Alfaxalone increases measured progesterone concentration in neutered male cats when determined by immunoreactivity.

OBJECTIVE: Alfaxalone is a commonly used anesthetic agent in small animals. In cats, alfaxalone can be administered as an IM agent to achieve clinically useful sedation or anesthesia, negating the need for IV injection in difficult patients. The molecular structure of alfaxalone is similar to the hormone progesterone (P4). It is hypothesized that alfaxalone would cross-react with the assay measuring progesterone causing a false elevation. ANIMALS: 8 healthy neutered male, domestic shorthair cats that were privately owned were enrolled in the study. METHODS: Male neutered cats were administered 3 mg/kg of alfaxalone IM. Blood samples were collected at set time points (baseline, 30 minutes, 60 minutes, 3 hours, 6 hours, and 10 hours after administration), and serum concentrations of progesterone immunoreactivity (IR) were determined using the Siemens Immulite 1000 automated immunoassay system. Statistical analysis was performed with repeated measures ANOVA and a Tukey-Cramer multiple comparisons test. A P value of < .05 was used for significance. RESULTS: Serum progesterone IR was significantly elevated at 30 minutes, 1 hour, and 3 hours (P < .05) when compared to baseline progesterone immunoreactivity. Progesterone immunoreactivity had returned to baseline by 6 hours. CLINICAL RELEVANCE: This study suggests that alfaxalone administered IM in cats may interfere with immunoassay measurement of serum progesterone for up to 6 hours. Caution should be used when interpreting serum progesterone immunoreactivity results in cats within 4 hours of alfaxalone.

Adeno-Associated Virus Gene Therapy in a Sheep Model of Tay-Sachs Disease.

Tay-Sachs disease (TSD) is a fatal neurodegenerative disorder caused by a deficiency of the enzyme hexosaminidase A (HexA). TSD also occurs in sheep, the only experimental model of TSD that has clinical signs of disease. The natural history of sheep TSD was characterized using serial neurological evaluations, 7 Tesla magnetic resonance imaging, echocardiograms, electrodiagnostics, and cerebrospinal fluid biomarkers. Intracranial gene therapy was also tested using AAVrh8 monocistronic vectors encoding the α-subunit of Hex (TSD α) or a mixture of two vectors encoding both the α and ß subunits separately (TSD α + ß) injected at high (1.3 × 1013 vector genomes) or low (4.2 × 1012 vector genomes) dose. Delay of symptom onset and/or reduction of acquired symptoms were noted in all adeno-associated virus-treated sheep. Postmortem evaluation showed superior HexA and vector genome distribution in the brain of TSD α + ß sheep compared to TSD α sheep, but spinal cord distribution was low in all groups. Isozyme analysis showed superior HexA formation after treatment with both vectors (TSD α + ß), and ganglioside clearance was most widespread in the TSD α + ß high-dose sheep. Microglial activation and proliferation in TSD sheep-most prominent in the cerebrum-were attenuated after gene therapy. This report demonstrates therapeutic efficacy for TSD in the sheep brain, which is on the same order of magnitude as a child's brain.

"π-Hole-π" Interaction Promoted Photocatalytic Hydrodefluorination via Inner-Sphere Electron Transfer.

We describe a metal-free, photocatalytic hydrodefluorination (HDF) of polyfluoroarenes (FA) using pyrene-based photocatalysts (Py). The weak "π-hole-π" interaction between Py and FA promotes the electron transfer against unfavorable energetics (ΔGET up to 0.63 eV) and initiates the subsequent HDF. The steric hindrance of Py and FA largely dictates the HDF reaction rate, pointing to an inner-sphere electron transfer pathway. This work highlights the importance of the size and shape of the photocatalyst and the substrate in controlling the electron transfer mechanism and rates as well as the overall photocatalytic processes.

A New Approach to Non-Coordinating Anions: Lewis Acid Enhancement of Porphyrin Metal Centers in a Zwitterionic Metal-Organic Framework.

We describe a new strategy to generate non-coordinating anions using zwitterionic metal-organic frameworks (MOFs). By assembly of anionic inorganic secondary building blocks (SBUs) ([In(CO2)4](-)) with cationic metalloporphyrin-based organic linkers, we prepared zwitterionic MOFs in which the complete internal charge separation effectively prevents the potential binding of the counteranion to the cationic metal center. We demonstrate the enhanced Lewis acidity of Mn(III)- and Fe(III)-porphyrins in the zwitterionic MOFs in three representative electrocyclization reactions: [2 + 1] cycloisomerization of enynes, [3 + 2] cycloaddition of aziridines and alkenes, and [4 + 2] hetero-Diels-Alder cycloaddition of aldehydes with dienes. This work paves a new way to design functional MOFs for tunable chemical catalysis.

Highly Porous Zirconium Metal-Organic Frameworks with ß-UH3-like Topology Based on Elongated Tetrahedral Linkers.

Two non-interpenetrated zirconium metal-organic frameworks (Zr-MOFs), NPF-200 and NPF-201, were synthesized via the assembly of elongated tetrahedral linkers with Zr6 and Zr8 clusters. They represent the first examples of MOFs to have the ß-UH3-like, 4,12,12T1 topology. Upon activation, NPF-200 exhibits the largest BET surface area (5463 m(2) g(-1)) and void volume (81.6%) among all MOFs formed from tetrahedral ligands. Composed of negative-charged boron-centered tetrahedral linkers, NPF-201 is an anionic Zr-MOF which selectively uptakes photoactive [Ru(bpy)3](2+) for heterogeneous photo-oxidation of thioanisole.

In Vivo Selection Yields AAV-B1 Capsid for Central Nervous System and Muscle Gene Therapy.

Adeno-associated viral (AAV) vectors have shown promise as a platform for gene therapy of neurological disorders. Achieving global gene delivery to the central nervous system (CNS) is key for development of effective therapies for many of these diseases. Here we report the isolation of a novel CNS tropic AAV capsid, AAV-B1, after a single round of in vivo selection from an AAV capsid library. Systemic injection of AAV-B1 vector in adult mice and cat resulted in widespread gene transfer throughout the CNS with transduction of multiple neuronal subpopulations. In addition, AAV-B1 transduces muscle, ß-cells, pulmonary alveoli, and retinal vasculature at high efficiency. This vector is more efficient than AAV9 for gene delivery to mouse brain, spinal cord, muscle, pancreas, and lung. Together with reduced sensitivity to neutralization by antibodies in pooled human sera, the broad transduction profile of AAV-B1 represents an important improvement over AAV9 for CNS gene therapy.

Widespread Central Nervous System Gene Transfer and Silencing After Systemic Delivery of Novel AAV-AS Vector.

Effective gene delivery to the central nervous system (CNS) is vital for development of novel gene therapies for neurological diseases. Adeno-associated virus (AAV) vectors have emerged as an effective platform for in vivo gene transfer, but overall neuronal transduction efficiency of vectors derived from naturally occurring AAV capsids after systemic administration is relatively low. Here, we investigated the possibility of improving CNS transduction of existing AAV capsids by genetically fusing peptides to the N-terminus of VP2 capsid protein. A novel vector AAV-AS, generated by the insertion of a poly-alanine peptide, is capable of extensive gene transfer throughout the CNS after systemic administration in adult mice. AAV-AS is 6- and 15-fold more efficient than AAV9 in spinal cord and cerebrum, respectively. The neuronal transduction profile varies across brain regions but is particularly high in the striatum where AAV-AS transduces 36% of striatal neurons. Widespread neuronal gene transfer was also documented in cat brain and spinal cord. A single intravenous injection of an AAV-AS vector encoding an artificial microRNA targeting huntingtin (Htt) resulted in 33-50% knockdown of Htt across multiple CNS structures in adult mice. This novel AAV-AS vector is a promising platform to develop new gene therapies for neurodegenerative disorders.

Facile control of the charge density and photocatalytic activity of an anionic indium porphyrin framework via in situ metalation.

An anionic indium porphyrin framework (UNLPF-10) consisting of rare Williams ß-tetrakaidecahedral cages was constructed using an octatopic ligand linked with 4-connected [In(COO)4](-) SBUs. Remarkably, the extent of indium metalation of porphyrin macrocycles in UNLPF-10 can be facilely tuned in situ depending on the M/L ratio during synthesis, resulting in a controllable framework charge density and photocatalytic activity toward the selective oxygenation of sulfides.

Direct X-ray observation of trapped CO2 in a predesigned porphyrinic metal-organic framework.

Metal-organic frameworks (MOFs) are emerging microporous materials that are promising for capture and sequestration of CO2 due to their tailorable binding properties. However, it remains a grand challenge to pre-design a MOF with a precise, multivalent binding environment at the molecular level to enhance CO2 capture. Here, we report the design, synthesis, and direct X-ray crystallographic observation of a porphyrinic MOF, UNLPF-2, that contains CO2-specific single molecular traps. Assembled from an octatopic porphyrin ligand with [Co2(COO)4] paddlewheel clusters, UNLPF-2 provides an appropriate distance between the coordinatively unsaturated metal centers, which serve as the ideal binding sites for in situ generated CO2. The coordination of Co(II) in the porphyrin macrocycle is crucial and responsible for the formation of the required topology to trap CO2. By repeatedly releasing and recapturing CO2, UNLPL-2 also exhibits recyclability.

Sustained normalization of neurological disease after intracranial gene therapy in a feline model.

Progressive debilitating neurological defects characterize feline G(M1) gangliosidosis, a lysosomal storage disease caused by deficiency of lysosomal ß-galactosidase. No effective therapy exists for affected children, who often die before age 5 years. An adeno-associated viral vector carrying the therapeutic gene was injected bilaterally into two brain targets (thalamus and deep cerebellar nuclei) of a feline model of G(M1) gangliosidosis. Gene therapy normalized ß-galactosidase activity and storage throughout the brain and spinal cord. The mean survival of 12 treated G(M1) animals was >38 months, compared to 8 months for untreated animals. Seven of the eight treated animals remaining alive demonstrated normalization of disease, with abrogation of many symptoms including gait deficits and postural imbalance. Sustained correction of the G(M1) gangliosidosis disease phenotype after limited intracranial targeting by gene therapy in a large animal model suggests that this approach may be useful for treating the human version of this lysosomal storage disorder.

A "pillar-free", highly porous metalloporphyrinic framework exhibiting eclipsed porphyrin arrays.

A "pillar-free", highly porous metalloporphyrinic framework (UNLPF-1) exhibiting eclipsed porphyrin arrays has been constructed from an octacarboxylate ligand connected with paddlewheel secondary building units. UNLPF-1 possesses two types of open metal sites and exhibits an efficient selectivity toward capture of CO2 over N2.

Application of the laryngeal mask airway for anesthesia in three chimpanzees and one gibbon.

Three pediatric chimpanzees and one pediatric gibbon were anesthetized for routine physical examination. Anesthesia was maintained with inhalant delivered via a laryngeal mask airway (LMA). The LMA was easy to insert, provided adequate control of the airway for ventilation, and caused no tracheal stimulation. No complications were observed. As compared with a face mask, the LMA has the advantage of a more secure airway; the ability to effectively ventilate the patient; less dead space, which leads to lower rebreathing of carbon dioxide; and less exposure of personnel to waste gases. As compared with an endotracheal tube, the LMA causes less airway trauma, is easier to place, and is less stimulating to the patient. The LMA should be considered for use in fasted non-human primates presented for procedures lasting less than 60 min where high peak inspiratory pressures are not needed.

Antinociceptive effects of butorphanol, buprenorphine, or both, administered intramuscularly in cats.

OBJECTIVE: To characterize the antinociceptive action of IM-administered butorphanol, buprenorphine, or a combination of both by use of a thermal threshold method in cats. ANIMALS: 2 male and 4 female domestic cats. PROCEDURES: In a controlled, masked, randomized, crossover study design, thermal thresholds were measured by use of a thermal threshold-testing device developed for cats. Each cat received 4 treatments 1 week apart, consisting of 2 simultaneous IM injections in a random order (butorphanol-saline [0.9% NaCl] solution, buprenorphine-saline solution, butorphanol-buprenorphine, and saline solution-saline solution). The tester was unaware of the treatment given. Thermal thresholds were measured prior to injection, at intervals up to 12 hours, and at 22 hours after injection. RESULTS: There was no significant change in threshold over time after saline solution administration. All 3 opioid treatment groups had significant increases in thermal threshold, compared with pretreatment values (butorphanol, from 50 minutes to 8 hours; buprenorphine, from 35 minutes to 5 hours; and butorphanol-buprenorphine, from 50 minutes to 8 hours). Thermal thresholds did not differ significantly among opioid treatments at any time points, and thermal thesholds of only 2 opioid treatments (butorphanol at 50 minutes and butorphanol-buprenorphine at 8 hours) were significantly different from that of saline solution. CONCLUSIONS AND CLINICAL RELEVANCE: All 3 opioid treatments provided similar antinociception, although there was considerable intercat variability in the response to the different opioid treatments. This emphasizes the importance of assessing each patient individually and applying the treatment that works best for that patient.