Use of alfaxalone in bearded dragons (Pogona vitticeps): optimizing pharmacodynamics and evaluating cardiogenic effects via echocardiography.

OBJECTIVE: Bearded dragons (Pogona vitticeps), a popular zoological companion species, frequently require sedation for procedures. A novel formulation of alfaxalone with preservatives was FDA approved for 28-day use after the vial is breached. Research has been performed in squamate species using alfaxalone without preservatives at various doses and routes of administration, but it is unknown whether preservatives affect quality of sedation or cardiac function. ANIMALS: 10 bearded dragons. PROCEDURES: This complete crossover study evaluated the pharmacodynamic effects of alfaxalone with preservatives administered to bearded dragons via intracoelomic (ICo; n = 10), SC (10), IM (9), and IV (9) injection at 15 mg/kg. RESULTS: Deep sedation was achieved in 9 of 10 ICo, 8 of 10 SC, 8 of 9 IM, and 9 of 9 IV administrations. Heart rate significantly decreased from baseline for ICo (P = .008; median heart rate, 46), IM (P = .018; 54), and IV (P = .033; 54) routes, but maintained within clinically acceptable limits. Respiratory rate significantly decreased from baseline for ICo (P = .011; median respiratory rate, 30), SC (P = .024; 12), IM (P = .028; 12), and IV (P = .043; 12) routes. Spontaneous ventilation was retained during all events. Time to first effects was significantly sooner with IV (0 min) administration compared with ICo (P = .02; 5 min) and IM (P = .008; 5 min). Time to loss and recovery of withdrawal, righting reflex, deep pain, and purposeful movement were not significantly different between routes of administration. End-systolic volume was the only echocardiographic parameter significantly affected by IV sedation. CLINICAL RELEVANCE: Sedation quality was most consistent via IV administration at 15 mg/kg, and minimal changes in cardiac function were observed.

Clinical disease and treatment of Leptospira kirschneri sv Grippotyphosa in a Sumatran tiger (Panthera tigris sumatrae).

CASE DESCRIPTION: A 12-year-old sexually intact male zoo-managed Sumatran tiger (Panthera tigris sumatrae) was evaluated for a 3-day history of vomiting, hyporexia, and lethargy. Radiographs were supportive of gastrointestinal obstruction, and an exploratory laparotomy was performed. CLINICAL FINDINGS: Diffuse tan foci were present on the liver parenchyma, and the tiger became icteric throughout the procedure. Hepatic histopathology and immunohistochemistry resulted in a diagnosis of leptospirosis. Serum microagglutination testing for Leptospira spp antibody titers were positive for L kirschneri serovar Grippotyphosa, rising from 1:400 to 1:3,200 in 2 days. TREATMENT AND OUTCOME: The tiger was treated with antimicrobials, ursodiol, and mirtazapine, and increased biosecurity measures were instituted. Free-ranging wildlife on grounds were trapped, euthanized, and submitted for necropsy to screen for disease vectors. The tiger's urine was intermittently opportunistically collected from the enclosure and remained PCR assay negative for Leptospira spp until being positive once again on day 595. Although the tiger was without clinical signs at that time, antimicrobial therapy and increased biosecurity protocols were instituted a second time until urinary Leptospira shedding was confirmed to have stopped. By 1,071 days after initial presentation, the tiger remained nonclinical, with no additional urinary shedding episodes. CLINICAL RELEVANCE: While domestic and nondomestic free-ranging felids have been reported as subclinical Leptospira spp carriers, this report indicates the clinical importance of leptospirosis when a tiger presents with generalized gastrointestinal signs and icterus. Due to the zoonotic potential, biosecurity measures are necessary. This patient had a clinically successful outcome with antimicrobial therapy and supportive care.

OPTIMIZING THE PHARMACODYNAMICS AND EVALUATING CARDIOGENIC EFFECTS OF THE INJECTABLE ANESTHETIC ALFAXALONE IN PRAIRIE RATTLESNAKES (CROTALUS VIRIDIS).

North American vipers are commonly housed in zoological institutions or studied as free-ranging populations. Because of their venomous predatory and defensive mechanism, sedation or anesthesia is frequently employed to facilitate safe handling and medical procedures, especially of the head. A new formulation of alfaxalone with proprietary preservatives was recently approved and indexed for 28-d use post-vial puncture. Pharmacodynamic effects of alfaxalone in its prior formulation have been researched in nonvenomous species, but the optimal dose and route of administration in vipers have not been reported. In part one, 10 prairie rattlesnakes (Crotalus viridis) participated in a complete four-route crossover study evaluating 20 mg/kg alfaxalone administered intracoelomically (ICo), SC cranial to the heart, IM cranial to the heart, and IV in the ventral coccygeal vein. HR significantly decreased from baseline during IV (P= 0.024), IM (P= 0.024), and SC (P= 0.028) administration. Respiratory rate significantly decreased following alfaxalone delivered IV (P = 0.027). Time to first effects was significantly faster in IV compared with IM (P= 0.01), SC (P= 0.001), and ICo (P= 0.036). All IV and IM administrations resulted in deep sedation, but 70% of the IV and 10% of the IM sedation events resulted in apnea and required intermittent positive ventilation via endotracheal tube. Fifty percent of the ICo sedation events and 10% of the SC sedation events did not result in sedation. One successful SC sedation event resulted in apnea. In part two, echocardiograms were performed in the same rattlesnakes at baseline and at maximum effect of sedation with 20 mg/kg alfaxalone administered IM. Cardiac contractility and output were unaffected. Administration of alfaxalone at 20 mg/kg IM cranial to the heart should facilitate safe handling and minimally invasive procedures in prairie rattlesnakes and related species.

EVALUATION OF THE INTER- AND INTRAINDIVIDUAL AGREEMENT OF A PODODERMATITIS SCORING MODEL IN GREATER FLAMINGOS (PHOENICOPTERUS ROSEUS).

Pododermatitis is an important cause of morbidity and mortality in flamingos under human care; management and treatment options vary widely based on subjective assessment from veterinarians or animal care staff (ACS). The objective of this study was to evaluate the agreement of pododermatitis severity scores assigned by veterinarians, ACS, and veterinary students when given a standardized rubric. Twenty-four greater flamingos (Phoenicopterus roseus) from a single zoo-managed flock were evaluated over time for pododermatitis. The individual feet of each bird were imaged, blinded, randomized, and scored for hyperkeratosis, fissures, nodules, papillomatous growth, and overall subjective score by seven evaluators (three veterinary specialists, two ACS, and two veterinary students) using a previously established flamingo pododermatitis scoring rubric. Interindividual reliability between evaluators and intraindividual agreement among specialists was determined. Reliable interindividual agreement was seen for fissures (Krippendorff's α [KA] = 0.807) between all seven evaluators, whereas the other individual lesions had very low reliability. Between the specialists, fissures had low interindividual reliability (KA = 0.782). Two specialists had strong intraindividual agreement for fissure score and one specialist had strong intraindividual agreement for overall subjective score (Cohen's κ [CK] 0.8-0.9, P < 0.01). Hyperkeratosis, papillomatous growth, nodules, and overall subjective score had low to moderate inter- and intraindividual reliability or agreement (KA, 0.06-0.49; CK, 0.02-0.8). In conclusion, the current scoring method for flamingo pododermatitis does not supply a reliable method for tracking foot health based on images alone across timepoints, except for fissures. Further analysis of the scoring system being used during a physical examination is warranted.

Managing antibiotics wisely: a quality improvement programme in a tertiary neonatal unit in the UK.

Microbial resistance to antibiotics is a serious global health problem compounded by antibiotic overuse and limited investment in new antibiotic research. Inappropriate perinatal antibiotic exposure is increasingly linked to lifelong adverse outcomes through its impact on the developing microbiome. Antibiotic stewardship may be the only effective preventative strategy currently available. As the first tertiary neonatal unit in the UK to collaborate in an international quality improvement programme (QIP) with Vermont Oxford Network (VON), we present the results of our antibiotic stewardship initiative. The QIP was officially launched in January 2016 and aimed to reduce antibiotic usage rate (AUR) by 20% of baseline by 31st December 2016 without compromising patient safety. A multidisciplinary team of professionals and parent representatives shared good practices and improvement strategies through international webinars and local meetings, devised uniform data collection methodology and implemented a number of carefully selected 'Plan-Do-Study-Act' cycles. Run charts were used to present data and, where appropriate, statistical analysis undertaken to compare outcomes. The QIP resulted in a sustained reduction in AUR from a baseline median of 347 to 198 per 1000 patient-days (a reduction of 43%). The proportion of culture-negative sepsis screens where antibiotics were stopped within 36-48 hours increased consistently from a baseline of 32.5% to 91%. The antibiotic days per patient at discharge reduced from a median of 3 to 2 days, and there was a reduction in practice variation. Our annual mortality and necrotising enterocolitis rates for the VON cohort (<30 weeks or <1500 g) were the best ever recorded, 5.5% and 1.4%, respectively. Audits confirmed a high level of staff and family awareness of the QIP. The QIP achieved a sustained reduction in antibiotic use without compromising patient safety. Our challenge is to sustain this improvement safely.

Diagnosis and treatment of esophageal foreign body or stricture in three ferrets (Mustela putorius furo).

CASE DESCRIPTION 3 ferrets (Mustela putorius furo), aged 1 to 2 years, were referred for evaluation of a 4-day to 2-week history of gastrointestinal signs, including anorexia, regurgitation, and vomiting. CLINICAL FINDINGS All 3 ferrets had clinical signs suggestive of dysphagia or esophagitis on initial examination. Esophagoscopy, barium-contrast esophagography, or both revealed foreign bodies with mucosal inflammation in 1 patient and an esophageal foreign body with stricture in 2 patients. One of the latter ferrets had a recent history of gastrotomy to remove a foreign body. TREATMENT AND OUTCOME 1 ferret was treated with endoscopic retrieval of the foreign bodies. Esophageal stricture was treated in 2 ferrets by means of endoscopic balloon dilation accompanied by placement of an esophageal stent in 1 ferret. After resolution of clinical signs and completion of all prescribed treatments, 2 of 3 ferrets successfully transitioned to a regular hard kibble diet; 1 ferret remained on a soft diet for 2 years. All owners were satisfied with the outcome of treatment. CLINICAL RELEVANCE Ferrets are prone to foreign body ingestion. Results of this small series of cases suggested that minimally invasive techniques may be useful for the management of esophageal disease in this species.

Reductive functionalization of a rhodium(III)-methyl bond by electronic modification of the supporting ligand.

Net reductive elimination (RE) of MeX (X = halide or pseudo-halide: Cl(-), CF3CO2(-), HSO4(-), OH(-)) is an important step during Pt-catalyzed hydrocarbon functionalization. Developing Rh(I/III)-based catalysts for alkane functionalization is an attractive alternative to Pt-based systems, but very few examples of RE of alkyl halides and/or pseudo-halides from Rh(III) complexes have been reported. Here, we compare the influence of the ligand donor strength on the thermodynamic potentials for oxidative addition and reductive functionalization using [(t)Bu3terpy]RhCl (1) {(t)Bu3terpy = 4,4',4''-tri-tert-butylpyridine} and [(NO2)3terpy]RhCl (2) {(NO2)3terpy = 4,4',4''-trinitroterpyridine}. Complex 1 oxidatively adds MeX {X = I(-), Cl(-), CF3CO2(-) (TFA(-))} to afford [(t)Bu3terpy]RhMe(Cl)(X) {X = I(-) (3), Cl(-) (4), TFA(-) (5)}. By having three electron-withdrawing NO2 groups, complex 2 does not react with MeCl or MeTFA, but reacts with MeI to yield [(NO2)3terpy]RhMe(Cl)(I) (6). Heating 6 expels MeCl along with a small quantity of MeI. Repeating this experiment but with excess [Bu4N]Cl exclusively yields MeCl, while adding [Bu4N]TFA yields a mixture of MeTFA and MeCl. In contrast, 3 does not reductively eliminate MeX under similar conditions. DFT calculations successfully predict the reaction outcome by complexes 1 and 2. Calorimetric measurements of [(t)Bu3terpy]RhI (7) and [(t)Bu3terpy]RhMe(I)2 (8) were used to corroborate computational models. Finally, the mechanism of MeCl RE from 6 was investigated via DFT calculations, which supports a nucleophilic attack by either I(-) or Cl(-) on the Rh-CH3 bond of a five-coordinate Rh complex.

Activation of carbon-hydrogen bonds and dihydrogen by 1,2-CH-addition across metal-heteroatom bonds.

The controlled conversion of hydrocarbons to functionalized products requires selective C-H bond cleavage. This perspective provides an overview of 1,2-CH-addition of hydrocarbons across d(0) transition metal imido complexes and compares and contrasts these to examples of analogous reactions that involve later transition metal amide, hydroxide and alkoxide complexes with d(6) and d(8) metals.

Carbon-oxygen bond formation via organometallic Baeyer-Villiger transformations: a computational study on the impact of metal identity.

Metal-mediated formation of C-O bonds is an important transformation that can occur by a variety of mechanisms. Recent studies suggest that oxygen-atom insertion into metal-hydrocarbyl bonds in a reaction that resembles the Baeyer-Villiger transformation is a viable process. In an effort to identify promising new systems, this study is designed to assess the impact of metal identity on such O-atom insertions for the reaction [(bpy)(x)M(Me)(OOH)](n) → [(bpy)(x)M(OMe)(OH)](n) (x = 1 or 2; bpy = 2,2'-bipyridyl; n is varied to maintain the d-electron count at d(6) or d(8)). Six d(8)-square-planar complexes (M = Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), and Co(I)) and eight d(6)-octahedral systems (M = Ir(III), Rh(III), Co(III), Fe(II) Ru(II), Os(II), Mn(I), and Tc(I)) are studied. Using density functional theory calculations, the structures and energies of ground-state and transition-state species are elucidated. This study shows clear trends in calculated ΔG(++)'s for the O-atom insertions. The organometallic Baeyer-Villiger insertions are favored by lower coordination numbers (x = 1 versus x = 2), earlier transition metals, and first-row (3d) transition metals.

Pt(II) and Pt(IV) amido, aryloxide, and hydrocarbyl complexes: synthesis, characterization, and reaction with dihydrogen and substrates that possess C-H bonds.

The Pt(II) amido and phenoxide complexes ((t)bpy)Pt(Me)(X), ((t)bpy)Pt(X)(2), and [((t)bpy)Pt(X)(py)][BAr'(4)] (X = NHPh, OPh; py = pyridine) have been synthesized and characterized. To test the feasibility of accessing Pt(IV) complexes by oxidizing their Pt(II) precursors, the previously reported ((t)bpy)Pt(R)(2) (R = Me and Ph) systems were oxidized with I(2) to yield ((t)bpy)Pt(R)(2)(I)(2). The analogous reaction with ((t)bpy)Pt(Me)(NHPh) and MeI yields the corresponding ((t)bpy)Pt(Me)(2)(NHPh)(I) complex. Reaction of ((t)bpy)Pt(Me)(NHPh) and phenylacetylene at 80 °C results in the formation of the Pt(II) phenylacetylide complex ((t)bpy)Pt(Me)(C≡CPh). Kinetic studies indicate that the reaction of ((t)bpy)Pt(Me)(NHPh) and phenylacetylene occurs via a pathway that involves [((t)bpy)Pt(Me)(NH(2)Ph)][TFA] as a catalyst. The reaction of H(2) with ((t)bpy)Pt(Me)(NHPh) ultimately produces aniline, methane, (t)bpy, and elemental Pt. For this reaction, mechanistic studies reveal that 1,2-addition of dihydrogen across the Pt-NHPh bond to initially produce ((t)bpy)Pt(Me)(H) and free aniline is catalyzed by elemental Pt. Heating the cationic complexes [((t)bpy)Pt(NHPh)(py)][BAr'(4)] and [((t)bpy)Pt(OPh)(py)][BAr'(4)] in C(6)D(6) does not result in the production of aniline and phenol, respectively. Attempted synthesis of a cationic system analogous to [((t)bpy)Pt(NHPh)(py)][BAr'(4)] with ligands that are more labile than pyridine (e.g., NC(5)F(5)) results in the formation of the dimer [((t)bpy)Pt(µ-NHPh)](2)[BAr'(4)](2). Solid-state X-ray diffraction studies of the complexes ((t)bpy)Pt(Me)(NHPh), [((t)bpy)Pt(NH(2)Ph)(2)][OTf](2), ((t)bpy)Pt(NHPh)(2), ((t)bpy)Pt(OPh)(2), ((t)bpy)Pt(Me)(2)(I)(2), and ((t)bpy)Pt(Ph)(2)(I)(2) are reported.

Net hydrogenation of Pt-NHPh bond is catalyzed by elemental Pt.

Synthesis and characterization of the monomeric complex ((t)bpy)Pt(Me)(NHPh) ((t)bpy = 4,4'-di-tert-butyl-2,2'-dipyridyl) has been accomplished. Mechanistic studies reveal that 1,2-addition of dihydrogen across the Pt-anilido bond to initially produce ((t)bpy)Pt(Me)(H) and free aniline is catalyzed by elemental Pt rather than through a pathway that involves direct activation of H(2) by Pt and 1,2-addition across the Pt-NHPh bond.

The binding of protein S and the protein S-C4BP complex to neutrophils is apoptosis dependent.

Vitamin K-dependent protein S and complement regulator C4b-binding protein (C4BP) form a high-affinity complex in plasma. We have previously shown that both free protein S and the C4BP-protein S complex can bind to apoptotic Jurkat cells. It has been demonstrated in the past that protein S and C4BP can bind to neutrophils. We now show that it is only the apoptotic neutrophil population that binds these proteins. In addition, we also show that binding is mediated through the Gla domain on protein S, which binds negatively charged phospholipids, since a monoclonal antibody directed against this domain blocks the binding. Thus, we conclude that binding of protein S and the C4BP-protein S complex to neutrophils is not cell specific, but rather apoptosis dependent.

Role of CCP2 of the C4b-binding protein beta-chain in protein S binding evaluated by mutagenesis and monoclonal antibodies.

Complement regulator C4b-binding protein (C4BP) and the anticoagulant vitamin K-dependent protein S form a high affinity complex in human plasma. C4BP is composed of seven alpha-chains and a unique beta-chain, each chain comprising repeating complement control protein (CCP) modules. The binding site for protein S mainly involves the first of the three beta-chain CCPs (CCP1). However, recently it has been suggested that CCP2 of the beta-chain also contributes to the binding of protein S. To elucidate the structural background for the involvement of CCP2 in the protein S binding, several recombinant beta-chain CCP1-2 variants having mutations in CCP2 were expressed and tested for protein S binding. Mutations were chosen based on analysis of a homology model of the beta-chain and included R60A/R101A, D66A, L105A, F114A/I116A and H108A. All mutant proteins bound equally well as recombinant wild type to protein S. Several monoclonal antibodies against the beta-chain CCP2 were raised and their influence on protein S binding characterized. Taken together, the results suggest that the role of CCP2 in protein S binding is to orient and stabilize CCP1 rather than to be directly part of the binding site.

Vitamin K-dependent protein S localizing complement regulator C4b-binding protein to the surface of apoptotic cells.

Apoptosis is characterized by a lack of inflammatory reaction in surrounding tissues, suggesting local control of complement activation. During the initial stage of apoptosis, cells expose negatively charged phospholipid phosphatidylserine on their surfaces. The vitamin K-dependent protein S has a high affinity for this type of phospholipid. In human plasma, 60-70% of protein S circulates in complex with C4b-binding protein (C4BP). The reason why protein S and C4BP form a high-affinity complex in plasma is not known. However, C4BP is an important regulator of the classical pathway of the complement system where it acts as a cofactor in degradation of complement protein C4b. Using Jurkat cells as a model system for apoptosis, we now show protein S to bind to apoptotic cells. We further demonstrate protein S-mediated binding of C4BP to apoptotic cells. Binding of the C4BP-protein S complex to apoptotic cells was calcium-dependent and could be blocked with Abs directed against the phospholipid-binding domain in protein S. Annexin V, which binds to exposed phosphatidylserine on the apoptotic cell surface, could inhibit the binding of protein S. The C4BP that was bound via protein S to the apoptotic cells was able to interact with the complement protein C4b, supporting a physiological role of the C4BP/protein S complex in regulation of complement on the surface of apoptotic cells.