Penile urethral resection and anastomosis augmentation with regional tissue tension relieving technique: A cadaveric mechanical study and clinical outcome in two dogs.

OBJECTIVE: To describe penile urethral augmentation anastomosis (PURAA) for resection anastomosis (RA) of the canine penile urethra by using autogenous tissue in two dogs and to determine the mechanical properties of the augmentation technique in cadaveric specimens. STUDY DESIGN: Cadaveric study and two case reports. ANIMALS: Sixteen canine cadavers and two dogs with urethral obstruction. METHODS: The lower urogenital system was harvested from cadavers and randomized into two groups: simple (S) and augmented (AUG) RA of the urethra. Tensile strength and peak load were compared between the two groups. Two dogs were treated with PURAA for urethral obstruction secondary to juxtaurethral neoplasms. RESULTS: Minimal tensile strength (MITS) and maximal tensile strength (MATS) were greater in the AUG group (MITS, 54.36 ± 24.0 N; MATS, 75.37 ± 34.79 N) compared with the S group (MITS, 11.78 ± 4.93 N, P = .0014; MATS, 13.74 ± 3.89 N, P = .0015). Both dogs recovered without complications. Histopathological examinations were consistent with a lipomatous mass in both cases. Both dogs had good medium-to-long-term outcomes. CONCLUSION: The augmentation technique improved the tensile properties of penile RA in normal cadavers and was associated with successful outcomes in two dogs. CLINICAL SIGNIFICANCE: Penile urethral augmentation anastomosis may help prevent stricture or leakage secondary to tension at the surgical site after penile urethral RA.

Defining Molecular Details of the Chemistry of Biofilm Formation by Raman Microspectroscopy.

Two protocols that allow for the comparison of Raman spectra of planktonic cells and biofilm formed from these cells in their growth phase have been developed. Planktonic cells are washed and flash-frozen in <1 min to reduce the time for metabolic changes during processing, prior to freeze-drying. Biofilm is formed by standing cells in 50 µL indentations in aluminum foil in an atmosphere of saturated water vapor for 24-48 h. The results for Escherichia coli type K12 cells, which do not readily form biofilm, are compared to those for Staphylococcus epidermidis cells, which prolifically synthesize biofilm. For E. coli, the Raman spectra of the planktonic and biofilm samples are similar with the exception that the spectral signature of RNA, present in planktonic cells, could not be detected in biofilm. For S. epidermidis, major changes occur upon biofilm formation. In addition to the absence of the RNA features, new bands occur near 950 cm-1 and between 1350 and 1420 cm-1 that are associated with an increase in carbohydrate content. Unlike the case in E. coli biofilm, the intensity of G base ring modes is reduced in but A and T base ring signatures become more prominent. For S. epidermis in the biofilm's amide III region, there is evidence of an increase in the level of ß-sheet structure accompanied by a decrease in α-helical content. The presence of biofilm is confirmed by microscope-aided photography and, separately, by staining with methyl violet.

Organometallic Ru(II) Photosensitizers Derived from π-Expansive Cyclometalating Ligands: Surprising Theranostic PDT Effects.

The purpose of the present study was to investigate the influence of π-expansive cyclometalating ligands on the photophysical and photobiological properties of organometallic Ru(II) compounds. Four compounds with increasing π conjugation on the cyclometalating ligand were prepared, and their structures were confirmed by HPLC, 1D and 2D (1)H NMR, and mass spectrometry. The properties of these compounds differed substantially from their Ru(II) polypyridyl counterparts. Namely, they were characterized by red-shifted absorption, very weak to no room temperature phosphorescence, extremely short phosphorescence state lifetimes (<10 ns), low singlet oxygen quantum yields (0.5-8%), and efficient ligand-centered fluorescence. Three of the metal complexes were very cytotoxic to cancer cells in the dark (EC50 values = 1-2 µM), in agreement with what has traditionally been observed for Ru(II) compounds derived from small C^N ligands. Surprisingly, the complex derived from the most π-expansive cyclometalating ligand exhibited no cytotoxicity in the dark (EC50 > 300 µM) but was phototoxic to cells in the nanomolar regime. Exceptionally large phototherapeutic margins, exceeding 3 orders of magnitude in some cases, were accompanied by bright ligand-centered intracellular fluorescence in cancer cells. Thus, Ru(II) organometallic systems derived from π-expansive cyclometalating ligands, such 4,9,16-triazadibenzo[a,c]napthacene (pbpn), represent the first class of potent light-responsive Ru(II) cyclometalating agents with theranostic potential.

In vitro multiwavelength PDT with 3IL states: teaching old molecules new tricks.

The purpose of the present investigation was to ascertain whether (3)IL excited states with microsecond lifetimes are universally potent for photodynamic applications, and if these long-lived states are superior to their (3)MLCT counterparts as in vitro PDT agents. A family of blue-green absorbing, Ru(II)-based transition metal complexes derived from the π-expansive dppn ligand was prepared and characterized according to its photodynamic activity against HL-60 cells, and toward DNA in cell-free media. Complexes in this series that are characterized by low-energy and long-lived (3)IL excited states photocleaved DNA with blue, green, red, and near-IR light. This panchromatic photodynamic effect translated to in vitro multiwavelength photodynamic therapy (PDT) with red-light cytotoxicities as low as 1.5 µM (EC50) for the parent complex and 400 nM for its more lipophilic counterpart. This potency is similar to that achieved with Ru(II)-based dyads containing long-lived (3)IL excitons located on appended pyrenyl units, and appears to be a general property of sufficiently long-lived excited states. Moreover, the red PDT observed for certain members of this family was almost 5 times more potent than Photofrin with therapeutic indices 30 times greater. Related Ru(II) complexes having lowest-lying (3)MLCT states of much shorter duration (≤1 µs) did not yield DNA photodamage or in vitro PDT with red or near-IR light, nor did the corresponding Os(II) complex with a submicrosecond (3)IL excited state lifetime. Therefore, metal complexes that utilize highly photosensitizing (3)IL excited states, with suitably long lifetimes (≫ 1 µs), are well-poised to elicit PDT at wavelengths even where their molar extinction coefficients are very low (<100 M(-1) cm(-1)). Herein we demonstrate that such unexpected reactivity gives rise to very effective PDT in the typical therapeutic window (600-850 nm).

Low Oxygen Concentration Reduces Neisseria gonorrhoeae Susceptibility to Resazurin.

Neisseria gonorrhoeae has developed resistance to every antibiotic currently approved for the treatment of gonorrhea, prompting the development of new therapies. The phenoxazine dye resazurin exhibits robust antimicrobial activity against N. gonorrhoeae in vitro but fails to limit vaginal colonization by N. gonorrhoeae in a mouse model. The lack of in vivo efficacy may be due to oxygen limitation as in vitro susceptibility assays with resazurin are conducted under atmospheric oxygen while a microaerophilic environment is present in the vagina. Here, we utilized broth microdilution assays to determine the susceptibility of N. gonorrhoeae to resazurin under low and atmospheric oxygen conditions. The minimal inhibitory concentration of resazurin for multiple N. gonorrhoeae clinical isolates was significantly higher under low oxygen. This effect was specific to resazurin as N. gonorrhoeae was equally susceptible to other antibiotics under low and atmospheric oxygen conditions. The reduced susceptibility of N. gonorrhoeae to resazurin under low oxygen was largely attributed to reduced oxidative stress, as the addition of antioxidants under atmospheric oxygen mimicked the reduced susceptibility to resazurin observed under low oxygen. Together, these data suggest oxygen concentration is an important factor to consider when evaluating the efficacy of new antibiotics against N. gonorrhoeae in vitro.

Ambulatory Pulmonary Artery Pressure Monitoring Reduces Costs and Improves Outcomes in Symptomatic Heart Failure: A Single-Centre Canadian Experience.

Background: Pulmonary artery pressure (PAP) monitoring reduces heart failure (HF) hospitalizations (HFHs) and improves quality of life in New York Heart Association (NYHA) class III HF. We evaluated the impact of PAP monitoring on outcomes and health spending in a Canadian ambulatory HF cohort. Methods: Twenty NYHA III HF patients underwent wireless PAP implantation at Foothills Medical Centre, Calgary, Alberta. Baseline, and 3-, 6-, 9-, and 12-month assessments of laboratory parameters, hemodynamics, 6-minute walk text and Kansas City Cardiomyopathy Questionnaire scores were collected. Healthcare costs 1 year pre- and post-implantation were collected from administrative databases. Results: Mean age was 70.6 years; 45% were female. Results were as follows: an 88% reduction in emergency room visits (P = 0.0009); an 87% reduction in HFHs (P < 0.0003); a 29% reduction in heart function clinic visits (P = 0.033), and a 178% increase in nurse calls (P < 0.0002). Questionnaire and 6-minute walk test scores at baseline vs last follow-up were 45.4 vs 48.4 (P = 0.48) and 364.4 vs 402.8 m (P = 0.58), respectively. Mean PAP at baseline vs follow-up was 31.5 vs 24.8 mm Hg (P = 0.005). NYHA class improved by at least one class in 85% of patients. Mean measurable HF-related spending preimplantation was CAD$29,814 per patient per year and postimplantation was CAD$25,642 per patient per year (including device cost). Conclusions: PAP monitoring demonstrated reductions in HFHs, and emergency room and heart function clinic visits, with improvements in NYHA class. Although further economic evaluation is needed, these results support the use of PAP monitoring as an effective and cost-neutral tool in HF management in appropriately selected patients in a publicly funded healthcare system.

Contexte: La surveillance de la pression artérielle pulmonaire (PAP) réduit les hospitalisations liées à l'insuffisance cardiaque (HIC) et améliore la qualité de vie des patients atteints d'insuffisance cardiaque (IC) de classe III de la New York Heart Association (NYHA). Nous avons évalué l'effet de la surveillance de la PAP sur les résultats et les dépenses en santé dans une cohorte de patients ambulatoires atteints d'IC au Canada. Méthodologie: Vingt patients atteints d'IC de classe III de la NYHA se sont fait implanter un dispositif sans fil de surveillance de la PAP au Foothills Medical Centre, à Calgary, en Alberta. Nous avons évalué les patients au départ et aux mois 3, 6, 9 et 12 en fonction des paramètres de laboratoire, de l'hémodynamique, des scores au test de marche de 6 minutes et au questionnaire de cardiomyopathie de Kansas City. Les coûts liés aux soins de santé un an avant et après l'implantation du dispositif ont été extraits de bases de données administratives. Résultats: L'âge moyen des patients est de 70,6 ans et 45 % sont des femmes. Les résultats vont comme suit : une réduction de 88 % des visites à l'urgence (p = 0,0009); une réduction de 87 % des HIC (p < 0,0003); une réduction de 29 % des visites à une clinique de fonction cardiaque (p = 0,033) et une augmentation de 178 % des appels du personnel infirmier (p < 0,0002). Les scores initiaux au questionnaire et au test de marche de 6 minutes par rapport à ceux du dernier suivi sont respectivement de 45,4 contre 48,4 (p = 0,48) et de 364,4 contre 402,8 m (p = 0,58). La PAP moyenne au départ par rapport au suivi est de 31,5 contre 24,8 mmHg (p = 0,005). La classe de l'IC selon la NYHA s'est améliorée d'au moins une position chez 85 % des patients. Les dépenses mesurables moyennes liées à l'IC avant l'implantation sont de 29 814 $ CA par patient par année et de 25 642 $ CA par patient par année après l'implantation (incluent le coût du dispositif). Conclusions: La surveillance de la PAP entraîne une réduction des HIC ainsi que des visites à l'urgence et à la clinique de fonction cardiaque, et une amélioration de la classe de l'IC selon la NYHA. Bien qu'une évaluation économique plus approfondie soit nécessaire, ces résultats militent en faveur de la surveillance de la PAP comme outil de prise en charge de l'IC sans incidence sur les coûts chez les patients adéquatement choisis dans un système de soins de santé financé par les fonds publics.

Escherichia coli pathotype contamination in raw canine diets.

OBJECTIVE: To investigate the prevalence of Escherichia coli contamination and E coli virulence gene signatures consistent with known E coli pathotypes in commercially available conventional diets and raw-meat-based diets (RMBDs). SAMPLE: 40 diets in total (19 conventionally cooked kibble or canned diets and 21 RMBDs) obtained from retail stores or online distributors. PROCEDURES: Each diet was cultured for E coli contamination in 3 separate container locations using standard microbiological techniques. Further characterization of E coli isolates was performed by polymerase chain reaction-based pathotype and virulence gene analysis. RESULTS: Conventional diets were negative in all culture based testing. In RMBDs, bacterial contamination was similar to previous reports in the veterinary literature, with 66% (14/21) of the RMBDs having positive cultures for E coli. Among the 191 confirmed E coli isolates from these diets, 31.9% (61/191) were positive for virulence genes. Categorized by pathotype, isolates presumptively belonging to the neonatal meningitis E coli pathotype (15.7% [30/191]) were the most common, followed by enterohemorrhagic E coli (10.5% [20/191]), enteropathogenic E coli (5.8% [11/191]), uropathogenic E coli (2.1% [4/191]), and diffusely adherent E coli (1.6% [3/191]). CLINICAL RELEVANCE: The results of this study reaffirmed the bacteriologic risks previously associated with RMBDs. Furthermore, potential zoonotic concerns associated with identified pathotypes in these diets may have significant consequences for owners in the animals' home environment. Potential risk associated with bacterial contamination should be addressed in animals fed RMBDs.

Escherichia coli probiotic exhibits in vitro growth-limiting effects on clinical feline uropathogenic E coli isolates.

OBJECTIVE: To characterize uropathogenic Escherichia coli (UPEC) in cases of clinical feline urinary tract infection (UTI) and subclinical bacteriuria and investigate the in vitro effects of E coli strain Nissle 1917 on isolate growth. ANIMALS: 40 cats with positive E coli culture results for urine collected during routine evaluation. PROCEDURES: Characterization of UPEC isolates was performed by PCR-based phylotype analysis and serotyping. Nissle 1917 effects on growth inhibition and competitive overgrowth against UPEC isolates were evaluated in vitro using a plate-based competition assay. RESULTS: Feline phylogroups were similar to previous human and feline UPEC studies, with most of the isolates belonging to phylogroup A (42.5%), B2 (37.5%), and D (15.0%). Fifty-two percent of isolates were found to be resistant to antimicrobials, with 19% of these being multidrug resistant (MDR). Nissle 1917 adversely affected the growth of 82.5% of all isolates and 100% of MDR isolates in vitro. The median zone of inhibition was 3.33 mm (range, 1.67 to 10.67 mm). Thirteen isolates were affected via competitive overgrowth and 20 via growth inhibition. CLINICAL RELEVANCE: UPEC isolates from cats were similar in phylogroup analysis to human and dog isolates. The in vitro effects of Nissle 1917 on UPEC warrant additional studies to determine if similar results can be duplicated in vivo.

Management strategies and clinical outcomes in breast cancer patients who develop left ventricular dysfunction during trastuzumab therapy.

BACKGROUND: Trastuzumab reduces risk of breast cancer recurrence but carries risk of cardiotoxicity that may be reversible upon treatment cessation and institution of left ventricular (LV) enhancement therapies (LVETx). We assessed management patterns of trastuzumab-induced cardiotoxicity (TIC) in a contemporary real-world setting. METHODS: We reviewed charts of all breast cancer patients who received adjuvant trastuzumab in British Columbia between January 2010 and December 2013, spanning the opening of a cardio-oncology clinic. LV dysfunction (LVD) was classified as minimal (LVEF nadir 45-49%), mild (40-44%) or moderate-severe (< 40%). Charts were reviewed for baseline characteristics, management strategies, and outcomes. Multivariable analysis was performed to identify patient characteristics associated with trastuzumab completion and cardiology referral. RESULTS: Of 967 patients receiving trastuzumab, 171 (17.7%) developed LVD, including 114 patients (11.8%) with LVEF declines of ≥10 to < 50%. Proportions of patients receiving cardiology referrals and LVETx increased and wait times to consultation decreased after a dedicated cardio-oncology clinic opened. LVETx was used more frequently in patients with moderate-severe LVD compared to minimal or mild LVD. Factors associated with completion of trastuzumab included mastectomy (OR 5.1, 95% CI 1.1-23.0) and proximity to quaternary care centre (OR 7.7, 95% CI 2.2-26.2). Moderate-severe LVD was associated with a lower probability of completing trastuzumab (OR 0.07 vs. minimal LVD, 95% CI 0.01-0.74). Factors associated with cardiology referral included heart failure symptoms (OR 8.0, 95% CI 1.5-42.9), proximity to quaternary care centre (OR 6.8, 95% CI 1.3-34.2), later year of cancer diagnosis (OR 2.4 per year, 95% CI 1.4-4.3), node-positive disease (OR 0.18, 95% CI 0.06-0.56), mastectomy (OR 0.05, 95% CI 0.01-0.52), and minimal LVD (OR 0.14, 95% CI 0.05-0.46). LVEF recovered to > 50% in 90.7% of patients. CONCLUSIONS: Management strategies in patients with TIC are associated with cancer characteristics and severity of cardiotoxicity. Access to dedicated cardio-oncology clinics may facilitate optimal care of this complex patient population.

Percutaneous Left Ventricular Assist Device in Cardiogenic Shock: A Five-Year Single Canadian Center Initial Experience.

BACKGROUND: Mechanical circulatory support in cardiogenic shock (CS) with percutaneous left ventricular assist devices (PVADs) has expanded rapidly, but there is a paucity of Canadian data. Conflicting observational reports have emerged regarding the benefit of PVADs in CS. We describe a 5-year experience with Impella CP for CS at a single Canadian tertiary care centre. METHODS: Consecutive adult patients with CS supported with Impella CP were included. Comprehensive clinical data and outcomes were retrospectively assessed. We evaluated patient characteristics, patterns of care, in-hospital outcomes, 6-month survival, and predictors of survival. RESULTS: Thirty-four patients were supported with Impella CP for CS over 5 years. A majority had acute myocardial infarction (94%) with advanced CS (68% Society for Cardiovascular Angiography and Intervention [SCAI] stage D or E). Survival to discharge was 58%. In patients who survived to discharge, 6-month survival was 100% with excellent functional status. SCAI CS stage and initial serum lactate showed significant associations with survival. There was also a trend towards improved survival with shorter door-to-PVAD time. Clinically significant bleeding was common (26%), and 3 patients had device-related vascular complications. CONCLUSION: Impella CP may have a role in carefully selected patients with CS. The SCAI shock classification and serum lactate may facilitate patient selection, and minimizing door-to-support time as well as bleeding complications are important considerations. Further clinical investigations, particularly in a Canadian setting, will be necessary to establish the role of this new technology in CS.

CONTEXTE: L'assistance circulatoire mécanique en cas de choc cardiogénique (CC) avec des dispositifs d'assistance ventriculaire gauche percutanée s'est rapidement développée, mais les données canadiennes restent rares. Des rapports d'observation contradictoires ont émergé concernant les avantages des dispositifs d'assistance ventriculaire gauche percutanée en cas de CC. Nous décrivons une expérience de cinq ans avec l'Impella CP pour les CC dans un seul centre de soins tertiaires canadien. MÉTHODES: Des patients adultes assistés par l'Impella CP, consécutivement à un CC, ont été inclus. Les données et les conclusions cliniques détaillées ont été évaluées rétrospectivement. Nous avons évalué les caractéristiques des patients, les modèles de soins, les bilans en milieu hospitalier, la survie à six mois et les indicateurs de survie. RÉSULTATS: Trente-quatre patients ont été pris en charge avec l'Impella CP pour un CC sur une période de cinq ans. Une majorité d'entre eux ont subi un infarctus aigu du myocarde (94 %) avec un CC avancé (68 % au stade D ou E sur l'échelle de la Society for Cardiovascular Angiography and Intervention [SCAI]). La survie jusqu'au congé hospitalier était de 58 %. Chez les patients qui ont survécu jusqu'à leur congé de l'hôpital, la survie à six mois était de 100 % avec un excellent état fonctionnel. Le stade de leur CC selon la SCAI et le lactate sérique initial ont montré des associations significatives avec le taux de survie. On a également constaté une tendance à l'amélioration de la survie avec un temps de porte à dispositifs d'assistance ventriculaire gauche percutanée raccourci. Des hémorragies importantes étaient fréquentes (26 %) et trois patients présentaient des complications vasculaires liées au dispositif. CONCLUSION: L'Impella CP pourrait avoir un rôle chez des patients atteints de CC soigneusement sélectionnés. La classification du choc selon la SCAI et le niveau de lactate sérique peuvent faciliter la sélection des patients, et la réduction du temps de « porte à assistance ¼ ainsi que les complications hémorragiques constituent des considérations d'importance. D'autres investigations cliniques, en particulier dans un contexte canadien, seront nécessaires pour établir le rôle de cette nouvelle technologie dans le CC.

The impact of implementation support on the use of a social engagement intervention for children with autism in public schools.

Several interventions have demonstrated efficacy in improving social outcomes for children with autism; however, few have been successfully implemented in schools. This study compared two implementation strategies to improve the use of a social engagement intervention for children with autism in public schools. In total, 31 children with autism in grades K-5 and 28 school personnel participated in a randomized controlled trial. Schools were randomized to (1) training in Remaking Recess, a social engagement intervention, or (2) training in Remaking Recess with implementation support. Linear regression with random effects was used to test the intervention effects on implementation fidelity and social outcomes (peer engagement, social network inclusion, and friendship nominations). In both groups, implementation fidelity improved after training but remained low. Children in the Remaking Recess with implementation support condition had significantly higher social network inclusion and received more friendship nominations than children in the Remaking Recess-only condition (p = 0.03). Children in both groups experienced reduced solitary engagement (p < 0.001) and increased joint engagement (p < 0.001). The results suggest that implementation supports may have an effect on outcomes above and beyond the intervention, and that further research is needed into the active intervention mechanisms.