The characterization of genetic variation at the level of DNA has generated significant advances in gene mapping and disease diagnosis, and forensic identification of individuals. It is now possible to identify individual DNA from various tissue specimens, like hair, using the PCR and oligonucleotide probes. To date, however, the number of hairs needed, the preservation conditions, and the kinds of hair suitable for DNA extraction have not been well known. We performed DNA extraction using hairs from different body sites, using different numbers of hairs, under various different preservation conditions to investigate the acquisition conditions of DNA data from hair using PCR and specific HLA-DQA probe. HLA-DQA genotyping of DNA extracted from peripheral blood was performed to compare the results of hair and blood HLA-DQA genotyping from individuals. The results are as follows: 1) The concentration of DNA extracted from a single strand of hair is 5.23+/-0.54 g/ml. It is possible to extract sufficient DNA for HLA-DQA genotyping from a single strand of hair. 2) DNA concentration is different according to body site. Concentrations are 7.01+/-0.33 g/ml in scalp hair, 6.28+/-0.29 g/ml in axillary hair, and 6.10+0.24 microgram/ml in pubic hair. 3) There is no difference between the electrophortic bands resulting from DNA extracted from the hair of an individual preserved under different conditions, such as room temperature, exposure to sunlight, exposure to low temperature (+4degrees C), or exposure to moisture. 4) There is no difference between the electrophoretic bands resulting from DNA extracted from hair of a single individual preserved for different lengths of time. 5) In an individual, the HLA-DQA genotype obtained from peripheral blood is identical to that obtained from hair. Even though the amout of DNA obtained from hair is limited, it is possible to identify the HLA-DQA genotype of an individual using a single strand of hair. This requires adequate extraction of DNA for PCR analysis using an allele specific probe. We believe that HLA-DQA genotyping using the PCR method on DNA extracted from hair is useful for disease diagnosis and forensic science.
Alleles ; Chromosome Mapping ; Diagnosis ; DNA* ; Forensic Sciences ; Genetic Variation ; Genotype ; Hair* ; Oligonucleotide Probes ; Polymerase Chain Reaction* ; Scalp ; Sunlight

PURPOSE: Three-dimensional(3-D) organization is critical for both the normal development and, tumor growth and progression. Cell-cell and cell-matrix interactions determine normal prostate development and its subsequent neoplastic transformation. To understand the epigenetic factors that lead to cell transformation, a 3-D human prostate cell culture was established with prostate epithelial cells grown in a rotating-wall vessel(RWV) under microgravity-simulated conditions with either alone or with prostate or bone stromal cells. We tested the hypothesis of whether phenotypic and genotypic alterations of LNCaP cells may be achived when grown as 3-D organoids. MATERIALS AND METHODS: LNCaP cells were seeded in RWV alone and with either human prostate or bone fibroblasts. After period of 2 months, RWV1, 2, and 3 cell lines were established from the prostate organoids and were characterized. RESULTS: While LNCaP cells injected orthotopically failed to form tumors in castrated mouse, RWV-derived cell lines formed PSA-producing tumors and metastasized to lymph node, bone, lung and liver, which stained positively by PSA antibody. RWV cells grew faster than parental LNCaP, especially in sex hormone-free conditions. Unlike parental LNCaP cells which respond positively to androgen and estrogen-induced growth and PSA expression, RWV cells are insensitive to sex steroid-induced growth, but remain sensitive to androgen for induction of PSA expression. Comparative genomic hybridization(CGH) results demonstrated that RWV cell lines have different chromosomal gain and loss each other as compared to those of LNCaP. CONCLUSIONS: Cell-cell and cell-matrix interactions of androgen-dependent and non-metastatic LNCaP cultured alone or with either prostate or bone fibroblasts in 3-D culture under microgravity-simulated conditions resulted in induction of androgen- independent and metastatic LNCaP sublines, RWV cell lines, meaning androgen- independent progression. Phenotype and genotype of RWV cell lines are definitely dissimilar to those of parental LNCaP. 3-D culture of prostatic epithelial cells under microgravity-simulated conditions could be novel approach to the study of normal development and cancer of prostate as an ideal in vitro model and, will be further exploited.
Animals ; Cell Culture Techniques ; Cell Line ; Epigenomics ; Epithelial Cells ; Fibroblasts ; Genotype ; Humans ; Liver ; Lung ; Lymph Nodes ; Mice ; Organoids* ; Parents ; Phenotype ; Prostate* ; Prostatic Neoplasms ; Stromal Cells

A clinical observation was done in 72 patients with renal cell carcinoma admitted to Department of Urology, St. Mary's Hospital, Catholic University Medical College from Jan. 1980 to Jun. 1988 retrospectively. There were 52 men and 20 women, giving a ratio of 2.6 to 1, with the highest incidence in the sixth to seventh decades (66%). The most common symptom and sign were hematuria, flank pain and palpable mass in orders, but classic symptom triad of renal cell carcinoma was present in only 11% of the patients. Symptoms secondary to metastasis were only initial presentations in more than half of the patients with stage IV disease, initially 11% (12 pts.) of all the patients with renal cell carcinoma. CT scan showed a high diagnostic accuracy (78%) as compared to pathologic examination. So recently angiography is not done routinely. In one patient, tumor was not detected by CT scan, but by ultrasonography and angiography. Liver scan is not indicated unless there are 2 or more abnormal values among liver function test including alkaline phosphatase, GOT and GPT. Bone metastasis is not correlated with the elevation of alkaline phosphatase and bone scan is indicated only when bone pain and/or gait disturbance are present. Radical nephrectomy is the choice of treatment and adjunctive therapy including. irradiation, chemotherapy, hormone therapy and immunotherapy were not effective. The majority of distant metastasis (86%) occurred within the first 2 years following nephrectomy and metastatic sites were lung, lymph nodes, liver and bone in orders. Of the 72 patients, it was possible for 36 patients to follow up more than 3 years. Patients with stage 1 disease showed 90%. (9/10) 3 year survival and there were no significant differences in 3 year survival between stage II (50%) and stage III (44%) disease. No patients with stage IV disease survived more than 3 years.
Alkaline Phosphatase ; Angiography ; Carcinoma, Renal Cell* ; Drug Therapy ; Female ; Flank Pain ; Follow-Up Studies ; Gait ; Hematuria ; Humans ; Immunotherapy ; Incidence ; Liver ; Liver Function Tests ; Lung ; Lymph Nodes ; Male ; Neoplasm Metastasis ; Nephrectomy ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed ; Ultrasonography ; Urology

PURPOSE: A cell-cell interaction in which in vivo inoculation of androgen-dependent, non-tumorigenic LNCaP and human bone fibroblast resulted in derivation of androgen-independent and metastatic LNCaP subline(C4-2) in castrated hosts. The purpose of this study is to evaluate if human prostate fibroblasts when grown together with LNCaP may promote androgen-independent growth and enhance metastatic potential. MATERIALS AND METHODS: LNCaP cells and human prostate fibroblasts derived either from peripheral or transition zone co-inoculated in athymic mice for 8 weeks, and then mice were castrated. The chimeric tumors were maintained for additional 4 weeks. The LNCaP sublines, designated P4 and T4, were established and characterized. These sublines were co-inoculated again in castrated mice with human prostate fibroblasts for 8-12 weeks. And then second generation LNCaP sublines, P4-2 and T4-2, were established and also characterized. RESULTS: Marked cytogenetic alterations were observed in P4-2, P4, T4-2 and T4 LNCaP sublines in comparison to parental LNCaP. Although LNCaP cells injected orthotopically did not form tumors in castrated hosts, LNCaP sublines formed PSA-producing tumors and had metastatic potentials to lymph node, lung, liver and bone. These P and T sublines had androgen-independent growth characteristics and metastatic potential. CONCLUSIONS: Cell-cell interactions between prostatic epithelium and their surrounding fibroblasts could contribute to androgen-independent characteristics and enhanced metastatic potential of localized prostate cancer in vivo.
Animals ; Cytogenetics ; Epithelium ; Fibroblasts* ; Humans* ; Liver ; Lung ; Lymph Nodes ; Mice ; Mice, Nude ; Parents ; Prostate* ; Prostatic Neoplasms

urothelial tumors of the upper urinary tract are relatively uncommon, mostly malignant. 25 patients with urothelial tumor of upper urinary tract seen at St. Mary`s hospital from Jan. 1975 to Dec. 1986 are reviewed retrospectively. The average age was 57.3 years, approximately half of them being more than 60 years old. Gross Hematuria was the most prevalent sign and cytologic examination of the urine has proved not to be diagnostic. The most common findings of excretory urography and retrograde urography were non- visualization of the kidney and filling defect. CT can help to evaluate the extent of disease by determining the invasion or metastasis to surrounding tissues or retroperitoneal node. All 25 patients were surgically explored, 23 patients underwent nephroureterectomy and 6 of them was also done lymphadenectomy, distal ureterectomy with ureteral reimplantation was done in 1 patient and simple nephrectomy was done under the misdiagnosis of renal tuberculosis in 1 patient. There were close relations between histological grade and pathologic grade. Of the 25 patients, stage A and grade II were the most common, each of them were 11 and 16 cases. The over-all 2 year survival rate of 13 patients followed up was seventy percent.
Diagnostic Errors ; Hematuria ; Humans ; Kidney ; Lymph Node Excision ; Middle Aged ; Neoplasm Metastasis ; Nephrectomy ; Prognosis ; Replantation ; Retrospective Studies ; Survival Rate ; Tuberculosis, Renal ; Ureter ; Urinary Tract* ; Urography

During the last one year. 23 selected cases of various glaucomatous eyes were received trabeculectom y ab extemo (5 X 3mm scleral-flap with a large peripheral iridectomy and only two scleral-flap corners sutures) under surgical microscope. Success cases which were controlled ocular tension to normal are 21 eyes(91.3%) out of total 23 glaucomatous eyes. Especially, all 11 eyes of simple open angle glaucoma were obtained excellent normotension after trabeculectomy, But one case of absolute glaucoma was failed because of vitreous prolapse, and another failed case was in group of acute closed glaucoma due to surgical complication of total hyphema(Table 1, 2).
Glaucoma ; Glaucoma, Open-Angle ; Intraocular Pressure ; Iridectomy ; Prolapse ; Trabeculectomy*

PURPOSE: To evaluate the usefulness of automated biopsy guns in image-guided biopsy of lung, liver. pancreas and other organs. MATERIALS AND METHODS: Using automated biopsy devices, 160 biopsies of variable anatomic sites were performed:Biopsies were performed under ultrasonographic(US) guidance in 95 and computed tomographic (CT) guidance in 65. We retrospectively analyzed histologic results and complications. RESULTS: Specimens were adequate for histopathologic diagnosis in 143 of the 160 patients(89.4%)-Diagnostic tissue was obtained in 130 (81.3%), suggestive tissue obtained in 13(8.1%), and non-diagnostic tissue was obtained in 14(8.7%). Inadequate tissue was obtained in only 3(1.9%). There was no statistically significant difference between US-guided and CT-guided percutaneous biopsy. There was no occurrence of significant complication. We have experienced mild complications in only 5 patients-2 hematuria & 2 hematochezia in transrectal prostatic biopsy, and 1 minimal pneumothorax in CT-guided percutaneous lung biopsy. All of them were resolved spontaneously. CONCLUSION: The image-guided biopsy using the automated biopsy gun was a simple, safe and accurate method of obtaining adequate specimen for the histopathologic diagnosis.
Biopsy* ; Diagnosis ; Firearms* ; Gastrointestinal Hemorrhage ; Hematuria ; Image-Guided Biopsy* ; Liver ; Lung ; Pancreas ; Pneumothorax ; Retrospective Studies

The rapid involution of the rat ventral prostate after castration is an active process initiated by removal of the inhibitory effects of androgen on prostatic cell death. The degradation of genomic DNA into nucleosomal-sized fragments if an early event in this process and is catalyzed by Ca2+Mg2+-dependent endonuclease activity which is dependent upon calcium ions. The morphologic correlation of the involution process involves a series of structural changes which are collectively referred to as apoptosis. Since the castration-induced endonuclease is dependent upon calcium ions for maximal activity, a potential involvement of a intracellular calcium in the castration-induced prostatic cell death was investigated. Acute disturbance in intracellular calcium homeostasis within the ventral prostate by means of a potent calcium channel blocker, nifedipine, simultaneous with castration resulted in a significant decrease in prostatic apoptosis. This result points to a potential role intracellular calcium levels in the mechanism of activation of castration-induced death of the androgen-dependent epithelial cells in the ventral prostate.
Animals ; Apoptosis* ; Calcium Channels* ; Calcium* ; Castration ; Cell Death ; Deoxyribonuclease I ; DNA ; Epithelial Cells ; Homeostasis ; Ions ; Nifedipine* ; Prostate* ; Rats*

PURPOSE: The mechanisms, which alter urinary bladder muscle function during infectious cystitis, are poorly understood. The objective of this study was to identify potential resident targets for endotoxin lipopolysaccharide(LPS) within normal bladder smooth muscle and test the hypothesis that LPS induces an inflammatory response within the bladder muscle and that this is associated with a decrease in muscle contractility. MATERIALS AND METHODS: Male rats were studied 24 hours after a single bolus i.p. injection of LPS(15mg/kg). Whole-mount preparations of the bladder muscle were immunohistochemically stained for neutrophils(myeloperoxidase), macrophages(ED2), activated leukocytes(LFA-1) and mast cells(FITC-Avidin). Contractile activity was assessed from muscle strips of the bladder in response to bethanechol(0.3-300microM). Voiding frequency and urine volume for 24 hours were measured using metabolic cage. Cystometry was performed to measure the intravesical bladder pressure. RESULTS: Using the resident macrophage marker ED2, dense network of macrophages were observed within the bladder muscle of control and LPS treated rats. Few neutrophils(myeloperoxidase-positive cells, 2.3 +/- 0.38 cells, x200) were detected in whole-mounts of bladder muscle of control rats, while LPS pretreatment resulted in a significant increase in the number of neutrophils which demonstrate inflammatory response(10.8 +/- 1.70 cells, x200, p<0.001). LFA-1 immunohistochemistry demonstrated an increased presence of LFA-1 positive cells in bladder muscle of LPS treated rats, which had a morphology similar to both neutrophils and resident macrophages. The expression of LFA-1 is known as a marker of cells that are in an activated state. LPS pretreatment resulted in a significant reduction in bladder muscle contractions in response to bethanechol(i.e. control = 0.049 +/- 0.010 vs. LPS= 0.029 +/- 0.003 gr/mm2/sec, 100microM, p<0.05). Voiding frequency of LPS treated rats was significantly increased compared to that of control rats. In LPS treated rats, voiding phase representing bladder contractility in cystometry was observed. CONCLUSIONS: These data demonstrate that a single intraperitoneal injection of LPS initiates an inflammatory response within the bladder muscle that is associated with a decrease in the functional activity of the bladder. We hypothesize that secretions from the resident macrophages and extravasated leukocytes within the muscle cause the observed suppression in bladder muscle activity in vitro.
Animals ; Cystitis ; Humans ; Immunohistochemistry ; Inflammation* ; Injections, Intraperitoneal ; Leukocytes ; Lymphocyte Function-Associated Antigen-1 ; Macrophages ; Male ; Muscle Contraction ; Muscle, Smooth ; Neutrophils ; Rats* ; Urinary Bladder*

PURPOSE: The growth and PSA secretion of prostate carcinoma is predominantly regulated by androgens. However, locally produced growth factors(GFs) also have been shown to play a crucial role in the proliferation of androgen-dependent prostatic tumor cells. Androgen has been proposed to stimulate the cell proliferation and PSA secretion by modulating the activity of some of these growth factors. The objectives of this study are to determine the effects of various GFs (epidermal growth factor; EGF, basic fibroblast growth factor; bFGF, keratinocyte growth factor; KGF, hepatocyte growth factor; HGF) on the growth and PSA secretion of androgen-dependent LNCaP prostate cancer cell line. MATERIALS AND METHODS: To determine the effects of EGF, bFGF, KGF and HGF on the growth and PSA secretion of LNCaP, LNCaP cells at a concentration of 3 x 103 cells/well, suspended in T-medium containing 2% TCM, were seeded in 96 well plates. Cells were exposed to six different concentrations (0.5, 1, 5, 10, 20 and 40 ng/ml) of GFs. Cell numbers were evaluated by crystal violet assay on day 3, 5 and 7, and PSA concentrations in conditioned medium were determined on day 5. RESULTS: EGF and HGF had minimal, not significant, stimulatory effects on the growth of LNCaP. However, bFGF and KGF had significant growth stimulatory effects (P<0.05). EGF, bFGF, KGF and HGF did not have any stimulatory effects on the PSA secretion of androgen-dependent LNCaP cell line. CONCLUSIONS: bFGF and KGF, not EGF and HGF, directly stimulate the growth of LNCaP cells. However, bFGF and KGF as well as EGF and HGF do not affect the PSA secretion of LNCaP. There seems to be another signal transduction pathway, which is not associated with GFs mentioned above, for PSA secretion of androgen-dependent LNCaP cell line.
Androgens ; Cell Count ; Cell Line* ; Cell Proliferation ; Culture Media, Conditioned ; Epidermal Growth Factor ; Fibroblast Growth Factor 2 ; Fibroblast Growth Factor 7 ; Gentian Violet ; Hepatocyte Growth Factor ; Intercellular Signaling Peptides and Proteins* ; Prostate* ; Prostatic Neoplasms* ; Signal Transduction