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This study aims to clarify the presently uncertain value of mass screening for thyroid cancer, which has been performed by physical examination along with mass screening for breast cancer in Zentsuji, Japan since 1983. Among 18,619 subjects, 36 individuals with thyroid cancer (0.19%) were found. The detection rate was 0.40% at the initial screening and 0.10% during subsequent periodic screening. All thyroid cancers were confirmed histologically as well differentiated carcinoma. The tumor size in the mass-screening group (14 +/- 6 mm) was significantly smaller than in patients presenting at an outpatient clinic during the same period (19 +/- 13 mm) (p < 0.05). The incidence of nodal metastases in the mass-screening group (38%) was significantly lower than in the outpatient group (68%) (p < 0.05). Thus mass screening seemed to find thyroid cancers in a relatively early stage. Mass screening for thyroid cancer was economic in this instance because it was performed together with screening for other cancers, such as breast cancer. Thyroid cancer screening required less than one additional minute per subject. The ultimate aim of mass screening is to reduce mortality. No improvement in prognosis from enforced mass screening for thyroid cancer was detected in this study. It cannot be demonstrated that there is sufficient value of mass screening for thyroid cancer to perform it independently despite early cancer detection.

The functional and phenotypic characteristics of lymphocytes separated from myasthenic thymoma (Th-L) were compared with those of lymphocytes separated from non-thymomatous thymus associated with thymoma (NTh-L) of the same patients and NTh-L of myasthenia gravis (MG) patients without thymoma. We examined whether Th-L and NTh-L of MG patients reacted to interleukin-2 (IL-2) to develop lymphokine-activated killer (LAK) activity and/or cytolytic activity against K562 (natural killer (NK) activity), and the phenotypic changes in such cells during incubation. Ten MG patients with thymoma and six MG patients without thymoma, and four non-MG thymoma patients were included in this study. Th-L and NTh-L of MG patients reacted with IL-2 to develop LAK and NK activities. The LAK activity developed from Th-L was significantly higher than that from NTh-L, and the LAK activity developed from Th-L was as high as that from peripheral blood lymphocytes (PBL) in MG patients without thymoma. The proportions of CD3+ cells, CD4+/CD8- cells, and CD4-/CD8+ cells in Th-L of MG patients increased significantly during incubation. On the other hand, the proportion of CD4+/CD8+ cells in Th-L of MG patients decreased significantly. The proportions of CD4+/CD8- cells, CD4-/CD8+ cells, and CD4+/CD8+ cells in NTh-L of MG patients with and without thymoma exhibited no change during incubation. These findings suggest that CD4+/CD8+ Th-L of MG patients may have a higher potential to react to IL-2 than NTh-L, and that the former cells might develop LAK activity like that of PBL on maturation to CD4+/CD8- cells and CD4-/CD8+ cells. Our findings also suggested that Th-L might play an important role in the pathogenesis of MG with thymoma.

We examined p53 mutations in 20 cancer samples from 19 chromate workers with lung cancer by Polymerase chain reaction-Single strand conformation polymorphism analysis and direct sequencing. Six missense mutations were identified in 4 (20%) of the 20 chromate lung cancer samples. Fewer mutations were found in the patients with lung cancers who had been exposed to chromate than in those who had not. However, the pattern of p53 mutations in lung cancer patients exposed to chromate differed from that of common lung cancers in 3 respects. There were no apparent G to T transversions, which are common base changes in lung cancers. Half of the mutational sites (3/ 6) had changes of AT base-pairs, and 2 of 4 mutational tumor samples had double missense mutations. Our results suggested that chromate exposure may induce point mutation of the p53 gene.

Azatyrosine is known to convert ras, raf or c-erbB-2-transformed NIH3T3 cells to a normal phenotype. We attempted to identify the signal-transduction process triggered by oncogenic c-ErbB-2 that was inhibited by azatyrosine. Azatyrosine did not suppress activation of Ras induced by introduction of c-ErbB-2. However, it inhibited increases in phosphorylation of c-Raf-1 induced by oncogenic c-ErbB-2. Furthermore, azatyrosine inhibited activation of the 12-O-tetradecanoylphorbol-13-acetate (TPA) response element in response to stimulation by oncogenic c-ErbB-2. These results suggest that this agent acts downstream of Ras in signal transduction from oncogenic c-ErbB-2 to nuclear factors. Moreover, we found that azatyrosine was incorporated into proteins instead of tyrosine. The simultaneous presence of a high concentration of tyrosine inhibited the conversion to a normal phenotype of transformed cells by azatyrosine. These results strongly suggest that incorporation of azatyrosine into proteins might convert the transformed cells in to cells with a normal phenotype.

Recently, it was suggested that splice variants of the surface glycoprotein CD44 (CD44v) were associated with tumor metastasis in some cancers. We examined the expression of variant forms of CD44 in 31 non-small cell lung carcinomas (NSCLCs) and in 8 normal lung tissue samples by reverse transcription-PCR (RT-PCR). CD44v3, CD44v5, CD44v6, and CD44v7 were not expressed or were weakly expressed in normal lung tissue (0 of 8). In contrast, CD44v3, CD44v5, CD44v6, or CD44v7 was expressed in 28 of 31 (90.3%) NSCLCs. Additionally, we examined the expression of CD44v6, which has been shown to be related to metastasis, in 5 normal lungs and 30 NSCLCs by RT-PCR and immunohistochemical analysis to clarify which cells express CD44v6 in NSCLC specimens. Thirty-six of 61 (59%) NSCLCs variably expressed CD44v6 by RT-PCR, and cancer cells were selectively immunostained by anti-CD44v6 antibodies in 23 of 30 (76.7%) NSCLCs. The results of immunohistochemical analysis almost correlated with those of RT-PCR. NSCLCs with lymph node metastasis expressed significantly more v6 exon than did those without lymph node metastasis [23 of 29 (79.3%) versus 13 of 32 (40.6%); P < 0.01]. There was a significant association between the intensity of v6 expression by RT-PCR and the frequency of cases showing lymph node metastasis (Cochran-Armitage's test, P < 0.002). In conclusion, this study demonstrated that in NSCLC, a number of variant forms of CD44 are frequently expressed, although these variants are infrequently expressed in normal lung tissue, and that the expression of CD44v6 is particularly associated with lymph node metastasis in NSCLC.

Total calcium is routinely measured after thyroidectomy in a clinical setting, while the measurement or calculation of the free calcium level is not generally performed. We reviewed total and free calcium levels in patients who underwent lobectomy (n = 15), subtotal thyroidectomy (n = 15) and total thyroidectomy (n = 15). Postoperative total calcium levels decreased significantly in comparison to preoperative levels in all thyroidectomies (p < 0.01), and this fall was significantly related to the extent of surgery (p < 0.01). In contrast, there was no significant difference between preoperative and postoperative free calcium levels in patients undergoing lobectomy, although we found a decrease in free calcium levels after both subtotal and total thyroidectomy. Total protein levels decreased regardless of the type of operation. Serum total calcium levels were thought to be altered by serum protein levels through the change of protein-bound calcium levels. When examined for free calcium levels, some patients were administered unnecessary calcium supplementation because hypocalcemia had been judged from the total calcium level. Since the wrong diagnosis may be given with regard to hypoparathyroidism by measurement of total calcium levels alone, we propose that free calcium levels should be routinely measured or calculated after thyroidectomy.

We report a case of pleuropulmonary blastoma occurring in the right upper lobe and pleura of a 29-month-old boy. Histologically, the tumor was composed of undifferentiated mesenchymal cells with occasional rhabdomyoblastic and chondroid differentiation. Immunohistochemical analysis showed vimentin immunoreactivity in most of the tumor cells, myoglobin, desmin, and actin in the rhabdoid cells, and S-100 protein in the chondroid cells. Fluorescence in situ hybridization (FISH) analysis showed trisomy 2 and normal chromosomal copy numbers for chromosomes 7, 12, 17, 18, and X in the majority of the tumor cell nuclei. The identification of trisomy 2 in the current pleuropulmonary blastoma confirms a previous cytogenetic finding of chromosome 2 abnormality with cultured metaphase cells of a similar case, indicating that the acquisition of an additional chromosome 2 copy in pleuropulmonary blastoma is a nonrandom chromosomal alteration and that pleuropulmonary blastoma is cytogenetically distinct from pulmonary blastoma in which chromosome 2 is bisomic. It is also suggested that pleuropulmonary blastoma may have an intimate tumorigenetic relationship with embryonal rhabdomyosarcoma.

The present study has demonstrated the influence of bile acids (BAs) on the development and growth of azoxymethane (AOM)-induced aberrant crypt foci (ACF). Male F344 rats were treated with two doses of AOM (15 mg/kg) at 7 days apart and fed either basal MF or MF plus 0.4% of cholic (CA), deoxycholic (DCA), chenodeoxycholic (CDCA), lithocholic (LCA) and ursodeoxycholic (UDCA) acid mixed diets for 8 weeks after the first AOM dose. The mean number of ACF/colon of the rats fed CA, DCA, CDCA and LCA were higher than that of MF-fed group and the differences were statistically significant (P < 0.005). But the mean number of ACFs/colon was significantly (P < 0.005) lower in UDCA diet-fed rats compared to MF. UDCA-fed rats also showed a significant decrease in average crypt multiplicity (number of crypts/focus) of ACF compared to MF alone. The mean number of ACF with > or =5 crypts was about 2.5-3.7 times higher in case of CA, DCA, CDCA and LCA and about 8.2 times lower in UDCA compared to the control MF diet group. In a parallel study, feeding for 18 weeks of the same BAs mixed diets without AOM administration did not significantly induce ACF. Therefore, these data suggest that dietary BAs by themselves do not induce ACF in F344 rats but enhance or, in the case of UDCA, suppress the development and growth of AOM-induced ACF.

A rare case of invasive thymoma with endobronchial polypoid growth is presented. A 68-year-old woman presented with coughing and sputum. The chest X-ray and computed tomography (CT) findings demonstrated a large mass with a calcified lesion in the left hilar region. A bronchoscopic examination showed a polypoid tumor in the left B3b bronchus. The tumor was surgically resected and revealed a unique polypoid endobronchial extension. This tumor was pathologically diagnosed to be invasive thymoma.

Thymic epithelial tumours are broadly classified into thymomas and thymic carcinomas. Although both tumours occasionally show invasive growth, they exhibit different clinical and biological findings. The oncogene and anti-oncogene in thymic epithelial tumours have not been evaluated fully. We investigated the expression of p53 protein by immunohistochemical analysis using the anti-p53 polyclonal antibody (CM-1) in 17 thymomas and 19 thymic carcinomas. We also examined p53 gene (exon 5-8) mutation in 18 thymic carcinomas by using polymerase chain reaction-single-strand conformation polymorphism methods and direct sequencing. Of the thymoma cases, only one invasive thymoma showed focal nuclear staining. Fourteen of the 19 thymic carcinomas (74%) showed nuclear staining. Point mutations of the p53 gene were recognized in only 2 of the 18 thymic carcinomas (11%). One was the mutation C to T transition in the first letter of codon 222 in exon 6, which results in the amino acid substitution from proline to serine. Another was a silent mutation. p53 protein accumulation is highly frequent in thymic carcinomas but not in thymomas, and gene mutation is uncommon in thymic carcinomas.

We have demonstrated the expression of thyrotropin receptor (TSH-R) in thyroid neoplasms (13 adenomas, 21 papillary carcinomas, two follicular carcinomas) and adjacent normal thyroid using the monoclonal antibody against human TSH-R and have also demonstrated a relationship between prognostic scores and the expression of TSH-R. Among the adenomas, eight showed an intensity similar to that of normal thyroid and five showed a higher intensity than normal. Two tumours exhibited heterogeneous distribution of TSH-R. Among the papillary carcinomas, seven showed similar intensity to normal tissue and four showed higher intensity and ten showed weaker intensity. Eight tumours showed heterogeneous distribution of the stain. Among the follicular carcinomas, one showed similar intensity to normal tissue and the other exhibited weaker intensity. Both cases showed homogeneous distribution of TSH-R. The adenomas never showed a weaker intensity than normal thyroid, but various intensities of TSH-R occurred in differentiated carcinomas. There was no significant relationship between the clinical data and the signal intensity in the adenomas. Among the papillary carcinomas, however, the group with weaker intensity had significantly poorer prognostic scores than the other two groups. Thus, we assume that low TSH-R may be expressed by the clinically high-risk group of patients with papillary thyroid carcinoma.

The functions of the transcription factor E2F-1 are regulated by the RB protein through the RB-binding domain of E2F-1 and this factor is considered to be an important molecule that functions downstream of the RB protein. In order to determine whether E2F-1 that cannot bind to RB might be associated with various human cancers, we searched for mutations in the RB-binding domain of E2F-1 using samples of DNA from various clinical specimens obtained from 406 cancer patients (with lung, pancreatic, stomach, colon, esophageal, and hepatic cancers) by analysis of polymerase chain reaction-mediated single-strand conformational polymorphism. No mutations or deletions were detected in genes for E2F-1 from any of the tumor tissues examined. These results suggest that a mutation or deletion in E2F-1 that might affect binding of the RB protein is not involved in human cancers.

Comparison of acquired mutations in the p53 tumor suppressor gene can illuminate factors contributing to carcinogenesis among cancer cohorts. Japan has an ethnically homogeneous population with a low incidence of breast cancer. Previously we reported an unusual frequency, allelic status, and clustering of mutations in breast cancers from the northern part of the main Japanese island. To extend these findings, exons 2-11 and adjacent intronic sequences were analysed in tumors of women from northern (Hokkaido) and southern (Tokushima) Japan. The frequency of breast cancers with p53 gene mutations in the Hokkaido group is the highest reported (81%) while that in Tokushima (28%) is similar to most other populations. Thirteen of the 19 mutations (68.4%) in the Hokkaido cohort were heterozygous, an unusually high frequency for p53 mutations in any tumor type. There were three missense mutations at codon 175, a known hotspot for alterations in the p53 gene, and three missense mutations at codon 179, a rare site for p53 changes. In addition, the patterns of p53 gene mutation differed between the two Japanese cohorts (P=0.04). The multiple differences in acquired p53 mutations suggest unsuspected biological differences among breast cancers in northern and southern Japan. In addition, the high frequency of p53 mutations in breast cancers from Hokkaido predicted a poorer prognosis for this population which was confirmed on examination of mortality data.

Rho proteins have been implicated in a variety of cytoskeletal functions, but it is unclear how Rho proteins regulate these cellular functions and how Rho proteins are regulated. In this study, we cloned the rhoB gene from the mouse genome and characterized its promoter region. The predicted amino acid sequence was identical to that encoded by the human rhoB gene. A site for initiation of transcription was found at position -376 relative to the site for initiation of translation. Deletion analysis of the 5'-flanking region of the rhoB gene revealed that the minimum region of the promoter was located between positions -507 and -376. Northern blotting analysis showed that the expression of the mouse rhoB gene was induced by serum, suggesting that expression of the rhoB gene might be controlled by some signal-responsive element(s).

Background: Since 1973 we have performed extended thymectomy for myasthenia gravis because of the presence of thymic tissue in the anterior mediastinal fatty tissue. Follow-up results were reviewed and influencing factors were investigated., Methods: Three hundred seventy-five patients with myasthenia gravis (286 nonthymomatous and 89 thymomatous) who have undergone extended thymectomies were reviewed. The status of the patients was evaluated as follows: A (remission), B (improvement), C (no change), D (deterioration), E (death due to myasthenia gravis). Evaluation was performed at 3 and 6 months, and at 1, 3, 5, 10, 15, and 20 years. The effectiveness of the operation was estimated by the remission rate (RR = A/Total number of patients evaluated) and the palliation rate (PR = A + B/Total number of patients evaluated) at each point., Results: Remission rates of the nonthymomatous patients were 15.2% (3 months), 15.9% (6 months), 22.4% (1 year), 36.9% (3 years), 45.8% (5 years), 55.7% (10 years), 67.2% (15 years), and 50.0% (20 years). Remission rates in the thymomatous patients were 13.6% (3 months), 17.5% (6 months), 27.5% (1 year), 32.4% (3 years), 23.0% (5 years), 30.0% (10 years), 31.8% (15 years), and 37.5% (20 years). Absence of thymoma, younger age, and short duration of the disease were favorable prognostic factors. Thymectomy was effective also in patients with ocular myasthenia gravis. Preoperative steroid administration did not improve the outcome., Conclusions: Extended thymectomy is an excellent operative procedure for myasthenia gravis in both nonthymomatous and thymomatous patients.

We examined the binding of various steroid hormones to DNA in vitro by means of 32P-postlabeling. Seventeen steroid hormones and cholesterol (CS) were incubated with human liver DNA at 37 degrees C for 1 h under aerobic conditions in the absence of catalysis. The reaction mixtures were analyzed by the nuclease P-1 version of 32P-postlabeling. The results showed that cortexolone (CX), prednisolone (PS), cortisone (CN), cortisol (CL), tetrahydrocortisol (TC), corticosterone (CC), 11-deoxycorticosterone (DC), dexamethasone (DX), dihydrocortisol (DL), and aldosterone (AL) covalently bound with DNA. However, progesterone (PG), 17 alpha-hydroxyprogesterone (HG), estrone (E1), estradiol (E2), estriol (E3), testosterone (TS), cortol (CR) and the original compound for biosynthesis, CS, did not form adducts. In absence of DNA, the steroids themselves did not give rise to any spot on TLC under the same conditions. The dose-responses of DNA binding by DC, DL, CC, CL and CN were linear. The relative adduct labeling of reactive steroids at a concentration of 2 mM were as follows: 68.8 (CX), 53.2 (PS), 39.6 (CN), 29.9 (CL), 20.9 (TC), 12.9 (CC), 12.3 (DC), 7.5 (DX), 4.7 (DL), 1.2 (AL) adducts per 10(8) nucleotides. Reactive and nonreactive steroids were distinguishable by the presence or absence of the carbonyl group (-CO-CH2OH) at carbon seventeen (C17) of the cholesterol skeleton. This implies that the electrophilic carbonyl or a neighboring group perhaps involved in the formation of covalent bond with DNA. To investigate the nature of target base(s) of these DNA reactive steroids, mononucleotides of all four bases of DNA were reacted with CN, CL, CC and cochromatographed with the obtained spots of DNA reactions. The results of which stated that these steroids and guanine reaction gave the same spots as observed in DNA reaction, indicating guanine is the main target of these DNA reactive steroids. Hep G2 human hepatocellular carcinoma cells were used as an alternative model. Although nine steroids (CL, DL, TC, PS, DX, PG, E2, TX, CR) did not react with intracellular DNA under our experimental conditions, our findings suggested that some hormonal steroids can form covalent DNA adducts in vivo.

The biological differences between lymphocytes separated from thymoma patients with myasthenia gravis (MG) and those separated from thymoma patients without MG was examined. We investigated whether lymphocytes in thymoma (Th-L) of patients with and without MG could react to interleukin-2 (IL-2) to develop cytolytic activity against K562 and lympho kine-activated killer (LAK) activity, and their phenotypic changes during incubation with IL-2. Sixteen thymoma patients who consisted of eight patients with MG and eight patients without MG were investigated. Th-L of all MG patients could react to IL-2 to develop LAK activity. Th-L of all of MG patients also developed cytolytic activity against K562 target cells. Cytolytic activity against K562 and LAK activity from Th-L of the MG patients were significantly higher than those from Th-L of the patients without MG (P < 0.01, P < 0.01). The proportions of CD3+ and CD4-/CD8+ cells of Th-L of MG patients increased significantly during the incubation period with IL-2 (P < 0.05, P < 0.05). The proportion of CD4+/CD8+ cells of Th-L of patients with MG decreased significantly (P < 0.05) during incubation with IL-2. On the other hand, there was no significant phenotypic change in Th-L of the patients without MG. These results indicate that MG induces the functional and phenotypic activation and functional and phenotypic maturation of Th-L.

We reported recently that azatyrosine inhibits the growth of c-Ha-ras, c-raf or c-erbB-2-transformed NIH3T3 cells and converts the transformed cells to cells with a normal phenotype. To analyze the mode of action of azatyrosine, we examined the effects of azatyrosine on the synthesis of macromolecules. Azatyrosine had no obvious inhibitory effects on the synthesis of DNA, RNA and protein in c-erbB-2-transformed cells. Furthermore, azatyrosine inhibited cell growth but did not interrupt the cell cycle at any specific stage. Thus, the mode of action of azatyrosine appeared to be different from that of typical anticancer drugs. Moreover, we found that azatyrosine was incorporated into proteins instead of tyrosine. The simultaneous presence of a high concentration of tyrosine inhibited the conversion to a normal phenotype of transformed cells by azatyrosine. These results strongly suggest that incorporation of azatyrosine into proteins might convert the transformed cells to cells with a normal phenotype. The analysis of azatyrosine-containing proteins by two-dimensional electrophoresis revealed that the mobilities of some proteins differed from those of the corresponding tyrosine-containing proteins. An alteration in molecular structure of the proteins that include azatyrosine residues might be associated with the ability of azatyrosine to convert transformed cells to cells with a normal phenotype.

Objective: TSH suppression therapy has been reported to be effective in some cases of papillary carcinoma of the thyroid. However, there has been little information regarding the status of expression of TSH-R mRNA in papillary carcinomas. Therefore, we examined the expression of TSH receptor (TSH-R) mRNA in thyroid tumours by in situ hybridization., Design: Surgically resected thyroid tumour specimens were used. The expression level of TSH-R mRNA was detected by in situ hybridization, and the difference between tumours and their adjacent normal thyroid tissues was assessed., Patients: Tumours and adjacent normal thyroid tissues were obtained from nine patients with differentiated papillary carcinomas and eight with adenomas., Measurements: We performed in situ hybridization on frozen sections of thyroid tissues. Digoxigenin-labelled oligonucleotide was used to hybridize and to detect TSH-R mRNA. We counted positively stained cells and non-stained cells in each section, calculated the positivity of stained cells and assessed the distribution of positively stained cells., Results: In normal thyroid tissue, the positivity was 95.3 +/- 2.3% (mean +/- SD) and the distribution of TSH-R mRNA was homogeneous. In adenoma, positivity was 93.5 +/- 4.4%. In most cases of adenoma, the distribution was homogeneous. In papillary carcinoma, the expression level of TSH-R mRNA was significantly lower and the positivity was 81.9 +/- 12.6%. Furthermore, in five cases of papillary carcinoma, the distribution was heterogeneous; four of these cases were stage 3, while the remaining case was stage 1. Three of the four cases with a homogeneous distribution were stage 1, while one case was stage 3., Conclusions: The heterogeneity of TSH-R mRNA distribution suggests the effect of TSH on carcinoma cells may be variable. Furthermore, papillary carcinomas of advanced clinical stage showed a tendency to low expression and heterogeneous distribution of TSH-R mRNA.

Between 1985 and 1988, the effect of using ftorafur (FT) or PSK (an immunotherapy agent) in combination with the conventional postoperative adjuvant therapy using mitomycin (MMC) plus tamoxifen (TAM) was assessed in stage II, oestrogen receptor-positive (ER+) breast cancer patients. Furthermore, in ER- breast cancer stage II patients, the effects of postoperative adjuvant therapy using MMC plus FT were compared with the effects of postoperative adjuvant therapy using MMC plus PSK. Patients had primary stage II breast cancer and had undergone total mastectomy plus axillary dissection or more radical surgery. On the day of surgery, MMC (13 mg/m2) was administered intravenously. Then, ER+ patients received one of three regimens of drug therapy, starting 2 weeks after surgery: regimen A (daily oral treatment with 30 mg of TAM), regimen B (daily oral treatment with 30 mg of TAM and 600 mg of FT) or regimen C (daily oral treatment with 30 mg of TAM and 3 g of PSK) [corrected]. ER- patients received either regimen D (daily oral treatment with 600 mg of FT) or regimen E (daily oral treatment with 3 g of PSK), starting 2 weeks after surgery. Of the 540 ER+ patients registered, 525 were evaluated. The 5-year overall survival rate for ER+ patients was higher for patients who received regimen B (94.2%) than for those who received regimen A (86.9%) or regimen C (89.9%) (P = 0.063). The 5-year relapse-free survival rate was higher for regimen B (88.9%) than for regimen A (78.6%) and regimen C (77.2%) (P = 0.010). Stratified analysis revealed better results with the FT-combined therapy in patients positive for lymph node metastasis and premenopausal patients. These results indicate the effectiveness of using FT in combination with TAM. Of the 376 ER- patients registered, 364 were evaluated. The 5-year overall and relapse-free survival rate for ER- patients did not differ significantly between patients who received regimen D and those who received regimen E.

Mutations of the p53 tumor suppressor gene are frequent in lung cancers. It is suggested that p53 mutations are associated with smoking-induced lung carcinogenesis. We examined p53 mutations in 53 lung cancers by analyzing reverse transcription-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) to ascertain the association between p53 mutations and smoking. Twenty-five (47%) of 53 lung cancers carried p53 mutations. A discriminant analysis showed that the Brinkman index (0.156) and gender (0.140) significantly influenced p53 mutations. Furthermore, there was a dose-response relationship between the quantity of cigarettes consumed and the frequency of p53 mutations in lung cancer patients (P < 0.001). In patients with adenocarcinoma, the frequency of p53 mutations correlated with the amount of the tobacco smoked (P < 0.05). We suggest that the p53 gene is a target of particular carcinogen in tobacco smoke.

In Brown Norway to Lewis rat lung transplantation, short-term administration of cyclosporine produces permanent adoption of allografts; however, the adopted grafts show symptoms of chronic rejection. To clarify the difference in cytokine production in acute an chronic rejection of the allografts, an immunohistochemical study was performed. In acute rejection, positive cells for respective cytokines were observed in infiltrating cells, increasing in number as the days after transplantation passed, and reaching a maximum on the fifth day. The strongest reactivity was observed perivenously. In chronic rejection, TNF-alpha positive cells were observed in the perivascular and peribronchial regions, especially around class II positive epithelia. The number of positive cells was, however, less than that in the vascular phase of acute rejection. Few cells were positive for IL-1 beta, IFN-gamma and, unexpectedly, for IL-4. These facts indicate the functional difference of infiltrating cells between acute and chronic rejection.

Azatyrosine [L-beta-(5-hydroxy-2-pyridyl)alanine] is known to convert ras- or raf- transformed NIH3T3 cells to a normal phenotype. We report herein that azatyrosine also inhibits the growth of c-erbB-2-transformed cells and induces normal morphology. We attempted to identify the signal-transduction process triggered by oncogenic c-ErbB-2 that was inhibited by azatyrosine. Azatyrosine neither affected the phosphorylation of the introduced oncogenic c-ErbB-2 nor did it suppress activation of Ras induced by introduction of c-ErbB-2. Thus, azatyrosine did not inhibit the signal transduction from oncogenic c-ErbB-2 to Ras. However, azatyrosine inhibited increases in phosphorylation of c-Raf-1 and c-Jun induced by oncogenic c-ErbB-2. Furthermore, azatyrosine inhibited activation of the 12-O-tetradecanoylphorbol-13-acetate (TPA) response element in response to stimulation by oncogenic c-ErbB-2. These results suggest that azatyrosine acts downstream of Ras in signal transduction from oncogenic c-ErbB-2 to nuclear factors.

We have reported recently that azatyrosine inhibits the growth of ras-, raf-, or c-erbB-2 transformed NIH3T3 cells and converts the transformed cells to cells with normal morphology. We show in this study that treatment of c-evbB-2-transformed cells with azatyrosine restore the capacity for the formation of actin stress fibers, which is closely associated with the phenotype of normal cells. Azatyrosine also restored the expression of the rhoB gene, the product of which regulates the formation of actin stress fibers. To investigate the roll of expression of rhoB in the morphological reversion, c-erbB-2 transformed cells were transfected with a rhoB expression vector. The cells that overexpressed rhoB were flatter than the parental transformed cells and the formation of actin stress fibers was observed in these flatter cells. Furthermore, the growth rate of these cells was lower than that of parental cells, although the flatter cells were still able to grow anchorage-independently. These results suggest that expression of rhoB is involved in the conversion of c-erbB-2-transformed cells to a normal phenotype that is induced by azatyrosine, but such expression is insufficient by itself for complete reversion.

Multiple sections of 40 consecutive cases with invasive ductal carcinoma of the breast, all of which bad wide intraductal cancerous extension, were examined by immunohistochemical analysis for evaluation of hormone dependency in several areas of breast cancer tissues. In this study, we examined the expression of pS2 protein in the central invasive area (CIV), central intraductal cancerous area (CDC) and forefront intraductal cancerous area (FDC). pS2 staining was positive in 52.5% (21/40) of CIV and a significant correlation was found between pS2 expression in CIV and the estrogen receptor status (ER). pS2 staining was positive in 77.5% of CDC and 85.0% of FDC, respectively. A majority (68.4%) of the cases that were negative pS2 in CIV were positive for pS2 in FDC. Moreover, the cases with noncomedo intraductal carcinoma in premenopausal status showed a higher positivity of pS2 expression in FDC than the cases with comedo-carcinoma, though the number of cases of comedo-carcinoma was limited. These findings suggest that endocrine therapy may be useful after breast conserving treatment regardless of the ER status of the primary tumor.

The role of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) in augmentation of lymphokine-activated killer (LAK) cell induction by interleukin-2 (IL-2) from pleural cavity mononuclear cells (PCMNCs) was examined in sixteen patients with resectable primary lung cancer not associated with malignant effusion. None of the patients had received any anticancer therapy prior to this study. Incubation of PCMNCs of patients without malignant effusion with GM-CSF for 4 days in the presence of IL-2 resulted in a significant increase in LAK activity against natural killer-resistant Daudi cells. This result was obtained by using the 4 h 51Cr-release assay. PCMNCs and blood mononuclear cells (BMNCs) were harvested simultaneously from pleural cavity lavage fluid and peripheral blood in lung cancer patients. The LAK activity developed from PCMNCs and BMNCs following incubation with IL-2 for 4 days, but the LAK activity from PCMNCs was significantly lower than that from BMNCs (P < 0.05). Incubation of PCMNCs with GM-CSF augmented the LAK activity from PCMNCs to a level as high as that from BMNCs. These results suggest that the combined use of GM-CSF with IL-2 may result in augmentation of LAK activity developed from PCMNCs of lung cancer patients without malignant effusion.

Thymidylate synthetase (TS) is a key enzyme as a methyl donor in the methylation reaction from dUMP to dTMP. TS activity was assessed in various tissue of mammary disorders. The descending order of TS activity was as follows: cancer-positive nodes, primary cancers, cancer-negative nodes, benign lesions, and normal parenchyma. Significant differences in TS activity were found between the positive nodes and each of the other tissues (p < 0.01). In node-positive cases, a significant correlation in TS activity was found between the primary cancers and positive nodes (r = 0.616, p = 0.033). There was no correlation between the nodal status and the TS activity in primary cancers. In 11 of 12 cases, the TS activity of positive nodes was higher than the 'calculated' TS activity of the primary cancer, which was defined as the TS activity per unit weight of cancer cells. A significant correlation was found between the calculated TS activity and the mitotic frequency in primary cancers (r = 0.697, p = 0.0001). On the other hand, a significant correlation could not be found between the TS activity and the mitotic frequency in positive nodes (r = 0.364, p = 0.244).

A 53-year-old female suffering from renal stones and hypercalcaemia was diagnosed as having primary hyperparathyroidism caused by hyperplasia of the parathyroid glands. She underwent total parathyroidectomy and implantation of parathyroid tissue. After one year, she underwent surgery for a jaw tumour. The pathological findings indicated it to be a cementifying fibroma. Jackson et al. (1990) reported the familial association of hyperparathyroidism with jaw tumours, and they suggested that this condition represents a new clinical syndrome. We believe that our case belongs to this syndrome.

The mechanism of mucosa-specific formation of DNA adducts, which was found recently in human intestines, was studied in male F344 rats treated with 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). There are three conceivable pathways for p.o. administered IQ to reach the target colonic mucosal cells: pathway 1, through the digestive canal which exposes from the lumenal direction; pathway 2, following enterohepatic circulation re-expose from the lumenal direction; and pathway 3, exposure via blood circulation. To investigate these possible pathways, the following surgical procedures were performed: (a) portal catheterization for IQ administration to eliminate pathway 1 and (b) choledochal catheterization for bile drainage to eliminate pathway 2. When both procedures are combined, only pathway 3 is active. Four types of IQ-DNA adducts were commonly observed in the colons of all experimental groups, with no qualitative difference between the mucosal and muscular layers. When IQ-HCl was administered by p.o. gavage at a dose of 100 mumol/kg body weight, approximately 70% of the IQ-DNA adducts in the colonic mucosa (13.1 +/- 4.3 adducts/10(7) nucleotides) was induced through pathway 1. Pathway 3 induced the remaining 30% of mucosal adducts, producing equal adduct levels in both layers. Pathway 2 did not work for adduct formation. The DNA adduct formation was unaffected in the presence of intestinal flora, indicating that detoxified IQ does not reactivate by floral enzymes. In conclusion, mucosa-specific DNA adduct formation in the colon is caused most likely by the absorption of carcinogens through the lumen.

We recently encountered a 6-year-old girl with a malignant phyllodes tumor of the breast. The patient's mother noticed a tumor on the right breast of her first doughter at 8 months after from her birth. The baby was brought to the Second Depertment of Surgery, University of Tokushima, at age 20 months. We recommended removal of the 3.5 x 2.9 cm tumor in the right breast, but it was not done. Next, she visited our department at 6 years of age because the tumor had gradually enlarged, reaching a size of 4.3 &timus; 4.0 cm. She underwent excisional resection of the tumor, and the tumor was diagnosed as a malignant phyllodes tumor of the breast coexisting with a borderline phyllodes tumor. To our knowledge, this is the youngest reported case of a malignant phyllodes tumor of the breast. Moreover, the malignant lesion was positive for estrogen receptor (ER) and showed strong proliferating cell nuclear antigen (PCNA) staining. On the other hand, the borderline part was negative for ER and showed weak PCNA staining. Thus, in the present case, the expression of ER, the exposure to estrogen (for example, the mother's estrogen during gestation) and increase in the proliferation rate may have played important roles in the mechanism of the transformation of the phyllodes tumor.

The patient was a 38-year-old woman who was diagnosed as having right breast cancer (T1aN1bM0, stage II) at the age of 36 when she underwent right mastectomy. Postoperative adjuvant therapy was performed using UFT and tamoxifen. After 2 years and 4 months, pulmonary metastasis was diagnosed from elevated tumor markers and chest X-P. After the relapse, treatment with goserelin acetate (Zoladex) and CPA was started. After 2 months of this treatment, the tumor in the lung had shrunk in size, and after 5 months partial remission (PR) was evaluated from improvements in chest X-P and normalization of tumor markers, This PR condition continues at present. A case report is presented of a patient with premenopausal recurrent breast cancer in whom good results were obtained by concomitant administration of Zoladex and CPA.

During screening for inhibitors of ras-mediated differentiation of PC12 cells, trichostatin A (TSA) was isolated from the metabolites of Streptomyces as a potent inhibitor. TSA blocked both oncogenic ras- and NGF-induced neurite outgrowth from PC12 cells. However, addition of TSA 1 h after NGF-stimulation did not inhibit neuronal differentiation, suggesting that TSA affects an early step in the NGF-signaling pathway mediated by ras. Northern blotting analysis showed that TSA prolonged the maximum expression period of c=fos mRNA triggered by NGF and delayed its return to the basal level. TSA reduced c-jun mRNA induction by NGF but greatly enhanced c-myc mRNA induced by NGF. Yoshida et al. (J. Biol. Chem, 265, 17174-17179, 1990) showed that TSA inhibits histone deacetylation, which might influence the gene expression involved in cellular differentiation. In this study, we also found that TSA prevents histone deacetylation in PC12 cells as well as other cell lines, suggesting that inhibition of histone deacetylation by TSA might affect the expression of early-response genes. We also demonstrated that TSA induced reversion of oncogenic ras-transformed NIH3T3 cells to a normal morphology, suggesting that inhibitors of ras-mediated differentiation of PC12 cells may be effective as anticancer agents.

A cDNA library was constructed from COLO 205 and 1056 clones randomly selected from this library were partially sequenced. Two hundred and two (38.4%) out of 526 independent genes had more than 80% similarity to the genes reported in GenBank. In Northern blot analysis, 96 out of 98 genes were shown to be expressed at the same level in colon and lung carcinoma cell lines and control fibroblasts. Only two clones, including human synovial phospholipase A-2 and a homologue to rabbit calgizzarin, were expressed at different levels among these cell lines. The full sequence of human calgizzarin was determined and its expression was remarkably elevated in colorectal cancers compared with that in normal colorectal mucosa.

We attempted to investigate the process of ischemia-induced disturbances in the rat liver, employing the electrical bio-impedance technique. The electrical bio-impedance was measured continuously over 6 h by the 4-electrode method, at various incubation temperatures, in six liver samples extracted from male Wistar rats. The electrical properties of biological tissues can be expressed in terms of three parameters: extracellular resistance (Re), intracellular resistance (Ri) and cell membrane capacitance (Cm). These three parameters were calculated from the measured values of the electrical impedance by the curve-fitting technique, using a computer program. The Re value increased rapidly after the rat livers were extracted, and then decreased slowly. The Re value reached a peak after about 13 min at 36 degrees C, and then decreased slowly, becoming constant after 3 h. There was a negative correlation between the Tmax of Re (the time when Re reached a maximum) and the incubation temperature (R = -0.973, P < 0.001). The Ri value decreased once in the early stage after extraction, followed by almost no change and then an increase after 4 h at 36 degrees C. The Cm showed a similar pattern of change to the Re value, and a negative correlation was also found between the Tmax of Cm and the incubation temperature (R = -0.969, P < 0.001). The increase in the Re and Cm values, and the decrease in the Ri value for quite long periods after the blood flow has stopped, suggest an increase in the resistance of extracellular fluid due to a decrease in its volume, an increase in cell membrane capacitance due to cell swelling, and a decrease in cellular fluid resistance due to an increase in its volume. The time when the Cm value decreases rapidly after an initial gradual decrease after the peak corresponds well with the time when the Ri value begins to increase, from which it is estimated that cell lysis proceeds and that the flow of extracellular fluid into the cell begins at this time. The findings of this study suggest the possibility of estimating the changes in liver tissue or the tissue structure due to ischemia.

With hyperthermia for treatment of superficial tumors in mind, a prototype applicator with two electrodes arranged concentrically on a disk was designed for efficient local heating, and a basic heating test was carried out. Frequencies as low as 200 kHz were used in order to simplify the configuration of the power device. The applicator consists of two electrodes, a circular inner electrode and another looped outer electrode, arranged concentrically. Water was passed through the applicator as a cooling mechanism; it was placed in direct contact with the target tissue to be heated and then charged with electricity. In the heating test using a phantom, oval hot spots were noted below the inner electrode. Using cooling water at 3 degrees C and 8.2 W, an isothermal line of 45 degrees C was located at a 5-mm radius circle around the central axis with 9 mm depth. A similar temperature distribution map was obtained in heating tests on the thigh muscle of a mongrel adult dog. The temperature distribution maps obtained from these tests corresponded closely with the results of theoretical analysis carried out according to the finite-element method. Since a comparatively low frequency was employed for this applicator the power device was simplified, which made adequate heating possible with low electric power. The temperature distribution map indicated that efficient local heating of superficial tumors could be achieved.

At age 35, the patient was diagnosed with left breast carcinoma (T4bN0M0, Stage IIIb) during her first pregnancy, and underwent mastectomy of the left breast and bilateral oophorectomy. Although CPA was administered as adjuvant chemotherapy after the operation, left pelvic pain developed about a year after the operation, and bone metastasis was detected from the imaging diagnosis. Then, the therapy was switched to UFT (600 mg/day) and tamoxifen (20 mg/day), which markedly relieved the pain along with bone scintiscan (NC) and simple X-P (NC). Five years have passed since the operation, and the progress is excellent. There is neither increase of pain nor adverse reactions to drugs. Furthermore, the patient received breast reconstruction operation at her strong insistence in the third year after operation, and has achieved a high quality of life (QOL).

The presence of several covalent DNA adducts in the human colon was demonstrated by 32P-postlabeling in a previous study. We demonstrated that DNA of all the colonic mucosa tested were selectively adducted by a single genotoxic agent and this modification was completely absent in the DNA of muscular layers. In this study, the DNA adducts of the small intestine are compared with those of the colon to understand the role of mucosa-specific DNA adduct (MSA) in intestinal carcinogenesis. The mucosal DNA of the small intestine from 19 adults undergoing surgery due to gastric carcinoma (seven cases), pancreatic carcinoma (four cases), colon carcinoma (four cases), small intestinal tumor (two cases), intestinal trauma (one case) and ectopic pancreas (one cases) were analyzed. The mucosa-adjacent muscular layer DNA of the corresponding samples was examined as a control. Several common DNA adducts were observed in both mucosal and muscular layers of all the adults. Besides these common background DNA adducts, two MSAs (Si1, Si2) were detected in most of the adults as in colon cases. Si1 was present in all adults examined (19/19 cases) at a level of 0.04-0.22 adducts/10(8) nucleotides (average 0.09) and Si2 was found in 13/19 patients at a level of 0.03-0.08 adducts/10(8) nucleotides (average 0.05). Si2 was the same adduct detected in the adult colonic mucosa. However, they were absent in the adjacent muscular layer as well as in the neonatal intestine tested as a control. The total level of the mucosa-specific DNA adducts of the small intestine was approximately 28-fold lower than that of the colon. Considering that the incidence of cancer in the small intestine is rather lower than that in the colon, these results may be relevant to the development of human intestinal cancer.

Late airway changes are frequently observed in long-term surviving rat lung allografts. In this present study, we investigate this problem using the mixed lymphocyte reaction, graft-versus-host assay (popliteal lymph node assay), skin grafting from the donor strain to the lung allograft recipient, and histology to evaluate this problem. The results show that spleen cells from recipient rats with long-term surviving lung allografts possessed alloreactivity in mixed lymphocyte reaction against both donor and third-party ACI antigens. Suppressor cell activity was not detected in spleen cells from these recipient rats. The popliteal lymph node assay showed that the response to recipient spleen cells was detectable but weaker than that of naive spleen cells from the same strain. Mean survival time of skin grafts from the donor strain to long-term allograft survivors was significantly longer than that between donor and recipient strains. Chest roentgenograms revealed opacification of long-term surviving lung allografts at 2 to 3 wk after skin transplantation that was coincident with the rejection of the skin grafts, and histologic examination of the lung allografts revealed changes compatible with acute rejection. We conclude that alloreactivity can be demonstrated in rats with long-term surviving lung allografts and suggest that late airway changes in these lungs are immunologically mediated.

We report a case of a 60 year-old man with an umbilical polyp composed of the aberrant pancreas and small intestinal mucosa. This is an extremely rare lesion which originates in remnants of omphalomesenteric duct and is usually diagnosed in the infant period. Microscopic examination disclosed the aberrant pancreas with exocrine glands and ducts, but no islets of Langerhans. The above findings suggested that the pancreas might have been HEINRICH II in type.

The in vitro experiment was carried out following 32P-postlabeling analysis to determine the DNA-reactive bile acids present in the human body. The unconjugated and conjugated forms of cholic (CA), chenodeoxycholic (CDCA), deoxycholic (DCA) and lithocholic acid (LCA) were added to calf thymus DNA followed by 1 h of incubation at 37 degrees C. After the incubation the mixture was analyzed by the nuclease P1 modification of 32P-postlabeling. Among the 12 bile acids tested, our results showed that unconjugated CDCA and LCA and the glycine and taurine conjugates of LCA (g-LCA, t-LCA) were able to bind covalently with naked DNA in vitro without intervention of any catalyst. It was also shown that bile acid alone did not give any spot on TLC. These binding reactions depended on the bile acid concentration in a linear manner. The data on the extent of binding at a concentration of 0.1 mg/ml showed values of 28.5 (t-LCA), 23.7 (g-LCA), 3.47 (LCA) and 1.32 (CDCA) adducts per 10(8) nucleotides. These reactive bile acids were also incubated with COLO 205 human colon carcinoma cells and Hep G2 human hepatocellular carcinoma cells for 24 h, but no specific DNA adduct was formed. Further, when LCA or CDCA was administered into male Fischer 344 rats by gavage at a dose of 10 mg/rat every 8 h for 3 days, no specific DNA adduct was detected in their liver or colon. Covalent DNA adducts are believed to cause alteration of the primary structure of genes, which is potentially linked to carcinogenesis. Though our preliminary data failed to detect any bile acid-related DNA adducts in the cultured cells or in rats, the results may provide a basis for assuming some of these bile acids to be potential initiators of colon cancer.

To identify the characteristics of cases with an wide intraductal cancerous extension (WICE), we examined the relationship between WICE and type IV collagen distribution, and the relationship between WICE and the content of the proliferation-associated proteins in human breast cancer. The immunohistochemical distribution of type IV collagen and proliferating cell nuclear antigen (PCNA) were investigated in formalin-fixed tissue sections from 21 breast cancer cases. We demonstrated a significant correlation (p=0.014) between the presence of WICE and immunostaining of type IV collagen in the cancerous ducts (ducts occupied by cancer cells) in the central invasive area of breast cancer. However, no correlation was found between the presence of WICE and the PCNA index (percentage of positive cells per 1000 tumor cells). These findings suggest that the lack of the process of the loss of type IV collagen in the duct wall is more important than the nature of the tumor cells in the development of WICE.

The electrical impedance of a pulmonary mass was measured in 53 patients of whom 44 had primary lung cancer, 5 had metastatic lung tumor, and 4 had organizing pneumonia. Because biologic tissue can be regarded, electrically, to consist of extracellular resistance (Re), intracellular resistance (Ri), and the electrical capacitance of the cell membrane (Cm), these three parameters were calculated from the measured electrical impedance of tissue by a curve-fitting technique using a computer program. The Re of lung tissue was significantly greater (p < 0.01) and the Cm of lung tissue was significantly less (p < 0.01) than that of a pulmonary mass. The Re of malignant tumors (both lung cancer and metastatic tumors) was significantly greater (p < 0.01) and the Cm of malignant tumors was significantly less (p < 0.01) than that of organizing pneumonia. With this information, we used a biopsy needle to diagnose nine intrathoracic lesions. This technique additionally allowed us to confirm the proximity of the needle tip of the mass. The electrical impedance of the lung mass was measured through the biopsy needle using a modified impedance analysis system before the biopsy was performed. There were no false-negative results, and one false-positive result. The rapid measurement of the electrical impedance of a pulmonary mass, preoperatively, may be of value in the clinical evaluation of a pulmonary mass both by facilitating needle guidance and by permitting diagnosis based on electrical impedance.

Seventy tumors of invasive ductal carcinoma of the breast were examined for p53 alteration by the RT-PCR-SSCP method. Sixty-five samples (92.9%) were investigated in the regions of codons 101-200 and 201-300. In total, 16 samples (24.6%) showed p53 gene alteration. We found that p53 gene alteration showed a correlation with (1) a negative ER status, (2) a negative PgR status, and (3) a high histologic grade (especially numerous mitotic figures) of the tumor. However, we found no correlation between p53 gene alteration and the lymph node status or clinical stage. Thus, p53 gene alteration in breast cancer may occur in highly malignant breast cancer other than advanced clinical stage cancer or node-positive cancer.

We described two patients of thyroid cancer with tracheal invasion. A 54-aged female visited our hospital with complaint of the neck pain. Echography showed a tumor in the right lobe of the thyroid. In operation the thyroid tumor invaded the trachea and right recurrent nerve. A 24-aged male visited our hospital with complaint of a mass of the left side neck. Echography showed a tumor in the left lobe of the thyroid. In operation the thyroid tumor invaded the trachea, lymph node and left recurrent nerve. In these two cases we could find thyroid tumors invaded into the trachea by echography. We re-evaluated the utility of echography.

A controlled cooperative study was carried out to assess the value of modified radical mastectomy for patients with stage II breast cancer. The data was analyzed from 11 institutions in the Shikoku District participating in a prospective clinical trial in which patients were randomly assigned either to a modified radical mastectomy group or an extended radical mastectomy group. These two groups of patients were similar to each other in terms of such background factors as age distribution, menopausal status, TNM classification, tumor size, location of the primary tumor, axillary nodal involvement, histological type, and estrogen receptor status. The median follow-up times in the modified and extended radical mastectomy groups were 4.7 and 4.5 years, respectively. The cumulative curves indicated no difference between the two groups in either disease-free survival or overall survival. The survival rates were classified according to the presence or absence of axillary nodal metastases. However, no significant difference was found between the two groups. These findings thus suggest that the routine removal of the grossly uninvolved major pectoral muscle and parasternal lymph nodes is not necessary in patients with stage II breast cancer.

To ascertain the cause of pleural fibrosis in lung allografts, pleural changes were investigated in rat syngeneic and allogeneic lung grafts. The pleura of lung syngeneic grafts showed no pathological changes except for mild edema on the first day after transplantation. In lung allografts, recipient cells migrated into the subpleural tissue early after transplantation (latent phase). In the vascuar phase, recipient lymphocytes in the subpleural tissue increased in number, while almost all alveolar structures were free from infiltration. Both CD4-positive and CD8-positive cells infiltrated in almost equal numbers with macrophages. The subsets of infiltrating cells were similar to those of the perivascular and peribronchial areas. In the late vascular or alveolar phase, fibroblasts were observed among the infiltrating cells, and fibrotic changes started. In the destructive phase, collagen formation with marked pleural thickening was dominant. Pulmonary acute rejection should be treated at least up to the late vascular phase to prevent pleural fibrosis.

The intraarterial arm injection of freshly isolated autologous lymphocytes to a patient with upper extremity secondary lymphedema brought about a rapid and remarkable reduction in arm swelling. The protein components in the edema fluid were analyzed by two-dimensional electrophoresis before and after lymphocyte injection. We observed the appearance of a novel protein spot, with an isoelectric point of 6.5, in an electropherogram as early as 30 minutes after the lymphocyte injection. Immunoblotting using antibody against human total serum proteins suggested that the novel protein was not derived from the serum. Because incubation in vitro of the lymphedema fluid with the isolated lymphocytes produced a new protein spot, corresponding to the novel protein observed in vivo, we suspect that the novel protein originated from limited hydrolysis of a unique protein present in the arm edema fluid. Significance of the novel protein and the role of limited proteolysis after lymphocyte injection in the management of lymphedema are examined.