Monitoring furosemide in racehorses participating in an EIPH program

Stevenson, A.; Weber, M. P.; Trudel, R.; Leavitt, R.; Woodard, Donna S.; Todi, F.; Mendonça, Monique Culturato Padilha; Robillo, V.; Young, Lee Mi; Kacew, Sam

doi:10.1111/j.1365-2885.1994.tb00229.x

Cited by 6 publications

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References 6 publications

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“…However, its expression is not limited to these tumors. CD99 immunoreactivity has been observed in RMS, 4 SS, 5-7 lymphoblastic lymphoma, 8 mesenchymal chondrosarcoma, 9,10 Merkel cell carcinoma (MCC) 11 and has been also reported in rare cases of Wilms' tumor, 12 small cell osteosarcoma 13 and DSRCT. 14,15 Thus, FLI-1 has been recently proposed as an additional immunohistochemical marker of ES/PNET, along side the traditional CD99.…”

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confidence: 99%

Utility of the immunohistochemical detection of FLI-1 expression in round cell and vascular neoplasm using a monoclonal antibody

et al. 2004

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FLI-1 nuclear transcription factor has been proposed as a useful tool in the differential diagnosis of small round cell sarcomas. Recently, FLI-1 has been reported as the first nuclear marker of endothelial differentiation. However, its clinical use has been hampered by major interpretation problems, due to the presence of background staining as well as staining variation between different lots of the same antiserum. In this study, a novel monoclonal antibody raised against the carboxyl terminal of the FLI-1 protein (clone GI146-222, BD Pharmingen) was tested in a series of small round cell and vascular neoplasms. Furthermore, in order to assess FLI-1 specificity, we analyzed its expression in a series of common epithelial and nonepithelial malignancies. In total, 15 Ewing's sarcomas, 10 rhabdomyosarcomas, 5 desmoplastic small round cell tumors, 10 synovial sarcomas, 10 high-grade pleomorphic sarcomas, 10 malignant melanomas, 5 Merkel's carcinomas, 10 colonic adenocarcinomas, 10 breast carcinomas, 10 lung adenocarcinomas, 20 angiosarcomas, 5 epithelioid hemangioendotheliomas, 10 Kaposi's sarcomas and 10 benign hemangiomas, were stained. A strong FLI-1 immunoreactivity was detected in all Ewing's sarcomas and vascular neoplasms, highlighting the high sensitivity of FLI-1 monoclonal antibody. However, 2/5 Merkel's carcinomas and 1/10 malignant melanomas showed a strong nuclear immunostaining, suggesting that FLI-1 may not be so helpful in the differential diagnosis of cutaneous Ewing's sarcoma. In addition, a weak immunoreactivity was found in 3/5 Merkel cell carcinomas, 3/10 synovial sarcomas, 5/10 malignant melanomas, 6/10 lung adenocarcinomas and in 1/10 breast carcinomas. In contrast, all the rhabdomyosarcomas, desmoplastic small round cell tumors, high-grade pleomorphic sarcomas and colonic adenocarcinomas tested were negative. Importantly, in contrast with previous studies, no background staining was observed. Our results indicate that FLI-1 monoclonal antibody can be reliably applied to the differential diagnosis of small round cell neoplasms of soft tissue, and confirm its important role as nuclear marker of endothelial differentiation, mainly helpful in those cases in which technical artifacts are seen by using the traditional membranous and cytoplasmic endothelial markers. 2 The EWS/FLI-1 chimeric protein binds DNA with the same affinity as FLI-1, but EWS/FLI-1 transcriptional activation of the target genes is much more efficient.3 ES/PNET belongs to a heterogeneous group of mesenchymal neoplasms featuring round cell morphology and therefore collectively known as small round cell sarcomas. Alveolar rhabdomyosarcoma (RMS), desmoplastic small round cell tumor (DSRCT), mesenchymal chondrosarcoma and poorly differentiated synovial sarcoma (SS) also belong to

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confidence: 99%

Utility of the immunohistochemical detection of FLI-1 expression in round cell and vascular neoplasm using a monoclonal antibody

et al. 2004

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“…The furosemide EIA used in the present study is a homologous-type and may exhibit a lower sensitivity compared with other immunological assay methods [15,17]. However, The lower limit of quantification of the EIA method is similar to that of the HPLC methods described in previous studies [3,6,7,13] and it appears to be sufficiently sensitive for the detection of furosemide in horse plasma. The precision and specificity of the EIA were validated to be acceptable within the limits of standardization of the method.…”

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confidence: 75%

“…Therefore, a pharmacokinetics study of furosemide in horses is seem to be important for an assessment of the effectiveness of the drug as a prophylactic for EIPH. High-performance liquid chromatography (HPLC) methods have generally been used for the quantitative analysis and pharmacokinetics study of furosemide [2,3,6,7,13]. However, using these methods it is necessary to extract and purify the furosemide from the plasma samples, and large sample volumes (from 0.5 to 1.0 ml) are also required.…”

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confidence: 99%

A Direct Enzyme Immunoassay for the Measurement of Furosemide in Horse Plasma

Nagata

Kurosawa

Kuwajima

2007

The Journal of Veterinary Medical Science

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ABSTRACT. A new enzyme immunoassay (EIA) for the measurement of furosemide in horse plasma is described. The lower limit of detection of this EIA method was 7.8 ng/ml. The intra-and inter-assay coefficients of variation ranged from 2.5% to 4.9% and 7.5% to 9.8%, respectively. Cross-reactivity with other compounds was not observed. There was a high correlation (r 2 =0.987) between the high-performance liquid chromatography and EIA results obtained for furosemide concentrations in horse plasma. These results indicate that the newly developed EIA method is useful for the quantitative analysis of furosemide in horse plasma. KEY WORDS: enzyme immunoassay, equine, furosemide.J. Vet. Med. Sci. 69(3): 305-307, 2007 Furosemide is the most widely used loop diuretic in veterinary medicine. It is generally used for the treatment of edema, ascites, hypertension, and, particularly in horses, is used as a prophylaxis for exercise-induced pulmonary haemorrhage (EIPH). EIPH is a frequent occurrence in strenuously exercised horses [8,14] and markedly affects their performance [4]. The failure of pulmonary capillaries, triggered by hypertension following exercise, has been recognized as a possible cause of EIPH [16]. It has been reported that furosemide attenuates exercise-induced pulmonary capillary hypertension, and that the magnitude of the effect is dose-dependent [9,10]. The excess of furosemide treatment, however, has the danger of dehydration and electrolyte imbalance. The research for the relationship between concentrations and effects of furosemide is necessary to take appropriate treatment protocols. Therefore, a pharmacokinetics study of furosemide in horses is seem to be important for an assessment of the effectiveness of the drug as a prophylactic for EIPH. High-performance liquid chromatography (HPLC) methods have generally been used for the quantitative analysis and pharmacokinetics study of furosemide [2,3,6,7,13]. However, using these methods it is necessary to extract and purify the furosemide from the plasma samples, and large sample volumes (from 0.5 to 1.0 ml) are also required. In the present study, our objective was to attempt a quantitative analysis of furosemide by using an immunological assay method. To this end, we have developed a new direct enzyme immunoassay (EIA) for the measurement of plasma furosemide concentrations.A furosemide and Keyhole limpet hemocyanin (KLH) conjugate was used as the immunogen. The conjugate was prepared according to the method described by Carlin et al.[1], which was modified as described below. Furosemide (15 mg) (Sigma, St. Louis, MO, U.S.A.), N-hydroxysuccinimide (NHS, 5.2 mg) (Sigma, St. Louis, MO, U.S.A.) and N, N-dicyclohexylcarbodiimide (DCC, 9.3 mg) (Pierce, Rockford, IL, U.S.A.) were mixed in 250 µl of N, N-dimethylformamide (DMF) (Sigma, St. Louis, MO, U.S.A.) and incubated for 1.5 hr at room temperature. The reaction mixture was purified by using a Sephadex LH20 column (Amersham Biosciences, Uppsala, Sweden) that was equilibrated with DMF. The fractions contain...

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“…A number of high plasma concentrations (greater than 125 ng/mL) were found in both groups of horses, which were considered outliers, in that they were much higher than expected, based on the distribution of concentrations around the mean (Uboh et al ., 1992). In a second study done under racing conditions, the mean plasma concentrations in horses administered 150–250 mg of furosemide 4 h prior to race time, was ≈ 20 ng/mL (Stevenson et al ., 1994). In this study, the highest outlier was 150 ng/mL.…”

Section: Use Of Plasma Concentration Of Furosemide To Regulate Its Admentioning

confidence: 99%

“…This method has not been adopted for the control of furosemide administration. Some jurisdictions are using a threshold plasma concentration of 100 ng/mL (Stevenson et al ., 1994). If it is above this concentration, the assumption is made that it was caused by the administration of a higher than permitted dose of furosemide, the dose was administered closer than 4 h before race time, or an additional dose was administered closer to race time than the 4 h window.…”

Section: Use Of Plasma Concentration Of Furosemide To Regulate Its Admentioning

confidence: 99%

Review of furosemide in horse racing: its effects and regulation

Soma

Uboh

1998

Vet Pharm & Therapeutics

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Furosemide has been used empirically and has been legally approved for many years by the US racing industry for the control of exercise-induced pulmonary haemorrhage (EIPH) or bleeding. Its use in horses for this purpose is highly controversial and has been criticized by organizations outside and inside of the racing industry. This review concentrates on its renal and extra-renal actions and the possible relationship of these actions to the modification of EIPH and changes in performance of horses. The existing literature references suggest that furosemide has the potential of increasing performance in horses without significantly changing the bleeding status. The pulmonary capillary transmural pressure in the exercising horse is estimated to be over 100 mmHg. The pressure reduction produced by the administration of furosemide is not of sufficient magnitude to reduce transmural pressures within the capillaries to a level where pressures resulting in rupture of the capillaries, and thus haemorrhage, would be completely prevented. This is substantiated by clinical observations that the administration of furosemide to horses with EIPH may reduce haemorrhage but does not completely stop it. The unanswered question is whether the improvement of racing times which have been shown in a number of studies are due to the reduction in bleeding or to other actions of furosemide. This review also discusses the difficulties encountered in furosemide regulation, in view of its diuretic actions and potential for the reduction in the ability of forensic laboratories to detect drugs and medications administered to a horse within days or hours before a race. Interactions between nonsteroidal anti-inflammatory drugs (NSAIDs) and furosemide have also been examined, and the results suggest that the effects of prior administration of NSAID may partially mitigate the renal and extra-renal effects which may contribute to the effects of furosemide on EIPH.

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Monitoring furosemide in racehorses participating in an EIPH program

Cited by 6 publications

References 6 publications

Utility of the immunohistochemical detection of FLI-1 expression in round cell and vascular neoplasm using a monoclonal antibody

Utility of the immunohistochemical detection of FLI-1 expression in round cell and vascular neoplasm using a monoclonal antibody

A Direct Enzyme Immunoassay for the Measurement of Furosemide in Horse Plasma

Review of furosemide in horse racing: its effects and regulation

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