Isoenzymes of alkaline phosphatase from purified extracts of liver, intestine, pancreas and bone of rats were determined by their isoelectric points and compared with those from serum. 1) The extracts obtained from homogenized tissues were centrifuged at 65,000xg and filtered through an Ultrogel AcA 34 column. Among the three major peaks obtained by gel filtration, the second peak fractions were further separated by isoelectric focusing. Isoenzymes of alkaline phosphatase were found only in the second peak. 2) Isoenzymes of alkaline phosphatase were distinguishable with pH 3.5-10 ampholytes. When pH 3-6 ampholytes were used, isoenzymes were more clearly separated, e.g., 4in serum, 5 in intestine and 2 each in the liver, pancreas, and bone. 3) Comparing the bands of the isoenzymes of alkaline phosphatase to those of serum, only the band with 5.04 pI was the same between serum and intestine. These results indicate that several forms of alkaline phosphatase, even though all are from the rat, may exist; and some of the isoenzymes of alkaline phosphatase found in the serum originated from the intestine.
Alkaline Phosphatase/analysis* ; Alkaline Phosphatase/blood ; Animal ; Isoenzymes/analysis* ; Isoenzymes/blood ; Rats

Macroenzymes are normal enzymes complexed with an immunoglobulin (usually IgG, rarely IgA or IgM). A number of macroenzymes have been reported in the literature. Among them, macro-AST has been detected in diseases such as acute and chronic hepatitis, various malignancies and autoimmune diseases, but usually not associated with any specific disease. We report a case of elevated AST activity in serum due to marco-AST formation in a female with chronic hepatitis C which was confirmed by AST isoenzyme electrophoresis. To our knowledge, this is the first report of macro-AST occurred in chronic hepatitis patient in Korea.
Aspartate Aminotransferases/*blood ; Female ; Hepatitis C, Chronic/*enzymology ; Humans ; Isoenzymes/blood ; Middle Aged

Alkaline Phosphatase ; blood ; Biomarkers ; blood ; Child, Preschool ; Female ; Humans ; Infant ; Isoenzymes ; blood ; Phosphorus Metabolism Disorders ; blood ; diagnosis ; Risk Factors

BACKGROUND: Intestinal alkaline phosphatase (ALP) is more prevalent in individuals of blood group B or O secretors and increases after a meal, especially, high-fat diet. The purpose of this study was to evaluate the prevalence and clinical significance of intestinal ALP in the sera of healthy adults. METHODS: Whole blood specimens were obtained from 42 healthy adults after fasting for at least 8 hours, and again at 2 hours after a regular meal. ALP was measured by TBA-200FR and analyzed for isoenzymes by Helena REP system. We also tested their ABO blood groups using GENEDIA anti-A and anti-B sera. RESULTS: The levels of fasting ALP, postprandial ALP, and the difference between the fasting and postprandial ALP (ALP difference) were 57.6+/-20.8 (12-111) IU/L, 62.3+/-17.4 (27-120) IU/L, and 4.6+/-15.4 (-8~63) IU/L, respectively. Delta (delta) ALP was 27.6+/-86.3 (-11.4~312.5)%. Among the 42 subjects, 6 were blood group A, 16 group B, 10 group AB, and 10 group O. Intestinal isoenzyme of ALP was detected in two subjects, both of whom were blood group O. The differences in fasting ALP, postprandial ALP, ALP difference, and delta ALP between ABO blood groups were statistically not significant. CONCLUSIONS: Intestinal ALP was detected in 5% of healthy adults, especially, in 20% of blood group O. Intestinal ALP has been known to be of no specific clinical significance. However, when ALP is measured in a non-fasting sample, the presence of intestinal ALP could result in an abnormally high ALP and subsequent unnecessary tests. Therefore, it is recommended that ALP should be measured only after fasting.
Adult* ; Alkaline Phosphatase* ; Blood Group Antigens ; Diet, High-Fat ; Fasting ; Humans ; Isoenzymes ; Meals ; Prevalence*

Endogenous nitric oxide(NO) plays an important role in the regulation of blood pressure. It has been known that the evoked NO-dependent dilator system may be impaired in various hypertensive models. The effects of NG-nitro-L-arginine(L-NNA), lipopolysaccharide(LPS) and tempol on mean arterial pressure(MAP) and the effects of L-NNA on isolated aorta tone were studied in order to elucidate potential alterations in resting vasodilator tone of NO in two-kidney, one clip(2K1C) hypertension. Plasma nitrite/nitrate levels were measured by colorimetric assay, and the expression of endothelial and inducible NO synthases(eNOS, iNOS) was determined by Western blot analysis. L-NNA caused an increase of MAP, while LPS produced a hypotensive effect in both 2K1C and control rats. The magnitude of the pressor or depressor response to L-NNA and LPS was comparable in the two groups. Tempol induced a sustained decrease in MAP in 2K1C rats, while it had no effects on MAP in control rats. Plasma concentrations of NO metabolites were significantly increased following the LPS-treatment in both 2K1C and control rats, while they were not affected by tempol-treatment. In endothelium-intact aortic rings precontracted with 25 mM KCl, L-NNA caused a dose-dependent contraction. The magnitude of the maximal contraction was attenuated in 2K1C rats as compared with control. An inhibition of contractile responses to L-NNA in the hypertensive group was also shown in rubbed rings, although the magnitude of contractions was markedly reduced. The vascular expression of both eNOS and iNOS was significantly decreased in 2K1C rats as compared with control. These results indicate that 2K1C hypertension is associated with a reduced basal vasodilator tone of NO and a decrease in the vascular expression of NOS isozymes.
Animals ; Aorta ; Blood Pressure ; Blotting, Western ; Hypertension ; Isoenzymes ; Nitric Oxide* ; Plasma ; Rats*

OBJECTIVE: To study the changes of serum creatinine kinase(CK) and its cardiac-specific isoenzyme compound(CK-MB) levels in crush injury rats. METHODS: Crush injury was produced in SD rats, the serum levels of CK and CK-MB were studied by automated biochemical analyzer. RESULTS: The levels of plasma CK and CK-MB were much higher in crush injury rats than those of the control group. CONCLUSIONS Cardiomyocyte injury may be induced in the early stage of crush injury rats.
Animals ; Creatine Kinase/blood* ; Creatine Kinase, MB Form ; Crush Syndrome/enzymology* ; Female ; Isoenzymes/blood* ; Male ; Rats ; Rats, Sprague-Dawley

Macroamylasemia is a condition of persistent, elevated serum amylase activity with no apparent clinical symptoms of a pancreatic disorder. In Korea, however, no such case has been reported to date. We report a case of a 17-year-old female diagnosed with macroamylasemia and acute appendicitis. One day earlier, she developed epigastric and right lower quadrant abdominal pain. She was characterized by high level of serum amylase, but normal lipase. Amylase isoenzyme analysis demonstrated increased fraction of salivary type and follow-up amylase level was persistently increased. Immunofixation disclosed the macroamylase binding with an immunoglobulin, consisting of IgA and kappa chain. The patient was treated by appendectomy, and the abdominal pain subsided.
Adolescence ; Amylases/blood* ; Appendectomy ; Appendicitis/enzymology ; Appendicitis/blood* ; Case Report ; Female ; Human ; IgA/blood ; Immunoglobulins, kappa-Chain/blood ; Isoenzymes/blood ; Protein Binding

OBJECTIVETo evaluate the diagnostic value of the changes in serum CK and its isozymes in the muscular infection and necrosis in electrical injured patients.

METHODSSeventeen patients were divided into A and B groups according to the causes, i.e. electrical injury as A and electrical arc flame burn as B groups. Obvious muscle necrosis was identified in A but not in B groups. The serum CK-MM concentration was determined after injury, before and after the operations. Simultaneously, the blood and urine routine, the hepatic and renal function and wound bacterial counting were determined and compared with those in 20 healthy people.

RESULTS1. The serum CK-MM in A group increased evidently after injury and 1 day after wound debridement to the 6 times that in normal control. The enzyme decreased to normal at 3 post-operative days in 15 cases and remained at relative high level in 2 cases due to the wound infection and lowered down to normal level after wound re-debridement. 2. The serum CK-MM in B group increased slightly before and after skin grafting.

CONCLUSIONCK-MM could be employed as the index for the infection and necrosis of the muscle in electrical injured patients due to its high specificity and sensitivity.

Adolescent ; Adult ; Burns, Electric ; blood ; enzymology ; Creatine Kinase ; blood ; Electric Injuries ; blood ; enzymology ; Humans ; Isoenzymes ; blood ; Male ; Middle Aged ; Skin Transplantation

The distribution of carbonic anhydrase (CA) isoenzymes I, II, IV, and IX was investigated in pancreatic islet of the rat using Western blotting analysis and immunohistochemistry. Western blotting analysis demonstrated strong CAI and II expression, but weak CAIV and no CAIX expression. Immunohistochemical reaction of pancreatic islet revealed no staining for CAI and II. CAIV was detected in the peripheral cells of the islet. CAIX was detected in the peripheral cells and occasional in the centrally located cells. Signals for CAIV were observed at the plasma membrane and/or in the cytoplasm of islet cells. Location of CAIV in the A cells was confirmed by subjecting serial sections of pancreas to staining for CAIV and glucagon, which showed colocalization in the A cells. Immunohistochemical staining of pancreatic acinus revealed abundant staining for CAI in interacinar blood vessels and CAII in ductal and acinar cells. These results demonstrate the differential distribution of CA isoenzymes in pancreatic islet, and suggest that A cells of pancreatic islet might contain both CAIV and IX.
Acinar Cells ; Animals ; Blood Vessels ; Blotting, Western ; Carbon* ; Carbonic Anhydrases* ; Cell Membrane ; Cytoplasm ; Glucagon ; Immunohistochemistry ; Islets of Langerhans* ; Isoenzymes* ; Pancreas ; Rats*

Studying effects of 50 Hz sinusoidal electromagnetic fields (SEMFs) with different intensities on peak bone mass (PBM) of rats may provide a theoretical basis for application of electromagnetic clinical field. 30 female SD rats, 6 weeks of age, were randomly divided into three groups: the control group, 0.1 mT electromagnetic field group (EMFs) and 0.6 mT EMFs. The EMFs groups were treated for 3 h/day. After 8 weeks, we examined their bone mineral densities (BMD) , measured their bone biomechanical properties, and made serum levels of osteocalcin (OC), tartrate-resistant acid phosphatase 5b (TRACP 5b), and histomorphometry. It was found that the BMD (P < 0.01), maximum mechanical load (P < 0.01) in the 0.1 mT group were significantly higher than those in the control group, and Yield strength (P < 0.05), the analyses of serum bone turnover markers and histomorphometric parameters were better than those in the control group (P < 0.05). However, the 0.6 mT group did not have significantly difference comparing with that in the control group. This study proved that 50 Hz 0.1 mT SEMFs can increased BMD, bone strength, and bone tissue microstructure. Therefore, 50 Hz 0.1 mT SEMFs can improve peak bone mass of rats.
Acid Phosphatase ; blood ; Animals ; Bone Density ; Bone and Bones ; physiology ; Electromagnetic Fields ; Female ; Isoenzymes ; blood ; Osteocalcin ; blood ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase