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Anti-Epstein-Barr virus (EBV) antibodies were tested in 11 children with chronic active EBV infection. Anti-virus capsid antigen (VCA)-IgG antibody titers ranged from 1:640 to 1:10,240. Anti-VCA-IgM antibody was consistently positive in 5 of the 11 patients; anti-VCA-IgA antibody was consistently positive in 6 of the 10 patients; anti-early antigen (EA)-IgG antibody was consistently positive in 10 of the 11 patients and anti-EA-IgA antibody was consistently positive in 4 out of the 7 patients. Anti-EBV nuclear antigen (EBNA) antibody was not detected in two patients. Consistently positive anti-VCA-IgA- and anti-EA-IgA- antibody may be a characteristic feature of abnormal antibody responses in severe chronic active EBV-infection in childhood.

Seventeen patients having extracardiac valved conduits placed between the right ventricle and pulmonary artery were followed for 7 to 87 months postoperatively (mean, 42 months), at the Heart Institute, Kenritsu Amagasaki Hospital, Japan. There were no late deaths in the study group. Three conduits have been replaced, all because of conduit stenosis. In two-dimensional echocardiographic examinations, commissural fusion and calcification of the valve were noted in 6 out of 16 xenograft valved conduits. Mechanical valve immobility was found in one patient. Neointimal peel of the dacron graft was noted in 6 out of 17 cases, and marked left ventricular deformity in the short axis view was found in 6. Late cardiac catheterization was done in 6 patients who were suspected of having valve failure and right ventricular hypertension by two-dimensional echocardiography. All 6 of these patients showed a high pressure gradient between the pulmonary artery and right ventricle and also had elevated right ventricular pressure. In conclusion, two-dimensional echocardiography is a simple, non-invasive and very accurate method for detecting conduit stenosis and valve failure. An echocardiographic series should be performed for a long-time postoperatively because obstructions of valved conduits may be progressive, and an operation may be advisable in order to prevent the development of advanced right ventricular hypertrophy and deterioration.

In order to improve the postoperative prognosis of gastric cancer patients we have performed preoperative endoscopic intratumoral administration of various biological response modifiers. In the present study we have investigated the kinetics and the immune response augmenting effect of intratumorally injected PSK, a protein-bound polysaccharide preparation, by immunohistochemical methods using anti-PSK antibody and various other antibodies. PSK-containing cells were located in the tumor tissues and follicular marginal zones of regional lymph nodes. Intratumorally administered PSK appeared to be phagocytized by the histiocytes and to cause them to become antigen-presenting cells. These cells may play a major role in augmenting immune responses in gastric cancer patients.

We report a case of unilateral hyperplasia of the adrenal medulla. The patient showed clinical features suggestive of pheochromocytoma. Removal of the hyperplastic adrenal gland resulted in complete disappearance of all prior symptoms, decrease of the plasma and urinary catecolamine levels and no high uptake in [133I] metaiodobenzylguanidine scintigraphy. A histological study revealed diffuse hyperplasia of the adrenal medulla. Up to now, there are relatively few reports of adrenal medullary hyperplasia in English literatures.

A new volatile derivative of taurine, N-isobutoxycarbonyltaurine methyl ester (methyl 2-(N-isobutoxycarbonylamino)ethanesulfonate), was prepared by a three-step procedure for the gas chromatographic determination of taurine in urine. First, taurine was converted to its silver salt by reaction with silver oxide; next the silver salt was reacted with isobutyl chloroformate to form the N-isobutoxycarbonyl derivative, and finally the derivative was reacted with methyl iodide to form N-isobutoxycarbonyltaurine methyl ester. The volatile derivative was analyzed by gas chromatography using a column of 3% OV-101 on Chromosorb W. When methyl 3-(N-isobutoxycarbonylamino) propanesulfonate was used as an internal standard, the calibration curve was linear between 0.5 and 5.0 mumol of taurine/ml and showed a good reproducibility. This method was applied to the determination of taurine in human urine. Recovery was 98.6 +/- 5.2%, when 1.25 to 5.0 mumol/ml of taurine was added to human urine.

Excretion of sulfate and taurine, two major metabolites of sulfur, was examined in rats to study the nutritional status of sulfur metabolism in the mammals. Rats maintained on a conventional laboratory diet excreted 1.83 +/- 0.14 mmol of free sulfate and 229.0 +/- 75.3 mumol of taurine/kg of body weight per day. When the diet was changed to a synthetic 25% casein diet, the taurine excretion decreased to 15% of the previous daily excretion, but sulfate excretion decreased only slightly. These decreased levels returned to the original levels when 5 mmol of L-cysteine/kg of body weight was administered into the stomach through a catheter. One week after the first L-cysteine administration, when sulfate and taurine excretion had returned to the original levels, 5 mmol of L-cysteine/kg of body weight was administered likewise. The rats excreted sulfur corresponding to about 95% of L-cysteine administered in the form of free sulfate and taurine within a few days following L-cysteine administration, and sulfate excretion was 3.5 times more than taurine excretion. These results seem to suggest that, in rats, sulfur metabolism is in a state of equilibrium and that sulfate is formed preferentially to taurine.

In order to clarify the origin of JC virus-induced brain tumors in rats, the development of tumors was sequentially analyzed histologically and immunohistochemically. Twenty-two of 30 rats (73%), which were intracerebrally inoculated with JC virus within 24 h of birth (group 1), developed, as a group, 45 brain tumors after 12 to 26 weeks. Seventeen of 27 rats (63%), which were inoculated on the 7th day after birth (group 2), developed 37 brain tumors as a group after a time 12 to 40 weeks. The tumors were found exclusively in the cerebrum. The microtumors, which were defined as tumors less than 2 mm in diameter, were located in the subependymal plate around the ventricular system. The microtumors and most part of the macrotumors consisted of cells of undifferentiated neuroectodermal nature, showing nuclear palisades and Homer-Wright-pseudorosette-like structures. Some tumor cells of macrotumors had an astrocytic nature and were positive for glial fibrillary acidic protein, S-100, Leu 7, and vimentin. In conclusion, the target cells of JC virus in rats may be undifferentiated subependymal cells of the cerebrum. The tumor cells show partial glial differentiation as they grow.

To analyze the possible major T cell recognition site(s) of chironomid antigens, we established human T cell lines and clones (CD3+ 4+ 8-) reactive to soluble extracts of the adult midge of Tokunagayusurika akamusi (TAA) and/or Chironomus yoshimatsui (CYA). All T cell lines and clones proliferated heavily in response to relatively large molecular weight fractions of TAA (MW greater than or equal to 15,000). Nine clones reactive to TAA were classified into 3 groups according to reactivity, indicating the existence of at least 3 distinct T cell recognition sites in TAA. Five T cell clones responded to both TAA and CYA, although the two chironomid antigens were serologically distinct. We conclude that T cell recognition sites of chironomid antigens are different from B cell recognition sites in humans.

The rescue of infectious virus from nonproducer BH RSV(-) cells by chick cellular DNA was attempted in order to investigate the functional state of endogenous and exogenous retroviral genes integrated within the cellular DNA. No infectious virus was rescued by transfection with DNAs of chick helper factor (chf)-negative chick embryo cells (CEC), chf-positive CEC or uninfected CEC producing endogenous Rous associated virus (RAV-0). On the other hand, infectious Rous viruses with the phenotype of RAV-0 and RAV-1 were rescued by transfection with DNAs of CEC which had been infected with RAV-0 and RAV-1. From these results, it seems that exogenous retroviral genes integrated in the cellular DNA are expressed rather easily by transfection while those present endogenously are not.

Chlorpyrifos, an organophosphorus insecticide, has been used to control termites since regulatory measures against the use of chlordanes were taken in September, 1986. We developed an improved gas chromatographic (GC) method for the assay of 3,5,6-trichloro-2-pyridinol (TCP) in the urine to use in the biological monitoring of exposure to chlorpyrifos. Urinary TCP was separated and determined accurately (C.V., 4%) with high sensitivity (detection limit, 10 ng/ml) and recovery (recovery greater than 90%) using a wide bore capillary column (WBC column). The accuracy and precision of the present GC method are satisfactory. The time course of urinary excretion of TCP was followed in workers. The urinary TCP level was low in the off-season and high in the busy season. Variation in the urinary TCP level corresponded to the termite control season and the length of the working period. The urinary TCP level showed a change reciprocal to the variations in the plasma cholinesterase activity. From these results, it is surmised that the urinary TCP level represents the extent of exposure to chlorpyrifos. The decrease in the level of cholinesterase activity is suggested to be due to exposure to chlorpyrifos. Determination of the urinary TCP level by GC using a WBC column is useful in the biological monitoring of chlorpyrifos in termite control workers and potentially has practical application to health care.

We performed a cytogenetic study on 140 nonpolymalformed patients with mental retardation of clinically undefined origin, using a high resolution banding technique, to determine how much chromosome abnormalities contribute to the etiology of this condition. A total of 15 patients (10.7%) were found to have autosomal or sex chromosomal abnormalities. Autosomal abnormalities included partial monosomy (5 cases), reciprocal translocation (one case), 13/14 robertsonian translocation (3 cases), unbalanced translocation (one case), inverted duplication of 15q (one case) and mosaic trisomy 21 (one case). Sex chromosomal abnormalities comprised structural rearrangement of the short arm of the X chromosome (one case) and 47, XXY in a pure or mosaic form (two cases). It should be noted that four out of the 5 cases of partial monosomy had subtle interstitial deletions, which might have been unidentified by the conventional G-banding method alone. In one case of the robertsonian translocation 46,XY,t(13;14)/45,XY,t(13;14), a small deletion was thought to have occurred in the cells with a chromosome number of 45. Comparison of clinical features of the 15 chromosomally abnormal patients with those of patients with normal karyotypes did not show any clinical parameter indicative of chromosome imbalance. These results suggest that a subtle chromosomal deletion is specific to mental retardation associated with few malformations. We believe that diagnostic evaluation of mentally retarded patients, even if nonmalformed, should include chromosome analysis using a high resolution banding technique.

A case of ovarian leiomyoma is reported, together with histologic, immunohistologic and electron microscopic findings. A solid firm tumor, measuring 6.5 X 5 X 5 cm, was found in the right ovary of a 65-year-old woman. The tumor had an obvious whorled pattern on the cut-surface. Well-differentiated, long spindle-shaped neoplastic cells revealed positive immunoreactivity for anti-desmin. Ultrastructural observations included numerous microfilaments with dense patches in the cytoplasm, micropinocytotic vesicles beneath plasma membranes and continuous basal laminae around neoplastic cells. These findings were compatible with leiomyoma. The possible histogenesis of ovarian leiomyoma was discussed.

A simple cell-free system for studying a priming factor involved in the repair of bleomycin-damaged DNA was established. The template-primer used for the repair DNA synthesis was prepared by treating the closed circular, superhelical form of pUC19 plasmid DNA with 2.2 microM bleomycin and 20 microM ferrous ions. Single-strand breaks were introduced into pUC19 DNA by the bleomycin treatment, and the DNA was consequently converted largely into the open circular form. A system for repair of this bleomycin-damaged DNA was constructed with a priming factor, DNA polymerase (DNA polymerase beta or Klenow fragment of DNA polymerase I), ATP, T4 DNA ligase and four deoxynucleoside triphosphates. After incubation, the conformation of the DNA was analyzed by agarose gel electrophoresis and electron microscopy. The open circular DNA was largely converted to the closed circular DNA, indicating that the single-strand breaks of DNA were repaired. When the priming factor was omitted, DNA repair did not occur. The present system seemed to be applicable to the study of priming factors involved in the repair of DNA with single-strand breaks caused not only by bleomycin but also by ionizing radiation or active oxygen.

Murine adrenal tumor cells (Y-1 clone) were stimulated by adrenocorticotropic hormone (ACTH) and cyclic adenosine 3',5'-monophosphate (cyclic AMP) to produce steroid hormone (delta 4, 3-keto steroids). The steroids were secreted into the medium immediately after synthesis. The optimum concentrations of ACTH and cyclic AMP for stimulation of steroid production were 10(-2) U/ml and 1.0 mM, respectively. In serum-free medium, ACTH and cyclic AMP stimulated steroidogenesis in Y-1 cells, but the amount of steroid hormone in the culture medium was low. However, a high level of steroid production was maintained with medium containing 10 mg/ml bovine serum albumin (BSA). In culture medium containing a higher concentration of BSA, Y-1 cells did not become spherical as is usually the case when steroid production is stimulated by ACTH or cyclic AMP. The morphological changes did not always correlate with steroid secretion by Y-1 cells.

It has been assumed that the in vivo reduction of 3-mercaptopyruvate, an intermediate of cysteine metabolism, to 3-mercaptolactate is catalyzed by lactate dehydrogenase (EC 1.1.1.27) though no definitive evidence has been presented. In order to examine this assumption, reduction of 3-mercaptopyruvate and its inhibition were studied using rat liver homogenate, lactate dehydrogenase purified from rat liver and anti-lactate dehydrogenase antiserum. Reduction of 3-mercaptopyruvate was actively catalyzed by rat liver homogenate and by the purified lactate dehydrogenase. This reducing activity was completely inhibited by anti-lactate dehydrogenase antiserum. These results indicate that the reduction of 3-mercaptopyruvate to 3-mercaptolactate in rat liver is catalyzed by lactate dehydrogenase.

To study the mechanism of DNA excision repair, a DNA repair system employing permeable mouse sarcoma (SR-C3H/He) cells was established and characterized. SR-C3H/He cells were permeabilized with a 0.0175% Triton X-100 solution. The permeable cells were treated with 1 mM ATP and 0.11 mM bleomycin, and then washed thoroughly to remove ATP and bleomycin. Repair DNA synthesis occurred in the bleomycin-damaged, permeable SR-C3H/He cells when incubated with ATP and four deoxyribonucleoside triphosphates. The repair nature of the DNA synthesis was confirmed by the BrdUMP density shift technique, and by the reduced sensitivity of the newly synthesized DNA to Escherichia coli exonuclease III. The DNA synthesis was optimally enhanced by addition of 0.08 M NaCl. Studies using selective inhibitors of DNA synthesis showed that aphidicolin-sensitive DNA polymerase (DNA polymerase alpha and/or delta) and DNA polymerase beta were involved in the repair process. The present DNA repair system is thought to be useful to study nuclear DNA damage by bleomycin, removal of the damaged ends by an exonuclease, repair DNA synthesis by DNA polymerases and repair patch ligation by DNA ligase(s).

The major gag protein (p34) of squirrel monkey retrovirus-H was purified in one chromatographic step by anion-exchange high performance liquid chromatography. The virus in a crude fraction was disrupted with Brij 35 in the presence of three kinds of protease inhibitors. The soluble virus lysate was injected into a Polyanion SI column, and p34 was eluted with a linear salt gradient. The recovery of the protein was about 60%. The purified p34 was nearly homogenous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining.

To clarify the initiation, development and recovery processes of disseminated intravascular coagulation (DIC), rat glomerular capillaries and fibrin thrombi were examined under transmission and scanning electron microscopes. DIC was induced in rats by a single intraperitoneal injection of endotoxin (Et., 7.5 mg/kg lipopolysaccharide:B, E. coli 026:B6). At 2 h after Et. injection, the endothelial surface of the glomerular capillary became irregular with projections like a sea anemone. At 4 h after Et. injection, agglomerated fibrin thrombi composed of fibrin fiber bundles with fine cross-striated fibriform structures were observed in the capillary lumen. The fibrin thrombi gradually changed into fine reticular systems suggesting a degradation process by 6 h after Et. injection, and formed a coarse granular agglomerate by 8 h after Et. injection. These fibrin thrombi disappeared within 12 h of Et. injection, but the endothelial surface remained edematous. At 24 h after Et. injection, the microstructure of the glomerular capillaries returned normal. Based on these observations, we concluded that DIC was primarily initiated by injury to the capillary endothelium, and that changes on the endothelial surface contributed to the development of DIC.

Three types of traditional Chinese herb medicine were used to treat 98 patients with advanced esophageal squamous cell carcinoma prior to surgical treatment. Forty-two patients with the same diagnosis were treated with these herbs plus cyclophosphamide (endoxan). One hundred similar patients received surgical treatment without herbs or endoxan treatment as controls. Histologic examinations of surgical specimens were made on all of these patients. Stromal lymphoid-cell infiltration and cancer tissue degeneration were more prominent in Menispernum dehuricum DC- or Chelidonium majus L-treated patients, and were less clear in patients treated with herbs plus endoxan and the controls. The antitumor action of herbs is thought to be brought about by the activation of an immunological rejection mechanism. Herbs plus endoxan may result in the masking of the immunological response of hosts without obviously damaging cancer tissues.

Sprague-Dawley rats, 6 with aminonucleoside nephrosis and 6 controls, were intravenously injected with human liver ferritin isolated from post mortem liver, and their 24-h urine samples were examined for human ferritin by immunoradiometric assay. In rats with aminonucleoside nephrosis, the amount of excreted ferritin in urine was forty times greater than in control rats. Much more monomeric ferritin was excreted than that of polymeric ferritin. We are the first to have utilized human liver ferritin as a tracer to measure a minor amount of ferritin by a commercially available kit. Our present study seems to indicate a critical role for glomerular basement membrane as a size barrier.