Are there histopathologic characteristics particular to fulminant hepatic failure caused by human herpesvirus-6 infection? A case report and discussion.

An 8-month-old boy was admitted to a neighboring hospital for severe liver dysfunction and drowsiness 4 days after a diagnosis of exanthem subitum. A diagnosis of fulminant hepatic failure was made, and liver biopsy was performed during the acute stage. The presence of human herpesvirus-6 variant B (HHV-6B) DNA was shown in liver tissue by polymerase chain reaction (PCR) and in the endothelium of the portal vein by in situ hybridization (ISH). Histologic examination showed microvesicular steatosis resembling that of Reye's syndrome, even though aspirin had not been prescribed. We considered HHV-6 to be the causative agent in this case and report what is perhaps the first precise histologic description of fulminant hepatic failure caused by HHV-6.

[MR hydrography of the abdomen: technical consideration of data acquisition and future prospects for clinical applications].

Recent advances in MR hydrography have enabled various clinical applications in the areas of abdominal and pelvic disorders, for example, MR cholangiopancreatography (MRCP), MR urography(MRU), and MR hydrography of the fetus. Although the two-dimensional(2D) single-slice method provides excellent information as to the global relationship between lesions and various anatomical structures, small lesions or detailed anatomical characteristics may not be visualized owing to the partial volume effect. Source images of the 2D multislice method are most suitable for the detection of small lesions and detailed evaluation of anatomical structures as "tomographic imaging", while the three-dimensional(3D) method is useful in obtaining data sets for 3D imaging. MR hydrography is a promising method for the noninvasive evaluation of various abdominal disorders, and it has the potential to play new roles in various anatomical regions. However, knowledge of the proper indications is essential for successful clinical application.

Revision of failed metal-backed patellar component of Miller/Galante-I total knee prosthesis.

Of 38 Miller/Galante-I total knee prostheses implanted between 1987 and 1990, 20 (53%) were revised. Failed metal-backed patella (15 knees; 40%) was the most frequent reason for revision. In most patients, the components were found well fixed to bone. The purpose of this study was to describe the revision technique we used for failed metal-backed patella. Total revision of all components would be ideal. However, removal of all components results in large bone loss, large blood loss, and a longer operation time. Because of the patients' relatively old age (78.7 years), less demanding activities of daily life (ADL), and the lower body weight of the Japanese patient, we chose a less invasive approach. The revision technique had the following features: (1) the grooved femoral component was not replaced, (2) the failed patellar component was replaced by an all-polyethylene patella, (3) synovectomy was performed, and (4) realignment of patellar tracking was done by lateral release and medial imbrication of the quadriceps mechanism. The mean follow-up after revision was 2 years, 4 months, and good short-term results were obtained. This less invasive approach would be beneficial for elderly patients whose ADL are less demanding.

Epitope mapping for the anti-rabbit cholesteryl ester transfer protein monoclonal antibody that selectively inhibits triglyceride transfer.

Among the monoclonal antibodies (Mab) against rabbit plasma cholesteryl ester transfer protein (CETP), Mab 14-8F cross-reacted with human CETP and selectively inhibited triglyceride transfer but not cholesteryl ester transfer (Ko, K. W. S., T. Ohnishi, and S. Yokoyama. 1994. J. Biol. Chem. 269: 28206;-28213). The epitope of this antibody was studied by using synthetic fragment peptides of rabbit and human CETP. Mab 14-8F reacted with the peptide R451-Q473 of human CETP near the carboxyl-terminal and not with the peptides representing any other regions, and inhibited the binding of human CETP to the goat antibody against its carboxyl-terminal peptide R451-S476. The experiments with a series of the fragment peptides in this region revealed that the epitope requires the segment 465-473 (EHLLVDFLQ) of human CETP or 485-493 (KHLLVDFLQ) of rabbit CETP (core epitope) though neither peptide by itself binds to the antibody. Both peptides needed extension at least by one residue beyond either amino- or carboxyl-end in order to show the reactivity to the antibody, but the effect was not highly residue-specific at least at the amino-end. Circular dichroism analysis demonstrated the increase of helical conformation by the extension of the "core epitope" peptides to either direction. Thus, the epitope is dependent on conformation of the core epitope induced by the presence of an additional residue(s) in either end. The core epitope occupies the central 64% of the reported linear epitope of Mab TP2, a widely used anti-human CETP monoclonal antibody that inhibits both cholesteryl ester and triglyceride transfer.Therefore, we conclude that the limited interaction of Mab with a common lipid interaction site causes selective inhibition of the transfer of triglyceride that has presumably lower priority than cholesteryl ester for the CETP reaction.

[The application of end user computing (EUC) for detection of lipoprotein lipase gene abnormality].

Lipoprotein lipase (LPL) is an enzyme digesting lipoprotein triglyceride (TG) in peripheral blood vessels. Most patients with LPL deficiency show very high plasma TG and low HDL-C. To establish an effective computer-based screening system to identify individuals with genetic LPL disorders, we selected 50 subjects whose plasma TG was over 350mg/dl and HDL-C was lower than 35mg/dl from patients at Hamamatsu University Hospital. We applied End User Computing (EUC) of our laboratory system to select high risk subjects with LPL gene abnormalities. Polymerase chain reaction (PCR) products from LPL gene exons 2-9 were screened by single-strand conformation polymorphism (SSCP), direct DNA sequence analysis and restriction fragment length polymorphism (RFLP). We found a novel missense mutation (1223C-->G, S323C) in LPL gene exon 7 from three subjects. By PCR-mediated site-directed mutagenesis and restriction digestion, the three subjects were found to be heterozygous. In addition, we identified two other common mutations in Japanese employing the RFLP method. One was the 1595C-->G (S447X) in exon 9 from six subjects, two homozygous and four heterozygous individuals. The other was a mutation of intron 3 (C-->T transition) from four heterozygous subjects. Using EUC screening method, we detected genetic LPL abnormalities more easily. The frequency of the LPL gene mutation in the 50 high-risk subjects was 26%, and was estimated to be one out of 2,000 patients at our clinic. Using the EUC system to screen for LPL mutations was established to be an effective computer-based screening system to identify individuals with genetic abnormalities.

[Analysis of the long-term prognosis for conjunctival malignant melanomas in Japan].

PURPOSE: To investigate the long-term prognosis for primary conjunctival malignant melanomas in Japan. MATERIALS & METHODS: We conducted a survey of 61 cases which had been reported in a 38-year period (1959 to 1996). We gathered information regarding the survival of patients, the post-operative follow-up period, the causes of death, and recurrences. Answers were obtained segarding 51 cases (84%). Detailed progress was identified in 23 of these cases. The survival rates were calculated using the Kaplan-Meier method. RESULTS: The survival rates were 95.1% after 1 year, 72.9% after 3 years, and 53.4% after 5 years. These values are relatively low compared with those reported in Europe and the United States.

Site-directed mutagenesis of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12.

The contribution of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12 to the catalytic reaction was studied by site-directed mutagenesis of these residues. A kinetic analysis of the purified mutant chitinases suggests the involvement of both the Asp-197 and Asp-202 residues in the catalytic events of this enzyme, although the effects of mutations of Asp-197 were less severe than those of the other mutations.

[Ultrasonography in primary malignant lymphoma of the thyroid].

We analyzed retrospectively the sonographic appearance of eight cases of primary malignant lymphoma of the thyroid. Sonography showed hypoechoic masses replacing thyroid parenchyma with posterior acoustic enhancement. A hypoechoic and/or nearly anechoic confluent nodular appearance characterized malignant lymphoma of the thyroid.

Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity.

Prokaryotic chitinases, class III plant chitinases, yeast chitinases, and endo-beta-N-acetylglucosaminidases share weak amino acid sequence similarities at the certain region of each enzyme. These regions have been assumed to be important for catalytic activities of the enzymes. To verify this assumption, three amino acid residues (Ser-160, Asp-200, Glu-204) in chitinase A1 of Bacillus circulans WL-12 were chosen, based on the amino acid sequence alignment of the regions sharing sequence similarity, and were replaced by site-directed mutagenesis. Kinetic parameters for 4-methylumbelliferyl-N,N',N"-triacetylchitotriose hydrolysis were determined with wild-type and seven mutant chitinases. Chitinases with Glu-204-->Gln mutation and Glu-204-->Asp mutation were essentially inactive and kcat values of these chitinases were approximately 1/5,000 and 1/17,000 of that of wild-type chitinase, respectively. Asp-200-->Asn mutation decreased the kcat value to approximately 1/350 of that of the wild-type enzyme, while the Km value decreased only slightly. On the other hand, neither the kcat value nor the Km value was affected by Asp-200-->Glu mutation. Thus, it appeared that Glu-204 and Asp-200 are directly involved in the catalytic events of chitinase A1. The role of the carboxyl group of Asp-200 can be fully substituted by that of Glu residue. The Ser-160-->Ala mutant retained 10% activity of the wild-type chitinase indicating that the hydroxyl group of Ser-160 is not absolutely required for the catalytic activity. These results indicate a lysozyme-type catalytic mechanism of the chitinase.

Effectiveness of lymphocytapheresis in a patient with ankylosing spondylitis.

Immunomodulation by lymphocytapheresis (LCA) was carried out in a patient with ankylosing spondylitis (AS) who suffered severe pain in joints and back. LCA was performed once a week and 3 x 10 9 lymphocytes were extracted each time. Fifteen courses of LCA were completed. The joint score was gradually decreased and the improvement was maintained 11 months after treatment. The population of NK cells detected by Leu-7 and Leu-11 monoclonal antibodies decreased and HLA-DR positive CD3 cells increased during and after the treatment. The effectiveness of LCA in AS may confirm the participation of immunological mechanisms in the pathogenesis of AS.

[Histochemistry of copper and copper-binding protein in liver with a comparison between histologic sections and biochemical evaluation].

A quantitative analysis of copper in liver tissue and a qualitative analysis of copper-binding protein in the histologic sections were subjected to correlative study. Liver sections, obtained from autopsies and surgical resections, were examined histopathologically by orcein staining, rhodanine staining, and other special stainings, in addition to hematoxylin eosin (HE) staining. The quantitative analysis was carried out by an atomic absorption spectrophotometry. Copper and copper-binding protein were not detectable by HE staining. The amount of orcein positive granules (OPGs) was graded semiquantitatively from negative (-) to three plus ( ). The average copper content was 24.9 micrograms/g dry liver weight (micrograms/g) in (-) group, 60.9 micrograms/g in (+) group, 158.9 micrograms/g in (++) group, and 299.3 micrograms/g in ( ) group respectively. Although the copper content and the amount of OPGs were relatively well correlated as mentioned above, standard deviations of copper contents were comparatively large in each group, especially in (++) or ( ) group. Based upon the comparative study of staining qualities of various dyes, rhodanine staining and orcein staining were specific for copper or copper-binding protein. Finally, orcein staining was thought to be the most excellent method from the viewpoints of sensitivity, specificity, contrast and simplicity. The distribution of excessive copper in pathologic conditions was fairly uneven. We recommend a histopathologic examination by using many liver sections stained by orcein, when the condition of copper overload is suspected.

[Radiotherapy for T1 and T2 carcinomas of the tongue: combination therapy using low-dose external irradiation and an interstitial implant].

Between January 1975 and October 1984, 114 previously untreated patients with squamous cell carcinoma of the tongue were treated by a radical course of irradiation at the Jikei University Hospital. Of these patients, eighty with T1 or T2 lesions are analyzed in this paper, some having been treated with an interstitial implant alone or with a combination of external irradiation and an interstitial implant. The two-year local control rate of the primary lesion reached 85% in T1 patients, and 68% for those graded T2. Late radiation injuries were seen in 18% of the cases with local control beyond one year. Most of these injuries, however, healed with conservative therapy. The cumulative 5-year survival rate was 75% in T1 patients, and 52% for those graded T2. The combination of low dose external irradiation (20-30 Gy) and an interstitial implant showed the same local control rate, the same incidence of late injury, and the same survival rate as those who received an interstitial implant alone. In cases that were given combination therapy, the sum dose of about 80 Gy was necessary for local control.

Tissue copper content in primary and metastatic liver cancers.

Tissue copper contents in 38 primary and 45 metastatic hepatic malignancies and 15 control livers were analyzed by atomic absorption spectrophotometry. The average copper content of 15 control livers was 23.1 +/- 13.0 micrograms/g dry weight (microgram/gdw). The copper content of five cholangiocellular carcinomas (CCCs) and 45 metastatic cancers was almost equal to the control level. Thirty three hepatocellular carcinomas (HCCs) contained a larger amount of copper (61.5 +/- 76.8 micrograms/gdw) than the control livers (p less than 0.05), but the copper content of HCCs showed a considerably wide variation. The average copper content of nine minute HCCs (126 +/- 112 micrograms/gdw) was significantly (p less than 0.05) higher than that of 24 large HCCs (37.2 +/- 39.9 micrograms/gdw). Histologically, orcein and paramethylaminobenzylidene rhodamine positive granules were seen in eight and four of nine minute HCCs, respectively. These granules were also found in some large HCCs, but were never found in CCCs and metastatic cancers. It was concluded that these excessive accumulations of copper and copper-binding proteins might present a helpful finding to distinguish some cases of HCC, especially small HCC, from CCCs, metastatic cancers and hypertrophic regenerative nodules of cirrhotic livers. The significance and possible pathogenesis of these copper accumulations in HCCs require further studies.

Role of lysosomes in coronary artery lesions of hypertensive rats.

Electron microscopically the role of acid phosphatase (acid P) positive lysosomes in the pathogenesis of intramyocardial coronary artery lesions of hypertensive rats with bilaterally constricted renal arteries was studied. At 3 postoperative weeks, acid P positive lysosomes were increased in endothelial cells of the coronary arteries, but at 5 weeks and thereafter, they were decreased in number. In the intima, intimal smooth muscle cells with acid P positive lysosomes appeared at 3 postoperative weeks, but their number remained small as late as 9 postoperative weeks. The internal elastic lamina was fragmented into amorphous masses from 3 postoperative weeks, at 5 weeks and thereafter the fragmentation and disruption became severer, and at 9 weeks the lamina disappeared because of marked disruption. At 3 postoperative weeks, acid P positive extracellular lysosomes were observed in the gaps of the fragmented internal elastic lamina. At 5 weeks and thereafter, extracellular lysosomes and attenuated processes of medial smooth muscle cells with lysosomes were seen extending through the gaps of the fragmented internal elastic lamina. Necrosis of these extending cell processes and lysosomes remaining after the necrosis were observed. The findings suggested that the fragmentation and lytic change of the internal elastic lamina observed in hypertensive rat intramyocardial coronary arteries might be induced by extracellular lysosomes discharged not only from endothelial cells and intimal smooth muscle cells but also from the extending processes of medial smooth muscle cells into the gaps of the internal elastic lamina.

A clinicopathological review of 12 autopsied cases of adult T-cell leukemia.

Twelve autopsied cases with adult T-cell leukemia (ATL) were reviewed clinicopathologically. The prognosis of three cases who had suffered from severe cutaneous lesions was much better than that of the other nine cases with no or negligible cutaneous lesions. The surface marker of leukemic cells from six cases was ordinary inducer/helper phenotype (OKT4+ and 8-), but in one case leukemic cells showed OKT4+ and 8+. In another case, a significant amount of leukemic cell infiltration was found in the thymic cortex. Calcium content in the bone of ATL cases was lower than that of the patients without ATL (control group), and six cases with ATL (50%) were complicated by severe hypercalcemia. Neither adenoma nor hyperplasia of the parathyroid glands was found in any case. In most severely hypercalcemic patients, bone trabeculae were actively absorbed by numerous osteoclasts and partly replaced by fibrous tissues. In two normocalcemic patients, skeletal calcium content was also markedly reduced by osteoporosis, but the activation of osteoclasts was inconspicuous. It was speculated that the manner of bone resorption in ATL cases was diverse and there were some clinicopathological subtypes in ATL from the viewpoints of cutaneous lesions, hypercalcemia, and bone lesions.

Endothelial changes of hypertensive rat mesenteric arteries.

The mesenteric arteries of hypertensive rats with bilaterally constricted renal arteries were observed by light, scanning electron and transmission electron microscopy. In the early stage of hypertensive arterial lesions, many leukocytes adhered to the endothelial surface, and large or small and polygonal endothelial cells were irregularly arranged. Sporadically brightened and atrophied endothelial cells were distributed on the injured surface. These atrophied cells were transmission electron microscopically confirmed to have undergone coagulation necrosis. In the surrounding of the atrophied cells were observed opened endothelial cell junctions and enlarged intercellular spaces to which many leukocytes and platelets adhered. Because of loosening of connection with neighboring cells, atrophied endothelial cells became solitary or being denuded. Opened endothelial cell junctions were also observed between the uninjured endothelial cells as well as in areas penetrated by leukocytic pseudopods. Where light microscopy disclosed the deposition of a large amount of fibrinoid substance in the intima and media, scanning electron microscopy showed opened endothelial cell junctions and denudation of endothelial cells. From the results of the present experimental study, it was suggested that in the genesis of the hypertensive rat arterial lesions, the degeneration, necrosis and denudation of endothelial cells, opened endothelial cell junctions between these cells, and insudation of blood plasma through the opened junctions or denuded areas might play important roles.

[A rapid method for determining urinary indoleacetic acid concentration and its clinical significance as the tumor-marker in the diagnosis of malignant diseases].

A rapid method for determining urinary indole-3-acetic acid (IAA) is introduced as the tumor-marker for the screening and diagnostic purpose of cancer patients by means of high performance liquid chromatography (HPLC). Its clinical significance is discussed along with a review of literatures. The IAA concentration and creatinine level of optionally collected urine samples were measured and used for the calculation of IAA amount per unit creatinine (microgram IAA/mg creatinine) in urine. Thus, an amount of 24-hours urinary IAA could be calculated without collecting a whole day's urine supply. Analysis of urinary IAA was performed within 10 minutes by HPLC. Urinary IAA level is usually high in the patients with the upper G-I tract cancers such as gastric cancer, esophageal cancer and hepato-biliary tract cancer, and also malignant hematopoietic disorders. But it is also high in non-cancer patients such as liver cirrhosis, diabetes mellitus and cholelithiasis occasionally. The patients with high urinary IAA level also showed high urinary levels of 5-hydroxy indoleacetic acid (5-HIAA) and monoamine oxidase activity (MAO). It was characteristic that hepatocellular carcinoma showed slight elevation of urinary IAA with normal levels of 5-HIAA and MAO. It is conclusive that the positive rate of elevated urinary IAA level was high in the patients with gastric cancer with ulcer-forming type in its morphological classification, and its level tends to elevate as the disease progresses. Therefore, the measurement of urinary IAA level in an optionally collected urine sample, as the tumor-marker, can be useful to check the progression and regression of gastric cancer.