Prevalence of androgen receptor gene mutations in latent prostatic carcinomas from Japanese men.

The incidence rate of clinically apparent prostatic carcinoma is 8-fold higher in the United States than in Japan, while the prevalence of latent prostatic carcinoma, a presumed precursor to clinical carcinoma, is similar in the two countries. The purpose of this study was to investigate the hypothesis that this profound difference in incidence rates of clinical carcinoma reflects distinct profiles of molecular genetic alterations in the latent precursor lesions that occur in the two countries. A significant fraction of latent carcinomas from Japanese men were found to contain inactivating mutations of the androgen receptor gene, while no such mutations were found in latent carcinomas from American men. No mutations were found in clinical carcinomas from either country. These data offer a potential molecular genetic explanation that may partially account for the distinct prostatic carcinoma incidence rates in these two populations.

Mutation analysis of the BRCA1 gene in ovarian cancers.

Germline mutations of the BRCA1 tumor suppressor gene on chromosome 17q are involved in a significant fraction of hereditary breast and ovarian cancers. Allelic deletions that include the BRCA1 locus are common in breast and ovarian cancers, implying that somatic mutations of this gene may play an important role in the more common sporadic forms of these tumors as well. The recent cloning of BRCA1 allows direct testing of this hypothesis. A combination of single strand conformation and sequencing analyses was used to examine the 22 coding exons and intronic splice donor and acceptor regions of BRCA1 for mutations in 115 unselected cases of epithelial ovarian carcinoma. Seven mutations were identified, all of which were present in the germlines of patients with remarkable family or medical histories of breast and/or ovarian cancer. Eighty-nine of these tumors were examined for loss of heterozygosity in the BRCA1 region of chromosome 17q, and 67% of the tumors studied exhibited allelic deletions that included this region. These data are consistent with the hypothesis that BRCA1 mutations are involved in the etiology of hereditary ovarian carcinomas but occur rarely in sporadic tumors, and that the frequent allelic loss on chromosome 17q in this cancer type reflects the involvement of an additional tumor suppressor gene(s).

Mutation of the Ki-ras protooncogene in human endometrial hyperplasia and carcinoma.

Previous studies have demonstrated that some human endometrial carcinomas contain an activating point mutation in codon 12 of the Ki-ras protooncogene. To examine the hypothesis that this mutation may occur at an earlier stage of neoplastic progression in the endometrium, we analyzed 89 samples of premalignant endometrial hyperplasia and an additional 84 samples of endometrial carcinoma for point mutations of Ki-ras codon 12. Mutations were found in all three types of endometrial hyperplasia, simple, complex, and atypical, with no clear evidence of a differential distribution in any particular type. Furthermore, the overall incidence of Ki-ras mutations in the hyperplasia specimens (16%) was similar to the incidence detected in carcinomas (18%), indicating that ras mutation may represent an early event in a subset of endometrial carcinomas. When the tissue samples were segregated as to country of origin, the frequency of this mutation was approximately 2-fold higher in hyperplasia and carcinoma samples from Japan than from the United States, where the incidence, clinicopathological characteristics, and risk factors for endometrial carcinoma differ dramatically. There was no apparent correlation, however, between ras mutation and any pathological, histological, or clinical parameter examined, except survival. The presence of a ras mutation was inversely associated with death from disease, suggesting that this molecular feature may characterize a subset of endometrial carcinomas with a good prognosis.

Mutation, allelotyping, and transcription analyses of the p73 gene in prostatic carcinoma.

A novel gene, p73, encoding a protein with significant homology to p53, was recently identified at 1p36. To investigate penetrance of p73 in prostatic carcinogenesis, mutation, allelotyping, and transcription analyses of p73 were performed in prostatic carcinoma. No types of mutation causing amino acid substitutions or frameshifts were found in 106 cases examined. Loss of heterozygosity in the gene was found in 2 of 38 cases (5.3%). Various expression levels of p73 alpha variant were observed in tumor compared with those in normal tissue. These data suggest that the p73 gene is not playing an essential role, but expression of p73 may associate with tumor growth in prostatic carcinogenesis.

Mutations of the BRCA2 gene in ovarian carcinomas.

Inherited mutations in the recently discovered BRCA2 gene are believed to be responsible for a significant fraction of early-onset hereditary breast cancers. Unlike BRCA1, however, which confers a high risk to both breast and ovarian cancer, the incidence of ovarian cancer appears to be much lower In BRCA2-linked families, causing uncertainty as to the relevance of BRCA2 to hereditary ovarian cancer. Numerous allelotype studies indicate that allelic deletions Including the BRCA2 locus on chromosome 13q are common in ovarian cancers in general, suggesting that somatic mutations of this gene may be involved in sporadic ovarian tumorigenesis. The purpose of this study was to test the hypothesis that germline or somatic mutations of BRCA2 are associated with hereditary and/or sporadic ovarian cancers, respectively. The entire 10.2-kb coding region of BRCA2 was screened for mutations in 130 consecutive ovarian tumors, the only selection criterion being a pathological diagnosis of epithelial ovarian carcinoma. Loss of heterozygosity at markers flanking BRCA2 was observed in 56% of the tumors. Four germline mutations and two somatic mutations were identified; two of the germline mutations are recurrent, having been previously described. Remarkably, the patients with germline mutations were late-onset cases with no medical or family histories suggestive of hereditary cancer. These data suggest that mutations of BRCA2 are rare in sporadic ovarian cancers, and that the proportion of ovarian cancers resulting from hereditary predisposition may be higher than previously suspected based on estimates derived from studies of highly penetrant genetic loci.

Polymerase chain reaction-single strand conformation polymorphism analysis of the p53 gene in paraffin-embedded surgical material from human renal cell carcinomas.

p53 tumour suppressor gene mutations were studied in 118 renal cell carcinomas using paraffin-embedded surgical material. Optimal results were obtained with analysis of exon lengths between 150 and 200 base pairs for polymerase chain reaction. Single strand conformation polymorphism and sequencing analysis revealed only two point mutations (2/118, 2%): one involving codon 135; TGC-->TTC (cysteine-->phenylalanine) and the other codon 175; CGC-->CAC (arginine-->histidine). Both of these cases were classified as granular cell subtype on microscopic observation. The data suggest that the p53 tumour suppressor gene is not related to tumour initiation, promotion, or progression of renal cell carcinomas. However, there is the possibility that granular cell type carcinomas may have a different genetic background from clear cell type renal neoplasms.

Prognostic status of p53 gene mutation in canine mammary carcinoma.

BACKGROUND: The p53 gene mutations have been associated with the development of human breast and canine mammary neoplasms; breast carcinoma patients with alterations of p53 gene are considered to have a poor prognosis. Mammary carcinoma represents the most common malignant tumor in female dogs. However, the prognostic significance of p53 gene mutation in the dog has been unclear. STUDY DESIGN: The alteration in exons 5-8 of p53 gene in 69 canine mammary carcinomas were investigated by PCR-SSCP with direct sequence analysis and statistically analyzed to compare with other clinicopathological parameters including age, neuter, tumor size, stage, histology, p53 expression, recurrence and death from carcinoma. RESULTS: 12 out of 69 (17%) carcinomas showed p53 gene mutations. After a follow-up period of 30 months, multivariate regression analysis revealed that p53 gene mutation was only an independent risk factor for increased risk of the recurrence and death from mammary carcinoma. CONCLUSION: The p53 gene alterations might contribute to the prognostic status in canine mammary carcinomas, in a way comparable to that of human tumors.

[Study on the establishment of erythropoietin producing human renal cell carcinoma heterotransplanted to nude mice].

We conducted the establishment of erythropoietin (Epo) producing human renal cell carcinoma heterotransplanted in nude mice (JRC 901) and analysed its histopathological and biological characteristics. Regarding to histopathological analysis, JRC 901 showed renal cell carcinoma with granular cell subtype, alveolar pattern and grade II malignancy. In an effort to the electron microscopic analysis, JRC 901 showed renal cell carcinoma with microvilli, rich lipid droplets and mitochondria. As to the tumour doubling time, the JRC 901 showed 14.81 days in a logarithmic phase. As to the karyotype, the JRC 901 showed human, 46, XY, -11, 8p+, 17q-, +mar. After tumour inoculation to the nude mice, the blood level of Epo increased at 5 weeks, and its level reached at 485.2 mU/ml at 12 weeks after tumour inoculation. Furthermore, immunohistochemical staining using anti-Epo showed positive staining within cytoplasm of JRC 901. Moreover, the production of Epo was observed in the level of mRNA (264 bp) using RT-PCR method. We conclude that the JRC 901 is a human renal cell carcinoma heterotransplantable to nude mice and this tumour produce the Epo after tumour inoculation to nude mice.

Establishment and characterization of human choriocarcinoma cell line derived from a metastatic focus of a testicular mixed germ cell tumor.

A human testicular choriocarcinoma cell line HKRT-II was established by the single-cell cloning method from a mixed cell culture system derived from a retroperitoneal metastatic germ cell tumor composed of a yolk-sac tumor, a choriocarcinoma, and an immature teratoma. Its primary tumor rose from the testis and was comprised of a seminoma, a yolk-sac tumor, a choriocarcinoma and an immature teratoma. The HKRT-II cells were spindle or polygonal in shape and contained multi-nucleated giant cells showing neoplasticity and pleomorphism. The cells proliferated in a stable manner, and the population doubling time was 42 hours. The chromosome numbers showed a wide distribution of aneuploidy, while the mode was in the hypertetraploid range. Double minute chromosomes and homogeneously staining regions were recognized in about 5% to 10% of the metaphase plates, respectively. Heterotransplantation was not difficult. Subcutaneous transplantation of 1 x 10(7) cells into nude mice formed a tumor composed of only a choriocarcinoma. The most noteworthy characteristics of the cell line were that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in in vivo grafted cells, and that the N-myc gene was amplified about 10 times.

Flow cytometric analysis of prostatic carcinoma with and without bone marrow metastasis.

We have analyzed 68 prostates obtained at autopsy, for DNA ploidy by means of flow cytometry from patients who had clinical prostatic carcinoma with (29 cases) and without (39 cases) bone marrow metastasis. Flow cytometric analysis revealed 42 diploid cases and 26 aneuploid cases in a total of 68 cases. Among the 26 cases of aneuploidy, 4 cases were tetraploid aneuploid and 22 cases were not tetraploid aneuploid. The highest incidence for the aneuploidy was found in stage D2 disease (19/29, 65.5%), while the highest incidence for diploid was seen in stage B disease (23/25, 92.0%). The correlation between ploidy and bone marrow metastasis was significant (P < 0.01). We thus reconfirm that flow cytometric analysis of paraffin embedded tissue is useful for prognostic evaluation of prostatic carcinoma.

Muscular composition of the gastric groove in the golden hamster.

The golden hamster possesses a forestomach and a glandular stomach. The gastric groove connects the cardia to the glandular stomach and is situated on the lesser curvature of the stomach. The constitution of the muscle fibers in the gastric groove was investigated. The gastric groove consisted of two lips and a groove floor. The muscle coat of the lips was composed of a mixture of smooth and striated muscle fibers. The smooth muscle fibers were components of the cardiac muscle loop. The striated muscle fibers were extensions from the esophageal inner circular muscle layer, and invaded about half the length of the lips. The muscle coat of the groove floor consisted of an inner circular muscle layer made up of smooth muscle fibers, and the outer longitudinal muscle layer of the striated muscle fibers extended from the esophageal outer longitudinal muscle layer. The present study revealed that the muscle coat of the gastric groove in the golden hamster was composed of smooth and striated muscle fibers, and that these striated muscle fibers were extensions of the esophageal muscle coat.

Confronting cisternae in canine testicular seminoma: special reference to appearance rate.

A number of confronting cisternae (CC) were found in canine testicular seminomas (SEM). The CC consisted of two or three closely attached cisternae separated by an electron-dense layer, but occasionally four or more cisternae were stacked in a similar fashion. They showed a short, straight, and direct continuity with the rER or the nuclear envelope. They were found in SEM cells of both interphase and mitotic stages. We statistically examined 18 cases of canine SEM concerning the CC appearance rate in interphase cells (CCARI) and in mitotic cells (CCARM), and the tumor mitotic index (TMI). The SEM were classified into three groups (intratubular SEM without invasion, intratubular SEM with invasion, and diffuse type SEM) using Nielsen and Lein's classification (1974). The CCARI and TMI of the diffuse type SEM were significantly higher than those of the intratubular SEM with invasion, and those of the latter group were significantly higher than those of the intratubular SEM without invasion. On the other hand, the CCARM were similar between these three groups, and this parameter showed a non-significant correlation with the TMI. The present study suggests that the increase in the number of CC in interphase canine SEM cells might be correlated with the tumor progression.

Nuclear bodies appearance rate in canine testicular Sertoli cell tumor.

A number of nuclear bodies (NB) were observed in the canine testicular Sertoli cell tumors (SCT). We statistically examined nineteen cases of canine SCT concerning the NB appearance rate (NBAR), and also examined the NBAR in four cases of the normal testicular Sertoli cells. The mean value of the total number of the NBAR of SCT was significantly higher than that of normal Sertoli cells. The SCT were classified into three groups according to the Nielsen and Lein's histological classification (1974): intratubular SCT without invasion, intratubular SCT with invasion, and diffuse type SCT. The mean value of NBAR of the diffuse type SCT was significantly higher than that of the intratubular SCT with and without invasion, and there was no significant difference between the mean values of NBAR of the latter two groups. The distribution of NBAR of the diffuse type SCT was significantly different from that of the intratubular SCT with and without invasion. On the other hand, the individual differences of NBAR of the diffuse type SCT and the intratubular SCT with invasion was significantly higher than that of the intratubular SCT without invasion. The present study suggests that the increase of NBAR in canine SCT might be correlated with the tumor invasive progression.

Effects of relative metabolic rate and heart rate variation on the performance of flight attendants.

The main work of the cabin attendants in an actual flight in service for passengers. The effects of flight attendant duties in flight differ from the effects of the same tasks performed on the ground. In this study, the relative metabolic rate (RMR) and heart rate (HR) of cabin attendants in a cruising aircraft galley and cabin are compared with those of a crew working in a mock-up apparatus on the ground. The types of work tested are: (a) oshibori (steamed towel) service, (b) soft drink service, (c) setting meal tray, (d) putting casserole on tray, (e) meal tray service, (f) walking on aisle. The RMR at each type of work during flight is as indicated: (a) 1.07-2.10, (b) 1.08-1.54, (c) 1.37-1.82, (d) 2.57-3.50, (e) 2.11-3.10 and (f) 1.84. The range of HR was: (a) 105-120, (b) 90-110, (c) 90-120, (d) 100-130 and 100-140 beats/min. In most cases, the RMR and HR levels of work done in the mock-up were lower than those recorded in flight. These results suggest that the oxygen intake of work done in flight is greater than that on a mock-up. One of the reasons might be that the cabin barometric pressure (ca. 660 torr or cabin altitude ca. 1,500 m) or an aisle inclination of about 3 degrees caused a decrease in the efficiency of oxygen intake during flight.

Hemangioma of the fingers.

Fingers often suffer trauma and the clinician is continuously faced with the difficult task of clarifying the distinction between a hemangioma and a traumatic lesion. This study was undertaken to examine ten cases in which a small skin mass located on a finger had been diagnosed preoperatively as hemangioma. Our results showed that seven masses were confirmed pathologically as hemangioma (five cavernous hemangiomas and two capillary hemangiomas), two as traumatic thrombosis and one varix. The clinical manifestations of the two cases of traumatic thrombosis were related to those of hemangioma. In the varix, endothelial proliferation was observed in the area of the thrombosis. This phenomenon is called "intravascular papillary endothelial hyperplasia", and can confuse the differential diagnosis between a vascular neoplasm and a traumatic thrombosis. Our findings demonstrate that since the traumatic lesions were firmer than the hemangiomas, hardness on physical examination may be a helpful indicator in the differential diagnosis of a hemangioma and a traumatic lesion.

[A long-term survival after partial nephrectomy in a case of pelvic tumor arising in a solitary kidney].

A 44-year-old man was admitted on January 21, 1975 because of asymptomatic hematuria. The patient had nephrectomy of his left kidney due to nephritis at the age of three. Cystoscopy revealed no abnormalities, and excretory urography showed an irregular filling defect and slight ectasia in right upper calyx. A clinical diagnosis of pelvic tumor of the right kidney was made and partial nephrectomy was performed on April 18, 1975. The resected kidney was 4.5 X 5.0 X 6.5 cm in greatest dimension and the tumor was well localized in the upper calyx. Pathological diagnosis was transitional cell carcinoma, papillary, grade 11, stage 1. About 2 years after the operation, the patient developed a rice-sized tumor in the bladder neck followed by transurethral resection. Otherwise he is in good condition to date, 7 years and 4 months after the partial nephrectomy.

[Prostatic involvement of bladder carcinoma].

BACKGROUND: The histological pattern of prostatic involvement by transitional cell carcinoma is still unclear. The present study was carried out in bladder carcinoma with prostatic involvement to clarify the histological invasion pattern and its association with primary lesions. METHODS: In the past 10 years, 83 cases of total cystectomy including prostatectomy underwent pathological diagnosis in our department. This study included 81 cases of transitional cell carcinoma (TCC), of which 11 showed prostatic involvement of bladder carcinoma. In these cases, the histological patterns of invasion were classified in relation to prostatic urethra, prostatic duct, stroma, lymphatic duct, capsule, vein and perineural invasion. Seventy cases without prostatic involvement were controls. The location, pathological grade, stage and lymphatic involvement of primary bladder carcinoma were compared in terms of prostatic involvement cases with control cases. RESULTS: Among those 11 cases, there were 3 cases in which only the prostatic duct was involved, 2 cases with invasion to only lymphatic duct, and involvement of both in 6 cases. One case of the prostatic duct involvement showed non-continuous invasion in the prostatic duct without prostatic urethra invasion, suggesting the possibility that non-continuous invasion could occur as a type of multicentric growth of TCC. CONCLUSIONS: Suspected routes of invasion of bladder carcinoma into the prostate were; 1 continuous transductal, 2 trans-lymphatic ductal, 3a combination of the two. It appears necessary to consider the possibility of TCC occurring in the prostate simultaneously with bladder carcinoma as a part of multicentric growth. There was a tendency of prostatic involvement cases include the bladder neck and trigone, and show lymphatic duct involvement more than non-prostatic involvement cases.

[Image cytometric DNA analysis in bladder tumors].

UNLABELLED: We studied image cytometric DNA analysis of bladder tumors to evaluate malignant potentials of bladder tumors. METHODS: Thirty nine samples were obtained by TUR from 37 patients. Nuclear DNA content of all samples were measured by image cytometer and were determined ploidy pattern by DNA histogram. RESULTS: Of 39 TCC non-diploid pattern was recognized in 50% of grade 1 cases, 73% of grade 2 cases and 100% of grade 3 cases. DNA ploidy was strictly correlated with histological grading in TCC. DNA non-diploid pattern was present in 67% of papillary tumors, 87.5% of non-papillary tumors and 100% in CIS. In diploid pattern 2 of 7 cases with grade 1 and 2 of 4 cases with grade 2 recurred. In non-diploid pattern 1 of 4 cases with grade 1, 4 of 10 cases with grade 2 and 4 of 6 cases with grade 3 recurred. There was no significant correlation between diploid and non-diploid pattern in grade 1, 2, 3. CONCLUSION: Image cytometric DNA analysis may be useful in addition to the classic and prognostic parameters of stage and grade, especially in TCC. The differences between image analysis system and flow cytometric analysis for DNA measurement were discussed.

[Histopathological effect and its predictors of neoadjuvant hormonal therapy by combined androgen blockade].

OBJECTIVES: Combined androgen blockade (CAB) uning LH-RH agonist and flutamide has been performed as neoadjuvant therapy for T 2, 3 prostate cancers (CaP). The histological effects of neoadjuvant CAB therapy and influential factors were investigated. METHODS: Materials were 20 CaP cases which were underwent radical prostatectomy (RP) after neoadjuvant CAB therapy. All cases were diagnosed by echo-guided sextant needle biopsies. RP was performed after serum PSA was decreased to undetectable level. Histological effect was evaluated by general rule for clinical and pathological studies on prostate cancer (Japanese Urological Association). All cases were divided 2 groups by histological effects as follows: Group A (poor effect group): G 0 and G 1, Group B (good effect group): G 2 and G 3. Immunostaining of p 53 (mutant type), bcl-2 and Chromogranin A (ChA) were performed for both pretreatment needle biopsy and RP specimen. In addition, pretreatment serum PSA and Gleason grade were also investigated. RESULTS: Down grading were found in 30%. Down staging were found in 35% (7 cases). All 7 cases were negative surgical margins and 5 of 7 were clinical T 3. Negative bcl-2 of biopsy specimen was correlation with down grading (p = 0.008). In the histopathological evaluation, G 0 was 1, G 1 were 10, G 2 were 6 and G 3 were 3 cases. Gleason 4 or 5 elements of biopsy were found in 9/11 cases in Group A but only 3/9 cases in Group B (p = 0.027). The bcl-2 positive cells of biopsy were found in 8/11 cases in Group A but only 1/9 cases in Group B (p = 0.006). The p 53 and/or bcl-2 positive cells of biopsy were found in 10/11 cases in Group A but only 3/9 cases in Group B (p = 0.007). Serum PSA and ChA were not correlation with histological effect of neoadjuvant CAB therapy. But, in 3 cases, ChA positive cell appeared after neoadjuvant therapy. CONCLUSIONS: We could not expect more than 50% cases to show the down grading and down staging. But, in T 3 case, surgical failure could be decrease. We could expect prostate cancer cases without positive bcl-2 cells, p 53 over expression and Gleason 4 x 5 to reveal the good histological effects of neoadjuvant CAB therapy.

[Analysis of tumor volume in latent prostatic carcinoma].

An assessment has been made of the histopathological characteristics of latent prostatic carcinoma and the tumor volume in 500 male Japanese patients who underwent dissection at The Jikei University since 1983. A microscopic observation was made of the prostatic glands extirpated totally at the necropsy, fixed with formalin and prepared as a step-section in a thickness of 3 mm. In the cases of latent carcinoma, after photographing the lesion in the same magnification and measuring the area of the carcinoma lesion with a digitizer, the volume was calculated by multiplying the thickness of 3 mm, and carcinoma volume was determined by integrating the value of each slice and adjusted by a conversion formula. The incidence of latent carcinoma was 104 cases out of 500 (22%). The incidence increased as the age layer becomes higher, and latent carcinoma was observed in 44% of the patients aged 80 and above. Complication of latent carcinoma with prostatic hyperplasia was presumed to be an independent phenomenon in so far as it is seen from the statistical aspect. The patients were classified histopathologically into well-differentiated type (64%), mixed type (27%) and poorly-differentiated type (9%), showing high incidence in the low-aged layer of well-differentiated lesions and in the high-aged layer of mixed type lesions and in the high-aged layer of mixed type lesions. The average tumor volume of latent carcinoma was so small as 231 mm3, but many of the lesions in the cases of well-differentiated type were small, being on average 103.9 mm3, but many of the lesions in the cases of poorly-differentiated type were large, being on average 642.2 mm3. Statistically, with a tumor size of 200 mm3 as the boundary, a difference was observed in the distribution of histological constitution between the group with smaller lesions and the group with larger lesions. As an application of this result to the clinical carcinoma of stage A, the value of volume of 200 mm3 was considered to be important as a diagnostic criterion in deciding the necessity of treatment.