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EN
Laryngopharyngeal reflux (LPR) is a common defect among laryngological and phoniatric patients. Although LPR is categorized as a superficial gastroesophageal reflux disease (GERD), differential diagnosis should treat these two diseases separately. LPR symptoms can be assessed in the interview using as a tool the reflux symptom index (RSI). In addition, changes in the larynx that occur during LPR might be seen during laryngoscopy and classified according to the reflux finding score (RFS). One of the main mucosal irritants in LPR is pepsin which digests proteins and impairs the functions of the upper respiratory tract cells by affecting carbonate anhydrase (CAIII) and the Sep 70 protein. Pepsin initiates inflammatory changes within the larynx, nasopharynx and nasal cavity. The use of pepsin detection in upper and lower throat secretions is a new direction in LPR diagnostics.
EN
Laryngopharyngeal reflux (LPR) is a common defect among laryngological and phoniatric patients. Although LPR is categorized as a superficial gastroesophageal reflux disease (GERD), differential diagnosis should treat these two diseases separately. LPR symptoms can be assessed in the interview using as a tool the reflux symptom index (RSI). In addition, changes in the larynx that occur during LPR might be seen during laryngoscopy and classified according to the reflux finding score (RFS). One of the main mucosal irritants in LPR is pepsin which digests proteins and impairs the functions of the upper respiratory tract cells by affecting carbonate anhydrase (CAIII) and the Sep 70 protein. Pepsin initiates inflammatory changes within the larynx, nasopharynx and nasal cavity. The use of pepsin detection in upper and lower throat secretions is a new direction in LPR diagnostics.
EN
INTRODUCTION: The aim of this study was to develop a method to assay the activity of pepsin in gastric juice based on the method described in the references, which was not entirely effective in laboratory tests. The study aims to improve the aforementioned method based on the digestion of bovine hemoglobin by pepsin and to introduce effective changes so that the results may be useful for diagnostic purposes. MATERIALS AND METHODS: To achieve this, tests were carried out using the hemoglobin of different species of animals and human hemoglobin, changing the incubation periods, concentrations and volumes of reagents. RESULTS: Tests based on the use of purified (crystalline) pepsin revealed that the most effective in the assay of enzyme activity is human hemoglobin. The originally proposed incubation periods were extended, and in order to inhibit the reaction, trichloroacetic acid was used in a higher concentration than suggested, while reducing by half the volume of reactants. In the tests carried out using a purified enzyme, a correlation between the known concentration of the enzyme and the absorbance of supernatant containing the reaction products of enzymatic digestion of hemoglobin was proved. CONCLUSIONS: Introducting changes leads to more effective diagnostic use.
PL
WSTĘP: Celem niniejszych badań było opracowanie metody oznaczania aktywności pepsyny w soku żołądkowym na podstawie metody opisanej w piśmiennictwie, która w badaniach laboratoryjnych okazała się nie w pełni efektywna. Podjęta praca jest próbą udoskonalenia metody polegającej na trawieniu hemoglobiny wołowej przez pepsynę i wprowadzenia skutecznych zmian, aby uzyskane wyniki mogły być użyteczne w diagnostyce klinicznej. MATERIAŁ I METODY: Przeprowadzono badania z wykorzystaniem hemoglobin różnych gatunków zwierząt i człowieka, zmiany czasów inkubacji, stężeń oraz objętości reagentów. WYNIKI: Próby oparte na wykorzystaniu roztworu pepsyny wykazały, że najskuteczniejsza w oznaczaniu aktywności enzymu jest hemoglobina ludzka. Pierwotnie proponowane czasy inkubacji zostały wydłużone, a do hamowania reakcji użyto kwasu trichlorooctowego o wyższym stężeniu, niż sugerowano, równocześnie zmniejszając o połowę objęto-ści reagentów. W próbach przeprowadzonych z użyciem oczyszczonego enzymu wykazano korelację między stężeniem enzymu a absorbancją supernatantu zawierającego produkty reakcji enzymatycznego trawienia hemoglobiny. WNIOSKI: Wprowadzone modyfikacje pozwalają na udoskonalenie procesu oznaczania aktywności pepsyny w soku żołądkowym i efektywnego wykorzystania w praktyce laboratoryjnej.
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