A novel strategy for NQO1 (NAD(P)H:quinone oxidoreductase, EC 1.6.99.2) mediated therapy of bladder cancer based on the pharmacological properties of EO9.

The indolequinone EO9 demonstrated good preclinical activity but failed to show clinical efficacy against a range of tumours following intravenous drug administration. A significant factor in EO9's failure in the clinic has been attributed to its rapid pharmacokinetic elimination resulting in poor drug delivery to tumours. Intravesical administration of EO9 would circumvent the problem of drug delivery to tumours and the principal objective of this study is to determine whether or not bladder tumours have elevated levels of the enzyme NQO1 (NAD(P)H:quinone oxidoreductase) which plays a key role in activating EO9 under aerobic conditions. Elevated NQO1 levels in human bladder tumour tissue exist in a subset of patients as measured by both immunohistochemical and enzymatic assays. In a panel of human tumour cell lines, EO9 is selectively toxic towards NQO1 rich cell lines under aerobic conditions and potency can be enhanced by reducing extracellular pH. These studies suggest that a subset of bladder cancer patients exist whose tumours possess the appropriate biochemical machinery required to activate EO9. Administration of EO9 in an acidic vehicle could be employed to reduce possible systemic toxicity as any drug absorbed into the blood stream would become relatively inactive due to an increase in pH.

The century for cytopathology.

By the end of the 19th century, exfoliated cancer cells had been described in all of the types of specimen in which we find them today. However, it was not until Drs. Papanicolaou and Traut published their account of the diagnosis of uterine cancer from exfoliated cells (1941 and 1943) that cytopathology acquired the momentum to develop into the powerful presence that it has in human medicine today. These and the subsequent publications by Papanicolaou stimulated the development and application of cytopathology worldwide, resulting in abundant literature on the subject and a galaxy of outstanding practitioners. The 1980s saw the development and widespread use of aspiration cytology. This was followed in the 1990s by the development of automated screening systems, marking the latest stage in the evolution of cytopathology. These and other events and achievements in cytopathology, from its meager beginnings in the early 20th century to its worldwide use and acceptance today, mark this century as the "century for cytopathology."

Amyloid in cytologic specimens. Differential diagnosis and diagnostic pitfalls.

OBJECTIVE: To describe and illustrate the characteristic features of amyloid in cytologic preparations and point out its diagnostic pitfalls. STUDY DESIGN: Five fine needle aspirates and one bronchial washing that contained amyloid were retrospectively reviewed. The aspirates were obtained from each of the five following sites: lung, occipital lymph node, thyroid gland, proximal humerus and subcutaneous soft tissue. Smears of all of the aspirates were stained with Papanicolaou stain, and in two cases they were also stained with Diff-Quik. Cell block sections were stained with hematoxylin and eosin. Congo red, CD45 and CD20 were used on selected cases. RESULTS: Amyloid appears as either flocculent material or irregularly shaped fragments with scalloped and pointed edges. The amorphous fragments are acellular and frequently associated with connective tissue cells. They stain eosinophilic to cyanophilic with Papanicolaou stain and deep blue with Diff-Quik. In two cases an exuberant giant cell reaction almost obscured the amyloid. In the thyroid aspirate, the amyloid was misinterpreted as colloid. In bronchial washings and lung aspirates, amyloid has to be distinguished from mucus, alveolar proteinosis, chondroid material and corpora amylacea. When circumferentially surrounded by lymphocytes or plasma cells, flocculent amyloid deposits may simulate adenoid cystic carcinoma. CONCLUSION: Amyloid can be easily overlooked or mistaken for other entities with similar staining qualities. Congo red staining can help to confirm the diagnosis.

Immunization of common marmosets with Epstein-Barr virus (EBV) envelope glycoprotein gp340: effect on viral shedding following EBV challenge.

Epstein-Barr virus (EBV), the cause of infectious mononucleosis, is involved in the pathogenesis of several human cancers, the highest frequency of association being found in undifferentiated nasopharyngeal carcinoma and endemic Burkitt's lymphoma. The development of animal models in which potential vaccines can be tested is important. EBV infection of the common marmoset, using the M81 strain originally derived from a patient with nasopharyngeal carcinoma, induces a carrier state in this animal. Persistent infection is characterized by the production of antibodies to viral antigens, and the secretion of EBV DNA into buccal fluids. Following immunization with envelope glycoprotein gp340 derived from a bovine papilloma virus expression vector, prior to EBV infection, viral DNA was detected significantly less frequently in the buccal fluids of immunized, than of nonimmunized, infected animals, indicating that although the carrier state had not been abolished, it had been altered. A reduction in virus load was also observed when offspring of seronegative, and on occasion seropositive, parents were immunized neonatally, before EBV challenge.

Immunisation of common marmosets with vaccinia virus expressing Epstein-Barr virus (EBV) gp340 and challenge with EBV.

Epstein-Barr virus (EBV) is the cause of infectious mononucleosis and is associated with a variety of life-threatening diseases in humans. Therefore the development of an effective vaccine is an important objective. Many of the initial studies of vaccine efficacy analyse the ability of vaccine preparations to prevent the induction of lymphomas in cottontop tamarins by the B95-8 strain of EBV. We used a vaccinia virus recombinant expressing gp340, vMA1, tested previously in the cotton-top tamarin, to evaluate a common marmoset model in which the challenge virus, M81, resembles more closely the wild-type strains of EBV in the general population than does the standard B95-8 strain. We characterised the M81 strain of EBV with respect to the sequence of its gp340/220 gene and in regard to the presence of a region deleted in B95-8. Replication of the challenge virus in the group vaccinated with vMA1 was decreased when compared to control groups.

Serum albumin and CA125 are powerful predictors of survival in epithelial ovarian cancer.

OBJECTIVE: To assess the prognostic significance of presentation serum albumin, clinical stage and CA125 levels in ovarian cancer. DESIGN: Retrospective analysis of data using a Cox proportional hazards model. SETTING: A district general hospital oncology unit. SUBJECTS: One hundred and fourteen consecutive patients with epithelial ovarian cancer. INTERVENTIONS: Cytotoxic chemotherapy and surgery. MAIN OUTCOME MEASURE: Survival. RESULTS: A linear increase in risk was observed with high log CA125 (P < 0.0001) and with low albumin (P < 0.0001). In late stage patients (III and IV) albumin is the best predictor of survival (P = 0.0006). The presence of ascites, blood transfusion, type of surgery or chemotherapy did not improve the predictive model. CONCLUSIONS: CA125 and albumin can be used to identify prognostic subgroups independently of stage. Albumin alone can also be used as a predictor of survival. A simple classification of patients into three groups based on serum albumin of 41 g/l or more, 35 to 40 g/l and 34 g/l or less provides a clear separation of survival curves in the present group of patients.

Clinical outcome in aggressively treated meningeal carcinomatosis.

OBJECTIVE: To critically evaluate the clinical outcome of patients with cytologically proved meningeal carcinomatosis and to identify factors associated with an improved outcome. DESIGN: A consecutive series of patients was identified from a cytopathology data base. The study period was from 1985 to 1990. Case records and results of radiologic investigations were reviewed; all patients were followed up until their deaths. SETTING: University hospital with a cancer center. PATIENTS: Thirty-six consecutive patients with cytologically proved meningeal carcinomatosis. Patients with lymphoma or leukemia were excluded. INTERVENTIONS: External beam radiation therapy, intrathecal chemotherapy via a ventricular catheter, and intravenous chemotherapy. MAIN OUTCOME MEASURES: Clinical assessment and survival. RESULTS: Symptoms improved in 15% of 33 treated patients. Response to treatment had no clear relationship with the age, site of involvement, or tumor type. Median survival for treated patients was only 9 weeks. Patients who received more than five intrareservoir chemotherapy treatments had a median survival of 23 weeks. Median survival for patients with breast carcinoma who received intravenous chemotherapy was 20 weeks, significantly better than that of patients not receiving intravenous chemotherapy. CONCLUSIONS: Survival is poor despite aggressive treatment. Intravenous chemotherapy may improve survival in patients with chemoresponsive primary tumors and deserves further study.

Analysis of the tumorigenic potential of common marmoset lymphoblastoid cells expressing a constitutively activated c-myc gene.

The respective roles of Epstein-Barr virus (EBV) and c-myc in the pathogenesis of endemic Burkitt's lymphoma (BL) are unclear. In order to help resolve the question whether constitutive expression of the c-myc gene in an EBV-immortalised B cell is sufficient to induce a tumorigenic phenotype, B cells from a common marmoset (Callithrix jacchus) were immortalised with EBV, transfected with a constitutively activated c-myc gene and inoculated into the host animals. Despite the cell line transfected with c-myc displaying enhanced growth characteristics, in vitro and in vivo experiments demonstrated that this was not sufficient to induce a tumorigenic phenotype. This supports our previous findings with EBV-immortalised human B cells transfected with an activated c-myc gene (Hotchin et al., 1990).

Cytomegalovirus infection of the cervix detected by cytology and histology: a report of five cases.

We present four cases of cytomegalovirus (CMV) infection detected by cervical cytology of asymptomatic young women. One patient who had undergone bone marrow transplantation was immunosuppressed, but no factors predisposing to CMV infection were identified in the other three patients. A cervical biopsy specimen from a fifth patient, who was also asymptomatic, demonstrated the locus of CMV infection to be in endocervical gland cells. Immunocytochemical and polymerase chain reaction studies on this biopsy specimen confirmed that the histological changes were caused by CMV. The finding of CMV-infected cells in cervical cytological or biopsy specimens is a rarity. Our observations in asymptomatic, non-immunocompromised women suggest their presence is an incidental finding, unlikely to have any clinical significance.

Postoperative apnea in infants.

This is a review of 127 neonates evaluated for postoperative apnea and bradycardia (A&B) after inguinal surgery. The patients could be divided into three groups based on postconceptional age (PCA) at operation. Ten of 29 patients operated on at PCA of 33 to 39 weeks developed episodes of A&B. Preoperative assessment was not reliable in their identification. In the second group of 54 patients at PCA 40 to 44 weeks, 8 developed A&B, whereas in the third group of 44 patients at PCA of 45 to 60 weeks 1 patient developed A&B. In the latter two groups preoperative assessment identified all patients at high risk. We conclude that after PCA of 40 weeks patients at risk for A&B can be identified preoperatively. Patients operated on up to 39 weeks PCA should all be observed in the hospital.

"Collagen balls" in peritoneal washings. Prevalence, morphology, origin and significance.

Peritoneal washings obtained at laparotomy from women undergoing surgery for neoplasms of the genital tract may contain "collagen balls," consisting of tissue fragments composed of collagen covered with mesothelial cells. Collagen balls were found in 19 (4.5%) of 418 peritoneal washings and were more prevalent in specimens labeled pelvic washings (17 of 294, or 5.8%) than in those labeled peritoneal washings (2 of 124, or 1.6%). In 15 of the 19 cases in which we found collagen balls, at least one ovary was available for microscopic examination. In 14 of the 15 cases minute nodular papillary stromal projections covered with mesothelium were found on the surface of the ovaries. We conclude that collagen balls, a nonspecific entity, most probably originate on the surface of the ovaries. Their significance lies in their being mistaken for mucin-distended cells exfoliated from a neoplasm or from detached fragments of a papillary ovarian neoplasm.

Clinical outcome in aggressively treated meningeal gliomatosis.

We reviewed 11 consecutive patients with an antemortem cytologic diagnosis of meningeal gliomatosis. In three patients, meningeal gliomatosis was diagnosed before surgical resection of the glioma. Three of five patients with anaplastic astrocytoma and one with oligodendroglioma improved with treatment. Patients with glioblastoma multiforme did not respond and had a median survival from the diagnosis of meningeal gliomatosis of only 8 weeks. Meningeal gliomatosis can be an early finding in gliomas. Treatment response is probably related to tumor histology and grade.

Cytological aspects of pleural, peritoneal and pericardial fluids from patients with systemic lupus erythematosus.

Serous effusions of nine of 33 patients with systemic lupus erythematosus contained lupus erythematosus (LE) cells, identifiable in Papanicolaou-stained smears, wet films stained with toluidine blue, and cell blocks stained with haematoxylin and eosin. Specimens in which LE cells were found contained at least a moderate number of polymorphonuclear neutrophilic leucocytes. Most specimens containing LE cells also contained cells that resembled LE cells (tart cells), which appeared to be small macrophages that had phagocytosed a non-homogenized nucleus of a cell that had undergone degeneration. In 34 years of cytologic practice we have recognized LE cells in serous effusions only from patients who were already diagnosed as having systemic lupus erythematosus.

Effects of fluorophore structure and hydrophobicity on the uptake and metabolism of fluorescent lipid analogs.

Cellular transport and metabolism of fatty acids are integral components of lipid metabolism, but the mechanisms and regulation involved are poorly understood. A variety of commercially available fluorescent analogs of fatty acids, are potentially useful probes for the study of lipid metabolism by such techniques as cell sorting and fluorescence microscopy. We have screened a series of fluorescent fatty acids to identify analogs that would reliably simulate the metabolic behavior of natural fatty acids; i.e., similar kinetics of transport, of intracellular movement, and of metabolic fate. The metabolic behavior of these analogs was compared with those of some naturally occurring fatty acids in HepG2 cells, which are a good model of some aspects of hepatic function. Fluorescent analogs containing polar fluorophores yielded the lowest rates of cellular uptake and conversion to acylated lipid products. Similarly, fluorescent analogs with the fluorophore located near the carboxylic acid group were poorly metabolized. Fatty acid analogs containing anthracene or pyrene at the n-terminus of the acyl chain were the most extensively incorporated into cellular lipids. The types and amounts of labeled lipid products formed from these analogs and from natural fatty acids were similar. Pyrene-labeled analogs have spectral properties that can be measured fluorometrically at very low concentrations. Therefore, we compared the cellular metabolism of 12-(1-pyrenyl)dodecanoic acid with those of palmitic and oleic acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Fine needle aspiration biopsy of the breast. Influence of the number of passes and the sample size on the diagnostic yield.

The sensitivity of fine needle aspiration (FNA) biopsy of the breast as a function of the number of aspirations performed on any given lesion was investigated. Four separate aspirations each were performed on over 400 lesions of the breast, 93 of which yielded a cytologic diagnosis. The incremental diagnostic yields of each subsequent aspiration were tabulated. The first aspirate of the sequence gave the greatest yield, with smaller incremental yields on the second through the fourth aspirates. Benign and malignant lesions gave similar results, as did palpable and nonpalpable lesions (the latter being usually smaller in size). A mathematical extrapolation of the data indicates that three or four aspirations of any given lesion provide the optimal yield within the limits of practicality. This performance of multiple FNA biopsies is particularly important when the pathologist does not perform the biopsy, or is unable to assist in the immediate interpretation of the specimen to assess its adequacy.

The pathognomonic cytologic picture of rheumatoid pleuritis. The 1989 Maurice Goldblatt Cytology award lecture.

Patients with rheumatoid arthritis of any degree of severity or duration may develop necrotizing granulomatous pleuritis, a morphologic replica of the inflammatory reaction characteristic of rheumatoid synovitis and rheumatoid nodules. The ensuing pleural effusion may show a unique cytologic picture consisting of slender, elongated multinucleated macrophages, round giant multinucleated macrophages and necrotic background material. A review of 24 such cases from our laboratory as well as the relevant literature clearly demonstrated that not only is this cytologic picture unique, but that it is pathognomonic of rheumatoid disease, even in patients who have not yet developed arthritis.

Curschmann's spirals in pleural and peritoneal effusions.

Curschmann's spirals were found in smears and cell block preparations of five spontaneously occurring pleural and peritoneal fluids. The spirals were similar to those seen in the respiratory tract, although generally much smaller. In three of the five cases, the fluids also contained mucus-secreting adenocarcinoma cells; it is postulated that the spirals formed from mucus secreted by these cells. In the other two cases, there was evidence of serosal inflammation; it is suggested that the spirals in these cases developed from submesothelial connective tissue mucosubstances that entered the serosal cavity through a mesothelium of increased permeability due to the inflammation. No simple explanation can be accepted as to the exact mode of spiral formation, which is presumed to be a complex physical and biochemical phenomenon.

Radiographically guided fine-needle aspiration of nonpalpable breast lesions.

Radiographically guided fine-needle aspiration (X-FNA) in 215 nonpalpable, mammographically detected breast lesions was performed by means of a coordinate-grid localization system. Aspirates were categorized either into four cytologic groups or as simple cysts. Based on the most stringent cytologic criteria, the maximum sensitivity for detection of carcinoma was 97% and the specificity was 94%. However, according to these strict cytologic criteria, only 46% of aspirates contained representative material. Based on less stringent cytologic criteria, the maximum sensitivity was 68% and the specificity was 97%. Forty-one of 74 lesions proved to be malignant at biopsy. Thirty-four patients did not complete adequate mammographic follow-up. High sensitivity and specificity can be achieved with X-FNA. However, management decisions ultimately require integration of mammographic findings with cytologic results. Close cooperation among mammographer, surgeon, cytopathologist, and patient is mandatory for successful results.