Murine lung carcinogenesis following exposure to ambient ozone concentrations.

Inbred strain A/J mice, responsive to the chemical induction of pulmonary adenomas, were used to assess any of several roles that the photochemical air pollutant ozone might play in lung carcinogenesis. In separate experiments, animals were exposed to two concentrations of ozone (0.31 +/- 0.01 and 0.50 +/- 0.02 ppm) intermittently for a 6-month period, to evaluate the potential of ozone to act as either a pulmonary carcinogen, a tumor promoter, or an inhalant capable of increasing lung tumor yield when exposure was in conjunction with a pulmonary carcinogen, urethane. Statistical analyses of results indicated that ozone exposure at both concentrations caused an increase in lung tumor number relative to clean air controls, but that ozone was not an effective tumor promoter under the conditions of our protocol. When ozone exposure immediately preceded treatment with urethane (CAS: 51-79-6), animals were at increased risk for the development of lung adenomas.

Frequency and distribution of occult micrometastases in lymph nodes of patients with non-small-cell lung carcinoma.

BACKGROUND: Accurate assessment of the presence and absence of tumor in the regional lymph nodes is critical in assessment of prognosis for patients with lung cancer. Development of sensitive immunohistochemical techniques and specific monoclonal antibodies has increased our capacity, in melanoma and breast cancer, to detect small groups of tumor cells or even single tumor cells in lymph nodes that appear to be tumor free in conventionally stained sections. PURPOSE: This retrospective study was designed to assess whether use of a polyclonal antikeratin reagent in immunohistochemical analysis offers any advantage over conventional histopathology in detection of regional lymph node metastases in non-small-cell lung cancer. METHODS: Paraffin-embedded tissue sections from regional lymph nodes of 65 patients with non-small-cell lung cancer were studied. We examined tissue from 588 nodes of 60 patients with a diagnosis of disease confined to the lung and from 72 nodes of five patients with a diagnosis of metastasis to some nodes. A polyclonal antikeratin antibody was applied to the lymph node tissue sections, using the avidin-biotin complex immunoperoxidase technique. RESULTS: Single tumor cells and small clusters of tumor cells (occult micrometastases) not visible on routine evaluation were readily detected in 38 (63%) of the 60 patients whose nodes appeared to be negative on examination of hematoxylin-eosin-stained slides. In the five patients with a diagnosis of node-positive non-small-cell lung cancer, five (10%) of 51 nodes that were tumor free on conventional examination contained metastases. Metastatic tumor cells were most often located in the subcapsular or medullary sinuses. The lymph nodes that contained occult tumor cells were those located nearest to the tumor, mainly in peribronchial and hilar locations. The median survival of patients with occult metastases (1977 days) was shorter than that of patients whose nodes contained no tumor (2456 days) but was longer than that of patients whose nodes contained metastases detectable on hematoxylin-eosin-stained slides (927 days). CONCLUSIONS: Our results suggest that, in patients with non-small-cell lung cancer, metastatic involvement of regional lymph nodes is more frequent than was previously determined by the conventional histologic method and substantially more frequent than in other tumor types, such as melanoma and breast cancer. IMPLICATIONS: The high frequency of occult nodal metastases in non-small-cell lung cancer makes it clear that, without immunohistochemistry, disease is understaged in many patients. Therefore, it seems essential that immunohistochemical evaluation of the lymph nodes be undertaken in clinical trials.

Hormone replacement therapy in the menopause: a suitable animal model.

Female CBA mice, aged 11 months, were treated cyclically with oral ethynyl oestradiol or oestrone sulphate for 3 months. The ovaries of all animals appeared to be atrophied. Target tissues throughout the genital tract showed a response to both oestrogens. Electron microscopy of both the endometrium and the urothelium demonstrated morphological changes characteristic of increased cellular metabolic activity in the treated mice. Endometrial hyperplasia developed in both treatment groups but more pronounced epithelial changes occurred with oestrone sulphate. This hyperplasia was accompanied by a doubling in the number of uterine cytoplasmic oestrogen receptors. A 50% fall in serum levels of luteinizing hormone in the treated mice revealed that the hypothalamic-pituitary system was still intact. Both oestrogens improved skeletal balance by changes in cortical-endosteal bone remodelling. The results suggest that the CBA strain of mouse is a suitable model for the study of the human climacteric and its response to hormone replacement therapy.

16 alpha-Iodo-testosterone: chemical synthesis and evaluation as a potential radiopharmaceutical.

The chemical synthesis and characterization, including 1H NMR, of 16 alpha-iodo-androstenedione and 16 alpha-iodo-testosterone are described. Each has been synthesized with 125I and tested in rats in vivo for accumulation in androgen dependent tissues over a 24 hr time course. Neither compound was accumulated in prostate against the blood gradient of normal or 24 hr castrate animals. The metabolism, subcellular distribution and binding of 16 alpha-[125I]iodo-testosterone to protein in prostate has also been examined. By comparison with data obtained after the administration of [3H]testosterone we conclude that the failure of this iodinated androgen to accumulate in androgen dependent tissues arises because of its low binding affinity for receptor protein.

17 alpha-Z-[125I]iodovinyloestradiol and its 3-acetate: chemical synthesis in vivo distribution studies in the rat. Comparison of tissue accumulation and metabolic stability with 17 alpha-E-[125I]iodovinyl and 16 alpha-[125I]iodo oestradiols.

Certain oestrogens on substitution with halogens, have been shown to retain receptor binding affinity and accumulate in target tissues and therefore could, when labelled with gamma-emitting halogen isotope, act as radiopharmaceuticals for the imaging of receptor positive breast tumours. In order to evaluate putative radiopharmaceuticals, 17 alpha-Z-iodovinyloestradiol ((17 alpha,20Z)-21-iodo-19-norpregna-1,3,5(10),20-tetraene-3,17 beta-diol) and its 3-acetate have been chemically synthesized and labelled with 125I. Tissue distribution studies in female rats have been compared to those obtained using 17 alpha-E-[125I]iodovinyloestradiol ((17 alpha,20E)-21-[125I]iodo-19-norpregna-1,3,5(10),20-tetraene-3,17 beta-diol) and its 3-acetate and to 16 alpha-[125I]iodo oestradiol (1,3,5(10)-estratien-16 alpha-[125I]iodo-3,17 beta-diol). All compounds showed a similar tissue distribution with the highest concentrations (cpm/g tissue) in the thyroid greater than liver greater than uterus greater than kidney greater than lung greater than blood greater than heart, spleen. The concentration in the uterus was highest after injection of 17 alpha-Z-iodovinyloestradiols or 16 alpha-iodo oestradiol. Target (uterus) to background (blood) tissue ratios were highest after injection of 17 alpha-Z-iodovinyloestradiol-3-acetate and 16 alpha-iodo oestradiol (6:1). Deiodination in vivo took place to a small degree, amounting to 1.6%, 0.9% and 0.35% of the injected dose after 4 hr with the 17 alpha-Z, 17 alpha-E and 16 alpha compounds, respectively. For reasons of chemical expediency and consideration of the biological results 17 alpha-Z-iodovinyloestradiol-3-acetate is the compound of choice for the detection of oestrogen receptor positive tissues.

Occult metastases in the axillary lymph nodes of patients with breast cancer node negative by clinical and histologic examination and conventional histology.

We examined axillary lymph nodes from 80 women with node-negative breast cancer, by immunohistochemistry, utilizing polyclonal antibodies to cytokeratins and carcino-embryonic antigen and monoclonal antibodies to cytokeratins and milk fat globulin. Occult metastatic tumor, undetectable in hematoxylin and eosin stained slides, but visible by immunohistochemistry, was detected in 23 of 80 patients (29 per cent). Occult tumor was observed in patients with invasive ductal carcinoma (21/76-28 per cent) and in individuals with invasive lobular carcinoma (2/4-50 per cent). In patients with occult metastases the primary tumors were slightly larger (mean 2.39 cm, range 1.00-5.00 cm) than those of patients whose nodes were negative for tumor cells (mean, 2.03 cm, range, 0.60-4.50 cm). Information concerning clinical outcome is available for 61 patients followed for between 1 and 7 years (mean 3.2 years). Three of 17 patients (18 per cent) who had occult tumor in the nodes developed distant metastases, all less than 3 years after initial surgery. One of the 44 patients (2 per cent) whose nodes were free of occult tumor developed distant metastases 5 years following surgery. Local recurrences in the area of the mastectomy occurred in one of 17 patients with occult nodal tumor (6 per cent), less than 1 year after surgery. Local recurrences were seen in three of 44 patients without occult metastases (7 per cent), in two patients 5 years after mastectomy and in one patient 7 years after mastectomy.

Intralesional BCG immunotherapy of pulmonary tumors.

A new method of baccile Calmette Guérin (BCG) immunotherapy has been evaluated in 25 patients with lung cancer. This technique involves the direct intratumor injection of BCG in patients with pulmonary tumors. Four patients underwent transbronchoscopic injections of tumor, with significant resolution of tumor in three. Twenty-one patients underwent percutaneous injection with the aid of fluoroscopic imaging. Twenty of these 21 underwent pulmonary resection. Toxicity was limited to fever and malaise. The severity of this toxicity was closely related to the magnitude of reactivity to PPD. There were no intraoperative or postoperative complications occasioned by BCG injection. The histologic features were characterized by granulomatous inflammatory reactions in the tumors and the regional lymph nodes. These histologic findings are identical to those seen in animal tumor models in which potent systemic antitumor immunity is induced. BCG intratumor injection is technically feasible and safe, and it is appropriate to begin phase 2 and phase 3 clinical trials.

Peritoneal gliomatosis from a gastric teratoma.

A case of gastric teratoma in a newborn is presented. Ten months after removal of the teratoma, peritoneal gliomatosis was discovered incidentally in inguinal hernia sacs. This is the first report of an association between peritoneal gliomatosis and a teratoma other than ovarian.

Bone tumor radiograph review by pathologists prior to pathologic diagnosis: a receiver operator curve analysis of diagnostic utility.

Radiologic-pathologic correlation has long been considered a prerequisite for the accurate diagnosis of bone lesions. We investigated using receiver-operating characteristic analysis the accuracy of histopathologic diagnoses with and without pathologist review of radiographs. While accuracy of diagnosis did improve when the radiographs were reviewed by the pathologist, this increase did not reach statistical significance (P=0.1311). Potential reasons for this finding including case selection and reviewer expertise are discussed. Based on our study, the review of radiographs did not result in a statistically significant difference in accuracy of the pathologic diagnosis of bone tumors. However, our data suggest that there may be a difference particularly in selected cases and among certain observers.

Effects of collagenase upon the intervertebral disc in monkeys.

The authors studied 5 rhesus monkeys for the effects of annulotomy, with or without the addition of a bacterially-derived collagenase into the nuclear-evacuated centrum of lumbar intervertebral discs. The animals were sacrificed from 3 weeks to 21 months after the single or staged double procedures. The earliest radiographic changes were loss of height of the interspace at 7 days, erosion of the vertebral body margins at 3 weeks, malalignment at 6 weeks, osteophytic formation at 3 1/2 months, sclerosis of vertebral body surfaces at 9 1/2 months, and fusion of adjacent vertebral bodies at 13 1/2 months. Pathological changes included early loss of disc convexity, focal new home formation, osteophytic new growth, through-and-through bone formation, irregular cystification, and disc replacement by hyaline fibrous tissue. Changes were reminiscent of those seen in humans with naturally acquired disc disease or aging of the disc. Injuries were produced in a tissue that probably has a limited scope of variation in reaction to insult.

An improved synthesis of 6beta-hydroxy-5beta-pregnane-3,20-dione.

6beta-Hydroxy-5beta-pregnane-3,20-dione, formerly prepared by hydroboration method, has been obtained in greatly improved yield by a simpler irradiation-hydrogenation procedure.

Specificity studies of monoclonal and rabbit antibodies raised against steroid glucuronides.

Monoclonal and rabbit antibodies raised against estrone-3-glucuronide and pregnanediol-3 alpha-glucuronide have been studied with respect to their ability to bind free estrone and its conjugates or free pregnanediol and its conjugates, respectively. High titre and high specificity were observed with monoclonal antibodies produced against pregnanediol-3 alpha-glucuronide, whereas the monoclonal antibodies produced against estrone-3-glucuronide were not so specific when compared with the corresponding rabbit antibodies. Both monoclonal and rabbit antibodies had affinity constants in the range of 10(9)--10(10) liter/mole.

The isolation of estriol-16 alpha-glucuronide and pregnanediol-3 alpha-glucuronide from late pregnancy urine.

A simple procedure is described, whereby estriol-16 alpha-glucuronide and pregnanediol-3 alpha-glucuronide have been isolated from late pregnancy urine. The conjugates isolated were used as haptens for raising antisera for radioimmunoassay.

Splenomegaly in mice following exposure to ambient levels of ozone.

Female strain A/J mice were exposed to 0.31 ppm (608 micrograms/m3) ozone continuously for 103 h every other week for 6 mth. Following an additional period of 5 mth in a filtered air environment, animals were killed and examined for evidence of altered spleen weight. It was observed that animals exposed to ozone had a greater spleen weight, and spleen to body weight ratio than air-breathing controls. In some of the ozone-exposed mice, pronounced splenomegaly was noted. Increased spleen weight appears to be another example of an extrapulmonary effect of ozone inhalation.

Pathological features of inflammatory bowel disease in childhood.

Inflammatory bowel disease (IBD) in childhood is realistically interpreted to mean ulcerative colitis (UC) and Crohn's disease of the colon. Their gross and microscopic features are discussed along with the differential diagnosis from other childhood conditions associated with bloody diarrhea.

Histopathology of cryptococcosis and other fungal infections in patients with acquired immunodeficiency syndrome.

OBJECTIVE: To gain insight into the histopathologic characteristics of fungal infection in acquired immunodeficiency syndrome (AIDS). METHODS: A review was conducted of the histopathology for 162 patients with evident fungal infection. RESULTS: The microscopic appearance of esophageal candidiasis that was common in patients with single organ involvement revealed necrotic debris containing proliferating hyphae at the site of mucosal erosions without fungal invasion of underlying tissue. The incidence of oral and esophageal candidiasis was followed by that of pulmonary aspergillosis and Candida infection. Eighteen patients had generalized cryptococcosis, representing the commonest generalized fungal disease. The essential histologic features of the disease consisted of yeast cell proliferation with a histiocytic response, but only minor lymphocytic and neutrophilic components. This was different from the manifestations of both Candida and Aspergillus infections. The two histologic patterns recognized in the pulmonary cryptococcal lesions could be graded with respect to the degree and type of inflammatory reaction. The milder one consisted of small scattered foci of intra-alveolar cryptococcal proliferation with a histiocytic response. Another pattern involved massive cryptococcal infection, which might be simply more extensive than that in the former. Capillary involvement of alveolar septa was an important common finding in all 18 patients.

Predicting the risk of cancer at the time of breast biopsy. Variation in the benign to malignant ratio.

The benign to malignant ratio (B:M ratio) among breast biopsies (number of benign breast lesions divided by number of breast cancers) is widely believed to be around 4:1 or 5:1. This belief appears to be based upon experience from the 1950's and 1960's. We investigated possible reasons for the current wide variability of the B:M ratio. A straight line relationship between the log B:M ratio and age exists in our data. The B:M ratio is sensitive to racial differences between patient groups but in only minimally affected by varied histologic inclusion criteria. The overall B:M ratio should not be used to counsel women about their breast cancer risk at the time of biopsy. Age-specific B:M ratios provide a more realistic risk assessment.

Mixed tumors of the salivary glands. Growth pattern and recurrence.

In 112 patients with mixed tumors of the salivary glands, 94 tumors were located in parotid glands, 11 in submandibular salivary glands, and 7 in monor palatal salivary glands. Capsular ingrowth, a characteristic growth pattern, may explain the frequency of multilobulation and capsular defects in these tumors, particularly those of long standing. The capsular thickness, the extent of fibrosis with stromal hyalinization, and the degree of lymphocytic infiltration within and around the capsule, were associated with cellularity of the tumors. Thus, 69% of the hypocellular neoplasms had incomplete capsules, and only 13% showed moderate to substantial lymphocytic infiltration; corresponding figures for hypercellular tumors were 22% and 42%, respectively. Of the recurrent lesions, 61% were hypocellular; 11% of the hypercellular tumors recurred. Incomplete resection, hypocellularity, and incomplete encapsulation were considered to be the major factors in recurrence.

[Deep-seated mycosis in patients with solid-organ transplantation--a study of autopsied cases in the United States].

Autopsies with solid-organ trans plantation (SOT) in UCLA Medical Center were reviewed, especially focussing on the deep-seated fungal infection and the incidence of fungal infections, causative fungi, and organs involved were evaluated. Deep-stated fungal infections were demonstrated in 21.0% of the patients with SOT. The incidence of fungal infections were 26.1% in kidney transplantation, which was the highest rate in SOT autopsies we reviewed, 25.0% in liver transplantation, 14.3% in lung transplantation, and 13.2% in heart transplantation. And the most common causative fungi were Aspergillus species, seen in 70.6% of SOT autopsies. In contrast, Candida species were observed in 25.5%. In addition, the incidence of deep-seated mycosis has been increasing since 1992, this was parallel to the increase of aspergillosis, most of which were found as a generalized spreading. In contrast, candidal lesions, were almost limited to the digestive tracts. The above suggests that from this standpoint, improvement of prophylactic and therapeutic technique against aspergillosis is the greatest problem in deep-seated mycosis in SOT patients.