Induction of retrovirus particles in human testicular tumor (Tera-1) cell cultures: an electron microscopic study.

The Tera-1 and Tera-2 cell lines, established from germ-cell tumors of the human testis, were examined by electron microscopy for particles with the morphology of retroviruses. Extracellular and budding particles were observed at low frequencies only in cultures of Tera-1 cells that had been treated with 5-iodo-2'-deoxyuridine and dexamethasone. No particles were detected in untreated cultures of Tera-1 cells or in any preparations of Tera-2 cells.

Production of carcinoembryonic antigen by human prostate epithelial cells in vitro.

The medium from all 16 human prostate epithelial cell cultures tested contained elevated levels of carcinoembryonic antigen (CEA) or CEA-like substance; 11 of 16 had greater than 20 ng CEA/ml. In contrast, medium from cultures of other human tissues (prostate fibroblasts, genitourinary tumors, melanoma, and nontumor tissue), as well as media controls, contained less than 1 ng CEA/ml. Results indicated that CEA determination may provide a way to identify human prostatic epithelium in culture.

Morphologic evidence for retrovirus production by epithelial cells derived from a human testicular tumor metastasis.

Ultrastructural examination of primary and subcultured epithelial cells established in vitro from an abdominal metastasis of a human testicular tumor revealed particles with the morphology of retroviruses. These structures, found only after extensive scanning of the cells, were observed budding from microvilli and from the outer cell membrane and as extracellular particles. Production of these virus particles was stimulated by the incubation of cells in culture medium containing 5-iodo-2'-deoxyuridine and dexamethasone.

In vitro cultivation of epithelial cells derived from tumors of the human urinary tract.

Finely minced explants from 54 TCC2 of the human urinary tract were cultured in vitro in an attempt to establish cell lines. Cells with epithelial morphology grew out from 48 tumor explants, and long-term cell cultures were established from 10. Six of the cell cultures have been maintained for over 18 months with 50 to 70 transfers and, therefore, are considered permanent cell lines. The epithelial cells in the established cultures are small, exhibit rapid doubling time, and show multilayering. The cells were examined both microscopically and by cultivation techniques, and they were found to be free from contaminating microorganisms, including Mycoplasma. The established cultures grow rapidly in roller bottles and, therefore, can be produced in large quantities. These cells also remain viable after being stored for 3 years in liquid nitrogen.

Cytogenetic evidence for premeiotic transformation of human testicular cancers.

To determine the point at which transformation of the germ cell occurs during meiosis in nonseminomatous testicular cancer, the sex chromosome compositions of 15 cell lines derived from primary tumors or metastases of 12 patients with testicular cancer were analyzed by trypsin G-banding analysis and Y-body staining. The simultaneous existence of both X- and Y-chromosomes in a single cell has been confirmed in 14 cell lines. This suggests that transformation of the cell occurs before the first meiotic division because it is known that segregation of X- and Y-chromosomes occurs during the first meiotic division. An incidental finding was the presence of Barr bodies in some cell lines containing more than one X-chromosome, which is consistent with the known primitive nature of testicular cancer and its ability to differentiate independently from the male host.

Nonrandom abnormalities in chromosome 1 in human testicular cancers.

Trypsin G banding was performed on metaphase chromosomes from 14 cell lines derived from primary tumors or metastases of 11 patients with testicular cancer. Most of the cell lines, 11 of 14, have a modal number between 51 and 61. All lines have numerical and structural changes involving chromosome 1 with trisomy of the q arm being the common aberration. Break points in chromosome 1 were nonrandom, being concentrated in the regions of p12, q12, p36, and p22, which resulted in morphologically identical marker chromosomes in different cases. These changes probably are not artifacts of cell culture. In one instance, three lines derived from the same patient, one from tissue removed at operation, and two from separate metastases removed at autopsy nearly 3 years later after unsuccessful radiotherapy and chemotherapy had identical chromosome compositions. In another case, lines derived from a primary tumor and a metastasis from the same patient also had identical marker chromosomes. The consistent involvement of chromosome 1 in aberrations may be associated with the highly malignant nature of testicular cancers.

Properties of cell lines established from transitional cell cancers of the human urinary tract.

Characterization studies have been carried out on eight cell lines (253J, 192B, 639V, 647V, 486P, 575A,743E, and 751G) established from transitional cell cancers of the human urinary tract. Although subtle morphological differences exist among individual lines, each has an epithelial morphology and exhibits multilayering. The doubling times for the cells range from 20 to 56 hr, and at least a 1-to-3 split can be achieved when they are subcultured every 4th day. Karyotypic analysis revealed a hyperdiploid stemline for each cell line, and presence of a Y chromosome was confirmed by Q banding in five of the lines. The tumorigenic nature of the cell lines was demonstrated by their production of tumors in hamsters and confirmed by colony formation in agar. The transitional cell cancer lines were shown to be free of Mycoplasma, and their glucose-6-phosphate dehydrogenase mobility patterns and their Karyotypes prove that they are not HeLa cells.

Cell line derived from a metastasis of a human testicular germ cell tumor.

A cell line, designated 833K-E, has been established from a metastasis of a human testicular germ cell tumor that consisted of four histological types of tumor cells. The 833K-E cells have morphological and ultrastructural characteristics of epithelial cells and a hyperdiploid karyotype indicative of their human male origin. The cells grow in agar cultures and produce in nude mice tumors which have the hstological features of embryonal carcinoma without differentiated elements. Many of the cells express a stage-specific mouse embryonic antigen, and low levels of the major histocompatibility antigens and beta 2-microglobulin also were detected on a large percentage of the cells. A lymphoblastoid cell line (833K-LC) established from the same tumor specimen expresses major histocompatibility antigens and beta 2-microglobulin but does not express the embryonic antigen.

Carcinoembryonic antigen-like substance derived from human prostate.

Carcinoembryonic antigen-like substance, previously detected in large amounts in the medium from cultures of human prostatic epithelial cells, also is present in extracts of benign and malignant human prostate. By column chromatography, the prostate-derived carcinoembryonic antigen-like substance derived from cultured prostate is the same as that in tissue extracts and is distinctly different from colon-derived carcinoembryonic antigen. The molecular weight of prostate-derived carcinoembryonic antigen-like substance is estimated to be greater than 5 x 10(5). Prostate-derived carcinoembryonic antigen-like substance may be a prostate-specific substance.

Replication error-type genetic instability at 1q42-43 in human male germ cell tumors.

The replication error phenotype, recognized as microsatellite sequence alterations, has recently been suggested to be associated with hereditary nonpolyposis colorectal cancer and other types of sporadic tumors. We examined paired tumor-normal DNAs from 69 human male germ cell tumors for somatic instability at the 1q42-43 region. Analysis of a variable number of tandem repeats marker (D1S74) and 3 (CA)n type microsatellite loci (D1S235, D1S180, and angiotensinogen) revealed genetic alterations in tumor DNAs of 26 (38.2%) cases. The changes observed comprised rearrangements with D1S74 detected by Southern blot analysis in 4 of 55 (7%) cases; replication error-type alterations with D1S235, D1S180, and angiotensinogen in 12 of 66 (18.2%) cases; and loss of heterozygosity in 12 of 67 (17.9%) cases with the same probes. The microsatellite sequence alterations were more common in histological subsets other than teratomas, while the loss of heterozygosity was significantly more frequent in teratomas compared to other histologies. These results suggest that microsatellite instability and loss of heterozygosity at 1q42-43 may be unrelated genetic events which may play a role in germ cell tumor development.

Frequent allelic deletions and loss of expression characterize the DCC gene in male germ cell tumors.

The DCC tumor suppressor gene has been shown to be frequently deleted or its expression reduced or absent in colorectal, gastro-intestinal, pancreatic, prostatic, and breast carcinomas, and glioblastomas. By allelotype analysis using the DCC-flanking polymorphic marker D18S5 we have previously shown that allelic deletions at 18q21 occur in 40% of male germ cell tumors (Murty et al., 1994). In order to further understand the role of DCC gene in germ cell tumorigenesis, we evaluated deletions by loss of heterozygosity (LOH) and mRNA expression by RT-PCR in tumor tissues and cell lines. Analysis of 61 paired normal-tumor DNAs using the probes D18S5, JOSH 4.4 (a polymorphism within the DCC locus) and a (CA)n polymorphism in an intron of DCC revealed that 45% of GCTs had allelic deletions. In addition, two homozygous deletions were found in the DCC gene among 91 (61 used in the LOH analysis and an additional 30) tumor DNAs when screened with the cDNA probes (pDCC 1.65, pDCC 1.9 and pDCC 1.0). By RT-PCR analysis of four normal testes, nine GCT cell lines and 14 tumor tissues, DCC gene expression was detected in all four normal testes, while four (45%) GCT cell lines and one (7%) tumor specimen showed lack of expression. In addition, DCC expression was highly reduced in three (21%) tumor tissues. The high frequency of LOH at 18q21 was characteristic of seminomas as well as all subsets of non-seminomas in primary as well as metastatic states. Frequent allelic loss in all histologic subsets, homozygous deletions, and loss of expression of DCC suggest that suppression of this gene's function is an early event in GCT development.

Smoking cessation counseling during periodic health examinations.

Periodic health examinations are an excellent opportunity to counsel smokers to quit. The impact of a specialized smoking cessation counselor on the smoking behavior of patients having periodic health examinations was studied in a general internal medicine practice. One hundred fifty-five smokers having periodic health examinations were randomly assigned to a control group who received usual physician advice or an intervention group who received usual physician advice and two counseling sessions with a smoking cessation counselor. The two groups were similar in all demographic variables, smoking-related baseline variables, and baseline levels of motivation and intention to quit smoking. The smoking status of 97% of the subjects was assessed 6 months later. In the intervention group, 46% made quit attempts and 19% quit, compared with 34% and 12%, respectively, in the control group. Intervention-group smokers made more quit attempts (mean +/- SD, 5.0 +/- 2.5 vs 1.8 +/- 0.2) and had a greater reduction in daily cigarette use (8.4 +/- 1.5 vs 3.5 +/- 1.3). Of the 74% of smokers with higher levels of motivation to quit smoking, more intervention-group smokers attempted to quit (70.8% vs 45.5%) and succeeded in quitting at the 6-month follow-up (27.1% vs 10.9%). Periodic health examinations are an excellent opportunity to counsel smokers to quit, especially those smokers with higher levels of motivation to quit smoking.

Jejunal ulceration and stricture due to wax-matrix potassium chloride tablets and amitriptyline.

A 68-year-old woman presented with abdominal distention of several weeks duration and an acute small bowel obstruction. For several years she had been prescribed amitriptyline 150 mg/d, L-thyroxine 100 micrograms/d, and digoxin 0.25 mg/d. For the previous year she had been taking hydrochlorothiazide 50 mg/d and wax-matrix KCl 20 mEq/d for hypertension. At surgery a "napkin-ring" stricture of the midjejunum was found. It had microscopic features consistent with KCl local toxicity. It is speculated that delayed gastrointestinal motility secondary to amitriptyline predisposed this patient to wax-matrix KCl toxicity and that this potential side effect be considered when prescribing wax-matrix KCl.

Preoperative risk factors for postoperative delirium.

The objective of this article was to estimate the incidence of delirium in a sample of patients undergoing elective surgery and to identify the preoperative factors most closely associated with developing this complication. Consecutive patients (n=500) underwent a full preoperative medical evaluation including assessment of cognitive and functional status. Daily evaluation on postoperative days 1 through 4 included medical record review and direct standardized patient interviews. Logistic regression was used to explore the associations between preoperative factors and postoperative delirium. Delirium was detected in 57 (11.4%) patients. Univariate factors associated with delirium included age> or =70 years (RR=3.1 [1.75,5.55]), preexisting cognitive impairment (RR=3.1 [1.73, 5.43]), greater preoperative functional limitations (RR=1.57 [1.27, 1.94]), and a history of prior delirium (RR 4.1 [1.98 to 8.27]. Adjusting for other factors, previous delirium (OR=4.08 [1.85, 9.0]), age> or =70 years (OR=3.2 [1.6, 6.0], and preexisting cognitive impairment (OR=2.16 [1.15, 4.0] remained predictive of delirium. Patients' perceptions that alcohol had affected their health (OR=6.53 [1.58 to 28.1]) and use of narcotic analgesics just prior to admission (OR=2.7 [1.37 to 5.3]) were also significantly associated with delirium postoperatively. Several easily obtained preoperative clinical factors can be used to identify patients at risk for postoperative delirium. This approach, when combined with specialized delirium teams using established guidelines, may be more effective in targeting patients at risk, thus reducing the number of episodes and days of delirium.

Recruitment in a primary care trial on smoking cessation.

The purpose of this study was to describe and compare the rates of recruitment during a randomized clinical trial on smoking cessation in two primary care practices. One site was a five-physician private family practice setting with about 15,000 patients. During 34 days, 576 patients were screened, of whom 22% were smokers. Among the smokers screened, 54% consented, 33% refused consent, and 13% were called in too early to consent. The other site was a six-physician academic medical practice with about 16,000 patients. During 53 days, 1,692 subjects were screened, of whom 16.2% were smokers. Among the smokers, 19% consented, 81% refused consent, and none were called in early. The enrollment of smokers was 3.3 times greater in the private practice than the academic practice. At the first site, study personnel screened 26.6 subjects per day, whereas the practice receptionist screened only 13.4 subjects per day (P less than .01). A randomized trial of having subjects read the informed consent versus having study personnel read it to them showed no differences in recruitment. The data suggest that private practices may have greater potential for subject recruitment than academic sites, that using study personnel improves recruitment, and that having study personnel actively involved in informed consent does not improve recruitment.

Evaluating cardiac risk in noncardiac surgery patients.

The cornerstones of the evaluation of cardiac risk in patients undergoing noncardiac surgery remain a thorough history and physical examination, and a resting electrocardiogram. However, new techniques to assess cardiac function allow more complete evaluation of high-risk patients.