Keisuke Watanabe

Measurement of serum levels of des‐γ‐carboxy prothrombin in patients with hepatocellular carcinoma by a revised enzyme immunoassay kit with increased sensitivity

Des‐γ‐carboxy prothrombin (DCP) is a useful tumor marker for hepatocellular carcinoma (HCC). The conventional enzyme immunoassay (EIA) kit for DCP lacks adequate sensitivity to detect small HCC. Thus, a revised EIA kit for DCP has been developed. In this revised DCP kit, the blank value has been reduced, making it now possible to obtain a normal value. The authors used this revised EIA kit for DCP with increased sensitivity and evaluated its usefulness as a tumor marker for HCC.

Apoptosis under hypercytokinemia is a possible pathogenesis in influenza-associated encephalopathy.

Abstract  Background : Influenza‐associated encephalopathy is reported to be frequent in Japan and East Asia. No evaluating markers except interleukin (IL)‐6 and tumor necrosis factor (TNF)‐α and no likely pathological mechanism for the disease have yet been elucidated.

Usefulness of ED036 kit for measuring serum PIVKA-II levels in small hepatocellular carcinoma

As a tumor marker for hepatocellular carcinoma (HCC), serum protein induced by vitamin K absence or antagonist-II (PIVKA-II) has high specificity, yet its sensitivity is relatively low, making it less suitable to serve as an adjunct in the diagnosis of small HCC, Recently, the ED036 kit (Eisai, Tokyo, Japan), whose detection limit is approximately ten times superior to that of a conventional kit (Eitest MONOP II, Eisai) has been developed. In this study, serum PIVKA-II levels in serum samples from 83 patients with benign chronic liver diseases (CLD) and 129 patients with HCC were measured with these two kits. With the ED036 kit, the cut-off value was set at 40 mAU/ml. For PIVKA-II measured with the ED036 kit, sensitivity was 45.0%, specificity 92.8%, and accuracy 63.7%, when we discriminated patients with HCC from those with CLD without HCC. While maintaining a high specificity, of 92.8%, the ED036 kit showed a significantly higher sensitivity than the conventional kit (45.0% versus 27.9%;P<0.0001). With patients who had HCC consisting of a single nodule 30mm or less in diameter, the positivity rate for serum PIVKA-II with the ED036 kit was significantly greater than the rate with the conventional kit (21.4% versus 9.5%;P<0.005). Thus, the ED036 kit was thought to be more useful than the conventional kit as a tumor marker for small HCC.

Cytochrome c is a possible new marker for fulminant hepatitis in humans

BackgroundCytochrome c is known as a substance related to apoptosis. We investigated serum cytochrome c levels in patients with fulminant hepatitis (FH) compared with these levels in patients with acute or chronic liver diseases.MethodsSerum cytochrome c was measured by an electrochemiluminescence immunoassay (ECLIA) method. The numbers of patients were as follows: fulminant hepatitis (FH; n = 15), acute hepatitis (AH; n = 12), chronic hepatitis (CH; n = 30), chronic hepatitis with acute aggravation (CHA; n = 6), liver cirrhosis (LC; n = 30), hepatocellular carcinoma (HCC; n = 30), and healthy volunteers (controls; n = 9).ResultsThe serum cytochrome c level in FH was 10 686 ± 7787 pg/ml, with a significant difference (P < 0.01) compared to levels in the other groups. In the FH patients, the serum cytochrome c level was significantly correlated to serum mitochondria (m)-GOT, hepatocyte growth factor (HGF), aspartate aminotransferase (AST), lactic dehydrogenase (LDH), and alkaline phosphatase (ALP), and it was negatively correlated to serum alpha-fetoprotein (AFP), and total bilirubin (T.Bil.) The serum cytochrome c level seemed to parallel the severity of hepatic coma. Immunohistochemical study indicated TdT mediated dUTP nick end labeling (TUNEL)-positive cells in the livers of patients with FH.ConclusionsThese results suggest that serum cytochrome c may be a possible new marker for acute liver failure.

Des‐gamma‐carboxy prothrombin (DCP) ratio, a novel parameter measured by monoclonal antibodies MU‐3 and 19B7, as a new prognostic indicator for hepatocellular carcinoma

Abstract: Background/Aim:  Recently, a novel des‐gamma‐carboxy prothrombin (DCP) antibody named 19B7 has been developed. It has been reported that the DCP ratio defined by DCP measured by MU‐3 antibody/DCP measured by 19B7 antibody demonstrated differences between hepatocellular carcinoma and vitamin K deficiency. The aim of this study was to clarify the relationship between the DCP ratio and tumour malignancy, as well as the prognostic significance of the DCP ratio after hepatectomy for hepatocellular carcinoma. Methods: From January 1996 to December 1999, 79 patients diagnosed with hepatocellular carcinoma underwent hepatectomy. To detect DCP in the plasma before surgery, we used a new EIA kit, Eitest PIVKA‐II kit (Eisai, Tokyo, Japan) with both MU‐3 and 19B7 antibody. The DCP ratio was calculated using the formula: DCP ratio = DCP level measured by MU‐3 antibody (mAU/ml)/DCP level measured by 19B7 antibody (mAU/ml). Pathologic findings using resected specimens were classified according to the Japanese General Rules for Primary Liver Cancer. Results: Among DCP‐positive patients, the DCP ratio was significantly higher in those with tumour thrombus in the portal vein (vp) or with a moderately or poorly differentiated hepatocellular carcinoma than in those without vp or with a well‐differentiated carcinoma (P = 0.0162, 0.0109, respectively). Patients with DCP ratio > 3 had a significantly higher incidence of vp than those with DCP ratio ≤ 3 (P = 0.0027). The survival rate was significantly higher in patients with DCP ratio ≤ 3 than in those with DCP ratio > 3 (P = 0.0002). This result was the same as in DCP positive cases (P = 0.0027). Conclusions: The DCP ratio can distinguish DCP produced by hepatoma cells from DCP produced by normal hepatocytes in vitamin K deficiency. The DCP ratio is a useful prognostic indicator that can be assessed before hepatectomy.

Usefulness of simultaneous determination of α-fetoprotein and des-γ-carboxy prothrombin in hepatocellular carcinoma

Serum γ-fetoprotein (AFP) and plasma des-γ-carboxy prothrombin (DCP), a protein induced by vitamin K absence or antagonist II (PIVKA-II) levels, were measured in 197 patients with primary hepatocellular carcinoma (HCC). DCP levels were determined by conventional enzyme immunoassay kit (E-1023) and a newly developed high-sensitivity kit using the avidin-biotin complex method. Cut-off levels of AFP and DCP by the E-1023 kit and of DCP by the high-sensitivity kit were put at 100 ng/ml, 0.1 arbitrary unit (AU)/ml, and 0.004 AU/ml, respectively. Positive rate of AFP and DCP by the E-1023 kit and the high-sensitivity kit for HCC was 48%, 44%, and 57%, respectively. The positive rate by combination assay with AFP and DCP by the high-sensitivity kit increased up to 73%. There was no correlation between serum levels of AFP and those of plasma DCP. A significant correlation between tumor size and DCP levels was observed, but not with AFP. The postoperative disease-free survival rates of patients in the group with elevated levels of AFP and DCP were lower than those with normal levels of AFP and DCP. There were various patterns of change in the AFP and DCP levels at the time of recurrence compared with preoperative patterns. The combination assay of AFP and DCP levels is useful for the diagnosis, prognosis, and postoperative monitoring for recurrence of HCC.

Focal epilepsy resulting from a de novo SCN1A mutation.

We found a DE NOVO missense mutation of the gene encoding the alpha1 subunit of the neuro-nal voltage-gated sodium channel, SCN1A, in a patient with repetitive focal seizures. At 5 months of age, the patient had a first seizure characterized by loss of consciousness and clonic convulsions in the left hand followed by secondary generalization lasting for 20 minutes in association with pyrexia. Although valproate was administered, she has had generalized seizures every month, mostly in association with elevated body temperature. Since 32 months of age, she also had a different type of seizure characterized by a fearful response followed by decreased consciousness, pallor, and salivation. Myoclonia or atypical absence seizures have never been observed until the last follow-up at 42 months of age. Genetic analysis showed a heterozygous missense mutation (c.5311A>T: I1771F) in the patient, which was not detected in her parents.

Characteristics of the PIVKA-II found in hepatocellular carcinoma, investigation using monoclonal antibodies MU-3 and 19B7

Abstract We classified PIVKA-II into two subtypes using a new monoclonal antibody (19B7) to PIVKA-II and a previously available monoclonal antibody (MU-3) and demonstrated differences in composition of PIVKA-II subtypes between HCC and benign liver diseases. These two monoclonal antibodies recognize almost the same site of PIVKA-II molecule, and the three-dimensional conformation of this site due to Gla formation might be important in differences of recognition by the two antibodies.