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Background: Consumer expenditures on their family pets are rapidly increasing, part of which can be attributed to prescription and OTC medications. In turn, community pharmacies are seeking and receiving an increased number of prescriptions for animals. Community pharmacists’ ability to safely care for animal patients is relatively unexplored. Human medications, their normal dosing and even medication excipients could be lethal in some animal patients. Objective: The overarching objective of this study was to assess pharmacists’ baseline knowledge of potential pet poisons. Methods: The sample consisted of licensed pharmacists registered with the North Carolina Board of Pharmacy. The Pet Toxins Survey (PTS), a survey consisting of 25 potential pet toxins, was administered during October and November 2015. Analyses consisted of calculating descriptive statics (including graphical summaries to test for normality), and inferential statistics (two-tailed t-tests and ANOVAs) to compare responses across demographic variables. Results: A 6.3% response rate was obtained. After selecting either a dog or a cat to establish a frame of reference, participants in this study were able to correctly identify 15 of the 25 listed items as toxic to a pet (60% accuracy). Participants did not express adequate concern for the ingestion of several potential toxins. This includes potential excipients found in medication formulations such as xylitol, tea tree oil and caffeine. Female participants and those age 50 years and older were more likely to indicate concern for each potential toxin. There was no significant difference observed in responses based on the pharmacists’ work setting. Conclusions: The findings of this investigation suggest that pharmacists are deficient in their understanding of veterinary toxicology. Given the rise of community pharmacists caring for animal patients, it’s paramount that pharmacists be able to confidently distinguish potential pet toxins from non-toxins. It is also important that pharmacists receive a better understanding of what exposures require immediate action and what action should be taken.

Protozoal diarrhea caused by Tritrichomonas foetus (blagburni) is a prevalent, lifelong, and globally distributed burden in domestic cats. Treatment is limited to the use of 5-nitroimidazoles and treatment failure is common. The repurposed gold salt compound auranofin has killing activity against diverse protozoa in vitro but evidence of efficacy in naturally occurring protozoal infections is lacking. This exploratory study investigated the efficacy and safety of auranofin for treatment of cats with naturally occurring, 5-nitroimidazole-resistant, T. foetus infection. The minimum lethal concentration (MLC) of auranofin against 5 isolates of feline T. foetus was determined under aerobic conditions in vitro. Healthy cats and cats with T. foetus infection were treated with immediate release auranofin (range, 0.5-3 mg/cat for 7 days) or guar gum-coated auranofin capsules (0.5 or 3 mg/cat for 7 days). Adverse effects were monitored by clinical signs and clinicopathologic testing. Efficacy was determined by fecal consistency score, bowel movement frequency, and single-tube nested PCR of feces for T. foetus rDNA. Fecal samples were assayed for concentrations of auranofin, known and predicted metabolites of auranofin, gold containing molecules, and total gold content using HPLC, LC-MS, ion mobility-MS, and ICP-MS, respectively. Auranofin was effective at killing isolates of feline T. foetus at MLC ≥ 1 μg/ml. Treatment of cats with T. foetus infection with either immediate release auranofin or a colon-targeted guar gum-coated tablet of auranofin did not eradicate infection. Treatment failure occurred despite fecal concentrations of gold that met or exceeded the equivalent MLC of auranofin. Neither auranofin, known or predicted metabolites of auranofin, nor any gold-containing molecules >100 Da could be detected in fecal samples of treated cats. Adverse effects associated with auranofin treatment were common but minor. These studies identify that in vitro susceptibility test results of auranofin may not translate to treatment effectiveness in vivo even when achieving gold concentrations equivalent to the MLC of auranofin in the target environment. These studies further establish the absence of any predicted or unpredicted gold containing metabolites in feces after oral administration of auranofin., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Reserpine is a popular drug in the equine industry for long-term tranquilisation. Clinical observations revealed that blood from horses receiving oral reserpine was hypercoagulable. No studies have documented the pharmacokinetics of orally administered reserpine nor the effects of reserpine on platelets in horses., Objectives: To evaluate the pharmacokinetics of oral reserpine in horses and the effects of clinically relevant concentrations of reserpine on platelet functionality in vitro., Study Design: Experimental controlled study., Methods: The pharmacokinetics of oral reserpine (2.5 mg/horse, once) were determined in six healthy adult horses. Plasma samples were collected and concentrations of reserpine were determined by UPLC-MS/MS. Using this data, the in vitro effects of reserpine on platelets were examined. Aggregation, adhesion and releasate assays for serotonin and thromboxane B 2 were performed on platelets exposed to varying concentrations of reserpine (0.01-10 ng/mL), aspirin (negative control) and saline (unexposed control)., Results: Oral reserpine administration demonstrated low plasma concentrations with a C max of 0.2 ± 0.06 ng/mL and a prolonged half-life of 23.6 ± 6.24 h. Simulations over a dose range of 2-8 μg/kg predicted C max at steady state between 0.06-0.9 ng/mL. Platelets exposed to these reserpine concentrations in vitro displayed increased aggregation and adhesion compared to unexposed or aspirin-exposed platelets as well as compared to higher concentrations of reserpine. These functional changes correlated with lower concentrations of serotonin and higher concentrations of thromboxane B 2 in the platelet suspension supernatant., Main Limitations: This study used a small number of horses and only in vitro platelet experiments., Conclusions: Oral reserpine demonstrates low plasma concentrations and a prolonged half-life in horses. At these concentrations, reserpine causes significant changes in platelet function, most likely due to serotonin release and re-uptake which primes platelets for activation and thromboxane B 2 release. These findings suggest that clinicians should harvest blood for biological processing prior to the onset of reserpine administration., (© 2018 EVJ Ltd.)

Background: To date, there is very limited data regarding pharmacists' preparedness to handle animal prescriptions. No previous studies exist examining the impact of a veterinary-pharmacy-focused educational intervention. Objective: To assess pharmacists' baseline knowledge of veterinary pharmacotherapy, as relevant to their professional responsibilities, and assess the impact of a piloted educational program. Methods: Two studies were conducted. The first study involved a statewide assessment of pharmacists' knowledge of veterinary pharmacotherapy; the second study assessed the impact of an educational intervention to improve pharmacists' veterinary pharmacotherapy knowledge base. Participants in the pilot study were assessed via pretest and posttest. Results: The statewide sample of participants (n = 602) received a mean score of 5.9 (SD = 2.6) on a 17-item questionnaire. There were no discernible differences in participants' knowledge based on the subject matter of the question (pathophysiology, dosing, counseling, compounding, legality, and toxicology). Using the same 17-item questionnaire, pilot study participants (n = 60) received a mean score of 5.2 (SD = 2.4) on the pretest and 16.6 (SD = 0.7) on the posttest. Conclusion: The findings of this study suggest that a substantial portion of pharmacists lack the knowledge needed to process and dispense the veterinary prescriptions most commonly encountered in community pharmacies. Furthermore, this study shows that implementation of an educational intervention can increase pharmacists' knowledge of core concepts necessary to safely care for animal patients., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2018.)

OBJECTIVE To investigate in vitro carboplatin release from 6 carrier media. SAMPLE 6 carboplatin-containing carrier media. PROCEDURES An in vitro release study was performed with 6 commercially available carrier media: a hemostatic gelatin sponge, a poloxamer copolymer gel, and 2 sizes (3 and 4.8 mm in diameter) of beads molded from each of 2 commercial calcium sulfate products. All carrier media contained 10 mg of carboplatin. Carrier media specimens were placed in 37°C PBS solution for 96 hours. Carboplatin concentrations in PBS solution were measured by use of high-performance liquid chromatography at 15 time points to calculate the amount and proportion of carboplatin released from each specimen. RESULTS Peak release of carboplatin from the poloxamer copolymer gel and hemostatic gelatin sponge were achieved after 4 and 20 hours, respectively. Maximum release did not differ significantly between the poloxamer copolymer gel and hemostatic gelatin sponge, but both released significantly more carboplatin within 96 hours than did both of the commercial calcium sulfate products. The poloxamer copolymer gel released 99% of the carboplatin, and the hemostatic gelatin sponge released 68.5% of the carboplatin. Peak release of carboplatin from the calcium sulfate beads was not reached within 96 hours. CONCLUSIONS AND CLINICAL RELEVANCE In this study, carboplatin release from the hemostatic gelatin sponge was incomplete. The poloxamer copolymer gel and hemostatic gelatin sponge released carboplatin rapidly in vitro, whereas calcium sulfate beads did not.

Administering chronic medications to feline patients without the daily battle of oral and injectable medications is the holy grail of veterinary pharmacotherapy. For some medications, the transdermal route may be the solution. However, there are many considerations for determining if a medication will be safe for the patient and caregiver as well as effective when administered transdermally. A comprehensive checklist to assess the appropriateness of transdermal therapy is provided.

Ronidazole (RDZ) is the only known effective treatment for feline diarrhea caused by Tritrichomonas foetus. This study aimed to develop guar gum-coated colon-targeted tablets of RDZ and to determine the pharmacokinetics of this delayed-release formulation in cats. Guar gum-coated tablets were administered orally once to five healthy cats (mean dose 32.3 mg/kg). The tablets were then administered once daily for 5 days to four cats (mean dose 34.5 mg/kg), and absorption studies repeated on day 5. Plasma was collected and analyzed for RDZ concentration, and pharmacokinetic noncompartmental and deconvolution analysis were performed on the data. There was negligible RDZ release until after 6 h, and a delayed peak plasma concentration (mean Cmax 28.9 μg/mL) at approximately 14.5 h, which coincides with colonic arrival in cats. Maximum input rate (mg/kg per hour) occurred between 6 and 16 h. This delayed release of ronidazole from guar gum-coated tablets indicates that release of RDZ may be delayed to deliver the medication to a targeted area of the intestine. Repeated dosing with guar gum tablets to steady-state did not inhibit drug bioavailability or alter the pharmacokinetics. Such targeted RDZ drug delivery may provide improved efficacy and reduce adverse effects in cats., (© 2012 John Wiley & Sons Ltd.)

Objective: To determine the concentration of doxycycline compounded from doxycycline hyclate tablets into liquid formulations for oral administration in veterinary species and stored for 28 days., Design: Evaluation study., Sample: Doxycycline hyclate tablets (100 mg) crushed and mixed with a 50:50 mixture of syrup and suspension vehicles for oral administration to produce 3 batches each of 2 doxycycline formulations: 33.3 and 166.7 mg/mL., Procedures: Formulations were stored, protected from light, at room temperature (22° to 26°C [71.6° to 78.8°F]) and at a controlled cold temperature (refrigerated 2° to 8°C [35.6° to 46.4°F]). Doxycycline was extracted from the formulations, and concentration was measured by high-pressure liquid chromatography on days 0 (date of preparation), 1, 4, 7, 14, 21, and 28. Concentrations were compared with those of a US Pharmacopeial Convention reference standard. Formulation quality at each point was also assessed through color change, formulation consistency, and suspension uniformity., Results: On days 0, 1, 4, and 7, the concentration of each formulation was within 90% to 110% of the reference standard (range, 93% to 109%), which was deemed acceptable. However, doxycycline concentrations had decreased dramatically by day 14 and remained low for the duration of the study period. Doxycycline concentrations on days 14, 21, and 28 were all < 20% (range, 14% to 18%) of the reference standard, and the quality of the formulations decreased as well. No effect of storage temperatures on doxycycline concentration was identified., Conclusions and Clinical Relevance: The concentration of doxycycline, compounded from commercial tablets in the vehicles evaluated to yield doses of 33.3 and 166.7 mg/mL, cannot be assured beyond 7 days.

Background: Progress in establishing if therapies provide relief to cats with degenerative joint disease (DJD)-associated pain is hampered by a lack of validated owner-administered assessment methods., Hypothesis: That an appropriately developed subjective owner-completed instrument (Feline Musculoskeletal Pain Index-FMPI) to assess DJD-associated impairment would have responsiveness and criterion validity., Animals: Twenty-five client-owned cats with DJD-associated pain., Methods: FMPI responsiveness (ability to detect the effect of an analgesic treatment) and validity (correlation with an objective measure) were explored through a stratified, randomized, double blinded, placebo-controlled, crossover 10-week clinical study. Meloxicam was administered to effect pain relief. A linear mixed model, backward stepwise regression, and Pearson correlations were used to assess responsiveness and criterion validity with the assumption that the NSAID would increase activity., Results: Positive responses of cats to placebo (P = .0001) and meloxicam treatment (P = .0004) were detected; however, the instrument did not detect any difference between placebo and meloxicam (linear mixed model), even for the high impairment cases. Percent meloxicam target dose administered, temperament, and total baseline FMPI score were covariates that most affected FMPI scores. Controlling for significant covariates, most positive effects were seen for placebo treatment. Positive treatment effects on activity were detected, but only for the cases designated as most highly impaired., Conclusions and Clinical Importance: Neither responsiveness nor criterion validity were detected by the inclusion criteria for cases in this study. The data suggest that further work is indicated to understand factors affecting activity in cats to optimize inclusion criteria., (Copyright © 2013 by the American College of Veterinary Internal Medicine.)

In this chapter, we argue that with careful attention to cell types in stationary-phase cultures of the yeast, S. cerevisiae provide an excellent model system for aging studies and hold much promise in pinpointing the set of causal genes and mechanisms driving aging. Importantly, a more detailed understanding of aging in this single celled organism will also shed light on aging in tissue-complex model organisms such as C. elegans and D. melanogaster. We feel strongly that the relationship between aging in yeast and tissue-complex organisms has been obscured by failure to notice the heterogeneity of stationary-phase cultures and the processes by which distinct cell types arise in these cultures. Although several studies have used yeast stationary-phase cultures for chronological aging, the majority of these studies have assumed that cultures in stationary phase are homogeneously composed of a single cell type. However, genome-scale analyses of yeast stationary-phase cultures have identified two major cell fractions: quiescent and non-quiescent, which we discuss in detail in this chapter. We review evidence that cell populations isolated from these cultures exhibit population-specific phenotypes spanning a range of metabolic and physiological processes including reproductive capacity, apoptosis, differences in metabolic activities, genetic hyper-mutability, and stress responses. The identification, in S. cerevisiae, of multiple sub-populations having differentiated physiological attributes relevant to aging offers an unprecedented opportunity. This opportunity to deeply understand yeast cellular (and population) aging programs will, also, give insight into genomic and metabolic processes in tissue-complex organism, as well as stem cell biology and the origins of differentiation.

Thirty-one dogs were randomised to receive intermittent wound infusion of bupivacaine or saline after surgery. Wound pressure sensitivity, pain scores, body temperature, heart rate, respiratory rate, analgesic drugs administered, time to walking and time to eating after surgery were recorded. Plasma bupivacaine concentrations were measured. The relative frequency distributions of the non-interventional and interventional pain scores, but not the relative frequency distributions of palpation pain scores or wound pressure sensitivity, were significantly different between groups following surgery. There was a significant difference between groups in the time to eating and in the amount and timing of analgesic drugs administered. Measured plasma bupivacaine concentrations demonstrated systemic absorption of the drug. Bupivacaine infusion into surgical wounds after surgery may improve post-operative recovery, but no effect on wound tenderness was demonstrated in this study., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Ronidazole (RDZ) is an effective treatment for feline Tritrichomonas foetus infection, but has produced neurotoxicity in some cats. An understanding of the disposition of RDZ in cats is needed in order to make precise dosing recommendations. Single-dose pharmacokinetics of intravenous (IV) RDZ and immediate-release RDZ capsules were evaluated. A single dose of IV RDZ (mean 9.2mg/kg) and a 95mg immediate-release RDZ capsule (mean 28.2mg/kg) were administered to six healthy cats in a randomized crossover design. Plasma samples were collected for 48 h and assayed for RDZ using high pressure liquid chromatography (HPLC). Systemic absorption of oral RDZ was rapid and complete, with detection in the plasma of all cats by 10 min after dosing and a bioavailability of 99.64 (±16.54)%. The clearance of RDZ following IV administration was 0.82 (±0.07) ml/kg/min. The terminal half-life was 9.80 (±0.35) and 10.50 (±0.82) h after IV and oral administration, respectively, with drug detectable in all cats 48h after both administrations. The high oral bioavailability of RDZ and slow elimination may predispose cats to neurotoxicity with twice-daily administration. Less frequent administration should be considered for further study of effective treatment of T foetus-infected cats., (Copyright © 2011 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.)

As yeast cultures enter stationary phase in rich, glucose-based medium, differentiation of two major subpopulations of cells, termed quiescent and nonquiescent, is observed. Differences in mRNA abundance between exponentially growing and stationary-phase cultures and quiescent and nonquiescent cells are known, but little was known about protein abundance in these cells. To measure protein abundance in exponential and stationary-phase cultures, the yeast GFP-fusion library (4159 strains) was examined during exponential and stationary phases, using high-throughput flow cytometry (HyperCyt). Approximately 5% of proteins in the library showed twofold or greater changes in median fluorescence intensity (abundance) between the two conditions. We examined 38 strains exhibiting two distinct fluorescence-intensity peaks in stationary phase and determined that the two fluorescence peaks distinguished quiescent and nonquiescent cells, the two major subpopulations of cells in stationary-phase cultures. GFP-fusion proteins in this group were more abundant in quiescent cells, and half were involved in mitochondrial function, consistent with the sixfold increase in respiration observed in quiescent cells and the relative absence of Cit1p:GFP in nonquiescent cells. Finally, examination of quiescent cell-specific GFP-fusion proteins revealed symmetry in protein accumulation in dividing quiescent and nonquiescent cells after glucose exhaustion, leading to a new model for the differentiation of these cells.

Salmonella enterica Serovar Typhimurium (S. Typhimurium) causes enterocolitis with diarrhea and polymorphonuclear cell (PMN) influx into the intestinal mucosa in humans and calves. The Salmonella Type III Secretion System (T3SS) encoded at Pathogenicity Island I translocates Salmonella effector proteins SipA, SopA, SopB, SopD, and SopE2 into epithelial cells and is required for induction of diarrhea. These effector proteins act together to induce intestinal fluid secretion and transcription of C-X-C chemokines, recruiting PMNs to the infection site. While individual molecular interactions of the effectors with cultured host cells have been characterized, their combined role in intestinal fluid secretion and inflammation is less understood. We hypothesized that comparison of the bovine intestinal mucosal response to wild type Salmonella and a SipA, SopABDE2 effector mutant relative to uninfected bovine ileum would reveal heretofore unidentified diarrhea-associated host cellular pathways. To determine the coordinated effects of these virulence factors, a bovine ligated ileal loop model was used to measure responses to wild type S. Typhimurium (WT) and a ΔsipA, sopABDE2 mutant (MUT) across 12 hours of infection using a bovine microarray. Data were analyzed using standard microarray analysis and a dynamic bayesian network modeling approach (DBN). Both analytical methods confirmed increased expression of immune response genes to Salmonella infection and novel gene expression. Gene expression changes mapped to 219 molecular interaction pathways and 1620 gene ontology groups. Bayesian network modeling identified effects of infection on several interrelated signaling pathways including MAPK, Phosphatidylinositol, mTOR, Calcium, Toll-like Receptor, CCR3, Wnt, TGF-β, and Regulation of Actin Cytoskeleton and Apoptosis that were used to model of host-pathogen interactions. Comparison of WT and MUT demonstrated significantly different patterns of host response at early time points of infection (15 minutes, 30 minutes and one hour) within phosphatidylinositol, CCR3, Wnt, and TGF-β signaling pathways and the regulation of actin cytoskeleton pathway.

To resolve the genetic heterogeneity within pediatric high-risk B-precursor acute lymphoblastic leukemia (ALL), a clinically defined poor-risk group with few known recurring cytogenetic abnormalities, we performed gene expression profiling in a cohort of 207 uniformly treated children with high-risk ALL. Expression profiles were correlated with genome-wide DNA copy number abnormalities and clinical and outcome features. Unsupervised clustering of gene expression profiling data revealed 8 unique cluster groups within these high-risk ALL patients, 2 of which were associated with known chromosomal translocations (t(1;19)(TCF3-PBX1) or MLL), and 6 of which lacked any previously known cytogenetic lesion. One unique cluster was characterized by high expression of distinct outlier genes AGAP1, CCNJ, CHST2/7, CLEC12A/B, and PTPRM; ERG DNA deletions; and 4-year relapse-free survival of 94.7% ± 5.1%, compared with 63.5% ± 3.7% for the cohort (P = .01). A second cluster, characterized by high expression of BMPR1B, CRLF2, GPR110, and MUC4; frequent deletion of EBF1, IKZF1, RAG1-2, and IL3RA-CSF2RA; JAK mutations and CRLF2 rearrangements (P < .0001); and Hispanic ethnicity (P < .001) had a very poor 4-year relapse-free survival (21.0% ± 9.5%; P < .001). These studies reveal striking clinical and genetic heterogeneity in high-risk ALL and point to novel genes that may serve as new targets for diagnosis, risk classification, and therapy.

Objective: To evaluate a technique for midhumeral peripheral nerve blockade in the dog., Study Design: Cadaveric technique development; in vivo placebo-controlled, prospective crossover study., Animals: Canine cadavers (n=38) and 8 clinically healthy, adult hound dogs., Methods: A technique for peripheral block of the radial, ulnar, musculocutaneous, and median nerves (RUMM block) was evaluated using cadaver limbs. Eight purpose-bred, research dogs were anesthetized; a RUMM block was performed on each thoracic limb. One limb from each dog randomly received 0.5% bupivacaine and the opposite limb was assigned to receive sterile saline solution as a control. After recovery from anesthesia, skin sensation at selected dermatomes was evaluated for 24 hours using a mechanical stimulus. Weight-bearing, conscious proprioception, and withdrawal reflex were also evaluated. One month after initial testing, each dog was reanesthetized and each limb received the opposite treatment., Results: Sensory thresholds were significantly increased over baseline measurements when compared with control limbs for all nerves. Complete sensory block was achieved in radial (15/16), ulnar (3/16), musculocutaneous (8/16), and median (11/16) nerves, using a mechanical stimulus of analgesia. Complete simultaneous block of all nerves was only obtained in 1 of 16 limbs., Conclusion: RUMM block resulted in desensitization of the skin in the associated dermatomes for 4-10 hours. Complete sensory block of the dermatomes supplied by the radial nerve was most consistent., Clinical Relevance: RUMM block may be an effective technique to provide adjunctive analgesia for dogs undergoing surgery of the distal aspect of the thoracic limb., (© Copyright 2010 by The American College of Veterinary Surgeons.)

Objective: An experiment was conducted to compare the effectiveness of individual versus group electronic brainstorming to address difficult, real-world challenges., Background: Although industrial reliance on electronic communications has become ubiquitous, empirical and theoretical understanding of the bounds of its effectiveness have been limited. Previous research using short-term laboratory experiments have engaged small groups of students in answering questions irrelevant to an industrial setting. The present experiment extends current findings beyond the laboratory to larger groups of real-world employees addressing organization-relevant challenges during the course of 4 days., Methods: Employees and contractors at a national laboratory participated, either in a group setting or individually, in an electronic brainstorm to pose solutions to a real-world problem., Results: The data demonstrate that (for this design) individuals perform at least as well as groups in producing quantity of electronic ideas, regardless of brainstorming duration. However, when judged with respect to quality along three dimensions (originality, feasibility, and effectiveness), the individuals significantly (p < .05) outperformed the group., Conclusion: When quality is used to benchmark success, these data indicate that work-relevant challenges are better solved by aggregating electronic individual responses rather than by electronically convening a group., Application: This research suggests that industrial reliance on electronic problem-solving groups should be tempered, and large nominal groups may be more appropriate corporate problem-solving vehicles.

Objective: To evaluate the stability and retention of viscous formulations of the antifungal drug clotrimazole in vitro and to evaluate retention times, absorption, and histologic response to these compounds when placed in the frontal sinus of dogs., Animals: 6 male Beagles., Procedures: 1% clotrimazole gels were formulated with hydroxypropyl cellulose, poloxamer, and carboxymethylcellulose sodium bases. Commercially available 1% clotrimazole creams were also evaluated. Each compound was incubated at 37 degrees C in a funnel. Volume retained and clotrimazole stability were evaluated for 4 weeks. Six compounds were then chosen for in vivo evaluation. The frontal sinuses of 6 dogs were filled with 1 of the 6 compounds. Computed tomographic evaluation was performed weekly for up to 4 weeks to evaluate gel retention. Blood samples were collected to evaluate clotrimazole absorption. Following euthanasia, sinuses were examined histologically., Results: Commercially available clotrimazole creams were not retained in funnels in vitro. In vivo, hydroxypropyl cellulose- and carboxymethylcellulose-based gels resulted in the most severe inflammatory response and were retained the longest. Poloxamer-based gels had a shorter retention time and were associated with less inflammation. Clotrimazole was minimally absorbed. Despite a marked inflammatory response to several of the clotrimazole-containing gels, no notable adverse clinical responses were observed., Conclusions and Clinical Relevance: Poloxamer gels had the most promise for improving drug contact within the frontal sinus of dogs, while limiting the inflammatory response. Poloxamer gels have the additional benefit of improved handling as a result of reverse gelation (ie, they gel when warmed to 37 degrees C).

Objective: To determine the effects of temperature and light over a 35-day period on stability of pergolide mesylate after compounding in an aqueous vehicle., Design: Evaluation study., Procedures: Pergolide was compounded into a formulation with a final target concentration of 1 mg/mL. Aliquots of the formulation were then stored at -20 degrees, 8 degrees, 25 degrees, or 37 degrees C without exposure to light or at 25 degrees C with exposure to light for 35 days. Samples were assayed in triplicate by means of high-pressure liquid chromatography immediately after compounding and after 1, 7, 14, 21, and 35 days of storage., Results: Mean+/-SD concentration of pergolide in the formulation immediately after compounding was 1.05+/-0.086 mg/mL. Samples exposed to light while stored at 25 degrees C had undergone excessive degradation by day 14, samples stored at 37 degrees C had undergone excessive degradation by day 21, and samples stored at 25 degrees C without exposure to light had undergone excessive degradation by day 35. The decrease in expected concentration corresponded with the appearance of degradation peaks in chromatograms and with a change in color of the formulation., Conclusions and Clinical Relevance: Results indicated that pergolide mesylate was unstable after compounding in an aqueous vehicle and that storage conditions had an effect on stability of the compounded formulation. Compounded pergolide formulations in aqueous vehicles should be stored in a dark container, protected from light, and refrigerated and should not be used >30 days after produced. Formulations that have undergone a color change should be considered unstable and discarded.

Anesthetization of large animals, such as the horse, is a challenge for veterinarians and compounding pharmacists. This article discusses the technique of intravenous anesthesia by using a combination of xylazine and ketamine in the horse for short-term unconsciousness; a combination of xylazine, diazepam, and ketamine, for improved sedation and muscle relaxation; and a combination of xylazine, ketamine, and guaifenesin, referred to as Triple Drip, for the maximum 1-hour duration of unconsciousness. Compounding pharmacists that have the facilities and training required to prepare sterile compounds can greatly assist equine surgeons in the treatment of veterinarians' most challenging patients.

Myasthenia gravis is a neruomuscular disease commonly encountered in veterinary medicine. With accurate diagnosis, treatment, and compliant caregivers, the prognosis for remission and a normal life is good. Unfortunately, drugs used to diagnose and treat myasthenia gravis are frequently unavailable due to manufacturer shortages. During these periods, compounding pharmacists can play a valuable role in maintaining a good quality of life for these patients by providing compounded versions of drugs that are commercially unavailable.

Dogs are utilized more and more to seek out illicit and hazardous substances under the guidance of humans. Therefore, veterinarians, dog handlers, and pharmacists must all collaborate to ensure adequate protection, treatment, and care for detection dogs in the line of duty. Ideally, veterinarians and pharmacists can work together to train and equip dog handlers with state of the art instructions and animal emergency kits that can provide life-saving decontamination at the site of exposure. Pharmacists can collaborate with veterinarians to provide animal emergency kits containing essential treatment aids such as apomorphine, hydrogen peroxide, activated charcoal/sorbitol cathartic suspension, ocular irrigants, and instructions and associated devices for administration. Immediate intervention provides the best prognosis for canine heroes that suffer exposure to occupational toxicological hazards.

Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and nonquiescent (NQ) fractions before entering stationary phase. To understand this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Eleven mutants that affected one or both phenotypes in Q or NQ fractions were identified. NQ fractions exhibit a high level of petite colonies, and nine mutants affecting this phenotype were identified. Microarray analysis revealed >1300 mRNAs distinguished Q from NQ fractions. Q cell-specific mRNAs encode proteins involved in membrane maintenance, oxidative stress response, and signal transduction. NQ-cell mRNAs, consistent with apoptosis in these cells, encode proteins involved in Ty-element transposition and DNA recombination. More than 2000 protease-released mRNAs were identified only in Q cells, consistent with these cells being physiologically poised to respond to environmental changes. Our results indicate that Q and NQ cells differentiate significantly, with Q cells providing genomic stability and NQ cells providing nutrients to Q cells and a regular source of genetic diversity through mutation and transposition. These studies are relevant to chronological aging, cell cycle, and genome evolution, and they provide insight into complex responses that even simple organisms have to starvation.

When peer reviewed scientific information is not available, veterinarians and pharmacists often turn to their colleagues seeking evidence based upon clinical experience in order to devise a treatment plan for a patient. Compounding pharmacists who provide care and compounds for veterinary patients have vast clinical experience of great educational value. Using specific guidelines, compounding pharmacists can prepare useful case reports and/or case series by proactively collecting clinical data, carefully selecting a case for presentation, and collating the details into a compelling story with a clear message that can survive editorial triage and contribute to the advancement of veterinary medical care.

Background: Autosomal recessive hereditary nephropathy (ARHN) in the English Cocker Spaniel is caused by a type IV collagen defect, but the underlying mutation is unknown., Animals: One hundred thirty-four English Cocker Spaniels (12 with ARHN, 8 obligate carriers, and 114 others), 3 mixed breed dogs with X-linked hereditary nephropathy (XLHN), and 7 other dogs without hereditary nephropathy were included., Methods: Diagnosis of ARHN was based on transmission electron microscopy and immunostaining of kidney. Quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR) was used to compare COL4A3, COL4A4, and COL4A5 mRNA concentrations in the renal cortex from ARHN-affected English Cocker Spaniels, XLHN-affected dogs, and dogs without hereditary nephropathy. The entire coding region of COL4A4 was sequenced in 2 ARHN-affected dogs, 2 obligate carriers, 2 English Cocker Spaniels of unknown status, and 2 healthy mixed breed dogs. The exon containing the mutation was sequenced for all 134 English Cocker Spaniels., Results: Quantitative real time RT-PCR implicated COL4A4 as the gene harboring the mutation, and sequencing identified a single nucleotide substitution at base 115 as the cause of ARHN in English Cocker Spaniels. This mutation, which causes a premature stop codon in exon 3 of COL4A4, was segregated with clinical status in all affected dogs and obligate carriers. The mutation also was identified in 39 of 114 other English Cocker Spaniels with previously unknown status., Conclusions and Clinical Importance: The cause of this disease has been identified, and use of a test for the mutation will permit eradication of ARHN in the English Cocker Spaniel.

Pharmacists can play a valuable role in providing care and preparations for all species of patients with diabetes. Practical knowledge of the differences in diabetes between animals and humans is essential to providing diabetic care to multiple species. Pharmacists should familiarize themselves with the clinical signs of hyperglycemia and hypoglycemia in various species and be prepared to provide appropriate counseling, information, preparations, devices, and referral when signs of inadequate glycemic control arise. Pharmacists also should develop awareness of potential problems associated with the use of human diabetic preparations in animal patients. By maintaining a current knowledge of care for diabetic pets, including the significance of the patient's dietary habits, medical history, and genetics, and species-specific drug reactions, the compounding pharmacist can contribute, through collaboration with veterinarians and pet caregivers, to optimal patient outcomes when providing care for pets with diabetes.

Tritrichomonas foetus, a microscopic single-celled flagellated protozoan parasite, traditionally identified as a cause of reproductive disease in cattle, has been demonstrated as an important cause of diarrhea in cats. Until recently, an effective antimicrobial treatment for feline Tritrichomonas foetus had not been idnetified. Since recommended dosages of antimicrobial drugs have failed in cats infected with Tritrichomonas foetus and in vitro studies have revealed multiple drug resistance, investigations continue in the effort to find an effective treatment. One particular study by Dr.Jody Gookin noted no clinicopathological abnormalities or adverse effects with the use of ronidazole, and the research concluded that ronidazole administered at 30 to 50 mg/kg orally twice daily for 2 weeks was capable of resolving diarrhea and eradicating infections of Tritrichomonas foetus in cats. Clinical use of ronidazole has revealed a reversible, possibly dose-related, neurotoxicity. Cats receiving ronidazole should be monitored for signs of nystagmus, ataxia, or behavior change, and ronidazole should be discontinued immediately if any of these signs are observed. Many compounding pharmacists have begun stocking pure ronidazole powder to prepare veterinarian prescriptions for the treatnment of Tritrichomonas foetus. Doses of ronidazole are usually prepared as capsules, the most desirable dosage form because (1)the capsules mask the extremely bitter taste of ronidazole, (2)exact dosing reduces the risk of neurotoxicity that has been reported at higher doses, and (3)the owner can be assured that the entire dose is ingested by the cat after administration of a capsule.

Pulmonary arterial hypertension is the most commonly recognized form of pulmonary hypertension in canines. Persistent pulmonary hypertension is a life-threatening condition in neonatal foals. Treatment of pulmonary hypertension in both canines and neonatal foals has focused on the correction of hypoxia and acidosis, not on the pulmonary hypertension itself. Identification of pulmonary hypertension in canines and foals has been made easier with the new diagnostic technologies that are being employed by veterinarians. Although retrospective reviews and studies have been published on the use of sildenafil, which is used primarily for the treatment of human erectile dysfunction, to treat pulmonary hypertension in human neonates, and the pharmacodynamics of sildenafil have been examined in various species, sildenafil has not been used widely in dogs or foals. Therefore, the adverse effects of sildenafil in canines and foals have not been determined. Until the stability of sildenafil in suspension has been determined, it is recommended that dose-specific capsules be prepared by compounding pharmacists utilizing lactose as a diluent and that veterinarians be encouraged to prescribe small quantities of capsules until a maintenance dose has been achieved.

Feline herpetic keratitis (infectious keratoconjunctivitis), an infection of the conjunctiva and cornea that is common in cats, involves a three-pronged treatment approach: (1)antiviral therapy, (2)immunomodulatory therapy, and (3)antibacterial therapy. Since very few agents have been approved for the treatment of feline herpetic keratitis, and many of the drugs that were approved at one time for this indication can no longer be purchased, compounding pharmacists can play a vital role in the prevention and treatment of this infection. Investigators at the Colorado State University College of Veterinary Medicine and Biomedical Science evaluated the effect of a new antiviral agent, cidofovir, for ophthalmic use in treating feline herpetic keratitis. Researchers concluded that application of cidofovir significatnly decreased the clinical signs of acute ocular infection in cats with feline herpesvirus type 1, one of the primary causes of feline herpetic keratitis. Working with veterinary ophthamologists, compounding pharmacists can develop other treatment options in patient-speciic dosages and dosage forms to ensure that treated cats have an excellent response to therapy.

To determine whether gene expression profiling could improve risk classification and outcome prediction in older acute myeloid leukemia (AML) patients, expression profiles were obtained in pretreatment leukemic samples from 170 patients whose median age was 65 years. Unsupervised clustering methods were used to classify patients into 6 cluster groups (designated A to F) that varied significantly in rates of resistant disease (RD; P < .001), complete response (CR; P = .023), and disease-free survival (DFS; P = .023). Cluster A (n = 24), dominated by NPM1 mutations (78%), normal karyotypes (75%), and genes associated with signaling and apoptosis, had the best DFS (27%) and overall survival (OS; 25% at 5 years). Patients in clusters B (n = 22) and C (n = 31) had the worst OS (5% and 6%, respectively); cluster B was distinguished by the highest rate of RD (77%) and multidrug resistant gene expression (ABCG2, MDR1). Cluster D was characterized by a "proliferative" gene signature with the highest proportion of detectable cytogenetic abnormalities (76%; including 83% of all favorable and 34% of unfavorable karyotypes). Cluster F (n = 33) was dominated by monocytic leukemias (97% of cases), also showing increased NPM1 mutations (61%). These gene expression signatures provide insights into novel groups of AML not predicted by traditional studies that impact prognosis and potential therapy.

Here we describe an automated, pressure-driven, sampling device for harvesting 10 to 30 ml samples, in replicate, with intervals as short as 10 s. Correlation between biological replicate time courses measured by microarrays was extremely high. The sampler enables sampling at intervals within the range of many important biological processes.

The U.S. Department of Energy recently announced the first five grants for the Genomes to Life (GTL) Program. The goal of this program is to "achieve the most far-reaching of all biological goals: a fundamental, comprehensive, and systematic understanding of life." While more information about the program can be found at the GTL website (www.doegenomestolife.org), this paper provides an overview of one of the five GTL projects funded, "Carbon Sequestration in Synechococcus Sp.: From Molecular Machines to Hierarchical Modeling." This project is a combined experimental and computational effort emphasizing developing, prototyping, and applying new computational tools and methods to elucidate the biochemical mechanisms of the carbon sequestration of Synechococcus Sp., an abundant marine cyanobacteria known to play an important role in the global carbon cycle. Understanding, predicting, and perhaps manipulating carbon fixation in the oceans has long been a major focus of biological oceanography and has more recently been of interest to a broader audience of scientists and policy makers. It is clear that the oceanic sinks and sources of CO(2) are important terms in the global environmental response to anthropogenic atmospheric inputs of CO(2) and that oceanic microorganisms play a key role in this response. However, the relationship between this global phenomenon and the biochemical mechanisms of carbon fixation in these microorganisms is poorly understood. The project includes five subprojects: an experimental investigation, three computational biology efforts, and a fifth which deals with addressing computational infrastructure challenges of relevance to this project and the Genomes to Life program as a whole. Our experimental effort is designed to provide biology and data to drive the computational efforts and includes significant investment in developing new experimental methods for uncovering protein partners, characterizing protein complexes, identifying new binding domains. We will also develop and apply new data measurement and statistical methods for analyzing microarray experiments. Our computational efforts include coupling molecular simulation methods with knowledge discovery from diverse biological data sets for high-throughput discovery and characterization of protein-protein complexes and developing a set of novel capabilities for inference of regulatory pathways in microbial genomes across multiple sources of information through the integration of computational and experimental technologies. These capabilities will be applied to Synechococcus regulatory pathways to characterize their interaction map and identify component proteins in these pathways. We will also investigate methods for combining experimental and computational results with visualization and natural language tools to accelerate discovery of regulatory pathways. Furthermore, given that the ultimate goal of this effort is to develop a systems-level of understanding of how the Synechococcus genome affects carbon fixation at the global scale, we will develop and apply a set of tools for capturing the carbon fixation behavior of complex of Synechococcus at different levels of resolution. Finally, because the explosion of data being produced by high-throughput experiments requires data analysis and models which are more computationally complex, more heterogeneous, and require coupling to ever increasing amounts of experimentally obtained data in varying formats, we have also established a companion computational infrastructure to support this effort as well as the Genomes to Life program as a whole.

We have assembled data from Caenorhabditis elegans DNA microarray experiments involving many growth conditions, developmental stages, and varieties of mutants. Co-regulated genes were grouped together and visualized in a three-dimensional expression map that displays correlations of gene expression profiles as distances in two dimensions and gene density in the third dimension. The gene expression map can be used as a gene discovery tool to identify genes that are co-regulated with known sets of genes (such as heat shock, growth control genes, germ line genes, and so forth) or to uncover previously unknown genetic functions (such as genomic instability in males and sperm caused by specific transposons).

Swollen, intensely eosinophilic glial cells are often observed in the vicinity of hemorrhagic lesions in post-mortem human brains. We sought to determine the nature of this change. Thirty adult human brains removed at autopsy and three surgical specimens were obtained 6h to 60 days following a hemorrhagic event. They were subjected to a battery of histochemical and immunohistochemical stains. The swollen cells, which were observed in the majority of autopsy specimens in which hemorrhage had occurred within the previous 9 days, stained intensely red with Masson stain and were immunoreactive for IgG, IgM, IgA, and fibrinogen. Some were also immunoreactive for glial fibrillary acidic protein, particularly in subpial and subependymal areas, if the lesions were more than 3 days old. In white matter some of the cells were immunoreactive for CNPase. There was no labeling with markers for macrophage/microglial cells. The absence of DNA fragment detection by TUNEL suggests that the cells were not dying. Mild glial cytoplasmic eosinophilia without swelling was observed in surgical specimens. No eosinophilic swollen glia were seen in perfusion-fixed rat brains with experimental intracerebral hemorrhage, although they were seen in rat brains that were not promptly fixed. We conclude that human macroglia, including astrocytes and oligodendrocytes, ingest plasma proteins that have been released into brain parenchyma. This likely represents a homeostatic mechanism that maintains the composition of the extracellular environment. If the tissue is not promptly fixed the cells become more swollen and eosinophilic.

This study shows that the incorporation of radiolabelled UTP into RNA in Percoll-gadient-purified potato mitochondria is regulated by the redox state of the mitochondrial electron transport chain. An early indication that there might be a redox effect on RNA synthesis was a decrease in UTP incorporation in incubates containing an oxidisable substrate, such as succinate or malate. Subsequent use of a variety of electron transport inhibitors acting at different points in the electron transport chain established that the redox state of the Rieske iron-sulphur protein was the major determinant of UTP incorporation. Inhibitors acting on the substrate side of the Rieske iron-sulphur protein, and causing oxidation of components on the oxygen side of their site of action, increased UTP incorporation into RNA. These included antimycin A, myxothiazole, and undecylhydroxydioxobenzothiazole at 500 nM. Inhibitors acting on the oxygen side of the Rieske iron-sulphur protein, and causing a reduction of components on the substrate side of the block, decreased UTP incorporation. These inhibitors were undecylhydroxydioxobenzothiazole at 25 nM and KCN. When phenazine methosulphate was present as an auto-oxidisable electron sink the effect of KCN was diminished. The conclusion from the inhibitor experiments that the redox state of the Rieske iron-suphur protein was important was supported when RNA synthesis was measured at a range of redox potentials. This gave a measured redox potential for the control of UTP incorporation into RNA of +270 mV and the slope of the curve indicated an n = 1 carrier. This value is close to the reported value of the Rieske iron-suphur protein. UTP incorporation was decreased by some 50% in the presence of low concentrations of okadaic acid (5 nM), an inhibitor of protein phosphatases PP1 and PP2A, and alpha-naphthyl acid phosphate, a broad-spectrum phosphatase inhibitor, indicating that the redox effect on RNA synthesis may be mediated via protein phosphorylation. We did not, however, detect an expected increase in RNA synthesis when protein kinase inhibitors were used, so the involvement of protein phosphorylation in the redox regulation of RNA synthesis is as yet uncertain.

A large facial vascular malformation was embolized with polyvinyl alcohol particles twice in 8 years. Resected tissue enabled long-term examination of this material, confirming its chemical inertness and revealing minimal tissue reaction to it apart from calcification. No particle migration, fragmentation, or absorption occurred. There was some recanalization of occluded vessels. Most vessels containing polyvinyl alcohol particles, and all of the larger vessels, were incompletely occluded, with particles becoming embedded in their walls.

A patient with dermatomyositis associated with carcinoma was treated with steroids and antibiotics for possible tuberculosis. Autopsy showed an overwhelming diffuse nongranulomatous infection of Mycobacterium tuberculosis involving only the skeletal muscles and one inguinal lymph node. The rare localization of tuberculosis to skeletal muscle in this case is possibly due to steroid immunosuppression and the humoral immune attack on muscle blood vessels that is a part of dermatomyositis.