The pharmacologic effects of isoxsuprine

Isoxsuprine is a therapeutic medication used to treat navicular disease and other lower limb problems in horses and is one of the more frequently detected therapeutic agents in racing horses. In crossover studies, horses were administered intravenous and oral isoxsuprine to determine the character and duration of pharmacological effects. Following intravenous administration, isoxsuprine significantly increased heart rate, spontaneous activity, and sweat production. There was an apparent, although statistically insignificant, increase in cutaneous blood flow. Skin temperature decreased below control values, and there was a significant decrease in core temperature. Isoxsuprine also reduced smooth muscle tone. In contrast, after oral dosing, there was no statistical difference between control and isoxsuprine-treated horses for any of the measured variables. It was concluded that the measurable physiologic effects of intravenous isoxsuprine are short-lived, since none of the above responses was apparent four hours or more after intravenous administration

Warrantless Arrest Jurisdiction In Texas: An Analysis And A Proposal

Within the State of Texas, there exist a great number of “peace officers” who are granted a wide range of power and authority. This includes the power to make warrantless arrests and searches pursuant to those arrests. Significant ambiguity exists regarding a peace officer’s jurisdiction. The confusion is largely due to imprecise statutory language and varying judicial interpretations. Article 998 of Texas Revised Civil Statutes, for example, bestowed on city police officers the same powers, authority, and jurisdiction as city marshals. The statute, though, neglected to define the extent of that jurisdiction, or even what “jurisdiction” meant in that context. Article 999, which defined the duties and powers of city marshals, endowed city marshals with “like power, authority, and jurisdiction as the sheriff.” A cursory reading of these articles could lead to the understanding that a city police officer had countywide jurisdiction. In Angel v. State, the court considered the arrest jurisdiction of Tomball city police officers who observed a suspect operating what was later determined to be a stolen road paving machine outside the city limits of Tomball, but within Harris County. The Texas Court of Criminal Appeals concluded that the interweaving of loosely worded statutes, specifically articles 998 and 999, evidences the legislature’s intent to expand the common law jurisdiction of city police officers to the entire county. A city police officer’s territorial jurisdiction for arrest purposes is an issue unduly and unnecessarily complicated by the court’s tracing of ambiguous statutory provisions, and the potential for a similar result exists for other kinds of peace officers

The Mare Reproductive Loss Syndrome and the Eastern Tent Caterpillar II: A Toxicokinetic/Clinical Evaluation and a Proposed Pathogenesis: Septic Penetrating Setae

Reviewing the mare reproductive loss syndrome (MRLS), it is proposed that the fundamental mechanism of this syndrome, which includes early fetal loss, late fetal loss, uveitis, pericarditis, and encephalitis, is tissue penetration by septic barbed setal fragments (septic penetrating setae) from Eastern tent caterpillars (Malacosoma americanum). Once ingested, these barbed setal fragments migrate through moving tissues, followed by rapid hematogenous spread of bacteria, bacterial emboli, and/or septic fragments of setae (septic penetrating setal emboli), collectively referred to as septic materials. Pathogenic bacteria, therefore, enter the horse as hitchhikers on or in the caterpillar setal fragments, and MRLS is caused by 1) the barbed setal fragments’ ability to penetrate moving tissues, including blood vessels, releasing septic materials, which rapidly distribute hematogenously; 2) the high sensitivity of the pregnant mare to bacteria from such septic materials introduced into the uterus, fetal membranes, or fetal fluids; 3) the unusually broad spectrum of bacterial pathogens carried on or in the setal fragments; and 4) the less effective antibacterial responses in certain susceptible extracellular fluids (e.g., fetal, ocular, pericardial, and cerebrospinal fluids). The driving force for MRLS pathology, including abortions, is septic material- induced bacterial proliferation, which provides a critical amplification step, enabling approximately 1-gram caterpillars to rapidly (32 hours) cause abortions in 680-kg (1,500- lb) mares. Calculations based on the unique eye data suggest that the actual number of distributing effective septic material quanta in field cases may be small—on the order of 10/horse/day—accounting for the lack of systemic clinical signs in affected horses. Therefore, it is proposed that MRLS starts with ingestion of Eastern tent caterpillars, followed by barbed setal fragments randomly penetrating intestinal tissues, including thinwalled venules and other blood vessels, with release of septic material that distributes hematogenously to all points in the body. Identification of abortigenic activity with the integument of the caterpillar and recent findings of large numbers of granulomatous lesions containing setal fragments in the intestines of pigs and rats directly supports the septic penetrating setal portion of the hypothesis. Analysis of the clinical syndromes and a toxicokinetic/ statistical analysis of MRLS suggest that setally-mediated introduction of septic material into blood vessels and other tissues may be key to understanding the very unusual toxicokinetics and pathogenesis of the unique group of syndromes that constitute MRLS. Like MRLS itself, this hypothesis is unique. The septic penetrating setal emboli portion is without precedent, is based on the unique clinical characteristics of MRLS, and appears well supported by ongoing experimental approaches

An Overview of the Methylxanthines and their Regulation in the Horse

Caffiene, theophylline and theobromine are naturally occurring members of the methylxanthine family;pentoxfylline, dyphylline and enprofylline are structurally related synthetic pharmaceuticals. Caffiene has predominantly central nervous system effects, theophylline, dyphylline and enprofylline have predominantly bronchodilator effects, while theobromine is associated with diuretic responses. Pentoxfylline is thought to increase red cell deformability and facillitate blood flow through capillary beds. The methylxanthines are not highly potent agents; they are typically administered in gram doses and they tend to have relatively long half-lives. They remain detectable in plasma and urine for relatively long periods. Similarly, traces of the naturally occurring members of this family are not uncommonly identified in forensic samples. In this review we report on the detection, actions, uses and regulatory control of this group of agents in performance horses

Frequency distribution of post race urine pH from Standardbreds compared with Thoroughbreds: research and regulatory significance

The concentration of drugs and drug metabolites in urine samples of racing horses is strongly influenced by urine pH(Tobin, 1981), depending on whether the drugs are weak acids or weak bases. Drugs that are weak acids tend to concentrate in besic urine. In contrast, drugs that are weak bases tend to concentrate in acidic urine. These relationships have a well-established theoretical basis (the Henderson-Hasselbalch relationship) and have been demonstrated repeatedly in experimental animals and man (Tobin, 1981). More recently, evidence suggests that these relationships also occur with clinically and forensically significant agents in equine urine (Wood, et al. 1990; Gerken et al.1991.

Clenbuterol in the horse: Confirmation and quantitation of serum clenbuterol by LC-MS-MS after oral and intratracheal administration

Clenbuterol is a β2 agonist/antagonist bronchodilator, and its identification in post-race samples may lead to sanctions. The objective of this study was to develop a specific and highly sensitive serum quantitation method for clenbuterol that would allow effective regulatory control of this agent in horses. Therefore, clenbuterol-d9 was synthesized for use as an internal standard, an automated solid-phase extraction method was developed, and both were used in conjunction with a multiple reaction monitoring liquid chromatography-tandem mass spectrometry (LC-MS-MS) method to allow unequivocal identification and quantitation of clenbuterol in 2 mL of serum at concentrations as low as 10 pg/mL. Five horses were dosed with oral clenbuterol (0.8 μg/kg, BID) for 10 days, and serum was collected for 14 days thereafter. Serum clenbuterol showed mean trough concentrations of ∼150 pg/mL. After the last dose on day 10, serum clenbuterol reached a peak of ∼500 pg/mL and then declined with a half-life of ∼7 h. Serum clenbuterol declined to 30 and 10 pg/mL at 48 and 72 h after dosing, respectively. By 96 h after dosing, the concentration was below 4 pg/mL, the limit of detection for this method. Compared with previous results obtained in parallel urinary experiments, the serum-based approach was more reliable and satisfactory for regulation of the use of clenbuterol. Clenbuterol (90 μg) was also administered intratracheally to five horses. Peak serum concentrations of ∼230 pg/mL were detected 10 min after administration, dropping to ∼50 pg/mL within 30 min and declining much more slowly thereafter. These observations suggest that intratracheal administration of clenbuterol shortly before race time can be detected with this serum test. Traditionally, equine drug testing has been dependent on urine testing because of the small volume of serum samples and the low concentrations of drugs found therein. Using LC-MS-MS testing, it is now possible to unequivocally identify and quantitate low concentrations (10 pg/mL) of drugs in serum. Based on the utility of this approach, the speed with which new tests can be developed, and the confidence with which the findings can be applied in the forensic situation, this approach, offers considerable scientific and regulatory advantages over more traditional urine testing approaches

Absence of detectable pharmacological effects after oral administration of isoxsuprine

Isoxsuprine is reported to be a peripheral vasodilator used in human and veterinary medicine to treat ischaemic vascular disease. In horses, it is generally administered orally to treat navicular disease and other lower limb problems. To deflne the scope and duration of its pharmacological responses after oral administration, 6 horses were dosed with isoxsuprine HCI (1.2 mg/kg bwt) q. 12 h for 8 days and then tested to assess the duration and extent of pharmacological actions. There was no significant difference between isoxsuprine and control treatment values for heart rate, spontaneous activity, sweat production, anal muscle tone, core and skin temperatures, and cutaneous blood flow. The lack of pharmacological effect following oral administration was in sharp contrast to the marked response following i.v. dosing reported in earlier experiments

Development of a method for the detection and confirmation of the alpha-2 agonist amitraz and its major metabolite in horse urine

Amitraz (N′-(2,4-dimethylphenyl)-N-[[(2,4-dimethylphenyl)imino] methyl]-N-methyl-methanimidamide) is an alpha-2 adrenergic agonist used in veterinary medicine primarily as a scabicide- or acaricide-type insecticide. As an alpha-2 adrenergic agonist, it also has sedative/tranquilizing properties and is, therefore, listed as an Association of Racing Commissioners International Class 3 Foreign Substance, indicating its potential to influence the outcome of horse races. We identified the principal equine metabolite of amitraz as N-2,4-dimethylphenyl-N′-methylformamidine by electrospray ionization(+)-mass spectrometry and developed a gas chromatographic-mass spectrometric (GC-MS) method for its detection, quantitation, and confirmation in performance horse regulation. The GC-MS method involves derivatization with t-butyldimethylsilyl groups; selected ion monitoring (SIM) of m/z 205 (quantifier ion), 278, 261, and 219 (qualifier ions); and elaboration of a calibration curve based on ion area ratios involving simultaneous SIM acquisition of an internal standard m/z 208 quantifier ion based on an in-house synthesized d6 deuterated metabolite. The limit of detection of the method is approximately 5 ng/mL in urine and is sufficiently sensitive to detect the peak urinary metabolite at 1 h post dose, following administration of amitraz at a 75-mg/horse intraveneous dose

The Mare Reproductive Loss Syndrome and the Eastern Tent Caterpillar: A Toxicokinetic/Statistical Analysis With Clinical, Epidemiologic, and Mechanistic Implications

During 2001, central Kentucky experienced acute transient epidemics of early and late fetal losses, pericarditis, and unilateral endophthalmitis, collectively referred to as mare reproductive loss syndrome (MRLS). A toxicokinetic/statistical analysis of experimental and field MRLS data was conducted using accelerated failure time (AFT) analysis of abortions following administration of Eastern tent caterpillars (ETCs; 100 or 50 g/day or 100 g of irradiated caterpillars/day) to late-term pregnant mares. In addition, 2001 late-term fetal loss field data were used in the analysis. Experimental data were fitted by AFT analysis at a high (P \u3c .0001) significance. Times to first abortion (“lag time”) and abortion rates were dose dependent. Lag times decreased and abortion rates increased exponentially with dose. Calculated dose × response data curves allow interpretation of abortion data in terms of “intubated ETC equivalents.” Analysis suggested that field exposure to ETCs in 2001 in central Kentucky commenced on approximately April 27, was initially equivalent to approximately 5 g of intubated ETCs/day, and increased to approximately 30 g/day at the outbreak peak. This analysis accounts for many aspects of the epidemiology, clinical presentations, and manifestations of MRLS. It allows quantitative interpretation of experimental and field MRLS data and has implications for the basic mechanisms underlying MRLS. The results support suggestions that MRLS is caused by exposure to or ingestion of ETCs. The results also show that high levels of ETC exposure produce intense, focused outbreaks of MRLS, closely linked in time and place to dispersing ETCs, as occurred in central Kentucky in 2001. With less intense exposure, lag time is longer and abortions tend to spread out over time and may occur out of phase with ETC exposure, obscuring both diagnosis of this syndrome and the role of the caterpillars

Detection and Confirmation of Ractopamine and Its Metabolites in Horse Urine after Paylean® Administration

We have investigated the detection, confirmation, and metabolism of the beta-adrenergic agonist ractopamine administered as Paylean to the horse. A Testing Components Corporation enzyme-linked imunosorbent assay (ELISA) kit for ractopamine displayed linear response between 1.0 and 100 ng/ml, with an 1-50 of 10 ng/ml, and an effective screening limit of detection of 50 ng/mL. The kit was readily able to detect ractopamine equivalents in unhydrolyzed urine up to 24 h following a 300-mg oral dose. Gas chromatography-mass spectrometry (GC-MS) confirmation comprised glucuronidase treatment, solid-phase extraction, and trimethylsilyl derivatization, with selected-ion monitoring of ractopamine-tris(trimethylsilane) (TMS) m/z 267, 250, 179, and 502 ions. Quantitation was elaborated in comparison to a 445 Mw isoxsuprine-bis(TMS) internal standard monitored simultaneously. The instrumental limit of detection, defined as that number of ng on column for which signal-to-noise ratios for one or more diagnostic ions fell below a value of three, was 0.1 ng, corresponding to roughly 5 ng/mL in matrix. Based on the quantitation ions for ractopamine standards extracted from urine, standard curves showed a linear response for ractopamine concentrations between 10 and 100 ng/mL with a correlation coefficient r \u3e 0.99, whereas standards in the concentration range of 10-1000 ng/mL were fit to a second-order regression curve with r \u3e 0.99. The lower limit of detection for ractopamine in urine, defined as the lowest concentration at which the identity of ractopamine could be confirmed by comparison of diagnostic MS ion ratios, ranged between 25 and 50 ng/mL. Urine concentration of parent ractopamine 24 h post-dose was measured at 360 ng/mL by GC-MS after oral administration of 300 mg. Urinary metabolites were identified by electrospray ionization (+) tandem quadrupole mass spectrometry and were shown to include glucuronide, methyl, and mixed methyl-glucuronide conjugates. We also considered the possibility that an unusual conjugate added 113 amu to give an observed m/z 415 [M+H] species or two times 113 amu to give an m/z 528 [M+H] species with a daughter ion mass spectrum related to the previous one. Sulfate and mixed methyl-sulfate conjugates were revealed following glucuronidase treatment, suggesting that sulfation occurs in combination with glucuronidation. We noted a paired chromatographic peak phenomenon of apparent ractopamine metabolites appearing as doublets of equivalent intensity with nearly identical mass spectra on GC-MS and concluded that this phenomenon is consistent with Paylean being a mixture of RR, RS, SR, and SS diastereomers of ractopamine. The results suggest that ELISA-based screening followed by glucuronide hydrolysis, parent drug recovery, and TMS derivatization provide an effective pathway for detection and GC-MS confirmation of ractopamine in equine urine