A soluble transforming growth factor beta type III receptor suppresses tumorigenicity and metastasis of human breast cancer MDA-MB-231 cells.

Transforming growth factor beta (TGF-beta) can promote late stage tumor progression in a number of model systems. In the present study, we have examined whether expression of a truncated soluble extracellular domain of TGF-beta type III receptor (sRIII) in human breast cancer MDA-MB-231 cells can antagonize the tumor-promoting activity of TGF-beta by sequestering active TGF-beta isoforms that are produced by the cancer cells. The secretion of sRIII reduced the amount of active TGF-beta1 and TGF-beta2 in the conditioned medium. This led to a significant reduction of the growth-inhibitory activity of the medium conditioned by sRIII-expressing cells on the growth of mink lung epithelial CCL64 cells in comparison with the medium conditioned by the control cells. The tumor incidence and growth rate of all of the three sRIII-expressing clones studied were significantly lower than those of the control cells in athymic nude mice. Four of five control cell-inoculated mice showed spontaneous metastasis in the lung, whereas none of the sRIII-expressing cell-inoculated mice had any lung metastasis. Thus, our results suggest that the sRIII may be used to antagonize the tumor-promoting activity of TGF-beta.

Ultrastructure of osteoclastoma-like giant cell tumor of thyroid.

An aggressive anaplastic tumor of the thyroid with osteoclastoma-like giant cells was studied by electron microscopy. The lack of junctional complexes or other obvious epithelial elements suggested mesenchymal origin. The giant cells resembled osteoclasts with respect to mitochondria, rough endoplastic reticulum, lysosomes, ribosome complexes and vacuoles, but the prominent nuclear golgi apparatus of giant cell tumors of bone was absent.

Role of natural killer cells in the pathogenesis of human acute graft-versus-host disease.

Clinical acute graft-versus-host disease (aGVHD) was correlated with alterations in PBL phenotype and skin immunohistology in 52 patients transplanted with HLA-identical bone marrow. Concurrent with the emergence of aGVHD, there was a profound decrease in absolute number of CD3- T cells and an increase in CD3-CD16+, CD56+ (a subset of which coexpress CD8+ "dim") NK cells in the PBL. CD4+ T and CD20+ B lymphocytes failed to recover within 90 days in the patients with grades II-IV aGVHD. Ex vivo partial T cell depletion, in itself, did not significantly impair T cell recovery as compared to that in non-T-depleted recipients unless aGVHD occurred. Although leukocytic cellular infiltration in the skin was generally sparse, CD16+ NK lymphocytes were significantly increased in grades II-IV aGVHD. By contrast, there was no significant increase in CD3+, CD4+, or CD8+ lymphocytes in these lesions as compared to skin biopsies obtained from BMT patients without aGVHD or from normal skin. Taken together, these findings suggest that NK cells may be important in the pathogenesis of human aGVHD.

Unusual immunofluorescence patterns for terminal deoxynucleotidyl transferase in blast crisis chronic myelogenous leukemia.

An unusual cytoplasmic distribution of terminal deoxynucleotidyl transferase (TdT) antigen in leukemic cells from two patients who had chronic myelogenous leukemia in blastic phase is described. In most leukemic cells that contain TdT, the intracellular location has been reported to be exclusively nuclear. The cells from these two patients demonstrated TdT staining in both the nucleus and the cytoplasm. The pattern is remarkably similar to that observed in thymocytes, in which bright cytoplasmic staining may also be seen. In the immunofluorescence procedures for detection of TdT in blasts from patients who have chronic myelogenous leukemia, significant cytoplasmic staining should not be mistaken for nonspecific absorption of immunoglobulins or specimen deterioration.

Evaluation of methods of detecting terminal deoxynucleotidyl transferase in human hematologic malignancies. Comparison of immunofluorescence and enzymatic assays.

Terminal transferase (TdT) activity and antigen have been measured in 267 specimens of human bone marrow and peripheral blood by using a biochemical assay for enzymatic activity and an immunofluorescence test for antigen. Oligo p(dA)50 and dGTP were used as reagents in the biochemical assay and either rabbit anti-calf TdT or rabbit anti-human TdT was used as the primary antibody for immunofluorescence. Because both false-positive and false-negative detection of TdT antigen occurs, the biochemical assay of TdT activity is considered the standard against which immunofluorescence assays must be measured. If specimens of cells contained TdT activity, then the immunofluorescence detected antigen in 91% of cases (rabbit anti-calf TdT) and 95% of cases (rabbit anti-human TdT). When no TdT activity was detected, the immunofluorescence test was positive in 7.8% of cases (rabbit anti-calf TdT) and 5.2% of cases (rabbit anti-human TdT). When air-dried slices were shipped by air mail to a distant location before being stained for immunofluorescence, TdT antigen was detected in only 33% of matched pair cases which contained TdT activity. From this study, the authors conclude that with current methodology, immunofluorescence tests for TdT antigen must be carried out on slides prepared in the testing laboratory and that such tests are reliable in more than 90% of cases. However, because a small percentage of results are false positives and false negatives, the authors suggest that if a patient's clinical response is not consistent with the immunofluorescence TdT result, an enzymatic assay for TdT activity be carried out.

Etiologies for non-correlating cervical cytologies and biopsies.

To investigate the etiologies for discrepancies between cervicovaginal smear and corresponding cervical biopsy results, 615 patients with cytologic diagnoses of dysplasia or malignancy during 1 year were reviewed. Sixty-nine patients (11%) were identified in which the cytologic and histologic diagnoses differed. Utilizing an algorithm developed for the study, these cases were assigned an etiologic category for discrepancy: colposcopic biopsy or cytologic sampling, cytologic screening, histotechnical processing, histologic or cytologic interpretation. The most common cause for a discrepancy was colposcopic biopsy sampling (36 cases, 51%). There were nine errors (13%) in biopsy interpretation, with seven underdiagnoses and two overdiagnoses. Eight errors (11%) in cytologic interpretation occurred with half of these representing underdiagnoses. The other causes for discrepancy were less common--cytologic sampling (6 cases), histotechnical processing (3 cases), cytologic screening (2 cases), and a combination of factors (5 cases). Use of this algorithm allows laboratories to identify problem areas and design specific corrective protocols to improve diagnostic accuracy and patient care.

Primary retroperitoneal cysts: report of an unusual case and a survey of the literature.

A case of a primary retroperitoneal cyst was associated with the chemical abnormalities of Cushing's disease and pheochromocytoma. Review of the literature failed to uncover a similar endocrinologically active retroperitoneal cyst. Primary retroperitoneal cysts, defined as those cysts lying in the retroperitoneal fatty tissue that have no apparent connections with any adult anatomical structure, are rare. These cysts are thought to arise from the mesonephros. They become symptomatic by virtue of their size or position. Diagnosis is suggested by an abnormal intravenous pyelogram and may be confirmed by arteriography. Treatment of choice is excision.

Pseudolymphoma of the stomach. A diagnostic and therapeutic dilemma.

Pseudolymphoma is an uncommon benign lesion of the stomach that poses a difficult problem in diagnosis and management. The clinical manifestations and endoscopic, radiologic, and biopsy findings are not generally helpful in making this diagnosis preoperatively. Histologic examination of the lesion is the only reliable method that distinguishes pseudolymphoma from true lymphoma. Distinguishing histologic features of pseudolymphomas are (1) formation of true germinal centers, (2) presence of a polymorphous inflammatory infiltrate, and (3) absence of lymph nodal involvement by lymphoma. We report four cases and review the literature to illustrate the features of pseudolymphoma. Subtotal gastric resection is done for diagnostic as well as for therapeutic purposes. Distinction of these benign lesions from malignant lymphomas is important so that unnecessary radical surgery and postoperative radiation therapy or chemotherapy are avoided.

Proliferating cell nuclear antigen in prostatic adenocarcinoma: correlation with established prognostic indicators.

OBJECTIVE: The utility of an antibody to proliferating cell nuclear antigen (PCNA), a growth-specific nuclear protein, was assessed as a prognostic variable for prostatic adenocarcinoma. Its expression was correlated with established prognostic indicators, including tumor grade, stage, prostatic-specific antigen (PSA), and percent of tumor in the gland at excision. METHODS: Forty archival needle biopsies containing a minimum of four hundred tumor cells were analyzed. Immunoperoxidase staining of paraffin sections was performed for PCNA (PC10) after pretreatment in antigen retrieval solution. A proliferative index (PI) for each case was derived using image analysis with measurement of at least four hundred twenty-five nuclei. RESULTS: PI values ranged from 2.4 to 31.3 percent. Mean PI values varied significantly (ANOVA, p = 0.005) among cases with dominant Gleason grade (DGG) of 3 (mean PI = 9.3%), 4 (mean PI = 13.7%), and 5 (mean PI = 18.8%). By t test, significant differences were noted for PI in cases with DGG 2 and 3 versus those with DGG 4 and 5 (p = 0.0065). PI for cases with DGG 3 versus 5 showed significant difference (p = 0.0017). Tumors of Gleason scores 5 to 7 differed significantly from those with scores 8 to 10 (p = 0.014). A statistical relationship for PI and PSA, clinical stage, and percent tumor at resection could not be established by linear regression. CONCLUSIONS: These findings suggest that additional study of the PI, as determined by PCNA immunohistochemistry and image analysis, may be warranted to determine its usefulness as an adjunctive parameter in prostate adenocarcinoma. This technique may be particularly useful in needle biopsies where limited tumor may render assessment of grade difficult.

Single focus of adenocarcinoma in the prostate biopsy specimen is not predictive of the pathologic stage of disease.

OBJECTIVES: To determine whether a very small focus of prostate cancer in a needle biopsy specimen correlates with organ-confined disease or with favorable disease parameters. METHODS: Of 598 needle biopsies of the prostate performed from January 1990 through June 1994, 49 specimens (8.2%) contained a microscopic focus (less than 2 mm in length of the entire biopsy core specimen) of adenocarcinoma. For these 49 patients, the clinical and pathologic features were correlated. RESULTS: Of these 49 patients, 27 (55.1%) underwent either radical prostatectomy, with or without pelvic lymph node dissection (26), or pelvic lymph node dissection alone (1). Seven of these 27 patients (25.9%) had extraprostatic disease: lymph node involvement (1), positive surgical margins (5), or seminal vesicle invasion (1). Ten of the 49 patients (20.4%) underwent radiotherapy, and 12 (24.5%) chose hormonal therapy. The pathologic stage for these 22 patients could not be ascertained. However, despite the limited amount of disease in the biopsy specimen, 2 patients treated with radiotherapy suffered a relapse (mean interval to recurrence, 11.5 months), and 3 patients treated with hormonal therapy (early or delayed) had bony metastasis at the time of diagnosis. Overall, 12 of the 49 patients (24.5%) had unfavorable disease (as defined by extraprostatic disease on pathologic specimen, relapse after radiotherapy, or bony metastasis at the time of diagnosis). CONCLUSIONS: These findings suggest that a microscopic focus of prostatic adenocarcinoma in a needle biopsy specimen, per se, does not predict the pathologic stage or the biologic behavior of a tumor.

Short-term treatment with novel ribonucleotide reductase inhibitors Trimidox and Didox reverses late-stage murine retrovirus-induced lymphoproliferative disease with less bone marrow toxicity than hydroxyurea.

We evaluated the ability of a short course of treatment with the ribonucleotide reductase (RR) inhibitor hydroxyurea (HU) and two novel RR inhibitors Trimidox (TX) and Didox (DX) to influence late-stage murine retrovirus-induced lymphoproliferative disease. LPBM5 murine leukaemia virus retrovirus-infected mice were treated daily with HU, TX or DX for 4 weeks, beginning 9 weeks post-infection, after development of immunodeficiency and lymphoproliferative disease. Drug effects on disease progression were determined by evaluating spleen weight and histology. Effects on haematopoiesis were determined by measuring peripheral blood indices (white blood cells and haematocrit) and assay of femur cellularity and femoral and splenic content of colony-forming units granulocyte-macrophage (CFU-GM) and burst-forming units-erythroid (BFU-E). HU, TX and DX partially reversed late-stage retrovirus-induced disease, resulting in spleen weights significantly below pre-treatment values. Spleen histology was also improved by RR inhibitor treatment (DX>TX>HU). However, as expected, HU was significantly myelosuppressive, inducing a reduction in peripheral indices associated with depletion of femoral CFU-GM and BFU-E. In contrast, although TX and DX were moderately myelosuppressive, both drugs were significantly better tolerated than HU. In summary, short-term treatment in late-stage murine retroviral disease with HU, TX or DX induced dramatic reversal of disease pathophysiology. However, the novel RR inhibitors TX and DX had more effective activity and significantly less bone marrow toxicity than HU.

Lymphangioleiomyomatosis.

Lymphangiomyomatosis (LAM) is a progressive disease afflicting women of childbearing age. LAM remains a rare disease, unfamiliar to many clinicians. It usually poses a diagnostic dilemma for the primary physician, possibly resulting in a delayed or missed diagnosis, unnecessary operative intervention, and inappropriate therapy. LAM is characterized microscopically by abnormal smooth muscle proliferation causing gradual obstruction of small airways, lymphatics, and vasculature. The proliferation eventually results in a characteristic clinical syndrome of progressive dyspnea, recurrent pneumothorax, chylous effusion, and hemoptysis. Despite a variety of treatment regimens developed since the first description of LAM, patient survival has not improved appreciably. Most patients die within 10 years of the time of diagnosis. This report presents a patient with LAM and a review of the literature.

Handling sentinel lymph node biopsy specimens.A work in progress.

This article reviews the "state of practice" with regard to sentinel lymph node biopsy, a new and evolving technique currently used most commonly for staging of malignant melanoma and adenocarcinoma of the breast. Sentinel lymph node biopsy has the potential to both increase the accuracy of lymph node sampling as a prognostic tool and to decrease the need for unnecessary and morbid extensive lymph node dissection in such patients. The need for close cooperation and planning involving the surgeon and pathologist is stressed, and gross room tissue handling, radiation safety, microscopic examination, and the use of ancillary diagnostic techniques are discussed.

Angiotrophic lymphoma. Presentation in uterine vessels with cytogenetic studies.

An unusual case of angiotrophic lymphoma diagnosed initially in the uterus is described. Immunohistochemical studies documented the intravascular tumor cells in the myometrium to be of B lymphocytic origin. The patient's bone marrow was hypercellular, and cytogenetic studies of the aspirate revealed a subpopulation of cells with multiple clonal chromosomal abnormalities. To our knowledge, this is the first case to document abnormal cytogenetic findings in this uncommon malignancy.

Adenocarcinoma of renal pelvis.

Adenocarcinoma accounts for a small percentage of neoplasms arising within the renal pelvis. We describe a mucinous adenocarcinoma of the renal pelvis that occurred in a 57-year-old woman. Investigation of the recent literature reveals an additional 12 cases of adenocarcinoma of the renal pelvis reported since 1980. These 13 cases are summarized in detail, for a total of 59 cases of adenocarcinoma of the renal pelvis documented in the English-language literature. These tumors can be subdivided into three major histologic types: tubulovillous, mucinous, and papillary non-intestinal. The tubulovillous and mucinous tumors are morphologically similar to intestinal tumors accounting for 71.5% and 21.5% of cases, respectively. They are believed to arise in foci of intestinal metaplasia. Only three cases (7%) were of the nonintestinal, nonmucinous, papillary subtype. These rare tumors are notable for their morphologic similarity to Bellini or collecting duct carcinoma, but a specific morphologic precursor has not been identified. Of the three subtypes, tumors of tubulovillous morphology confer the worst prognosis with 70% of patients dying within 5 years. Thirty-three percent of mucinous tumors and none of the papillary nonintestinal tumors were fatal.

Expression of HER-2/neu oncoprotein and epidermal growth factor receptor and prognosis in gastric carcinoma.

Fifty-six specimens of gastric carcinoma were examined for the localization of HER-2/neu oncoprotein (HER-2/neu) and epidermal growth factor receptor (EGFR) by immunohistochemistry using polyclonal antibodies on paraffin-embedded material. Strong membrane staining for HER-2/neu was noted in 14 cases (25%), all of which were of the intestinal type. Only cytoplasmic staining was found in an additional 21 cases (37.5%), including seven diffuse tumors. Twenty-four cases (nine diffuse and 15 intestinal) showed cytoplasmic staining with accentuation on the cell membrane for EGFR. Patchy staining was common for HER-2/neu, while EGFR immunoreactivity was always diffuse. Twenty cases (35.7%) showed positive staining for both, 15 cases (26.8%) for HER-2/neu only, four cases (7.1%) for EGFR only, and 17 cases (30.4%) for neither. Expression of HER-2/neu was more commonly associated with intermediate-grade and high-stage tumors. Cases with positive (either membrane or cytoplasmic) staining for HER-2/neu showed poorer overall mean survival (308 days) than cases that failed to stain (763 days). The EGFR-positive cases showed shorter mean survival (387 days) than the negative cases (547 days), but this difference did not reach statistical significance. The EGFR positivity did not further reduce survival in HER-2/neu-positive cases (362 days). The results of this study support the hypothesis that the expression of HER-2/neu may be a significant predictor of prognosis in patients with gastric carcinoma. Our findings also suggest that expression of these two closely related protooncogenes in malignant and benign gastric tissues is independent of each other and that EGFR does not potentiate the oncogenic effect of HER-2/neu.

Gleason histologic grading in prostatic carcinoma. Correlation of 18-gauge core biopsy with prostatectomy.

Histologic grading of adenocarcinoma of the prostate gland is a reliable predictor of extension and metastasis. Studies involving correlation of grade between biopsy and prostatectomy specimens have traditionally involved biopsies using a large-bore (14-gauge) cutting needle. However, common practice has shifted to the use of biopsy cores with a smaller caliber (18 gauge). This study was undertaken to determine the degree of correlation of tumor grade between 18-gauge core biopsy samples and excised glands. Sixty-seven patients with stage A or B adenocarcinoma of the prostate gland who had previously undergone 18-gauge core biopsy, who underwent radical prostatectomies, were studied. The Gleason score was determined by referred consensus among three pathologists. There was exact agreement between biopsy and excision in 39 cases (58%), whereas 24 cases (36%) differed by one digit. Three cases (4.5%) were undergraded, and one case (1.5%) was overgraded by two or more points. Only six tumors (8.9%) would have been incorrectly specified by the degree of differentiation. Discrepancies in grade of two points or more were not more frequent in cases with a small tumor volume (< or = 10%) in the biopsy specimens. We concluded that with careful histologic evaluation, the grade of tumor identified in these smaller biopsy cores correlates well with that seen at prostatectomy.

Skin contraction with pulsed CO2 and erbium:YAG laser.

The purpose of this study was to assess the physical response of skin to laser resurfacing in a real-time, quantitative fashion. The study was designed to assess skin contraction from two opposite standpoints. First, change in tension was measured during laser application while samples were held at constant length. Second, change in length of a sample under no tension was measured during laser treatment. These two disparate analyses represent the two possible extremes of the clinical situation in which skin exists under some tension with some laxity to allow for decrease in length. A custom apparatus with digital interface for skin tension measurements was used to produce single sample tracings of change in skin tension with laser treatment. Length change was measured for individual samples by continuous sonomicrometer readings. Individual sample data were then plotted in a time versus tension/length graph. Skin contracts immediately to a peak level and then relaxes to a sustained plateau level for both CO2 and erbium:YAG lasers. Increased contraction was noted when the beam penetrated into the dermis. Greater peak and plateau contraction is observed after the beam has penetrated into the dermis. Skin contraction varies directly with energy for CO2 and erbium:YAG laser. Findings were similar when skin tension was measured with the sample held at constant length and when length change was measured with the sample under no tension. Char left on the skin after a pass with CO2 laser substantially decreases skin contraction. High-density settings with CO2 laser yield pulse stacking, which effectively irradiates the same portion of tissue with char on it. Skin contraction varies inversely with computer pattern density settings for CO2 laser due to this pulse stacking effect. Density has little effect on skin contraction for the erbium:YAG laser because little char is generated. Histologic analysis identified a zone of coagulated dermis that correlates linearly with skin contraction.

Resolution of Ga-67 citrate uptake in the left neck mass of Hodgkin's disease and reversion of double scoliosis of cervical-thoracic and lower lumbar vertebrae.

A 6-yr-old boy underwent a total body Ga-67 citrate imaging study because of a large mass of Hodgkin's lymphoma in the left neck and the left anterior chest wall region. The images showed intense uptake in the left neck extending anteroinferiorly to the left upper chest wall corresponding to the left neck and chest region. In addition, there was mild cervical-upper thoracic scoliosis with convexity to the right and mild scoliosis of the lower lumbar scoliosis with concavity to the left. After three cycles of chemotherapy, in the follow-up Ga-67 citrate total body images seven months after his first Ga-67 citrate imaging, the intense uptake in the left neck and the left upper chest wall had been resolved and the scoliosis of the cervical-thoracic and lower lumbar spine had also been reversed to normal. This case shows that a Ga-67 citrate imaging study is useful for first diagnosis and subsequent monitoring of the therapeutic effects in a follow-up imaging. Also Ga-67 citrate imaging provided evidence that the scoliosis had been reversed.

c-Abl and Arg are activated in human primary melanomas, promote melanoma cell invasion via distinct pathways, and drive metastatic progression.

Despite 35 years of clinical trials, there is little improvement in 1-year survival rates for patients with metastatic melanoma, and the disease is essentially untreatable if not cured surgically. The paucity of chemotherapeutic agents that are effective for treating metastatic melanoma indicates a dire need to develop new therapies. Here, we found a previously unrecognized role for c-Abl and Arg in melanoma progression. We demonstrate that the kinase activities of c-Abl and Arg are elevated in primary melanomas (60%), in a subset of benign nevi (33%) and in some human melanoma cell lines. Using siRNA and pharmacological approaches, we show that c-Abl/Arg activation is functionally relevant because it is requiredfor melanoma cell proliferation, survival and invasion. Significantly, we identify the mechanism by which activated c-Abl promotes melanoma invasion by showing that it transcriptionally upregulates matrix metalloproteinase-1 (MMP-1), and using rescue approaches we demonstrate that c-Abl promotes invasion through a STAT3 → MMP-1 pathway. Additionally, we show that c-Abl and Arg are not merely redundant, as active Arg drives invasion in a STAT3-independent manner, and upregulates MMP-3 and MT1-MMP, in addition to MMP-1. Most importantly, c-Abl and Arg not only promote in vitro processes important for melanoma progression, but also promote metastasis in vivo, as inhibition of c-Abl/Arg kinase activity with the c-Abl/Arg inhibitor, nilotinib, dramatically inhibits metastasis in a mouse model. Taken together, these data identify c-Abl and Arg as critical, novel, drug targets in metastatic melanoma, and indicate that nilotinib may be useful in preventing metastasis in patients with melanomas harboring active c-Abl and Arg.