteria (HB) were cultured on marine agar while total Vibrio spp. and V aestuarianus were selectively numerated using thiosulfate citrate bile salts sucrose agar (TCBS agar) and the species-specific hybridisation method, respectively. PCR was performed to detect V aestuarianus in sediment when it became unculturable. Both total Vibrio spp. and V aestuarianus had a seasonal trend. The highest concentrations were recovered in the warmest months. Its abundance ranged from 10(2) to 4 x 10(5) CFU ml(-1) in haemolymph and from 10(3) to 1 x 10(4) CFU g(-1) in the sediment. Temperature was the main factor influencing the concentration of Vibrio spp. and V. aestuarianus in the sediment. Thus V aestuarianus might subsist during the cold seasons in the sediment, from which it can emerge when environmental conditions became favourable.


Subject(s)
Crassostrea/microbiology , Geologic Sediments/microbiology , Seasons , Vibrio/isolation & purification , Animals , Aquaculture , France , Oceans and Seas , RNA, Ribosomal, 16S/genetics , Time Factors , Vibrio/classification , Vibrio/genetics
16.
Appl Environ Microbiol ; 76(24): 8011-8, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20935127

ABSTRACT

There is considerable interest in the use of psychrotrophic bacteria for food biopreservation and in the understanding of cold adaptation mechanisms. The psychrotrophic biopreservative Lactococcus piscium strain CNCM I-4031 was studied for its growth behavior and proteomic responses after cold shock and during cold acclimation. Growth kinetics highlighted the absence of growth latency after cold shock, suggesting a very high promptness in cold adaptation, a behavior that has never been described before for lactic acid bacteria (LAB). A comparative proteomic analysis was applied with two-dimensional gel electrophoresis (2-DE), and upregulated proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both cold shock and cold acclimation triggered the upregulation of proteins involved in general and oxidative stress responses and fatty acid and energetic metabolism. However, 2-DE profiles and upregulated proteins were different under both conditions, suggesting a sequence of steps in cold adaptation. In addition, the major 7-kDa Csp protein was identified in the L. piscium CNCM I-4031 genome but was not cold regulated. The implication of the identified cold shock proteins and cold acclimation proteins in efficient cold adaptation, the possible regulation of a histidyl phosphocarrier protein, and the roles of a constitutive major 7-kDa Csp are discussed.


Subject(s)
Bacterial Proteins/analysis , Cold Temperature , Lactococcus/physiology , Proteome/analysis , Stress, Physiological , Chromatography, Liquid , Electrophoresis, Gel, Two-Dimensional , Lactococcus/chemistry , Lactococcus/growth & development , Tandem Mass Spectrometry
17.
Syst Appl Microbiol ; 31(5): 358-65, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18718730

ABSTRACT

Eleven Vibrio isolates invading the hemolymph of live and moribund oysters (Crassostrea gigas) collected in the field and from a hatchery in France, were characterized by a polyphasic approach. Phylogenetic analysis of 16S rRNA, gyrB and toxR genes indicated high homogeneity between these strains and the Vibrio aestuarianus type strain (ATCC35048(T)), and confirmed previous 16S rRNA analysis. In contrast, DNA:DNA hybridization was from 61% to 100%, while phenotypic characters and virulence tests showed a large diversity between the strains. Nevertheless, several common characters allowed the isolates to be distinguished from the reference strain. On the basis of several distinct phenotypic characteristics, it is proposed to establish two subspecies within the V. aestuarianus spp. group, V. aestuarianus subsp. aestuarianus [D. Tison, R. Seidler, Vibrio aestuarianus: a new species from estuarine waters and shellfish, Int. J. Syst. Bacteriol. (1983) 699-702] and V. aestuarianus subsp. francensis for these French isolates. The characters that differentiate the new strains from V. aestuarianus subsp. aestuarianus(T) are virulence (positive for 63% of the isolates) and 12:0 fatty acid content. The colonies were smaller and uncoloured, whereas no growth occurred at 35 degrees C or on TCBS, and the strains did not utilize several substrates, including L-serine, alpha-cyclodextrin, D-mannitol, alpha-glycyl-L-aspartic acid, L-threonine and glucose-1-phosphate.


Subject(s)
Bacterial Typing Techniques , Crassostrea/microbiology , Vibrio , Animals , Bacterial Typing Techniques/methods , DNA, Bacterial/analysis , DNA, Bacterial/genetics , France , Genotype , Hemolymph/microbiology , Nucleic Acid Hybridization , Phenotype , Phylogeny , RNA, Ribosomal, 16S/genetics , Species Specificity , Vibrio/classification , Vibrio/genetics , Vibrio/isolation & purification , Vibrio/pathogenicity , Virulence
18.
J Med Chem ; 46(2): 222-36, 2003 Jan 16.
Article in English | MEDLINE | ID: mdl-12519061

ABSTRACT

Cyclin-dependent kinases (CDKs) regulate the cell cycle, apoptosis, neuronal functions, transcription, and exocytosis. The observation of CDK deregulations in various pathological situations suggests that CDK inhibitors may have a therapeutic value. In this article, we report on the identification of 6-phenyl[5H]pyrrolo[2,3-b]pyrazines (aloisines) as a novel potent CDK inhibitory scaffold. A selectivity study performed on 26 kinases shows that aloisine A is highly selective for CDK1/cyclin B, CDK2/cyclin A-E, CDK5/p25, and GSK-3 alpha/beta; the two latter enzymes have been implicated in Alzheimer's disease. Kinetic studies, as well as the resolution of a CDK2-aloisine cocrystal structure, demonstrate that aloisines act by competitive inhibition of ATP binding to the catalytic subunit of the kinase. As observed with all inhibitors reported so far, aloisine interacts with the ATP-binding pocket through two hydrogen bonds with backbone nitrogen and oxygen atoms of Leu 83. Aloisine inhibits cell proliferation by arresting cells in both G1 and G2.


Subject(s)
Antineoplastic Agents/chemical synthesis , CDC2-CDC28 Kinases , Cyclin-Dependent Kinases/antagonists & inhibitors , Enzyme Inhibitors/chemical synthesis , Glycogen Synthase Kinase 3/antagonists & inhibitors , Protein Serine-Threonine Kinases/antagonists & inhibitors , Pyrazines/chemical synthesis , Adenosine Triphosphate/chemistry , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Binding, Competitive , CDC2 Protein Kinase/antagonists & inhibitors , Cell Division/drug effects , Cyclin-Dependent Kinase 2 , Cyclin-Dependent Kinase 5 , Cyclin-Dependent Kinases/chemistry , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Glycogen Synthase Kinase 3/chemistry , Humans , Hydrogen Bonding , Models, Molecular , Molecular Conformation , Neurons/cytology , Neurons/drug effects , Protein Serine-Threonine Kinases/chemistry , Pyrazines/chemistry , Pyrazines/pharmacology , Structure-Activity Relationship , Tumor Cells, Cultured
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