Anti-interferon-α neutralizing antibody is associated with nonresponse to pegylated interferon-α plus ribavirin in chronic hepatitis C.

Pegylated interferon-α (PEG-IFN-α) plus ribavirin (RBV) treatment fails to achieve a sustained virological response (SVR) in approximately 20-50% of patients with chronic hepatitis C virus (HCV) infection. We assessed the contribution of an anti-IFN-α neutralizing antibody (NAb) on the nonresponse to treatment. NAbs were detected using an antiviral assay that assessed the neutralizing effects of serum samples against IFN. Serum samples were obtained at the end of the treatment and evaluated for the presence of NAbs using recombinant IFN-α as a standard. We studied 129 PEG-IFN-α/RBV-treated patients. In the 82 end-of-treatment responders, no NAbs were detected. Of the 47 patients who did not respond, seven (15%) were positive for NAbs. We also examined an additional 83 patients who had not responded to PEG-IFN-α treatment, and detected 12 with NAbs. Patients with good IFN-responsive characteristics, including HCV genotype 2/3 and major allele homozygotes for interleukin-28B, were included in the 19 patients with NAbs. No NAbs interfered with the antiviral activity of natural human IFN-ß (nIFN-ß) and re-treatement of patients with NAbs with nIFN-ß/RBV achieved SVR. Our analyses revealed that the emergence of anti-IFN-α NAbs was a candidate causal factor of PEG-IFN-α-treatment failure. Therefore, these antibodies should be assayed in patients who do not respond to PEG-IFN-α therapy, and if detected, other effective treatments, i.e., medications that are not neutralized by anti-IFN-α NAbs, should be considered.

Necrotizing fasciitis caused by Cryptococcus neoformans in a patient with pemphigus vegetans.

Cryptococcosis occurs most often in immunocompromised people. The cutaneous features of cryptococcosis include papules, pustules, nodules, subcutaneous swelling, abscesses, molluscum contagiosum-like or tumour-like lesions, cellulitis, blisters, ulcers and very rarely, necrotizing fasciitis (NF). NF is a destructive soft-tissue infection that is most typically caused by group A streptococci or by a combination of facultative and anaerobic bacteria. We present the case of a 55-year-old woman with pemphigus vegetans, who developed cryptococcal NF in the legs. She had been treated with immunosuppressants including plasmapheresis and pulse therapy with steroid and cyclophosphamide. Cryptococcal NF localized to the legs is very rare. Because diagnosis and treatment of cryptococcal infection is often delayed, clinicians should be aware of the possibility of cryptococcal infection when antibacterial therapy is not effective in an immunocompromised patient.

Annular leucocytoclastic vasculitis.

Leucocytoclastic vasculitis (LV) is characterized by necrotizing inflammation around small blood vessels, composed mainly of neutrophils and their debris. The skin lesions of LV are polymorphous, but an annular variant is rare. Four previous reports have named this variant 'annular LV' (ALV) and in this paper, we report two additional cases. Patient 1 was a 80-year-old man, who presented with pruritic, erythematous target lesions on his legs, soles, abdomen and axillae; histological examination showed typical LV throughout the entire dermis. Oral prednisolone 15 mg daily rapidly resolved the patient's symptoms. Patient 2 was a 64-year-old man, who was referred to our hospital because of multiple purpuric target lesions on the buttocks, right thigh, lower legs, upper arms and forearms. Histopathology revealed LV throughout the entire dermis. Oral prednisolone 20 mg daily rapidly resolved the lesions. We categorized these cases morphologically as ALV and clinically as small-vessel vasculitis. ALV is not a distinct condition, but includes a broad range of small-vessel vasculitides.

Epithelioid hemangioendothelioma of the liver associated with thrombocytopenia and coagulopathy.

Epithelioid hemangioendothelioma of the liver is a rare vascular neoplasm with intermediate malignant potential. The prognosis is highly unpredictable. We report the case of a 59-year-old woman who had the tumor radically resected, but multiple metastases of the liver developed associated with thrombocytopenia and consumption coagulopathy, as observed in Kasabach-Merritt syndrome. The patient did not respond to any treatment and the behavior of the tumor was very aggressive. The patient died 15 months after radical resection of the tumor.

Idiopathic hemochromatosis with the mutation of Ala176Val heterozygous for HFE gene.

Most patients with hereditary hemochromatosis are homozygous for C282Y in the HFE gene in populations of Celtic origin, but the genetic cause of this disease is unknown in Japan because of its rarity. A 48-year-old Japanese patient was recently diagnosed with idiopathic hemochromatosis. Analysis of the entire coding region of the patient's HFE by RT-PCR showed a heterozygous nucleotide substitution at nucleotide 527 from C to T, which resulted in A176V amino acid substitution. Another mutation at nucleotide 942 from T to C was observed, but this was a nonsense mutation. C282Y and another mutation, H63D, were not found in the patient. The mutation may have a possible role on the cause of hemochromatosis in this Japanese case.

Development of orally active nonpeptidic inhibitors of human neutrophil elastase.

5-Amino-2-phenylpyrimidin-6-ones, some of their desamino derivatives, and miscellaneous derivatives were synthesized and biologically evaluated on both in vitro activity and oral activity in an acute hemorrhagic assay. These compounds contained an alpha-keto-1,3,4-oxadiazole moiety to bind covalently to the Ser-195 hydroxy group of human neutrophil elastase (HNE). Among those tested, compounds 11a-c,e,i-l(F), 11d,e,k(H), 21d,e,k(F), and 21d,e(H) showed a good oral profile. RS-Mixture 3(H) was selected for clinical evaluation based on its oral potency, duration of action, enzyme selectivity, safety profile, and ease of synthesis. Structure-activity relationships (SARs) are discussed.

Investigation of TTV by in situ hybridization in patients with chronic hepatitis.

To clarify whether TT virus (TTV) was present in liver tissues, 12 liver tissue samples from patients with chronic hepatitis positive for TTV in their serum and 11 samples from serum-negative patients were obtained by needle biopsies and investigated using in situ hybridization. Positive staining was observed in nine (75%) of 12 cases positive for TTV (serum-positive group) and three (27.3%) of 11 cases negative for TTV (serum-negative group) (P=0.061). Three kinds of staining patterns were observed: nuclear, cytoplasmic and both. In 58.3% (7/12) of the patients positive for TTV staining, the stained areas were found in both the nucleus and cytoplasm. Only cytoplasmic staining was observed in three cases from the serum-positive group. Only nuclear staining was observed in two cases from the serum-negative group. No significant differences were found in the clinical background between the in situ hybridization-positive and -negative groups, and between the serum-positive and -negative groups. The present study shows that TTV exists in the liver tissue, especially in hepatocytes, of chronic hepatitis patients and that the localization of TTV in the cell is different from case to case, although why this is so remains to be clarified.

The tumor-derived fetal-intestinal alkaline phosphatase cDNA is identical in sequence to the adult intestinal alkaline phosphatase isozyme gene.

The alkaline phosphatase (AP) of Caco-2 cells, a cell line derived from a human adenocarcinoma of the colon, is quite similar to fetal intestinal AP in its enzymatic properties. The nucleotide sequence of a cDNA encoding AP produced in Caco-2 cells was examined. The sequence was identical to one of the three sequences of adult intestinal AP reported previously. We further investigated the entire nucleotide sequence of cDNA of intestinal-type AP produced in cancer cell lines such as HuH-7 cells, FL-amnion cells, and HuG-1 cells. The sequence of these cell APs was identical to that of Caco-2 cell AP. These results indicate that cancer cells producing intestinal-type AP have the same nucleotide sequence as that of adult intestinal AP, and suggest that the differences in electrophoretic mobilities of these cell APs compared with adult intestinal AP may be due to post-translational modifications.

Tumor necrosis factor alpha alters the cytotoxic effect of hydrogen peroxide in cultured hepatocytes.

We examined whether tumor necrosis factor-alpha (TNF) affects the cytotoxic capacity of reactive oxygen species on rat hepatocytes in culture. Both TNF and reactive oxygen species are involved in many inflammatory events including hepatic ischemia/reperfusion injury and endotoxic shock. Synchronous treatment of hepatocytes with both TNF and H2O2 demonstrated that TNF (2000 ng/ml) enhanced the cytotoxic effect of H2O2 (500 microM). By contrast, pretreatment with TNF (2000 ng/ml) for 24 h followed by exposure to H2O2 (1000 microM) reduced the reactive oxygen-induced cytotoxicity. We conclude that TNF increases the effects of reactive oxygen-induced cytotoxicity when exposed synchronously, whereas TNF pretreatment induces a cytoprotective effect to reactive oxygen species, presumably by up-regulation of the reduced form of glutathione levels in hepatocytes.

Familial hypocholinesterasemia found in a family and a new confirmed mutation.

A 45-year-old man was hospitalized because of acute hepatitis. His serum cholinesterase (ChE) was below 10 IU/l (normal range: 105-240 IU/l) during the disease course and after his recovery. The patient was suspected of having familial hypocholinesterasemia. His family members were healthy except that his father had hypertension and gall stones. Analysis of ChE gene in the propositus and his family revealed three point mutations at nucleotides 298 (CCA to TCA), 1,410 (CGT to CGG) and 1,615 (GCA to ACA). The first mutation caused an amino acid change at codon 100 from proline to serine, which was a new mutation not previously reported, but the second one was a silent mutation. The third mutation resulted in an amino acid alteration from alanine to threonine at codon 539 in exon 4 of the ChE gene. The mode of transmission of these mutations is described.

Reduced gastric surface mucus layer in experimental portal hypertension.

The pathogenesis of portal hypertensive gastropathy has not yet been thoroughly elucidated. Changes in the gastric surface mucus layer in prehepatic portal hypertensive and cirrhotic rat models were studied by observing frozen sections fixed with formaldehyde vapor and stained with hematoxylin and eosin. We produced prehepatic portal hypertensive rats by partial ligation of the portal vein, and cirrhotic rats by prolonged administration of carbon tetrachloride (CCl4) and phenobarbital sodium. The thickness of the corporal and antral gastric surface mucus was significantly reduced in prehepatic portal hypertensive and cirrhotic rats compared with the values obtained in control rats for portal hypertension (subjected to sham operation) and control rats for cirrhosis (treated with phenobarbital but not CCl4). These results indicate that the hemodynamic changes associated with portal hypertension reduce the thickness of the gastric surface mucus layer and may be one of the causes of the gastropathy associated with portal hypertension.

Alzheimer-type pathology in a patient with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS).

A 53-year-old Japanese woman with a point mutation in mitochondrial DNA (tRNALeu(UUR), nt3243) consistent with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) and Alzheimer-type brain pathology is reported. This woman had suffered myopathy and psychosis without any clinical evidence of, stroke-like episodes during the last 10 years of her life, and had died after an accident. At autopsy 30 h post mortem, a part of the brain was snap frozen for biochemical and histochemical studies, and the remaining part was processed for a routine examination and electron microscopy. In the brain there were no ischemic lesions. Instead, primitive/diffuse senile plaques were found throughout the brain, predominantly in the frontal and temporal lobes, while Alzheimer neurofibrillary tangles were found only in the parahippocampal gyrus. These plaques were positive for beta-protein and mostly negative for tau protein, ubiquitin, neurofilaments, alpha-choline acetyltransferase, and acetylcholinesterase. Mutations in codon 331 of the ND2 gene as well as codons 693, 713 and 717 of the beta-amyloid precursor protein gene, known to be responsible for some cases of familial Alzheimer disease, were not found. Furthermore, coincidental Down syndrome was ruled out by chromosome analysis. The results suggest a possible correlation between this mitochondrial DNA abnormality and Alzheimer-type pathology.

Management of new hepatic nodules detected by intraoperative ultrasonography during hepatic resection for hepatocellular carcinoma.

BACKGROUND: During hepatic resection for hepatocellular carcinomas (HCCs) it is not uncommon that intraoperative ultrasonography detects "new nodules" that were not found by preoperative examinations. Because the operative procedure may have to be changed if the new nodule is another HCC lesion, differential diagnosis of such nodule is critical. This study examines ultrasonographic findings and clinical features of new nodules and discusses how to cope with such nodules in the operating room. METHODS: Fifty-one new nodules detected in 92 liver resections were analyzed. Intraoperative ultrasonography was performed by using 5.0 or 7.5 MHz probes after mobilization of the liver. Histologic diagnosis of the new nodules was made by means of enucleation, resection with the primary lesions, thick-needle biopsy, or additional partial resection of the liver. RESULTS: New nodules were detected in 27 (29.3%) of 92 resected cases. Internal echoic pattern of the nodules were type I, hypoechoic (29 nodules); type II, hyperechoic (19); and type III, mosaic (3). Ten HCC nodules (17.9%) were included, and chance of being malignant for each type was 24.1%, 0%, and 100%, respectively. Of the seven patients with malignant new nodules, three underwent additional systematic resection and all were alive without recurrence 49, 13, and 11 months after the operation. Others were treated by use of enucleation (two cases), intraoperative ethanol injection (one case), and intraarterial chemotherapy (one case). CONCLUSIONS: Although most of the new nodules lacked specific findings for HCC, hypoechoic nodules, 24.1% of which were HCCs, should not be overlooked. Histologic confirmation of the new nodules is necessary especially when the number of lesions detected before operation is multiple or the interval between lipiodol computed tomography and the operation is longer than 2 months. Once the diagnosis of HCC has been made for the new nodule, systematic additional resection to remove the new lesion is recommended.

A fetal intestinal-type alkaline phosphatase produced in Caco-2 cells.

Enzymic, immunological and lectin-binding properties of alkaline phosphatase (AP) produced in Caco-2 cells, a human colon carcinoma cell line, were investigated. The enzyme was very similar to fetal intestinal (meconium) AP in the enzymic and immunological properties, but different from fetal intestinal AP in lectin-binding properties; expression of the galactose moiety was altered in AP of Caco-2 cells, compared to that of fetal intestinal AP. These results indicate that AP of Caco-2 cells can be used in place of fetal intestinal AP when the enzymic properties of an AP of unknown origin are investigated, but cannot be used instead of fetal intestinal AP in the structural study of AP.