Toshiaki Yoshida

Autoantibodies against a 210kDa glycoprotein of the nuclear pore complex as a prognostic marker in patients with primary biliary cirrhosis

It has been reported that the presence of anti‐nuclear antibody against a 210kDa glycoprotein of nuclear pore complex (anti‐gp210) is highly specific for primary biliary cirrhosis (PBC). The aim of the present study was to investigate the significance of anti‐gp210, especially as a prognostic marker. The presence of anti‐gp210 was ascertained in 113 patients with PBC and 162 controls by indirect immunofluorescence assay using HepG2 cells and immunoblotting analysis using nuclear extracts from HeLa cells. Anti‐gp210 was detected in 25 of the 113 (22.1%) patients. None of the 162 controls was positive for anti‐gp210. The appearance and titre of anti‐gp210 in the patients with PBC did not vary from the time of diagnosis and through their clinical course. Anti‐mitochondrial antibodies (AMA), including antibodies against pyruvate dehydrogenase complex, branched chain α‐ketoacid dehydrogenase complex and α‐ketoglutarate dehydrogenase complex, were not detected by enzyme‐linked immunosorbent assay in five of the 113 (4.4%) patients with PBC. However, anti‐gp210 alone was positive in one of these five patients. The difference in prognosis was statistically significant; patients with PBC positive for anti‐gp210 died from hepatic failure more frequently than those who were negative (P < 0.01), although there were no statistically significant differences in the frequency of jaundice and the histological stage at the time of diagnosis between the two groups. We suggest that the presence of anti‐gp210 is one of the independent prognostic markers able to predict, at the time of diagnosis, a poor outcome in patients with PBC.

Two patients with severe alcoholic hepatitis accompanied by hypercytokinemia and granulocytic hyperelastasemia, successfully treated by intravenous infusion of urinastarine (Miraclid)

Abstract Severe alcoholic hepatitis (SAH) is not simply a disease of the liver, but it also causes infection and multiple organ failure, and therefore carries an extremely poor prognosis. We report the successful treatment of two patients with SAH. Case 1: The patient was a 55‐year‐old man. He was a heavy drinker whose alcohol intake had increased for some 3 years to 1.8 L sake a day. Slight clouding of consciousness, fever, and jaundice were evident on his admission to our hospital. Laboratory data showed leukocytosis with a predominance of polymorphonuclear leukocytes, and a decline in coagulability. He tested negative for various hepatitis virus markers. With the diagnosis of SAH made, steroid pulse therapy and bilirubin adsorption therapy were administered. The jaundice improved and the interleukin‐8 (IL‐8) level decreased. Continuous intravenous infusion of urinastarine (Miraclid) normalized the granulocyte elastase level. Improvement was also seen in coagulability, ascites, icterus and consciousness. Case 2: The patient was a 49‐year‐old man. He was a heavy drinker whose alcohol intake had increased for 1 month. Fever, jaundice, ascites, and mild disturbance of consciousness were evident at the time of admission. Examination on admission diagnosed SAH. Bilirubin adsorption and continuous intravenous infusion of urinastarine were initiated. As a result, circulating IL‐8 level was decreased and coagulability was improved. Therapy for granulocytic hyperelastasemia and hypercytokinemia supervening on SAH is a new effective approach on improvement in the disease.

Utility of spermidine in PCR amplification of stool samples.

When nucleic acid is amplified from clinical samples by PCR, it is necessary to suppress the action of any inhibitory substances in the amplification reaction. Inhibitory substances of PCR are present in clinical samples such as stool, and in some cases inhibitors of PCR cannot be removed (Wilson 1997; Verweij et al. 2000). Extraction or purification of nucleic acids from stool has been reported in a variety of articles describing the use of an ion-exchange column (Olive 1989), immunomagnetic separation (Widjojoatmodjo et al. 1992), dilution (Wilde et al. 1990; Machiels et al. 2000), cellulose fiber paper (Wilde et al. 1990), anion-binding resin (Dowd et al. 1998), magnetic bead-based hybridization capture (Ahlquist et al. 2000), and electrophoretic capture affinity protocol (Kent Moore et al. 2008). In addition, several substances, such as bovine serum albumin (McKeown 1994; Kreader 1996; Abu Al-Soud and Radstrom 2000) and T4 gene 32 protein (Panaccio and Lew 1991; Kreader 1996), have been used to alleviate the effects of inhibitory substances during the PCR steps. Ahokas and Erkkila (1993) reported that spermidine shows a concentrationdependent effect on the yield and specificity of PCR in plant material. Spermidine is a type of polyamine that interacts with DNA phosphate groups (Ouameur and