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The purpose of this study was to search for chromosomal susceptibility loci for comitant strabismus. Genomic DNA was isolated from 10mL blood taken from each member of 30 nuclear families in which 2 or more siblings are affected by either esotropia or exotropia. A genome-wide search was performed with amplification by polymerase chain reaction of 400 markers in microsatellite regions with approximately 10 cM resolution. For each locus, non-parametric affected sib-pair analysis and non-parametric linkage analysis for multiple pedigrees (Genehunter software, http://linkage.rockefeller.edu/soft/) were used to calculate multipoint lod scores and non-parametric linkage (NPL) scores, respectively. In sib-pair analysis, lod scores showed basically flat lines with several peaks of 0.25 on all chromosomes. In non-parametric linkage analysis for multiple pedigrees, NPL scores showed one peak as high as 1.34 on chromosomes 1, 2, 4, 7, 10, 15, and 16, while 2 such peaks were found on chromosomes 3, 9, 11, 12, 18, and 20. Non-parametric linkage analysis for multiple pedigrees of 30 families with comitant strabismus suggested a number of chromosomal susceptibility loci. Our ongoing study involving a larger number of families will refine the accuracy of statistical analysis to pinpoint susceptibility loci for comitant strabismus.</P>

Cultured rat liver cells which were cloned from a single cell were transformed into malignant cells by a chemical carcinogen, 4-dimethylaminoazobenzene (DAB). The DAB-transformed cells produced tumors when back-transplanted into new-born rats but the carcinogen-untreated control cells did not. Characteristics of the transformed liver cells were compared to those of DAB-untreated control cells in regard to the morphology, the consumption of DAB from the culture medium by the cells, the incorporation of 3H.DAB into the cells, and the aggregate.forming ability of the cells in rotation culture. The results showed that no significant parameter of malig. nant transformation in culture was detectable except the tumorigenicity of the transformed cells upon the inoculation into animals.

A case of left atrial myxoma accompanied by peculiar symptoms is reported. A 15-year old boy had progressive congestive heart failure and three episodes of acute attacks of panctyopenia. The anemia was accompanied by helmet-shaped, broken red blood cells, erythroid hyperplasia and elevation of indirect bilirubin. The thrombocytopenia gave rise to hemorrhagic tendency of the skin and mucous membrane. The leukocytopenia was seen at the same time. The patient also had general constitutional disturbances showing generalized malaise, persistent fever, elevation of erythrocyte sedimentation rate, positive C-reactive protein, pulmonary infection and anginal attacks. Postmortem examinations revealed a left atrial myxoma and intricated pulmonary changes. There was obliterative endarteritis of the left coronary branch and pulmonary arteries. The interstitial pulmonary fibrosis was also prominent. The pancytopenia should have been induced by the mechanical damage of circulating blood cells by the left atrial myxoma. The pathological findings of the lungs were highly suggestive of pulmonary hypertension, which was assumed to be due to mitral block caused by the atrial myxoma.

To obtain a useful rat liver cell line for in vitro carcinogenesis, two rat diploid epithelial cell lines were established from a 7-day-old male rat by the repeated colonial clone method. More than 80% of cells from each cell line have maintained normal diploid karyotype for over 30 months in vitro. The diploid cells were identi. fied as normal diploid karyotype by conventional Giemsa and trypsin. Giemsa techniques. They showed little difference in morphology and growth rate between early and late passages. Without cloning, they tended to be heterogenous in cell morphology, became heteroploid in chromosome and showed increased growth potential with time. Highly heteroploid cells which were derived from one of the lines produced ascites and solid tumors when inoculated into syngeneic rats intraperitoneally. Histologically, the tumors were diagnosed as poorly differentiated hepatocarcinomas. One of these diploid epithelial cell lines in early passage contained some activity of tyrosine transaminase and liver type aldolase and .glycokinase. Therefore, it is suggested that these epithelial cell lines represent liver parenchymal cells.

A new model of status epilepticus (SE), which was induced by intermittent electrical stimulation (20 Hz for 20 sec every min for 180 min) of the deep prepyriform cortex, has been developed in the conscious rat. SE was induced in 9 of 16 rats in the drug-free group. The number of stimulation trains required to induce SE in this status subgroup was 125.6 +/- 12.7 (mean +/- SEM) and the mean duration of self-sustained seizure activity (SSSA) occurring after cessation of the stimulation session was 295.4 +/- 111.4 min. Some animals showed secondary generalized seizures. Significant cell loss was observed in the hippocampal CA3 pyramidal cell layer ipsilateral to the stimulation site and bilateral CA1 areas in the status subgroup compared with the group subjected to sham operation. In addition, there was a significant negative correlation between the duration of SSSA subsequent to the stimulation session and the total number of intact pyramidal neurons observed in the bilateral CA1 and ipsilateral CA3 subfields of the status subgroup. There were significant differences between the status and non-status subgroups with respect to the number of afterdischarges (ADs) and the total AD duration during the stimulation session. Pretreatment with phenobarbital (30 mg/kg) prevented the development of SE and hippocampal cell loss completely. Pretreatment with MK-801, a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist (0.25 or 1 mg/kg), also prevented hippocampal cell loss, although it did not block SE generation completely, which suggests dissociation of the mechanisms underlying the development of SE and hippocampal damage. These results indicate that prolonged SSSA actually causes hippocampal damage and it is critically dependent upon NMDA receptor participation.

Indocyanine green (ICG) was injected into rat liver nodules induced by 2-acetylaminofluorene (2-AAF) via portal vein. The relationship between ICG staining and cell atypism of liver nodules was examined by means of histology and DNA flow cytometry. After 2-AAF administration, many small nodules appeared on the liver surface. All hyperplastic nodules were ICG stained until 10 weeks after the administration, but some nodules were not stained after 14 weeks. ICG-stained nodules histologically consisted of benign tissues and borderline lesions, and many of them showed "diploidy" in DNA cytometry. ICG-unstained nodules consisted of hepatocellular carcinoma (HCCs) and borderline lesions, and many of them showed "aneuploidy". In this way, it has been suggested that HCC could derive from hyperplastic nodules and that they might lose an ability to take up ICG in the process of hepatocarcinogenesis. Immunohistochemical staining for glutathione-S-transferase alpha (GST-alpha), a carrier protein of ICG in hepatocytes, was well correlated with ICG staining in the nodules, suggesting that the loss of ICG uptake in HCC was partly due to the decrease of GST-alpha. Moreover, the appearance of ICG unstained and aneuploid nodules was significantly inhibited in rats which were fed on diet containing Syosaiko-to after the administration of 2-AAF. Chemopreventive effect of Syo-saiko-to on hepatocarcinogenesis was identified.

A cell strain having low tumor-producing capacity was exposed in culture to 3'-methyl-N,N-dimethyl-4-aminoazobenzene (3'-Me-DAB) in the presence or absence of liver microsomes, and whether or not the cells will progress to those having high tumor-producing capacity was examined. When transplanted into rats, the cells treated with 3'-Me-DAB four (Exp-I) or thirteen times (Exp-II) formed larger tumors than untreated control cells, the latter treatment being more efficient in this regard. Furthermore, the tumors formed by the cells treated with 3'-Me-DAB in the presence of liver microsomes were considerably larger than those formed by the cells treated with 3'-Me-DAB alone. The subcutaneous tumors produced by the cells treated with 3'-Me-DAB with S-15 Mix showed poorly differentiated histology compared with those produced by control and 3'-Me-DAB-treated cells. The frequency of lung metastasis tended to increase by 3'-Me-DAB with S-15 Mix. The cells treated with 3'-Me-DAB in the presence or absence of liver microsomes differed from untreated control cells in vitro in some properties, including the size of aggregates in rotation culture, plating efficiency in liquid medium and morphology. These observations suggest that cell malignancy was promoted by 3'-Me-DAB alone as well as by 3'-Me-DAB in the presence of liver microsomes.

We studied the pathways of complement activation associated with the islet cell surface antibody (ICSA) obtained from sera of 7 patients (age less than 15 years) with insulin dependent diabetes mellitus (IDDM). The target cells were 51CR labelled rat islet cells and the complement source was human AB serum. Complement-dependent antibody mediated cytotoxicity (CAMC activity) was obtained using the percentage of cytotoxicity. CAMC activity of untreated sera was significantly inhibited by treating with EGTA or EDTA (p less than 0.001). The CAMC activity of EDTA-treated sera was significantly lower than that of EGTA-treated sera (p less than 0.001). In the inactivated human AB serum, it was lower than that of EGTA-treated sera (p less than 0.05), but not different from that of EDTA-treated sera. These results show that the complement activation associated with ICSA in patients occurred not only via the classical pathway but also via the alternative pathway.

We evaluated the respiratory functions of patients with pulmonary emphysema who underwent lung volume reduction surgery (LVRS) by the mean transit time (MTT) with Xe-133 lung ventilation scintigraphy, forced expiration volume in 1 sec (FEV1.0), residual volume (RV), distance walked in 6 min (6-min walk), and the Hugh-Jones classification (H-J classification) before and after LVRS. In 69 patients with pulmonary emphysema (62 men, 7 women; age range, 47-75 years; mean age, 65.4 years +/- 6.1, preoperative H-J classification, III (two were II)-V) who underwent LVRS, all preoperative and postoperative parameters (MTT 3 weeks after LVRS and the others 3 months after LVRS) were judged statistically by the Wilcoxon signed-ranks test and Odds ratio. Every postoperative parameter was improved with a significant difference (P < 0.05) compared to preoperative parameters. MTT at 3 weeks after LVRS was not associated with %FEV1.0 and the H-J classification at 3 months after LVRS, but was associated with RV and a 6-min walk at 3 months after LVRS. MTT was useful for the clinical evalution of aerobic capability after LVRS.

Previous in vitro and animal experiments have shown that sulforaphane, which is abundant in broccoli, inhibits Helicobacter pylori (H. pylori) infection and blocks gastric tumor formation. This suggests that broccoli consumption prevents chronic atrophic gastritis (CAG) introduced by H. pylori infection and, therefore, gastric cancer. For an epidemiological investigation of the relationship between the broccoli consumption and CAG, a cross-sectional study of 438 male employees, aged 39 to 60 years, of a Japanese steel company was conducted. CAG was serologically determined with serum cut-off values set at pepsinogen I < or = 70 ng/ml and a ratio of serum pepsinogen I/pepsinogen II < or = 3.0. Broccoli consumption (weekly frequency) and diet were monitored by using a 31-item food frequency questionnaire. The prevalence of CAG among men who ate broccoli once or more weekly was twice as high as that among men who consumed a negligible amount (P < 0.05). Multiple logistic regression analysis indicated that broccoli consumption once or more weekly significantly increased the risk for CAG (odds ratio, 3.06; 95% confidence interval, 1.12-8.38; P < 0.05), after controlling for age, education, cigarette smoking, and alcohol consumption. The present study failed to show an expected association between frequent broccoli consumption and a low prevalence of CAG.

We studied the differentiation of thyroid nodules using fine-needle aspiration biopsy (FNA) and Tl-201 scintigraphy quantitative analysis. One-hundred and thirty-one thyroid nodules were examined: 83 follicular lesions (58 benign and 25 malignant lesions) and 48 non-follicular lesions (8 benign and 40 malignant lesions). During Tl-201 scintigraphy examinations, an early and a delayed image were acquired 10 and 120 min after an intravenous injection, respectively. The T/N ratio (counts of nodular lesion/counts of contralateral normal thyroid tissue) of each image was calculated quantitatively. We assessed the ability of the Tl-201 scintigraphy and of the FNA analysis to differentiate benign and malignant lesions and determined the cut-off levels for the assays. For the follicular lesions, the area under the ROC (Receiver Operating Characteristic) curve (Az) for the Tl-201 scintigraphy data was greater than that for the FNA data. For the non-follicular lesions, the Az for the FNA data was greater than that for the Tl-201 scintigraphy data. We set cut-off levels at 1.370 for follicular lesions, and 1.070 for non-follicular lesions. The sensitivity and specificity were 76% and 82.7% for follicular lesions, and 90% and 87.5% for non-follicular lesions, respectively. The overall accuracy of the analysis was 84.0%.

The mechanism of cell dissociation with trypsin and EDTA was examined. Cell dissociation was possible when trypsin and EDTA were given simultaneously, when trypsin was given after EDTA treatment, but not when trypsin was given before EDTA treatment.

Albumin was highly purified from a commercially available rat albumin preparation (Fraction V) using disc electrophoresis. The purified albumin had the same antigenicity as Fraction V. A monospecific anti-rat albumin rabbit serum was obtained. The antiserum was used in a double antibody quantitative method for determining rat albumin.

Clinical trials of immuno-chemotherapy were conducted on malignant lymphoma patients. Patients during the period from 1972 through 1977 were allocated to two groups retrospectively according to the mode of treatment, i.e., chemotherapy alone (historical control group, 35 patients) and chemotherapy with OK-432 (treated group, 15 patients). Comparisons were made of the two groups, which were homogeneous with regard to induction chemotherapy, maintenance chemotherapy, stage and histologic type of disease. The treated group had a higher remission rate, and a longer remission duration and survival than the control groups, especially in patients with Hodgkin's disease but the difference was not statistically significant owing to the limited number of cases.

We investigated the diagnostic capabilities of I-131, Tl-201, and Tc-99m-MIBI (hexakis-2-methoxyisobutyl- isonitrile) scintigraphy for thyroid cancer metastases after total thyroidectomy over the entire body and for every locus before and after thyroid bed ablation. After total thyroidectomy of thyroid cancer, 36 cases were subjected to I-131 treatment 64 times. They consisted of 17 men and 19 women with 31 papillary carcinomas and 5 follicular carcinomas. Their ages were 22--75(an average of 60.5+/-12.3) years. I-131 scintigraphy(I-131), Tl-201 scintigraphy(Tl-201), and Tc-99m- MIBI scintigraphy (Tc-99m-MIBI) were performed. We defined the metastases as those cases in which serum thyroglobulin (Tg)increased significantly or in which we were able to prove the lesions on CT (computed tomography), MRI (magnetic resonance imaging) or bone scintigram. Three radiology medical specialists visually evaluated each scintigram and calculated the sensitivity, specificity, and likelihood ratio. For whole-body sensitivity, both Tl-201 and Tc-99m-MIBI were high before ablation and I-131 was high after ablation. Before ablation, the negative likelihood ratio was less than 0.1 for Tl-201 and Tc-99m-MIBI, while the positive likelihood ratio was more than 10 for Tl-201. After ablation, the positive likelihood ratio for I-131, Tl-201, and Tc-99m-MIBI was more than 10. The sensitivity of the mediastinum was appropriate, except for I-131 before ablation, and the sensitivity of the lung before and after ablation was inferior for either tracer. The specificity of the cervix for I-131 before ablation was markedly deteriorated, but it increased after ablation.

We investigated the usefulness of helical computed tomography(CT)in the morphological diagnosis of pulmonary vein stenosis, particularly that in infants and small children. In total, 20 helical CT examinations were performed in 10 post-operative cases of Total Anomalous Pulmonary Venous Drainage(TAPVD), 3 cases of single right ventricle, and 1 case of single left ventricle. In all cases, distinct morphological imaging was possible. Pulmonary vein stenosis could be categorized into three types: (1)stenosis from the anastomosis of the common pulmonary vein (CPV)-the left atrium (LA) to the peripheral pulmonary vein; (2) stenosis only at the anastomosis of CPV-LA; and (3) stenosis due to compression by nearby organs. Coronal views by multiplanar reconstruction (MPR) provided morphological information along the up-down direction of the body axis. Morphological diagnosis of pulmonary vein stenosis is important in deciding prognosis and therapeutic regimens, and helical CT was considered useful for such diagnosis in our 14 young patients.

The elimination of fibroblast-like cells from primary cultures of fetal human livers was studied. A fibroblast-like cell line (HuF), which was obtained by subculturing fetal human liver cells 4 or more times, was briefly treated with hydrocortisone (HC) or putrescine (PUT). The growth of HuF cells was inhibited by HC at a concentration of 10(-2) M and by PUT at a concentration higher than 10(-3) M. Long-term treatment of HuF cells with 10(-3) M HC inhibited the growth of the cells. Primary cultures of fetal human livers were made in medium containing HC or PUT, and morphological and functional examinations were made. The cultures were predominantly composed of epithelial-like cells, with few fibroblast-like cells, when the HC concentration was 10(-5)M to 10(-3) M. A high amount of albumin was secreted at these concentrations of HC. On the other hand, at 10(-3) M PUT, many epithelial-like cells were seen, but albumin was undetectable. The present results indicate that albumin-producing epithelial-like cells can be selectively maintained in medium containing HC, in primary cultures of fetal human livers.

The replicative responses of suckling and adult rat hepatocytes in primary culture to growth-stimulating factors were compared. By addition of L-proline alone, the [3H]-thymidine labeling of suckling rat hepatocytes was dramatically enhanced, but that of adult ones was not. Epidermal growth factor (EGF), insulin, triiodothyronine (T3) and glucagon also enhanced the labeling of suckling rat hepatocytes regardless of the presence or the absence of L-proline. On the other hand, in the absence of L-proline, only EGF enhanced the labeling of adult rat hepatocytes, and, in the presence of L-proline, insulin as well as EGF enhanced the labeling. In the presence of growth factors and L-proline, the number of suckling rat hepatocytes increased up to about 143%, whereas that of adult rat hepatocytes hardly increased. Thus, a remarkable difference in replicative responses to growth factors and L-proline was observed between suckling and adult rat hepatocytes in primary culture.

Primary cultures of surgically obtained noncancerous portions of human liver tissues were made. Liver tissues were poorly dissociated with collagenase, but well dissociated with dispase. The yield and viability of cells were improved somewhat when dissociated with collagenase followed by dispase. The mean cell yield was 1.1 X 10(6) cells/g liver. The epithelial-like morphology of the dissociated liver cells was maintained for about one week, but thereafter degenerative alteration of cells was observed. In liver explant culture, an active outgrowth of cells was observed for more than one month. Albumin production in culture fluids from dissociated livers was detectable for about 2 weeks, but later became undetectable, while that from explant culture was detectable for at least one month. These data demonstrate that adult human hepatocytes can be isolated from noncancerous portions of livers with relatively high yield, and that albumin production of the dissociated cells is detectable for several days.

With the purpose of revealing the biological effects of the X-ray irradiation the authors extracted phospholipids from the liver of irradiated animals and proved that this substance has the action to inhibit the growth of the bone marrow cells, the motility of pseudo-eosinophilis and the erythropoiesis in tissue culture, suggesting that the injury will mainly be induced by the toxic substances produced by irradiation.