Enantio- and Diastereoselective Mannich Reactions of ß-Dicarbonyls by Second Stage Diastereoconvergent Crystallization.

The synthesis of chiral α-monosubstituted-ß-dicarbonyls is a challenging task in asymmetric catalysis due to the rapid, typically uncontrolled, product racemization or epimerization under most reaction conditions. For this reason, diastereoselective additions of unsubstituted ß-dicarbonyls to π-electrophiles are unusual. Herein, we disclose a simple catalytic crystallization-driven enantio- and diastereoselective Mannich reaction for the synthesis of stereodefined α-monosubstituted-ß-keto esters, dissymmetric ß-diesters, dissymmetric ß-diketones, and ß-keto amides that productively leverages product epimerization in solution. Mechanistic studies suggest a scenario where the initial enantioselective, diastereodivergent skeletal assembly is catalyzed by a chiral tertiary amine organocatalyst, which then facilitates second stage crystallization-induced diastereoconvergence to provide the challenging α-stereocenter in excellent stereoselectivity.

Rhodium-Catalyzed Asymmetric Arylation-Induced Glycolate Aldol Additions of Silyl Glyoxylates.

(Diene)Rh(I) complexes catalyze the stereoselective three-component coupling of silyl glyoxylates, arylboronic acids, and aldehydes to give glycolate aldol products. The participation of Rh-alkoxides in the requisite Brook rearrangement was established through two component Rh-catalyzed couplings of silyl glyoxylates with ArB(OH)2 to give silyl-protected mandelate derivatives. The intermediacy of a chiral Rh-enolate was inferred through enantioselective protonation using a chiral Rh-catalyst. Diastereoselective three-component couplings with aldehydes as terminating electrophiles to give racemic products were best achieved with a bulky aryl ester on the silyl glyoxylate reagent. Optimal enantioselective couplings were carried out with the tert-butyl ester variant using an anisole-derived enantiopure tricyclo[3.2.2.02,4 ]nonadiene ligand.

Dearomative Synthesis of Chiral Dienes Enables Improved Late-Stage Ligand Diversification.

An efficient synthesis of chiral nonracemic diene ligands is facilitated by an enantioselective dearomative intermolecular arene cyclopropanation of anisole. The functionality of the resulting cycloheptatriene engenders distinct chemical environments in a downstream tricyclic bis(enol) triflate that permits selective late-stage functionalization. The synthesis of diverse C1- and pseudo-C2-symmetric dienes is therefore viable by iterative palladium-catalyzed cross-coupling reactions. The ligands provide moderate to high selectivities in known Rh(I)-mediated asymmetric transformations.

Stereodivergent Nucleophilic Additions to Racemic ß-Oxo Acid Derivatives: Fast Addition Outcompetes Stereoconvergence in the Archetypal Configurationally Unstable Electrophile.

Additions of carbon nucleophiles to racemic α-stereogenic ß-oxo acid derivatives that deliver enantiomerically enriched tertiary alcohols are valuable, but uncommon. This article describes stereodivergent Cu-catalyzed borylative cyclizations of racemic ß-oxo acid derivatives bearing tethered pro-nucleophilic olefins to deliver highly functionalized cyclopentanols containing four contiguous stereogenic centers. The reported protocol is applicable to a range of ß-oxo acid derivatives, and the diastereomeric products are readily isolable by typical chromatographic techniques. α-Stereogenic-ß-keto esters are typically thought to have extreme or spontaneous configurational fragility, but mechanistic studies for this system reveal an unusual scenario wherein productive catalysis occurs on the same time scale as background substrate racemization and completely outcompetes on-cycle epimerization, even under the basic conditions of the reaction.

Further studies on the application of vinylogous amides and ß-halovinylaldehydes to the regiospecific synthesis of unsymmetrical, polyfunctionalized 2,3,4- and 1,2,3,4- substituted pyrroles.

Highly functionalized pyrroles with appropriate regiochemical functionality represent an important class of marine natural products and potential drug candidates. We describe herein a detailed study of the reaction of α-aminoacid esters with vinylogous amides and also ß-halovinylaldehydes for the regiospecific synthesis of 2,3,4-trisubstituted and 1,2,3,4-tetrasubstituted pyrroles. Since the vinylogous amides and ß-halovinylaldehydes are readily available precursors, rapid access to a wide variety of unsymmetrically substituted pyrroles is accomplished via this methodology.

Application of vinylogous carbamates and vinylogous aminonitriles to the regiospecific synthesis of uniquely functionalized pyrroles and quinolones.

Pyrroles and quinolones represent core structures, which are routinely found in both natural and synthetic bioactive substances. Consequently, the development of efficient and regiospecific methods for the preparation of such heterocycles with unique functionality is of some importance. We describe herein the regiospecific synthesis of 1,2,3,4-tetrasubstituted pyrroles containing polar substituents and such products are prepared from vinylogous carbamates and vinylogous aminonitriles. We also describe the regiospecific synthesis of 3-aryl containing 1,3,6trisubstituted quinolones from vinylogous carbamates. The use of an amine exchange reaction to prepare precursors for the pyrrole and quinolone forming cyclizations represents a key factor in the strategy.

Outcome of prolonged acute vena cava occlusion after iatrogenic transection and repair in a dog.

A 12-year-old castrated male Airedale terrier dog was diagnosed with a hepatocellular carcinoma in the right medial liver lobe. During tumor resection, inadvertent stapling and transection of the caudal vena cava occurred. Complete caval occlusion was required for 18 minutes and primary anastomosis was completed. The dog received 2 blood transfusions and developed mild pelvic limb edema after surgery. Computed tomography evaluation 9 months after surgery showed collateral circulation and suspected stricture of the vena cava with an absence of clinical effect. The dog remained alive and asymptomatic more than 1 year after surgery.

Résultat d'une occlusion aiguë et prolongée de la veine cave caudale après transsection iatrogénique et réparation, chez un chien. Un Airedale terrier mâle stérilisé de 12 ans fut diagnostiqué avec un carcinome hépatocellulaire du lobe hépatique médian droit. Durant l'excision de la tumeur, la veine cave caudale fut malencontreusement agrafée et sectionnée. Une occlusion cavale complète s'est avérée nécessaire pour un total de 18 minutes, le temps de faire l'anastomose du vaisseau sanguin. Deux transfusions sanguines furent administrées et le chien développa de l'œdème léger des membres pelviens. Après huit jours d'hospitalisation, le patient reçut son congé. Plusieurs réévaluations ont pu confirmer l'absence de dysfonction d'organes ou de formation de thrombus. Une tomodensitométrie effectuée 9 mois après la procédure chirurgicale a pu mettre en évidence la formation d'une circulation collatérale en réponse à une stricture suspectée de la veine cave caudale. Le chien demeura en vie et asymptomatique plus d'un an après la chirurgie en question.(Traduit par Dre Marie-Chantal Halwagi).

Ortho group activation of a bromopyrrole ester in Suzuki-Miyaura cross-coupling reactions: Application to the synthesis of new microtubule depolymerizing agents with potent cytotoxic activities.

New microtubule depolymerizing agents with potent cytotoxic activities have been prepared with a 5-cyano or 5-oximino group attached to a pyrrole core. The utilization of ortho activation of a bromopyrrole ester to facilitate successful Suzuki-Miyaura cross-coupling reactions was a key aspect of the synthetic methodology. This strategy allows for control of regiochemistry with the attachment of four completely different groups at the 2, 3, 4 and 5 positions of the pyrrole scaffold. Biological evaluations and molecular modeling studies are reported for these examples.

Biofilm-Associated Gene Expression in Staphylococcus pseudintermedius on a Variety of Implant Materials.

OBJECTIVE: To evaluate the expression of biofilm-associated genes in Staphylococcus pseudintermedius on multiple clinically relevant surfaces. STUDY DESIGN: In vitro experimental study. SAMPLE POPULATION: Two strains of methicillin-resistant S. pseudintermedius isolated from clinical infections representing the most common international isolates. METHODS: A quantitative polymerase chain reaction (qPCR) assay for expression of genes related to biofilm initial adhesion, formation/maturation, antimicrobial resistance, and intracellular communication was developed and validated. S. pseudintermedius biofilms were grown on 8 clinically relevant surfaces (polymethylmethacrylate, stainless steel, titanium, latex, silicone, polydioxanone, polystyrene, and glass) and samples of logarithmic and stationary growth phases were collected. Gene expression in samples was measured by qPCR. RESULTS: Significant differences in gene expression were identified between surfaces and between bacterial strains for most gene/strain/surface combinations studied. Expression of genes responsible for production of extracellular matrix were increased in biofilms. Expression of genes responsible for initial adhesion and intracellular communication was markedly variable. Antimicrobial resistance gene expression was increased on multiple surfaces, including stainless steel and titanium. CONCLUSION: A method for evaluation of expression of multiple biofilm-associated genes in S. pseudintermedius was successfully developed and applied to the study of biofilms on multiple surfaces. Variations in expression of these genes have a bearing on understanding the development and treatment of implant-associated biofilm infections and will inform future clinical research.

Computed tomography diagnosis of a thoracic and abdominal penetrating foreign body in a dog.

A 1.5-year-old, spayed female, mixed-breed dog was presented for hemoabdomen associated with an abdominal mass. Upon presentation bicavitary effusion was diagnosed. A penetrating intra-abdominal wooden foreign body was identified using computed tomography. This case describes a thoracic penetrating wooden foreign body causing bicavitary effusion following migration into the retroperitoneal space.

Diagnostic d'un corps étranger thoracique et abdominal pénétrant par tomodensitométrie chez un chien. Une chienne de race croisée stérilisée âgée de 1,5 ans a été présentée pour un hémoabdomen associé à une masse abdominale. Sur présentation, une effusion bicavitaire a été diagnostiquée. Un corps étranger en bois intra-abdominal pénétrant a été identifié par tomodensitométrie. Ce cas décrit un corps étranger en bois pénétrant dans le thorax et causant une effusion bicavitaire après la migration dans l'espace rétropéritonéal.(Traduit par Isabelle Vallières).

Impact of polymethylmethacrylate additives on methicillin-resistant Staphylococcus pseudintermedius biofilm formation in vitro.

OBJECTIVE: To evaluate the impact of gentamicin, silver, or both additives in polymethylmethacrylate (PMMA) beads on methicillin-resistant Staphylococcus pseudintermedius (MRSP) biofilm formation in vitro. SAMPLE: 4 preparations of PMMA beads (formed with no additive [control], gentamicin, silver, and gentamicin and silver). PROCEDURES: Beads from each group were exposed to 10 MRSP isolates known to be strong biofilm formers. Following incubation, the beads were rinsed to remove planktonic bacteria, then sonicated to dislodge biofilm-associated bacteria. Resulting suspensions were serially diluted, plated on blood agar, and incubated overnight; CFUs were counted. Variance of mean CFU counts following log10 transformation was analyzed among PMMA groups. RESULTS: None of the PMMA additives tested completely inhibited MRSP biofilm formation. There was a significant effect of gentamicin and gentamicin plus silver on this variable, compared with controls, but not of silver alone. There was no difference between gentamicin and gentamicin plus silver. When only isolates not susceptible to gentamicin were evaluated, there were no significant differences among PMMA additive groups. Within gentamicin-susceptible isolates, there was an impact of gentamicin and gentamicin plus silver, but no impact of silver alone and no difference between gentamicin and gentamicin plus silver. CONCLUSIONS AND CLINICAL RELEVANCE: Gentamicin-impregnated PMMA was effective at reducing biofilm formation of gentamicin-susceptible MRSP isolates but had no effect on isolates not susceptible to gentamicin. Silver-impregnated PMMA had no effect on MRSP biofilm formation. Results suggested that gentamicin-impregnated PMMA may not be effective in vivo against MRSP isolates not susceptible to gentamicin. Antibacterial efficacy of silver should not be assumed without proper testing of the target bacteria and specific silver compound.

Laparoscopic-assisted extirpation of falciform ligament hemangiosarcoma in a dog.

An 8-year-old, spayed female, bichon frisé dog had incidental nodules within its falciform ligament identified on routine abdominal ultrasonography. A laparoscopic-assisted technique provided both a diagnostic and a therapeutic treatment option. A histopathological diagnosis of hemangiosarcoma was made. This is the second case reporting hemangiosarcoma of the falciform fat.

Extraction assistée par laparascopie d'un hémangiosarcome du ligament falciforme chez un chien. Une chienne Bichon frisé stérilisée âgée de 8 ans avait des nodules secondaires dans le ligament falciforme qui ont été identifiés lors d'une échographie abdominale de routine. Une technique assistée par laparascopie a fourni un diagnostic et une option de traitement thérapeutique. Un diagnostic d'hémangiosarcome a été posé à l'histopathologie. Il s'agit du deuxième cas d'hémangiosarcome du gras falciforme signalé.(Traduit par Isabelle Vallières).

Identification of appropriate reference genes for qPCR studies in Staphylococcus pseudintermedius and preliminary assessment of icaA gene expression in biofilm-embedded bacteria.

BACKGROUND: Quantitative PCR is rapidly becoming the standard method for analyzing gene expression in a wide variety of biological samples however it can suffer from significant error if stably expressed reference genes are not identified on which to base the analysis. Suitable reference genes for qPCR experiments on Staphylococcus pseudintermedius have yet to be identified. RESULTS: Three reference genes in S. pseudintermedius were identified and validated from a set of eight potential genes (proC, gyrB, rplD, rho, rpoA, ftsZ, recA, sodA). Two strains of S. pseudintermedius were used, and primer specificity and efficiency were confirmed and measured. Ranking of the genes with respect to expression stability revealed gyrB, rho and recA as the best reference genes. This combination was used to quantify expression of a single biofilm associated gene, icaA, in logarithmic, stationary and biofilm growth phases, revealing that expression was significantly upregulated in the biofilm growth phase in both strains. CONCLUSION: Three reference genes, gyrB, rho and recA, were identified and validated for use as reference genes for quantitative PCR experiments in S. pseudintermedius. Also, the biofilm associated gene icaA was shown to be significantly upregulated in biofilm samples, consistent with its role in biofilm production.

Distannoxane speciation during esterification catalysis: revealing insights provided by electrospray ionization mass spectrometry.

Dimeric tetraalkyldistannoxanes are have been reported to catalyze esterification reactions, but are difficult to investigate in detail due to the lack of suitable spectroscopic handles. Electrospray ionization mass spectrometry (ESI-MS), in conjunction with a tethered charge on a tin atom, reveals that immediate decomposition to mono-tin carboxylate compounds occurs in the presence of carboxylic acid.

Implementing a Web-based content management system.

The Children's Hospital of Philadelphia transformed its web site to enhance patient satisfaction and attract new patients, as well as meet the needs of clinicians and the hospital's business plans. They accomplished this by implementing a content management system that would allow content creation and updating to be delegated to appropriate department staffers, thereby eliminating bottlenecks and unnecessary steps and ensuring that the web site receives fresh content much more quickly.

Lipopolysaccharide O-antigen antibody-based detection of the fish pathogen Flavobacterium psychrophilum.

Several yellow-pigmented species within the family Flavobacteriaceae are commonly associated with diseases in fish and are difficult to speciate due to their fastidious, slow-growing nature and cross-reactive antigens. Here we report the development of specific, antibody-diagnostic tests for Flavobacterium psychrophilum, the aetiological agent of rainbow trout fry syndrome and bacterial cold water disease. A unique antigen from F. psychrophilum, the lipopolysaccharide (LPS) O-polysaccharide (O-PS), formed the basis for the antibody test. LPS O-PS was purified and conjugated to keyhole limpet haemocyanin and bovine serum albumin for the generation of rabbit immune sera and the development of antibody-based diagnostic tests. Rabbit polyclonal anti-O-PS serum was highly specific for F. psychrophilum, without the need for prior cross-absorption with related bacteria and was the basis of an effective ELISA diagnostic test. Antibodies were purified from rabbit anti-O-PS serum and adsorbed onto coloured latex beads for the development of a specific, bead agglutination assay for F. psychrophilum.