SummaryPregnancy is associated with depressed fibrinolysis as judged from the decreased fibrinolytic response to venous occlusion. In order to elucidate if this decreased response is due to an increase in plasminogen activator inhibitors 1 and 2 (PAI-1, PAI-2), and/or to decreased release of tissue-type plasminogen activator (t-PA) antigen during venous occlusion, 36 women (18 women with normal pregnancy and 18 with gestational hypertension without proteinuria) were followed during pregnancy and puerperium. In each woman a 20 min venous occlusion was performed in the second and in the third trimester of pregnancy and 3 days after delivery. The increase in t-PA antigen after venous occlusion relative to basal value was in the second trimester of pregnancy on average 3.7 fold, in the third trimester 4.4 fold, and so not reduced compared to non-pregnant women (3.7 fold increase). After delivery the increase in t-PA antigen was significantly enhanced (8.5 fold, p <0.005). The fibrinolytic response to venous occlusion measured by euglobulin and t-PA activity was significantly decreased in the third trimester compared to non-pregnant values (both p <0.005) and returned to somewhat higher (euglobulin clot lysis) or significantly higher (t-PA activity, p <0.01) values 3 days after delivery. Decreased euglobulin and t-PA activity after venous occlusion in the third trimester coincided with significant increases in basal PAI activity, PAI-1 antigen and PAI-2 antigen (2.9, 2.5 and >30 fold increase relative to non-pregnant values, respectively, all p <0.001). No significant differences in fibrinolytic variables were observed between nor-motensive and hypertensive pregnant women. It was concluded that t-PA antigen release during venous occlusion is not decreased during pregnancy and puerperium, and that decreased fibrinolytic response measured by global methods should be attributed to increased t-PA inhibitors. Gestational hypertension without proteinuria is not characterized by changes in fibrinolytic responses different from those observed in normal pregnancy.
The possibility of using the immunoferment analysis for determination of lysozyme of mucus and fish tissues has been investigated. This method was adapted by reducing the dilution of the supernatant of biological material with buffer from 1:5 to 1:1. Clinically healthy fish (10 samples of pike and 10 samples of carp) were taken from fish ponds at water temperature of 14-16ºC. Biological material (blood, spleen, mucus of superficial integuments) was selected concerning the commonly used methods in ichthyopathology. The determination of lysozyme activity was carried out using high sensitivity RIDASCREEN ® FAST Lysozym test kit. When diluting the supernatant of biological material with buffer in a ratio of 1:5 (according to the method), it was not possible to calculate the data by the calibration curve, since the lysozyme parameters of the samples were lower than the lysozyme standard (0,050 mkg/g). Therefore, the final dilution was reduced and the experiment was continued with dilution of the supernatant with a protein extraction buffer in a ratio of 1:1. The data of the statistical processing of the results have led to conclusion the use of RIDASCREEN ® FAST Lysozym test kit in the proposed modification for the immunoferment analysis of lysozyme activity in tissues and mucus of fish is possible. A high content of lysozyme was noted in the mucus of the superficial integuments of fish, the lowest -in the blood serum. The content of lysozyme in the tissues and organs of pike is higher than in carp. Досліджена можливість використання методу імуноферментного аналізу для визначення лізоциму слизу і тканин риб. Для цього методику імуно-ферментного аналізу для визачення лізоцимної активності було адаптовано за рахунок зменшення розведення супернатанту біологічного матеріалу буфером з 1:5 до 1:1. Для досліджень брали клінічно здорову рибу ( по 10 екземплярів щуки та коропа з рибницьких ставів) за температури води 14-16ºС. Біологічний матеріал (кров, селезінку, слиз поверхневих покривів) відбирали згідно загальноприйнятих в іхтіопатології методів. Визначення лізоцимної активності проводили методом імуноферментного аналізу за допомогою тест-набору RIDASCREEN ® FAST Lysozym, перевагою якого є висока чутливість. За розведення супернатанту біологічного матеріалу буфером 1:5 (згідно методики) дані не піддавались обрахунку за калібрувальною кривою, оскільки показники лізоциму зразків були нижчі за показники стандарту лізоциму (0,050 мкг/г) на калібрувальній кривій. Тому було зменшено кінцеве розведення і дослід продовжили із розведенням супернатанту буфером екстракції білку у співвідношенні 1:1. Дані статистичного опрацювання результатів дозволили зробити висновок про придатність використання тест-набору RIDASCREEN ® FAST Lysozym в запропонованій модифікації для імуноферментного аналізу лізоцимної активності в тканинах та слизу риб. Найвищий вміст лізоциму відмічали у слизу поверхневих покривів риб, найнижчий -у сироватці крові. Вміст лізоциму у тканинах та органах щуки вищий ніж у коропа.Ключові слова: риба, щука, короп, гумора...
<p>The pro-antioxidant balance in the liver and muscles of sterlet under the influence of artificial carbonic dioxyde hibernation and anaesthesia have been studied. In experiments were used two-years old sterlet weighing 170–210 g which were grown under conditions of a closed water supply. Four groups of five copies of each fish were formed: Control group I (fish remained intact); Group II (the clove oil was used for fish anaesthesia, which was added to the water); Group III (fish were hibernated by carbon dioxide); Group IV - control of hibernation (after complete recovery from carbon dioxide hibernation, these groups of fish were returned to the pool of incubation shop for her follow–up). The research of intensity of the formation products of peroxide oxidation of lipids (POL) in the body of sterlet in the experimental conditions were conducted by determining the content of thiobarbituric acid active products (TBA–active products) in the muscles and liver that are generated at the final stages of lipid peroxidation. The content of malonic dialdehyde (MDA), one of the intermediate products of lipid peroxidation of membrane was determined photometrically by the concentration of colored complex formed by its reaction in the acidic environment of the two molecules of thiobarbituric acid (TBA). Catalase activity was determined by the ability of hydrogen peroxide to form the stable colored complex with Molybdenum salts. The results support the adaptive character in fluctuations in the antioxidant protection system by the actions of carbon dioxide hibernation and anaesthesia on the sterlet body. This also contributes to usage of these conditions in fish transportation at long distances without stress factors.</p>
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