Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum.

Hoof, V O Van; Lepoutre, L G; Hoylaerts, Marc; Chevigné, R; Broe, Marc E. De

doi:10.1093/clinchem/34.9.1853

Cited by 65 publications

(20 citation statements)

References 21 publications

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“…6 Furthermore, the resistance to heat treatment of the ALP isoenzyme fraction of TH is greater than that of a normal bone fraction. 5 Taking this into account, we speculate that the fraction may be of liver origin.…”

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confidence: 88%

“…1-3 TH is considered to be present when serum ALP is >1000 IU/L; 1 furthermore, the elevation is often accompanied by elevation of both liver and bone ALP isoenzymes. 4,5 We report the case of an infant with Epstein-Barr virus (EBV)-associated TH (EBVATH).A male infant was found to have hepatomegaly at his routine health check-up at 18 months of age, and was referred to hospital by his physician. On physical examination, he was in good general condition, but the liver was palpable 5 cm below the right costal margin, and the spleen was enlarged to 3 cm below the left costal margin.…”

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confidence: 99%

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Benign transient hyperphosphatasemia associated with Epstein–Barr virus infection

Koike

Aoki

2013

Pediatrics International

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Elevated serum alkaline phosphatase (ALP) is sometimes observed in children during routine blood chemistry analysis or in patients with various illnesses, and returns to normal within several months. This condition is known as benign transient hyperphosphatasemia (TH). 1-3 TH is considered to be present when serum ALP is >1000 IU/L; 1 furthermore, the elevation is often accompanied by elevation of both liver and bone ALP isoenzymes. 4,5 We report the case of an infant with Epstein-Barr virus (EBV)-associated TH (EBVATH).A male infant was found to have hepatomegaly at his routine health check-up at 18 months of age, and was referred to hospital by his physician. On physical examination, he was in good general condition, but the liver was palpable 5 cm below the right costal margin, and the spleen was enlarged to 3 cm below the left costal margin. Laboratory findings were as follows: white blood cell count, 9900/μL; aspartate aminotransferase, 49 IU/L; alanine aminotransferase, 22 IU/L; lactate dehydrogenase, 359 IU/L; ALP, 3697 IU/L; glucose, 94 mg/dL; calcium, 10.0 mg/dL; phosphorus, 5.3 mg/dL; ferritin, 5.8 ng/mL; neuron-specific enolase, 23 ng/mL; EBV-specific antibody testing including viral capsid antigen (VCA)-IgG and -IgM, and EBV nuclear antigen (EBNA), were all negative. Serum ALP isoenzymes measured on agarose electrophoresis (Bio Medical Laboratories, Tokyo, Japan) had a slightly diffuse pattern with bimodal peaks between Fig. 1 Serum alkaline phosphatase (ALP) isoenzyme patterns in children with hyperphosphatasemia. (a) Present case: bimodal peaks between ALP 2 and 3 indicate benign transient hyperphosphatasemia. (b) Fourteen-month-old boy with serum ALP = 1319 IU/L: a single peak between 2 and 3 indicates normal bone growth in a healthy infant. (c) Seven-year-old girl with serum ALP = 1007 IU/L and Epstein-Barr virus infection: liver dysfunction with cholestasis. Serum ALP isoenzymes were measured on agarose electrophoresis (Bio Medical

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confidence: 88%

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Benign transient hyperphosphatasemia associated with Epstein–Barr virus infection

Koike

Aoki

2013

Pediatrics International

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“…Bone alkaline phosphatase activity was quantified by means of agarose electrophoresis with reagents from Analis (Suarl ee, Belgium). (33) Osteocalcin, carboxy-terminal collagen cross-links (CTX), and serum type 1 procollagen (P1NP) were measured on a Modular EE instrument (Roche Diagnostics, Vilvoorde, Belgium) using reagents from the same manufacturer.…”

Section: Biochemical Serum Parametersmentioning

confidence: 99%

A Novel Domain-Specific Mutation in a Sclerosteosis Patient Suggests a Role of LRP4 as an Anchor for Sclerostin in Human Bone

Fijałkowski

Geets

Steenackers

et al. 2016

Journal of Bone and Mineral Research

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Mutations in the LRP4 gene, coding for a Wnt signaling coreceptor, have been found to cause several allelic conditions. Among these, two are characterized by a strong skeletal involvement, namely sclerosteosis and Cenani-Lenz syndrome. In this work, we evaluated the role of LRP4 in the pathophysiology of these diseases. First, we report a novel LRP4 mutation, leading to the substitution of arginine at position 1170 in glutamine, identified in a patient with sclerosteosis. This mutation is located in the central cavity of the third b-propeller domain, which is in line with two other sclerosteosis mutations we previously described. Reporter assays demonstrate that this mutation leads to impaired sclerostin inhibition of Wnt signaling. Moreover, we compared the effect of this novel variant to mutations causing Cenani-Lenz syndrome and show that impaired membrane trafficking of the LRP4 protein is the likely mechanism underlying Cenani-Lenz syndrome. This is in contrast to sclerosteosis mutations, previously shown to impair the binding between LRP4 and sclerostin. In addition, to better understand the biology of LRP4, we investigated the circulating sclerostin levels in the serum of a patient suffering from sclerosteosis owing to a LRP4 mutation. We demonstrate that impaired sclerostin binding to the mutated LRP4 protein leads to dramatic increase in circulating sclerostin in this patient. With this study, we provide the first evidence suggesting that LRP4 is responsible for the retention of sclerostin in the bone environment in humans. These findings raise potential concerns about the utility of determining circulating sclerostin levels as a marker for other bone-related parameters. Although more studies are needed to fully understand the mechanism whereby LRP4 facilitates sclerostin action, it is clear that this protein represents a potent target for future osteoporosis therapies and an interesting alternative for the antisclerostin treatment currently under study.

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“…It was recently reported that this band is HIALP. In 1988 Van Hoof et al (1) employed neuraminidase treatment and polyclonal and monoclonal antibodies against the intestinal type and confirmed that the band overlapping bone ALP on agarose electrophoresis is an intestinal variant of ALP. In 1998 Hoshino et al (2) confirmed that this band is a high-molecular band by polyacrylamide gel electrophoresis (PAGE), and Matsushita et al (3) reported that the band could be separated from bone ALP by conversion to ALP5 by subtilisin treatment.…”

Section: Introductionmentioning

confidence: 99%

High‐molecular intestinal alkaline phosphatase in chronic liver diseases

Ooi

Shiraki

Morishita

et al. 2007

Clinical Laboratory Analysis

186

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The presence of high-molecular intestinal alkaline phosphatase (HIALP) different from bone ALP detected in the alpha(2)beta region was recently clarified. In this study we used a novel method in which HIALP was detected after conversion to ALP(5) by protease to investigate the clinical significance of the appearance of HIALP in patients with chronic liver disease. The subjects were 241 patients with chronic liver disease. When a decrease in ALP(3) in the alpha(2)beta region and an increase in ALP(5) in the beta region were noted, the patient was judged HIALP-positive. In the patients with chronic liver disease, the total ALP activity (T-ALP) increased with progression of the pathology in the order of chronic hepatitis (CH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). HIALP appeared in 22.4% and 49.3% of patients with CH and LC, respectively, but the positivity rate decreased to 30.4% in HCC. As autoimmune liver diseases, primary biliary cirrhosis (PBC) and autoimmune hepatitis (AIH) were investigated. T-ALP was lower in PBC+AIH than in LC and HCC, but the HIALP-positive rate was high (44.4%). The HIALP-positive rate was dependent on ABO blood groups, and was high in blood groups B and O. In conclusion, the HIALP-positive rate was particularly high in patients with chronic liver disease, and was related to the pathological progression, which suggests that the method is clinically useful.

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Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum.

Cited by 65 publications

References 21 publications

Benign transient hyperphosphatasemia associated with Epstein–Barr virus infection

Benign transient hyperphosphatasemia associated with Epstein–Barr virus infection

A Novel Domain-Specific Mutation in a Sclerosteosis Patient Suggests a Role of LRP4 as an Anchor for Sclerostin in Human Bone

High‐molecular intestinal alkaline phosphatase in chronic liver diseases

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