Amylase in human milk.

Amylase activity and isoenzyme pattern were determined in human milk from various stages of lactation and were compared with that in duodenal juice. The activity is high in colostrum and somewhat lower in milk from day 15 to day 90 after delivery. In this period of lactation, human milk contains higher amounts of amylase than duodenal juice from infants aged 1 to 6 months. Low activity was found in milk from 90 days or more after delivery. The amylase is of the salivary type. A pH of 5.3 does not inactivate the amylase; there is considerable human milk amylase activity in duodenal juice after a meal of human milk. Human milk amylase could thus contribute to the breast-fed infant's ability to digest starch.

Demonstration and semiquantitative determination of complexes between various proteases and human alpha2-macroglobulin.

Complex formation in vitro between human alpha2-macroglobulin and the human proteases cationic trypsin, chymotrypsin, plasmin and granulocyte elastase and collagenase was clearly visualized by the use of thin-layer electrofocusing in polyacrylamide gel followed by electrophoresis in agarose gel containing antibodies against human alpha2-macroglobulin. The technique permits semi-quantitative determination of the amount of complex and can demonstrate the formation of complexes between alpha2-macroglobulin and protease in vivo in ascitic fluid in peritonitis.

Lactate dehydrogenase and its isoenzymes in vaginal fluid in vaginitis/vaginosis cases and in healthy controls.

The objective of the study was to measure the lactate dehydrogenase (LD) activity in vaginal lavage fluid of women with vaginitis/vaginosis and in healthy pre- and post-menopausal controls. Also to analyse the LD isoenzyme patterns in such samples and compare the influence on the LD activity by different storage and sampling methods. Twenty of the women studied, who had no signs of inflammation as evidenced from vaginal wet smears, were pre-menopausal and 8 post-menopausal. Fifty-eight non-pregnant patients with vaginitis/vaginosis or non-inflammatory gynaecological conditions were analysed for LD isoenzyme patterns. The LD activity was correlated to vaginal pH. Furthermore, the LD activity was determined in another 100 women screened for Chlamydia trachomatis. Finally, the influence on the LD activity when sampling was made by a cytological brush vs vaginal lavage and analysed after different storage periods, as studied. The LD activity was elevated, i.e. >2 micro kat/L, in all but two of the women with leucorrhoea. Only women with bacterial vaginosis (BV) without leucorrhoea, had an increased LD activity. An increased vaginal pH correlated to the LD concentration. The LD activity was elevated in cases with vulvovaginal candidiasis, trichomoniasis, chlamydial cervicitis and senile colpitis. Storage of samples for up to six hours had no influence on the test outcome. Brush and lavage fluid samples did not differ with regard to the rate of positive LD tests. In healthy women, the LD activity is low and predominated by slow-migrating isoenzymes, i.e. LD 4 and 5. The LD activity is generally increased in cases of vaginitis and in women with BV and chlamydial cervicitis. In trichomoniasis, particularly high concentrations of LD 5, are found.

Transvaginal ultrasound and lactate dehydrogenase isoenzyme activity profile in uterine aspirate for diagnosis of endometrial carcinoma in women with postmenopausal bleeding.

Postmenopausal patients with vaginal bleeding (n = 72) were evaluated with the combination of transvaginal ultrasonography (TVS) and analysis of the lactate dehydrogenase (LD) isoenzyme activity profile in uterine fluid aspirates. TVS evaluation of the endometrium was classified as <5 mm, > or =5 mm, or poorly defined. The LD isoenzyme activity profile was characterized as abnormal or normal. Pathologic findings were further evaluated with diagnostic curettage. TVS found the endometrium to be > or =5 mm or poorly defined in 44 patients (61%). Endometrial carcinoma was found in 6 of 72 patients (8%). They appeared in the TVS groups endometrium >5 mm (n = 2) and endometrium poorly defined (n = 4) but not in the endometrium <5 mm. The LD isoenzyme activity profile was abnormal in the six malignant cases and in ten benign cases. Thus, the need for further evaluation with hysteroscopy and curettage was reduced to 16 cases. Since TVS had 100% sensitivity but only 42% specificity, it is suitable for first-level examination in patients with postmenopausal bleeding. The second-level method should have similarly high sensitivity but much higher specificity. The LD isoenzyme activity profile in uterine fluid aspirates had 100% sensitivity and 85% specificity. Another important feature is that the method is not sensitive to endometrial thickness, amount of sample, sampling device, or dilution. Thus, it is more reliable than aspiration histology. For every hysteroscopy or curettage that can be replaced by LD analysis, the cost is reduced by approximately EUR 720 or 540, respectively.

Electrophoretic separation, detection, and variation of amylase isoenzymes.

An electrophoretic technique for demonstrating amylase isoenzymes is described. After separation in an agarose gel containing a linear polyacrylamide polymer to reduce electroendosmotic flow, the amylase fractions are visualized by incubation with a commercially available dye-starch polymer (Phadebas Amylase Test). The technique detects amylase fractions with activities below 10 U/l. Some characteristic changes in such diseases as acute and chronic pancreatitis, cystic fibrosis of the pancreas, macroamylasemia and inherited variants as well as after maxillofacial surgery are mentioned.

Sources of the serum isoamylases and their normal range of variation with age.

The isoamylases in various human tissue homogenates and body fluids were separated by agarose gel electrophoresis. Nothing suggested any significant production of amylase in the liver. Minute amounts of amylase belonging to the pancreatic group of isoamylases might be produced by the glands of the proximal duodenum. The specific group of isoamylases produced in the female genital tract could not be demonstrated in serum or urine. The activity of amylase in serum was derived from two groups of isoenzymes, one group originating from the salivary glands, the other from the pancreatic gland. The contribution of each of these two sources to the total serum amylase was determined from early foetal life to adult age. A very low activity of the salivary isoamylases was regularly found in serum from 14-week-old foetuses. The activity increased steadily with age and reached the normal adult level, about 80 U/l, at the age of 5 years. The pancreatic group of isoamylases in serum developed later; the majority of children below 3 months had no demonstrable pancreatic isoamylase activity. The activity rose slowly to reach adult level, about 80 U/l, at the age of 10 to 15 years. The activity did not vary with sex, and the diurnal variation of the isoamylase was negligible. In children with cystic fibrosis of the pancreas the activity of pancreatic isoamylases in serum was low.

Micromodification of the Phadebas amylase test.

A micromodification of the original Phadebas Amylase Test has been adapted for the LKB Calculating Absorptiometer. This test requires only 15 mul of serum and reduces the amount of reagent used by a factor of ten. The results of this semiautomatic micromethod have a coefficient of variation of about 4% and correlate well (r = 0.996) with the method as originally described.

Effect of ultrasound guided core biopsy of prostate on serum concentration of prostate specific antigen and acid phosphatase activity.

Forty-three patients had their serum concentrations of prostate specific antigen and activity of tartrate inhibited acid phosphatase measured before and after digital rectal examination, transrectal ultrasonography and transrectal core biopsy. Transrectal core biopsy significantly increased the values for both tumor markers but rectal examination and ultrasonography without biopsy had no such effect. The measurements returned to normal within one week of biopsy in all but four patients who still had slightly increased concentrations of prostate specific antigen. We recommend that the concentration of prostate specific antigen and activity of tartrate inhibited acid phosphatase are checked before biopsy of the prostate is carried on.

Changes in amylase, hepatic enzymes and bilirubin in serum upon initiation of alcohol abstinence.

The serum levels of bilirubin, aspartate aminotransferase (ASAT), gamma-glutamyltransferase (GT), and pancreatic isoamylase were determined in 156 alcohol addicts during the first 9 days upon initiation of alcohol abstinence. During this period decreasing activities were recorded for S-bilirubin and S-ASAT but patients possessing moderatly increased serum activity of GT showed no decrease. Increased activities of S-ASAT and S-GT were still frequently seen at the end of the observation period. For S-pancreatic isoamylase, both decreases and increases from initial activities were frequently found. In patients with low activities initially, the S-pancreatic isoamylase activity increased during the observation period and thus, pathologically increased activities were recorded more often 9 days after alcohol withdrawal than on arrival at the hospital.

Serum ethanol, hepatic enzymes and length of debauch in chronic alcoholics.

The serum concentration of ethanol and the activities of aspartate aminotransferase, alanine aminotransferase and gamma-glutamyltransferase (GT) in 40 male chronic alcoholics were determined on admission to hospital. The serum activities of the enzymes were highest in patients with established alcoholism for less than 5 years. The serum concentration of ethanol, however, was lowest among these patients and gradually increased with the duration of alcoholism. No correlation was found between the serum ethanol level and the activity of any of the enzymes. The duration of the current debauch, which was shortest in cases of long-standing alcoholism, showed a positive correlation with the S-GT activity.

Relation of amylase to starch and Lycasin metabolism in human dental plaque in vitro.

Acid production activity (APA) in plaque suspensions from glucose, boiled soluble starch and hydrogenated starch hydrolysate (Lycasin) was studied in 11 subjects. Amylase (alpha-1,4-glucan 4-glucanohydrolase, EC 3.2.1.1) activity was measured in plaque and whole saliva samples from the same persons. Lycasin was found to be hydrolyzed by salivary amylase under the formation of di- and oligosaccharides, however, with a lower rate than starch. A high correlation was found between APA from glucose and from soluble starch and between APA from soluble starch and plaque amylase activity. No correlation was found between amylase activity in saliva and APA from soluble starch or between amylase activity in saliva or plaque and APA from Lycasin. APA from Lycasin was about 62% and from soluble starch about 76% of the APA from glucose. 0-25% of the total number of cultivable microorganisms from the plaque produced extracellular starch-degrading enzymes. No correlation was found between number of starch-degrading microorganisms and APA from soluble starch or between these numbers and the plaque amylase activity. By electrophoreses only amylase fractions of human origin were found in whole saliva, plaque supernatants and plaque suspensions, indicating that the microbial amylase activity in the plaque is low compared with that of salivary origin.

Isoamylases in pancreatic carcinoma and chronic relapsing pancreatitis.

Pancreatic isoamylases were determined in serum and in duodenal aspirate in 7 patients with pancreatitis. The serum activities were abnormally low in 3 cases with cancer and in 7 cases with pancreatitis. In 2 cases with normal pancreatic isoamylase activities in serum, pancreatic disease was reflected by an abnormal isoamylast pattern. Thus the serum amylast determination was abnormal in 12 of the 20 patients.

Does hypokalaemia precede delirium tremens?

Serum-potassium was determined daily in 37 chronic alcoholics when alcohol was withdrawn on admission to hospital. Serum-potassium was normal in all patients at admission but in 26 patients in whom delirium tremens developed a contiuing decrease in serum-potassium led to hypokalaemia, (mean 2.9 mmol/l) when delirium tremens started. At the end of the attack serum-potassium rapidly returned to normal. The change in serum-potassium was not associated with any variation in serum electrolytes or acidbase balance. In the 11 patients in whom delirium tremens did not develop serum-potassium concentration remained unchanged.

Improved electrophoretic resolution of human serum alkaline phsophatase isoenzymes in agarose gel by Triton X-100.

Triton X-100 caused the liver isoenzymes of alkaline phosphatase in some serum samples to separate into two fractions on agarose gel electrophoresis. One of these fractions migrated at the rate of the original one, and one migrated more slowly. The latter fraction corresponded to a fast-moving component on electrophoresis in Cellogel and seems to be identical with a slowly moving isoenzyme in starch and polyacrylamide gel electrophoresis. The migration rates of the bone, placental, and intestinal isoenzymes were unaltered, but the fractions appeared sharper.

Serum isoamylases in cystic fibrosis.

Salivary and pancreatic isoamylases were determined in the sera of 33 patients with cystic fibrosis (CF) and in 34 CF-parents. Pancreatic serum isoamylase activities were greatly decreased in patients with CF, and only two values from this group fell within the normal range. Salvary isoamylases, however, were markedly increased so that the total serum amylase activities were normal. We interpret the low pancreatic isoamylase levels in serum to reflect reduced exocrine masses in the pancreas of CF-children. In heterozygotes, the mean activity of the pancreatic isoamylase was significantly higher than in the reference group, while salivary isoamylases were within the normal range. Due to a genetic polymorphism pancreatic isoamylase may occur in two main fractions. This variant type of pancreatic isoamylase appeared more frequently in CF-heterozygotes than was expected from a reference group of blood donors.