Levels of vitamin K, immunoreactive prothrombin, des-γ-carboxy prothrombin and γ-glutamyl carboxylase activity in hepatocellular carcinoma tissue

Abstract To clarify the mechanism of production of des-γ-carboxy (abnormal) prothrombin (DCP) by hepatocellular carcinoma (HCC), we measured the levels of vitamin K, DCP, immunoreactive prothrombin and the activity of γ-glutamyl carboxylase in liver tissues from HCC patients and in the medium of cultured human hepatoma cells. There was no significant difference in vitamin K (K₁, MK-4) contents between HCC and non-HCC cirrhotic liver tissues. The activity of γ-glutamyl carboxylase per unit amount of endogenous microsomal prothrombin precursor was decreased in HCC tissue compared with non-HCC liver tissue (positive plasma DCP: 335 ± 72 vs 372 ± 67, negative plasma DCP: 370 ± 84 vs 393 ± 56 nmol/min per mg prothrombin precursor, P > 0.05), although the total incorporation of ¹⁴COOH into microsomal precursor protein was higher in the former. By contrast, levels of DCP and immunoreactive prothrombin in HCC tissue were greater ( P > 0.05) than those in non-HCC cirrhotic liver tissue. Furthermore, production of large amounts of immunoreactive prothrombin was observed in human hepatoma cells huH-1 and huH-2, which produced large amounts of DCP. These results suggest that there was excessive synthesis of prothrombin precursors by human HCC tissue and hepatoma cell lines huH-1 and huH-2. Thus, excessive synthesis of prothrombin precursors seems to be the main mechanism of DCP production by HCC.     

Epstein-Barr virus infection,Hodgkin's disease,non-Hodgkin's lymphoma,and reactive follicular hyperplasia in Japanese children: Evaluation of paraffin-embedded specimens using polymerase chain reaction and immunohistochemistry

Epstein-Barr virus (EBV) infections are common in Japanese children, with infections by EBV type 1. The relationships between EBV infection and lymphadenopathies in Hodgkin's disease (HD), non-Hodgkin's lymphomas (NHL), reactive follicular hyperplasia (RFH), and infectious mononucleosis (IM) in 37 Japanese children were evaluated. Formalin-fixed, paraffin-embedded lymph node specimens that were obtained at surgical resection or biopsy were evaluated for the presence of EBV DNA and the latent membrane protein-1 (LMP-1) using polymerase chain reaction (PCR) and immunohistochemical staining. The PCR detected EBV DNA in nine of 13 (69.2%) patients with RFH, including a case of IM, all three (100%) patients with HD, and one of 21 (4.8%) patients with NHL. All EBV-positive samples contained EBV type 1. Reed-Sternberg's cells in HD were immunohistochemically positive for LMP-1, whereas all cases of RFH and NHL were negative for LMP-1. Results suggest that EBV infection may be related to HD. Although no proof exists that EBV infection contributes to the transformation of cells, thus causing RFH or NHL, the present authors suggest that the EBV-positive cases in Japanese children demonstrate a relationship between the clinical and histopathological features of the lymphadenopathy and EBV-type 1 infection.     

Three novel and six common mutations in 11 patients with methylmalonic acidemia

BACKGROUND: Patients with a defect in methylmalonyl-coenzyme A mutase (MCM) are classified as having methylmalonic acidemia, which is divided into two subclasses: mut(0) and mut(-). Fifty-five disease-causing mutations have been identified. Although most are private mutations, only three (E117X, G717V, and N219Y) are reportedly common in Japanese, Black, and Caucasian populations, respectively. Here we identified mutations in 11 Japanese patients with MCM deficiency. METHODS: Mutational analysis was performed in 11 unrelated Japanese patients with MCM deficiency using polymerase chain reaction and direct sequencing. RESULTS: Three novel (L494X, R727X, and 449_461del) and six previously reported (R93H, E117X, N219Y, R369H, G648D and IVS2 + 5G>A) mutations were identified. The L494X mutation was found in three unrelated patients, and the R93H, E117X, R369H, G648D, and IVS2 + 5G>A mutations occurred more than once. Two of the patients were classified as mut(-) phenotype because of residual [(14)C]-propionate incorporation in the presence of a high concentration of hydroxocobalamin. The two mut(-) patients were heterozygous for the G648D mutation and presented with lethargy and metabolic acidosis after 2 years of life. Their psychomotor development has been documented as normal. The patients with the R727X or c.374_385del [corrected] mutations clinically exhibited mut(0) phenotype. Two patients with mut(0) phenotype died in infancy. One presented early in the neonatal period; the other was symptomatic in the late infantile period. CONCLUSIONS: The L494X, R93H, E117X, R369H, G648D, and IVS2 + 5G>A mutations are found in more than two unrelated families in the Japanese population. The short-term outcome was generally poor in patients with mut(0), and therefore alternative treatments should be considered.     

Transvenous Dual Chamber Pacing via a Unilateral Left Superior Vena Cava

A 74-year-old woman with a unilateral left superior vena cava required dual chamber permanent pacing after a radical cardiac operation for an incomplete form of endocardial cushion defect. An active fixation ventricular lead was used to prevent the instability induced by the strange course of the electrode. For atrial pacing, a ventricular passive fixation lead was used. A transvenous dual chamber pacemaker was successfully inserted via a unilateral left superior vena cava.     

Vagal Enhancement as Evidence of Residual Ischemia After Inferior Myocardial Infarction

Background: Acute inferior myocardial infarction (MI) often induces transient sinus bradycardia through vagal enhancement, known as Bezold-Jarisch reflex, which is explained by preferential distribution of vagal nerve in the inferior wall. We examined vagal activity in relation to the occurrence of residual ischemia in patients with old inferior MI and assessed its diagnostic usefulness.
Methods: Exercise myocardial scintigraphy was performed in 15 patients with old inferior MI, 19 angina pectoris (AP) patients with inferior ischemia but no MI, and 32 control subjects who had no evidence of cardiac disease. We analyzed the connection of residual ischemia in old MI with ST-segment response to exercise and with vagal activity as determined by coefficient of component variance of high frequency (CCV_HF).
Results: Exercise-induced percentage change in CCV_HF was higher in patients with old MI and residual ischemia (18.8 ± 13.5%) and AP (5.5 ± 9.7%) than old MI but no residual ischemia (–24.1 ± 4.9%) or control (–22.8 ± 4.5%, P = 0.006). Percentage change in CCV_HF > –5% had a good diagnostic value for the detection of residual ischemia in patients with old inferior MI with sensitivity of 83%, specificity of 89%, accuracy of 87%, and positive likelihood ratio of 7.50, which was higher than that of ST-segment depression (67%, 50%, 56%, and 1.33).
Conclusions: Vagal enhancement was associated with residual ischemia in old inferior MI as well as inferior AP. Measurement of CCV_HF is useful in improving the diagnostic reliability of exercise electrocardiography in patients with old inferior MI.     

Further studies on the resolution of human mercapt- and nonmercaptalbumin and on human serum albumin in the elderly by high-performance liquid chromatography

High-performance liquid chromatographic (HPLC) analysis of human serum albumin (HSA) on Asahipak GS-520 showed at least two peaks, the principal component corresponding to human mercaptalbumin (HMA) and the secondary one to nonmercaptalbumin (HNA). HPLC analysis of HSA on Asahipak ES-520 N showed three peaks, the principal component corresponding to HMA, the secondary one to HNA having mixed disulfide with cysteine or glutathione and the tertiary one to HNA oxidized higher than mixed disulfide. Two kinds of rapid HPLC for the resolution of HSA into HMA and HNA were developed by the present authors. Using these HPLC, the present authors found a significant decrease in the fraction of HMA in the elderly.     

Overall pesticide effects on growth and emergence of two species of Ephemeroptera in a model stream carrying pesticide- polluted river water

The overall pesticide effects on mortality, growth and emergence of two dominant species of Ephemeroptera in Japanese rivers, Epeorus latifolium and Ecdyonurus yoshidae, were assessed using an outdoor channel carrying water from the Kokai River. Young larvae collected from another river were introduced into cages in the channel after their body lengths were measured. The concentrations of 17 pesticides were measured three times a week from April to August 1993. A shrimp mortality test on water samples was conducted concurrently. A relationship between the high mortality of the shrimp in the water samples and of E. latifolium in the channel was recognized. The mortality of E. yoshidae increased only when the shrimp mortality increased drastically in early June, reflecting the difference in insecticide susceptibility between the two mayfly species. Almost all the larvae which had been introduced into the channel in winter and/or early spring, when pesticides had cleared from the river, emerged in spring, although their growth rate during the winter was low. The shrimp mortality in the river water samples was caused by the overall pesticide toxicity. The increase in the mortality of the mayfly larvae in the channel might be due to the overall pesticide toxicity, although their concentrations were low and varied independently