Dr. Trepanier is a Professor of Medicine at the University of Wisconsin School of Veterinary Medicine. She is Board Certified by the American College of Veterinary Internal Medicine and the American College of Veterinary Clinical Pharmacology. Her research interests include idiosyncratic drug toxicity, pharmacogenetics, and comparative drug biotransformation among species.
Biochemical, functional, and histopathologic characterization of lomustine-induced liver injury in dogs.
Am J Vet Res. 2020 Oct;81(10):810-820
Authors: Dedeaux AM, Flesner BK, Reinhart JM, Langohr IM, Husnik R, Geraci SN, Taboada J, Rademacher N, Thombs LA, Bryan JN, Trepanier LA, Boudreaux BB
OBJECTIVE: To characterize the biochemical, functional, and histopathologic changes associated with lomustine-induced liver injury in dogs.
ANIMALS: I0 healthy purpose-bred sexually intact female hounds.
PROCEDURES: Dogs were randomly assigned to receive lomustine (approx 75 mg/m2, PO, q 21 d for 5 doses) alone (n = 5) or with prednisone (approx 1.5 mg/kg, PO, q 24 h for 12 weeks; 5). For each dog, a CBC, serum biochemical analysis, liver function testing, urinalysis, and ultrasonographic examination of the liver with acquisition of liver biopsy specimens were performed before and at predetermined times during and after lomustine administration. Results were compared between dogs that did and did not receive prednisone.
RESULTS: 7 of the I0 dogs developed clinical signs of liver failure. For all dogs, serum alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities, bile acid concentrations, and liver histologic score increased and hepatic reduced glutathione content decreased over time. Peak serum ALT (r = 0.79) and ALP (r = 0.90) activities and bile acid concentration (r = 0.68) were positively correlated with the final histologic score. Prednisone did not appear to have a protective effect on histologic score.
CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, liver enzyme activities, particularly ALT and ALP activities, should be closely monitored during lomustine treatment and acute increases in those activities may warrant discontinuation of lomustine to mitigate liver injury. Nonspecific ultrasonographic findings and abnormal increases in liver function tests were not detected until the onset of clinical liver failure. Glutathione depletion may have a role in lomustine-induced hepatopathy and warrants further investigation.
PMID: 32969725 [PubMed - as supplied by publisher]
Glutathione S-transferase theta genotypes and environmental exposures in the risk of canine transitional cell carcinoma.
J Vet Intern Med. 2019 Apr 22;:
Authors: Luethcke KR, Ekena J, Chun R, Trepanier LA
INTRODUCTION: Transitional cell carcinoma (TCC) in humans is associated with environmental exposures and variants in glutathione S-transferase (GST) genes. Scottish Terriers have a high breed risk for TCC, but the relationship between genetic and environmental risk in dogs is not fully understood.
HYPOTHESES: Scottish Terriers have a higher frequency of GST-theta variants compared to lower risk breeds. Dogs with TCC of any breed have a higher frequency of GST-theta variants along with higher environmental exposures, compared to controls.
ANIMALS: One hundred and five Scottish Terriers and 68 controls from lower risk breeds; 69 dogs of various breeds with TCC, and 72 breed- and sex-matched unaffected geriatric dogs.
METHODS: In this prospective case-control study, dogs were genotyped for 3 canine GST-theta variants: GSTT1 I2+28 G>A, a GSTT1 3'UTR haplotype, and GSTT5 Asp129_Gln130del. Owners of dogs with TCC and unaffected geriatric controls completed a household environmental questionnaire.
RESULTS: The GSTT1 3'UTR haplotype and GSTT5 Asp129_Gln130del variants were significantly underrepresented in Scottish Terriers (minor allele frequency [MAF] = 0.000 for both), compared to dogs from lower risk breeds (MAF = 0.108 and 0.100; P ≤ .0002). Dogs with TCC did not differ from unaffected geriatric controls across the 3 investigated loci. Transitional cell carcinoma was associated with household insecticide use (odds ratio [OR] = 4.28, 95% confidence interval [CI] = 1.44-12.33, P = .02), and was negatively associated with proximity to a farm (OR = 0.49, 95% CI = 0.25-0.99, P = .04).
CONCLUSIONS AND CLINICAL IMPORTANCE: Low-activity GST-theta loci are unlikely contributors to TCC risk in dogs. Increased risk is associated with household insecticide use, and possibly with less rural households.
PMID: 31008543 [PubMed - as supplied by publisher]
Prospective crossover clinical trial comparing transdermal with oral phenobarbital administration in epileptic cats.
J Feline Med Surg. 2019 Jan 28;:1098612X18823577
Authors: Barnes Heller HL, Trepanier LA, Robertson M, Mei C
OBJECTIVES: The aim of this study was to compare serum phenobarbital concentrations, adverse events and client satisfaction during 14 weeks of transdermal vs oral phenobarbital administration to epileptic cats.
METHODS: This was a prospective, fixed-order, crossover pilot study. Nine client-owned cats with presumptive or diagnosed idiopathic epilepsy were enrolled. Oral phenobarbital (PO-PB) was administered for weeks 1-14 (median starting dosage of 3.8 mg/kg [2.0-5.4 mg/kg/day] q12h); transdermal phenobarbital (TD-PB) was administered for weeks 14-28 (median starting dosage 18.8 mg/kg/day [17.6-24.0 mg/kg/day] q12h). Serum phenobarbital concentrations (S-PB) were measured at weeks 2, 14, 16 and 28. Client satisfaction questionnaires and biochemistry were evaluated at 14 and 28 weeks.
RESULTS: Median S-PB concentrations during oral administration were 21 µg/ml (observed range 11-40 µg/ml) at week 2 and 22 µg/ml (8-35 µg/ml) at week 14, and at the higher TD dosage were 18 µg/ml (0-42 µg/ml) at week 16 and 17 µg/ml (7-50 µg/ml) at week 28. Phenobarbital concentrations were significantly correlated with PO dosage at week 2 ( r = 0.75, P = 0.03) but not at weeks 16 and 28. Significantly more dose adjustments were needed during the TD phase, ( P = 0.03) but 6/9 owners (67%) still preferred TD to PO administration. Adverse effects were mild and comparable in both groups.
CONCLUSIONS AND RELEVANCE: Therapeutic S-PB concentrations were achievable in some cats using TD-PB at 18 mg/kg/day q12h. Poor correlation between TD dosage and S-PB concentrations was observed and more dosage adjustments were required during TD administration. These findings necessitate close therapeutic drug monitoring if TD-PB is prescribed.
PMID: 30688552 [PubMed - as supplied by publisher]
RNA expression profiling in sulfamethoxazole-treated patients with a range of in vitro lymphocyte cytotoxicity phenotypes.
Pharmacol Res Perspect. 2018 Apr;6(2):e00388
Authors: Reinhart JM, Rose W, Panyard DJ, Newton MA, Liebenstein TK, Yee J, Trepanier LA
The lymphocyte toxicity assay (LTA) is a proposed surrogate marker of sulfonamide antibiotic hypersensitivity. In the LTA, peripheral blood mononuclear cells (PBMCs) undergo apoptosis more readily in hypersensitive versus tolerant patients when exposed to drug-hydroxylamine metabolites in vitro. The purpose of this study was to identify key gene transcripts associated with increased cytotoxicity from sulfamethoxazole-hydroxylamine in human PBMCs in the LTA. The LTA was performed on PBMCs of 10 patients hypersensitive to trimethoprim-sulfamethoxazole (HS) and 10 drug-tolerant controls (TOL), using two cytotoxicity assays: YO-PRO (n = 20) and MTT (n = 12). mRNA expression profiles of PBMCs, enriched for CD8+ T cells, were compared between HS and TOL patients. Transcript expression was interrogated for correlation with % cytotoxicity from YO-PRO and MTT assays. Correlated transcripts of interest were validated by qPCR. LTA results were not significantly different between HS and TOL patients, and no transcripts were found to be differentially expressed between the two groups. 96 transcripts were correlated with cytotoxicity by YO-PRO (r = ±.63-.75, FDR 0.188). Transcripts were selected for validation based on mechanistic plausibility and three were significantly over-expressed by qPCR in high cytotoxicity patients: multi-specific organic anion transporter C (ABCC5), mitoferrin-1 (SLC25A37), and Porimin (TMEM123). These data identify novel transcripts that could contribute to sulfonamide-hydroxylamine induced cytotoxicity. These include SLC25A37, encoding a mitochondrial iron transporter, ABCC5, encoding an arylamine drug transporter, and TMEM123, encoding a transmembrane protein that mediates cell death.
PMID: 29511567 [PubMed - in process]
Evaluation of potential serum biomarkers of hepatic fibrosis and necroinflammatory activity in dogs with liver disease.
J Vet Intern Med. 2018 Feb 27;:
Authors: Raghu C, Ekena J, Cullen JM, Webb CB, Trepanier LA
BACKGROUND: Serum interleukin 6 (IL-6), chemokine ligand 2 (CCL2), C-reactive protein (CRP), and the ratio of aspartate transaminase to alanine transaminase (AST:ALT) have been correlated with fibrosis and necroinflammatory activity in humans with various hepatopathies.
HYPOTHESIS/OBJECTIVES: To determine whether increases in serum IL-6, CCL2, CRP, or AST:ALT were associated with moderate to severe fibrosis or necroinflammatory activity in dogs with various hepatopathies.
ANIMALS: Forty-four client-owned dogs with clinical evidence of liver disease and 10 healthy purpose-bred dogs, all undergoing liver biopsies by laparoscopy or laparotomy.
METHODS: Measurement of serum IL-6, CCL2, CRP, AST, and ALT before scheduled liver biopsy and evaluation of liver histopathology using the METAVIR scoring system used in human medicine, blinded to clinical presentation.
RESULTS: Median serum IL-6 was approximately twice as high in dogs with high fibrosis scores (15.5 pg/mL; range, 1.4 to 235 pg/mL) compared to dogs with low fibrosis scores (7.6 pg/mL; range, 1.4 to 148.1 pg/mL), with marginal significance (P = .05). Median serum CCL2 was significantly higher in dogs with active necroinflammation (444 pg/mL; range, 144 to 896 pg/mL) compared to dogs without detectable necroinflammation (326 pg/mL; range, 59 to 1692 pg/mL; P = .008), but with considerable overlap between groups. Neither serum CRP nor AST:ALT ratios were significantly different based on fibrosis or necroinflammatory scores.
CONCLUSIONS AND CLINICAL IMPORTANCE: Because of substantial variability among dogs, single measurements of IL-6 and CCL2 have limited diagnostic utility for identifying fibrosis or necroinflammation, respectively, in dogs with various chronic liver diseases. The value of these biomarkers should be explored further in monitoring response to treatment in individual dogs with chronic hepatopathies.
PMID: 29485210 [PubMed - as supplied by publisher]
Research Directions in Genetic Predispositions to Stevens-Johnson Syndrome / Toxic Epidermal Necrolysis.
Clin Pharmacol Ther. 2017 Nov 06;:
Authors: Manolio TA, Hutter CM, Avigan M, Cibotti R, Davis RL, Denny JC, Grenade L, Wheatley LM, Carrington MN, Chantratita W, Chung WH, Dalton AD, Hung SI, Lee MTM, Leeder JS, Lertora JJL, Mahasirimongkol S, McLeod HL, Mockenhaupt M, Pacanowski M, Phillips EJ, Pinheiro S, Pirmohamed M, Sung C, Suwankesawong W, Trepanier L, Tumminia SJ, Veenstra D, Yuliwulandari R, Shear NH
Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) is one of the most devastating of adverse drug reactions (ADRs) and was, until recently, essentially unpredictable. With the discovery of several risk alleles for drug-induced SJS/TEN and the demonstration of effectiveness of screening in reducing incidence, the stage is set for implementation of preventive strategies in populations at risk. Yet much remains to be learned about this potentially fatal complication of commonly used drugs.
PMID: 29105735 [PubMed - as supplied by publisher]
Immunogenicity of trimethoprim/sulfamethoxazole in a macaque model of HIV infection.
Toxicology. 2016 Aug 10;368-369:10-18
Authors: Wong YY, Rakasz EG, Gasper DJ, Friedrich TC, Trepanier LA
BACKGROUND: Sulfonamide hypersensitivity has a high incidence in HIV infection and correlates with low CD4+ counts, but the mechanisms are not understood. The aims of this study were to determine whether trimethoprim/sulfamethoxazole (TMP/SMX) led to SMX adduct formation, immunogenicity, or signs of drug hypersensitivity in SIV-infected rhesus macaques, and whether differences in antioxidants, pro-inflammatory mediators, or SMX disposition were predictive of drug immunogenicity.
METHODS: Nine macaques chronically infected with SIVmac239 and 7 non-infected controls were studied. Baseline blood ascorbate, glutathione, IFN-γ, LPS, sCD14, and cytochrome b5 reductase measurements were obtained, macaques were dosed with TMP/SMX (120mg/kg/day p.o. for 14days), and SMX metabolites, lymph node drug adducts, drug-responsive T cells, and anti-SMX antibodies were measured.
RESULTS: Four of 9 of SIV-positive (44%), and 3 of 7 SIV negative (43%) macaques had drug-responsive T cells or antibodies to SMX. Two macaques developed facial or truncal rash; these animals had the highest levels of lymph node drug adducts. Antioxidants, pro-inflammatory mediators, and SMX metabolites were not predictive of drug immunogenicity; however, the Mamu DRB1*0401/0406/0411 genotype was significantly over-represented in immune responders.
CONCLUSIONS: Unlike other animal models, macaques develop an immune response, and possible rash, in response to therapeutic dosages of TMP/SMX. Studying more animals with CD4+ counts <200cells/μl, along with moderately restricted ascorbate intake to match deficiencies seen in humans, may better model the risk of SMX hypersensitivity in HIV-infection. In addition, the role of Mamu-DRB1 genotype in modeling drug hypersensitivity in retroviral infection deserves further study.
PMID: 27565715 [PubMed - in process]
Genome-Wide Association Study in Immunocompetent Patients with Delayed Hypersensitivity to Sulfonamide Antimicrobials.
PLoS One. 2016;11(6):e0156000
Authors: Reinhart JM, Motsinger-Reif A, Dickey A, Yale S, Trepanier LA
BACKGROUND: Hypersensitivity (HS) reactions to sulfonamide antibiotics occur uncommonly, but with potentially severe clinical manifestations. A familial predisposition to sulfonamide HS is suspected, but robust predictive genetic risk factors have yet to be identified. Strongly linked genetic polymorphisms have been used clinically as screening tests for other HS reactions prior to administration of high-risk drugs.
OBJECTIVE: The purpose of this study was to evaluate for genetic risk of sulfonamide HS in the immunocompetent population using genome-wide association.
METHODS: Ninety-one patients with symptoms after trimethoprim-sulfamethoxazole (TMP-SMX) attributable to "probable" drug HS based on medical record review and the Naranjo Adverse Drug Reaction Probability Scale, and 184 age- and sex-matched patients who tolerated a therapeutic course of TMP-SMX, were included in a genome-wide association study using both common and rare variant techniques. Additionally, two subgroups of HS patients with a more refined clinical phenotype (fever and rash; or fever, rash and eosinophilia) were evaluated separately.
RESULTS: For the full dataset, no single nucleotide polymorphisms were suggestive of or reached genome-wide significance in the common variant analysis, nor was any genetic locus significant in the rare variant analysis. A single, possible gene locus association (COL12A1) was identified in the rare variant analysis for patients with both fever and rash, but the sample size was very small in this subgroup (n = 16), and this may be a false positive finding. No other significant associations were found for the subgroups.
CONCLUSIONS: No convincing genetic risk factors for sulfonamide HS were identified in this population. These negative findings may be due to challenges in accurately confirming the phenotype in exanthematous drug eruptions, or to unidentified gene-environment interactions influencing sulfonamide HS.
PMID: 27272151 [PubMed - in process]
Therapeutic serum phenobarbital concentrations obtained using chronic transdermal administration of phenobarbital in healthy cats.
J Feline Med Surg. 2015 Apr;17(4):359-63
Authors: Delamaide Gasper JA, Barnes Heller HL, Robertson M, Trepanier LA
Seizures are a common cause of neurologic disease, and phenobarbital (PB) is the most commonly used antiepileptic drug. Chronic oral dosing can be challenging for cat owners, leading to poor compliance. The purpose of this study was to determine if the transdermal administration of PB could achieve serum PB concentrations of between 15 and 45 μg/ml in healthy cats. Nineteen healthy cats were enrolled in three groups. Transdermal PB in pluronic lecithin organogel (PLO) was applied to the pinnae for 14 days at a dosage of 3 mg/kg q12h in group 1 (n = 6 cats) and 9 mg/kg q12h in group 2 (n = 7 cats). Transdermal PB in Lipoderm Activemax was similarly applied at 9 mg/kg q12h for 14 days in group 3 (n = 6 cats). Steady-state serum PB concentrations were measured at trough, and at 2, 4 and 6 h after the morning dose on day 15. In group 1, median concentrations ranged from 6.0-7.5 μg/ml throughout the day (observed range 0-11 μg/ml). Group 2 median concentrations were 26.0 μg/ml (observed range 18.0-37.0 μg/ml). For group 3, median concentrations ranged from 15.0-17.0 μg/ml throughout the day (range 5-29 μg/ml). Side effects were mild. One cat was withdrawn from group 2 owing to ataxia and sedation. These results show therapeutic serum PB concentrations can be achieved in cats following chronic transdermal administration of PB in PLO at a dosage of 9 mg/kg q12h. More individual variation was noted using Lipoderm Activemax. Transdermal administration may be an alternative for cats that are difficult to medicate orally.
PMID: 25098448 [PubMed - in process]
Polymorphisms in the carcinogen detoxification genes CYB5A and CYB5R3 and breast cancer risk in African American women.
Cancer Causes Control. 2014 Nov;25(11):1513-21
Authors: Blanke KL, Sacco JC, Millikan RC, Olshan AF, Luo J, Trepanier LA
PURPOSE: Cytochrome b 5 (encoded by CYB5A) and NADH cytochrome b 5 reductase (encoded by CYB5R3) detoxify aromatic and heterocyclic amine mammary carcinogens found in cigarette smoke. We hypothesized that CYB5A and CYB5R3 polymorphisms would be associated with breast cancer risk in women.
METHODS: We characterized the prevalence of 18 CYB5A and CYB5R3 variants in genomic DNA from African American (AfrAm) and Caucasian (Cauc) women from the Carolina Breast Cancer Study population (1,946 cases and 1,747 controls) and determined their associations with breast cancer risk, with effect modification by smoking.
RESULTS: A CYB5R3 variant, I1M+6T (rs8190370), was significantly more common in breast cancer cases (MAF 0.0238) compared with controls (0.0169, p = 0.039); this was attributable to a higher MAF in AfrAm cases (0.0611) compared with AfrAm controls (0.0441, p = 0.046; adjusted OR 1.41, CI 0.98-2.04; p = 0.062). When smoking was considered, I1M+6T was more strongly associated with breast cancer risk in AfrAm smokers (adjusted OR 2.10, 1.08-4.07; p = 0.028) compared with never smokers (OR = 1.21; 0.77-1.88; p for interaction = 0.176). I1M+6T and three additional CYB5R3 variants, -251T, I8-1676C, and *392C, as well as two CYB5A variants, 13G and I2-992T, were significantly more common in AfrAms compared with Caucs.
CONCLUSIONS: CYB5R3 I1M+6C>T should be considered in future molecular epidemiologic studies of breast cancer risk in AfrAms. Further, variants in CYB5A and CYB5R3 should be considered in the evaluation of other tumors in AfrAms that are associated with aromatic and heterocyclic amine exposures, to include prostate, bladder, and colon cancers.
PMID: 25225034 [PubMed - in process]
Cutaneous pythiosis in two dogs from Wisconsin, USA.
Vet Dermatol. 2014 Feb;25(1):52-e21
Authors: Oldenhoff W, Grooters A, Pinkerton ME, Knorr J, Trepanier L
BACKGROUND: Pythium insidiosum is an oomycete that causes cutaneous lesions or infiltrative gastrointestinal disease in dogs, cats, humans, horses and other mammals, primarily in tropical and subtropical climates.
HYPOTHESIS/OBJECTIVES: We report the clinicopathological findings associated with cutaneous pythiosis in two dogs from a Northern temperate climate zone.
ANIMALS: A 3-year-old intact male Chesapeake Bay retriever was presented with an ulcerated soft-tissue swelling over the left eye. A 4-year-old spayed female German shepherd dog was presented with a soft-tissue swelling overlying the right hock. Both dogs lived in northern latitudes (between 43 and 45°N) and neither had travelled outside of Wisconsin or Michigan's upper peninsula, USA.
METHODS: Histopathological examination and culture of affected tissues on specialized media, serology for anti-P. insidiosum antibodies, P. insidiosum-specific PCR and ribosomal RNA gene sequencing were carried out.
RESULTS: Histopathological examination revealed pyogranulomatous and eosinophilic inflammation associated with wide, poorly septate hyphae.
CONCLUSIONS AND CLINICAL IMPORTANCE: Even clinicians who practice in temperate climates should consider pythiosis as a differential diagnosis for young to middle-aged adult dogs presented with ulcerated cutaneous nodules or infiltrative gastrointestinal disease.
PMID: 24372864 [PubMed - indexed for MEDLINE]
Idiosyncratic drug toxicity affecting the liver, skin, and bone marrow in dogs and cats.
Vet Clin North Am Small Anim Pract. 2013 Sep;43(5):1055-66
Authors: Trepanier LA
Idiosyncratic drug toxicity reactions are, by definition, uncommon, but can lead to serious or even fatal organ toxicity. The liver, skin, and peripheral blood cells/bone marrow are common targets. Most of these reactions are the result of reactive metabolites, which may cause local cell or organelle damage, or may be amplified by a systemic immune response. Individual risk may depend on differences in drug biotransformation, levels of oxidative stress, or antigen presentation.
PMID: 23890238 [PubMed - indexed for MEDLINE]
Applying pharmacokinetics to veterinary clinical practice.
Vet Clin North Am Small Anim Pract. 2013 Sep;43(5):1013-26
Authors: Trepanier LA
This article describes clinical examples in which pharmacokinetic parameters can be used to optimize veterinary patient care. Specific applications include extrapolating drug dosages, optimizing therapy with therapeutic drug monitoring, interpreting pharmacokinetic information provided by drug labels and pharmaceutical companies, and adjusting drug dosages in patients with hepatic or renal failure.
PMID: 23890235 [PubMed - indexed for MEDLINE]
Challenges in exploring the cytochrome P450 system as a source of variation in canine drug pharmacokinetics.
Drug Metab Rev. 2013 May;45(2):218-30
Authors: Martinez MN, Antonovic L, Court M, Dacasto M, Fink-Gremmels J, Kukanich B, Locuson C, Mealey K, Myers MJ, Trepanier L
The cytochrome P450 (CYP) superfamily constitutes a collection of enzymes responsible for the metabolism of a wide array of endo- and xenobiotic compounds. Much of the knowledge on substrate specificity and genetic identification of the various CYP isoforms is derived from research in rodents and humans and only limited information has been captured in the dog. Currently, there exist many gaps in our knowledge of canine CYP diversity as a result of the paucity of studies focusing on canine CYPs, canine CYP polymorphisms, and the therapeutic consequences of these genetic variants. Challenges engendered by this lack of information is further amplified by inter- and intraspecies differences in the specificity and affinity of substrates and inhibitors, prohibiting a simple extrapolation of probe substances used in human CYP research. This creates a need to develop and validate canine-specific CYP probes. Failure to understand this potential metabolic and pharmacogenomic diversity can also influence the interpretation of data generated in dogs to support human drug development. It is with these objectives in mind that we provide an overview of what is currently known about canine CYPs with the hope that it will encourage further exploration into this important area of research.
PMID: 23432217 [PubMed - indexed for MEDLINE]
Evaluation of polymorphisms in the sulfonamide detoxification genes NAT2, CYB5A, and CYB5R3 in patients with sulfonamide hypersensitivity.
Pharmacogenet Genomics. 2012 Oct;22(10):733-40
Authors: Sacco JC, Abouraya M, Motsinger-Reif A, Yale SH, McCarty CA, Trepanier LA
OBJECTIVE: To determine whether polymorphisms in the sulfonamide detoxification genes, CYB5A (encoding cytochrome b(5)), CYB5R3 (encoding cytochrome b(5) reductase), or NAT2 (encoding N-acetyltransferase 2) were over-represented in patients with delayed sulfonamide drug hypersensitivity, compared with control patients who tolerated a therapeutic course of trimethoprim-sulfamethoxazole without adverse event.
METHODS: DNA from 99 nonimmunocompromised patients with sulfonamide hypersensitivity who were identified from the Personalized Medicine Research Project at the Marshfield Clinic, and from 99 age-matched, race-matched, and sex-matched drug-tolerant controls, were genotyped for four CYB5A and five CYB5R3 polymorphisms, and for all coding NAT2 SNPs.
RESULTS: CYB5A and CYB5R3 SNPs were found at low allele frequencies (<3-4%), which did not differ between hypersensitive and tolerant patients. NAT2 allele and haplotype frequencies, as well as inferred NAT2 phenotypes, also did not differ between groups (60 vs. 59% slow acetylators). Finally, no difference in NAT2 status was found in a subset of patients with more severe hypersensitivity signs (drug reaction with eosinophilia and systemic symptoms) compared with tolerant patients.
CONCLUSION: We found no evidence of a substantial involvement of these nine CYB5A or CYB5R3 polymorphisms in sulfonamide hypersensitivity risk, although minor effects cannot be completely ruled out. Despite careful medical record review and full resequencing of the NAT2 coding region, we found no association of NAT2 coding alleles with sulfonamide hypersensitivity (predominantly cutaneous eruptions) in this adult Caucasian population.
PMID: 22850190 [PubMed - indexed for MEDLINE]
Dapsone-associated methemoglobinemia in a patient with slow NAT2*5B haplotype and impaired cytochrome b5 reductase activity.
J Clin Pharmacol. 2012 Feb;52(2):272-8
Authors: Abouraya M, Sacco JC, Hayes K, Thomas S, Kitchens CS, Trepanier LA
PMID: 21422237 [PubMed - in process]
Cholinergic crisis after neostigmine administration in a dog with acquired focal myasthenia gravis.
J Vet Emerg Crit Care (San Antonio). 2011 Oct;21(5):547-51
Authors: Foy DS, Trepanier LA, Shelton GD
OBJECTIVE: To describe the presentation and successful management of a dog experiencing a cholinergic crisis after neostigmine administration.
CASE SUMMARY: An 18-month-old neutered male Maltese-crossbred dog was diagnosed with acquired focal myasthenia gravis based on history and clinical signs of dysphagia and regurgitation, multiple series of thoracic radiographs showing focal to generalized megaesophagus, and an increased acetylcholine receptor antibody titer. After this diagnosis, the dog was initially treated with a single oral dose of pyridostigmine and later injectable neostigmine due to difficulty swallowing. Within 15 minutes of receiving a single dose (0.05 mg/kg) of subcutaneous neostigmine, the dog began showing muscarinic cholinergic signs of salivation and defecation, which progressed to nicotinic cholinergic signs of weakness and tachypnea. Within 30 minutes the dog experienced respiratory arrest and required ventilation. After 16 hours of ventilation, the dog recovered uneventfully and subsequently achieved a clinical and serologic remission from myasthenia gravis without further treatment.
NEW OR UNIQUE INFORMATION PROVIDED: Cholinergic crisis and differentiation from a myasthenic crisis is described in the human literature. This case represents the first report in the veterinary literature of a cholinergic crisis in a dog treated with neostigmine.
PMID: 22316202 [PubMed - indexed for MEDLINE]
Individual variability in the detoxification of carcinogenic arylhydroxylamines in human breast.
Toxicol Sci. 2011 Jun;121(2):245-56
Authors: Rhoads K, Sacco JC, Drescher N, Wong A, Trepanier LA
Cytochrome b(5) (b5) and NADH cytochrome b(5) reductase (b5R) detoxify reactive hydroxylamine (NHOH) metabolites of known arylamine and heterocyclic amine mammary carcinogens. The aim of this study was to determine whether NHOH reduction for the prototypic arylamine 4-aminobiphenyl (4-ABP) was present in human breast and to determine whether variability in activity was associated with single nucleotide polymorphisms (SNPs) in the coding, promoter, and 3'untranslated region (UTR) regions of the genes encoding b5 (CYB5A) and b5R (CYB5R3). 4-ABP-NHOH reduction was readily detected in pooled human breast microsomes, with a K(m) (280μM) similar to that found with recombinant b5 and b5R, and a V(max) of 1.12 ± 0.19 nmol/min/mg protein 4-ABP-NHOH reduction varied 75-fold across 70 individual breast samples and correlated significantly with both b5 (80-fold variability) and b5R (14-fold) immunoreactive protein. In addition, wide variability in b5 protein expression was significantly associated with variability in CYB5A transcript levels, with a trend toward the same association between b5R and CYB5R3. Although a sample with a novel coding SNP in CYB5A, His22Arg, was found with low reduction and b5 expression, no other SNPs in either gene were associated with outlier activity or protein expression. We conclude that b5 and b5R catalyze the reduction of 4-ABP-NHOH in breast tissue, with very low activity, protein, and messenger RNA expression in some samples, which cannot be attributed to promoter, coding, or 3'UTR SNPs. Further studies are underway to characterize the transcriptional regulation of CYB5A and CYB5R3 and begin to understand the mechanisms of individual variability in this detoxification pathway.
PMID: 21447608 [PubMed - indexed for MEDLINE]
Evaluation of sulfonamide detoxification pathways in haematologic malignancy patients prior to intermittent trimethoprim-sulfamethoxazole prophylaxis.
Br J Clin Pharmacol. 2011 Apr;71(4):566-74
Authors: Abouraya M, Sacco JC, Kahl BS, Trepanier LA
AIMS: Patients with haematologic malignancies have a reportedly high incidence of sulfamethoxazole (SMX) hypersensitivity. The objective of this study was to determine whether deficiencies in sulfonamide detoxification pathways, to include glutathione (GSH) and ascorbate (AA), and cytochrome b(5) (b5) and cytochrome b(5) reductase (b5R), were prevalent in these patients. A secondary pilot objective was to determine whether the incidence of drug hypersensitivity following intermittent trimethoprim-SMX (TMP-SMX) prophylaxis approached that reported for high dose daily regimens.
METHODS: Forty adult patients with haematologic malignancies (HM) and 35 healthy adults were studied; an additional 13 HM patients taking ascorbate supplements (HM-AA) were also evaluated. Twenty-two of 40 HM patients were prescribed and were compliant with TMP-SMX 960 mg three to four times weekly.
RESULTS: There were no significant differences between HM and healthy groups in plasma AA (median 37.2 µm vs. 33.9 µm) or red blood cell GSH (1.9 mmvs. 1.8 mm). However, plasma AA was correlated significantly with leucocyte b5/b5R reduction (r= 0.39, P= 0.002). Deficient b5/b5R activities were not found in HM patients. In fact, patients with chronic lymphocytic leukaemia or myeloma had significantly higher median activities (80.7 µmol mg(-1) min(-1)) than controls (18.9 µmol mg(-1) min(-1), P= 0.008). After 3-4 weeks of treatment, no patients developed SMX-specific T cells and only one patient developed rash.
CONCLUSIONS: Deficiencies of blood antioxidants and b5/b5R reduction were not found in this population with haematologic malignancies, and the development of skin rash and drug-specific T cells appeared to be uncommon with intermittent TMP-SMX prophylaxis.
PMID: 21204907 [PubMed - indexed for MEDLINE]
Antifungal treatment of small animal veterinary patients.
Vet Clin North Am Small Anim Pract. 2010 Nov;40(6):1171-88
Authors: Foy DS, Trepanier LA
Antifungal therapy has progressed significantly with the development of new drugs directed at various processes in fungal cell metabolism. Within veterinary medicine, treatment options for systemic mycoses remain limited to amphotericin B, ketoconazole, fluconazole, and itraconazole. However, newer triazoles, echinocandins, and lipid-based formulations of amphotericin B are now approved for use in humans. This article provides a comprehensive review of the antifungal medications available for veterinary patients, and includes a brief discussion of the newer, presently cost-prohibitive, antifungal therapies used for systemic mycoses in humans.
PMID: 20933143 [PubMed - indexed for MEDLINE]
Combined ascorbate and glutathione deficiency leads to decreased cytochrome b5 expression and impaired reduction of sulfamethoxazole hydroxylamine.
Arch Toxicol. 2010 Aug;84(8):597-607
Authors: Bhusari S, Abouraya M, Padilla ML, Pinkerton ME, Drescher NJ, Sacco JC, Trepanier LA
Sulfonamide antimicrobials such as sulfamethoxazole (SMX) have been associated with drug hypersensitivity reactions, particularly in patients with AIDS. A reactive oxidative metabolite, sulfamethoxazole-nitroso (SMX-NO), forms drug-tissue adducts that elicit a T-cell response. Antioxidants such as ascorbic acid (AA) and glutathione (GSH) reduce SMX-NO to the less reactive hydroxylamine metabolite (SMX-HA), which is further reduced to the non-immunogenic parent compound by cytochrome b (5) (b5) and its reductase (b5R). We hypothesized that deficiencies in AA and GSH would enhance drug-tissue adduct formation and immunogenicity toward SMX-NO and that these antioxidant deficiencies might also impair the activity of the b5/b5R pathway. We tested these hypotheses in guinea pigs fed either a normal or AA-restricted diet, followed by buthionine sulfoximine treatment (250 mg/kg SC daily, or vehicle); and SMX-NO (1 mg/kg IP 4 days per week, or vehicle), for 2 weeks. Guinea pigs did not show any biochemical or histopathologic evidence of SMX-NO-related toxicity. Combined AA and GSH deficiency in this model did not significantly increase tissue-drug adduct formation, or splenocyte proliferation in response to SMX-NO. However, combined antioxidant deficiency was associated with decreased mRNA and protein expression of cytochrome b (5), as well as significant decreases in SMX-HA reduction in SMX-NO-treated pigs. These results suggest that SMX-HA detoxification may be down-regulated in combined AA and GSH deficiency. This mechanism could contribute to the higher risk of SMX hypersensitivity in patients with AIDS with antioxidant depletion.
PMID: 20221587 [PubMed - indexed for MEDLINE]
Acute vomiting in cats: rational treatment selection.
J Feline Med Surg. 2010 Mar;12(3):225-30
Authors: Trepanier L
PRACTICAL RELEVANCE: The control of nausea and vomiting in cats is important in order to prevent the development of food aversion, anorexia (with its associated complications of weight loss and dehydration), and hepatic lipidosis.
CLINICAL CHALLENGES: There are several antiemetic drugs that are clinically effective in cats. Making a rational choice from the available options requires knowledge of the likely cause of the vomiting, and the mechanisms of action and side effects of each drug. For example, a drug such as prochlorperazine, which can cause sedation, may be a useful first-line choice in a hospitalized cat that requires mild sedation to be handled, but would be undesirable in a critically ill cat.
AUDIENCE: For companion animal and feline practitioners, the vomiting cat is a common presentation.
EVIDENCE BASE: The guidance provided in this review draws on the findings of clinical trials in humans, experimental studies in cats, some clinical trials in cats, and clinical experience.
PMID: 20193913 [PubMed - indexed for MEDLINE]
Cytochrome b5 and NADH cytochrome b5 reductase: genotype-phenotype correlations for hydroxylamine reduction.
Pharmacogenet Genomics. 2010 Jan;20(1):26-37
Authors: Sacco JC, Trepanier LA
OBJECTIVES: NADH cytochrome b5 reductase (b5R) and cytochrome b5 (b5) catalyze the reduction of sulfamethoxazole hydroxylamine (SMX-HA), which can contribute to sulfonamide hypersensitivity, to the parent drug sulfamethoxazole. Variability in hydroxylamine reduction could thus play a role in adverse drug reactions. The aim of this study was to characterize variability in SMX-HA reduction in 111 human livers, and investigate its association with single nucleotide polymorphisms (SNPs) in b5 and b5R cDNA.
METHODS: Liver microsomes were assayed for SMX-HA reduction activity, and b5 and b5R expression was semiquantified by immunoblotting. The coding regions of the b5 (CYB5A) and b5R (CYB5R3) genes were resequenced.
RESULTS: Hepatic SMX-HA reduction displayed a 19-fold range of individual variability (0.06-1.11 nmol/min/mg protein), and a 17-fold range in efficiency (Vmax/Km) among outliers. SMX-HA reduction was positively correlated with b5 and b5R protein content (P<0.0001, r=0.42; P=0.01, r=0.23, respectively), and expression of both proteins correlated with one another (P<0.0001; r=0.74). A novel cSNP in CYB5A (S5A) was associated with very low activity and protein expression. Two novel CYB5R3 SNPs, R59H and R297H, displayed atypical SMX-HA reduction kinetics and decreased SMX-HA reduction efficiency.
CONCLUSION: These studies indicate that although novel cSNPs in CYB5A and CYB5R3 are associated with significantly altered protein expression and/or hydroxylamine reduction activities, these low-frequency cSNPs seem to only minimally impact overall observed phenotypic variability. Work is underway to characterize polymorphisms in other regions of these genes to further account for individual variability in hydroxylamine reduction.
PMID: 19997042 [PubMed - indexed for MEDLINE]
Clinical presentation of 26 anaplasma phagocytophilum-seropositive dogs residing in an endemic area.
J Am Anim Hosp Assoc. 2010 Nov-Dec;46(6):405-12
Authors: Mazepa AW, Kidd LB, Young KM, Trepanier LA
Anaplasma (A.) phagocytophilum, the etiological agent of canine granulocytic anaplasmosis, is capable of inciting moderate to severe clinical disease in a variety of mammals and is endemic in the upper midwest. The purpose of this study was fourfold: to describe the range of clinical signs in dogs seropositive to A. phagocytophilum; to examine the prevalence of immune-mediated hemolytic anemia (IMHA) in this population; to evaluate whether specific clinical signs were associated with coexposure to Borrelia (B.) burgdorferi in actively infected dogs; and to determine whether clinical response to doxycycline was complete in treated dogs. Medical records of dogs seropositive for A. phagocytophilum were reviewed retrospectively. Peripheral blood smears were also reviewed retrospectively for granulocytic Anaplasma morulae. Lethargy (81%), inappetence (58%), and lameness (50%) were the most common clinical signs, followed by fever (46%). Thrombocytopenia was the most common laboratory abnormality, and IMHA was diagnosed in three dogs. Dogs that were thrombocytopenic and had antibodies to both A. phagocytophilum and B. burgdorferi had a median platelet count of 51,000/μL (range 20,000 to 171,000/μL), which was significantly lower than the count in dogs with antibodies only to A. phagocytophilum (P=0.04). Some dogs had an apparent relapse of clinical signs after an appropriate course of doxycycline. Testing for A. phagocytophilum by polymerase chain reaction, serum antibody assays, and/or blood smear evaluation should be considered in dogs with IMHA, cough, or epistaxis and that reside in A. phagocytophilum-endemic areas. If moderate to severe thrombocytopenia is present, testing for concurrent B. burgdorferi infection may be warranted.
PMID: 21041333 [PubMed - indexed for MEDLINE]