• Title/Summary/Keyword: trypsin-like immunoreactivity (TLI)

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Preparation of Monoclonal Antibodies for Canine Trypsin-Like Immunoreactivity (개 트립신양(樣) 면역반응성 단클론 항체의 제작)

  • Kim, Sung-Soo;Kang, Ji-Houn;Cheong, Kwang-Myun;Yoo, Jai-Cheol;Chong, Chom-Kyu;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.25 no.5
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    • pp.317-323
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    • 2008
  • Canine trypsin-like immunoreactivity (cTLI), which is a mirror of the concentration of trypsin and trypsinogen, is a pancreas-specific enzyme and a suitable marker for canine pancreatitis and especially exocrine pancreatic insufficiency (EPI). To develop the immunochromatographic test kit, monoclonal antibodies that recognize cTLI were prepared. Anionic trypsin, cationic trypsin, and chymotrypsin from canine pancreas were successfully purified to homogeneity, using ammonium sulfate fractionation and benzamidine-affinity chromatography. The purification fold for anionic trypsin was 108 times when compared with that of the homogenation of pancreas. The molecular weights by SDS-PAGE analysis were approximately 23 kDa for chymotrypsin and approximately 20 kDa for cationic trypsin and anionic trypsin, respectively. Using the purified trypsin-like proteins, ten hybridomas which secret canine trypsin-specific monoclonal antibody were prepared. Klotz plot indicated that hybridomas, 5G2H10G4 and 2F4A11, have high affinity constant (Ka) of $4.1\;{\times}\;10^{9}$ and $1.8\;{\times}\;10^{9}$, respectively. Especially, 5F9H3 showed the cationic typsin-specific binding pattern and its Ka was determined to $4.5\;{\times}\;10^{9}$. The development of immunochromatographic test kit using these monoclonal antibodies against cTLI will be very useful in the diagnosis of canine EPI or canine pancreatitis.

Effects of Pancreatectomy on Serum Trypsin-like Immunoreactivity (TLI) in Dogs (개의 췌장적출이 혈청 Trypsin-like Immunoreacitivity (TLI)에 미치는 영향)

  • 곽우연;윤화영;오태호;윤영민;이창우;최희인
    • Journal of Veterinary Clinics
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    • v.18 no.1
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    • pp.7-13
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    • 2001
  • Depending on the degree of exocrine pancreatic insufficiency, there are inconsistent values in the serum trypsin-like immunoreactivity (TLI), hematology, and serology. To determine the correlation between those values and complete pancreatic insufficiency, 10 pancreatectomized dogs as the treated group and 10 sham operation dogs as control group were used. The entire treated group showed significant decrease in serum TLI level from 2.63$\pm$0.20 ng/ml (day 0) to 1.81$\pm$0.43 ng/ml at day 1 after the pancreatectomy (p<0.05) and the decline maintained till the end of the experiment. Blood glucose level gradually increased, but albumin level showed significant decrease (p<0.05) at day 1 and maintained the decline. Three clinical signs were observed such as depression, vomiting, and diarrhea. The entire treated group showed diarrhea throughout the experiment, but depression and vomiting were gradually diminished. The activity of serum alkaline phosphatase in the treated group increased till day 2 then decreased, on the other hand, control group showed continuous increase till day 4, and then decreased. Only experimental group showed the increased alanine aminotransferase activity at day 1. Serum lipase activity in the treated group jumped up at day 1 and then dropped down, which was even lower than the control. Regarding serum amlyase activity gradually decreased. According to these results, the assay of trypsin-like immunoreactivity showed the consistent result, so it suggests that TLI is an useful tool to determine the pancreatic exocrine function and possible diagnosis of exocrine pancreatic insufficiency compared to the simple assay of plasma amylase and lipase.

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Suspected Case of Exocrine Pancreatic Insufficiency in a Bengal Tiger (Panthera tigris tigris)

  • Rhim, Haerin;Han, Jae-Ik
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.240-242
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    • 2018
  • A 1-year-old, female Bengal tiger (Panthera tigris tigris) presented signs of weight loss and dark browncolored diarrhea. On fecal examination, numerous intact and broken red blood cells were found, but both parasites and inflammatory signs were absent. No significant findings were seen in serum biochemistry profiles, including electrolytes, with negative feline pancreatic lipase immunoreactivity (fPLI). Diagnostic kits using feces or peripheral blood were negative for feline parvovirus, feline coronavirus, feline immunodeficiency virus, and feline leukemia virus. Based on the result of feline trypsin-like immunoreactivity (fTLI) concentration ($4.6{\mu}g/L$), the tiger was provisionally diagnosed to have exocrine pancreatic insufficiency (EPI). After this diagnosis, pancreatic enzymes were prescribed. The feces of the tiger returned to normal form and her weight was increasing. EPI is uncommon and not described extensively in Felidae, including domestic cats. Feline EPI is associated with a variety of non-specific signs and it should be considered in the differential diagnosis of cases presenting with weight loss, diarrhea, and other gastrointestinal signs. In this case, the patient was strongly suspected to have EPI based on the very low fTLI concentration, though the concentration of fTLI in tigers has not yet been determined. This is the first report to present a suspected EPI case in Bengal tigers.