Molecular genetic evidence supporting the neoplastic nature of stromal cells in 'fibrosis' after chemotherapy for testicular germ cell tumours.

A residual retroperitoneal mass containing only fibrosis and necrosis is present in 40-52% of patients with advanced testicular germ cell tumours after chemotherapy. The biological nature and genetic characteristics of the stromal cells in these residual masses have not been adequately investigated. Laser-microdissected stromal cells from 27 patients who underwent retroperitoneal lymph node dissection after chemotherapy for metastatic testicular germ cell tumour were analysed. Allelic loss in the stromal cells of fibrosis was present at one or more of the ten microsatellite DNA loci examined in 23 (85%) of the cases. Chromosome arm 12p anomalies, the hallmark of germ cell neoplasia, were present in nine (33%) cases. The high frequency of allelic losses and chromosome arm 12p anomalies in the stromal cells from residual retroperitoneal fibrous masses after chemotherapy for testicular germ cell tumours suggests that the stromal cells are derived from the same tumour progenitor cells as the pre-existing metastatic germ cell tumour.

The S100 proteins for screening and prognostic grading of bladder cancer.

The S100 gene family, which is composed of at least 24 members carrying the Ca2+ binding EF-hand motif, has been implicated in both intracellular and extracellular functions, including enzyme activities, immune responses, cytoskeleton dynamics, Ca2+ homeostasis, cell growth and cell differentiation. Altered S100 protein levels are associated with a broad range of diseases, including cardiomyopathy, inflammatory and immune disorders, neurodegenerative disorders and cancer. Although the precise role of S100 protein in carcinogenesis is poorly understood, it seems that formation of homo- and hetero-dimers, binding of Ca2+ and interaction with effector molecules are essential for the development and progression of many cancers. Several studies have suggested that S100 proteins promote cancer progression and metastasis through cell survival and apoptosis pathways. In animal models of bladder cancer, several S100 proteins are differentially expressed in bladder tumors relative to normal urothelium. In human bladder cancer, overexpression of S100A4, S100A8 or S100A11 are associated with stage progression, invasion, metastasis and poor survival. This review summarizes these findings and evaluates their implications for human bladder cancer management.

Down-regulation of GATA-2 transcription during Pneumocystis carinii infection.

Differences in gene expression between Pneumocystis carinii-infected and noninfected rats were examined. Total RNA was isolated from homogenized rat lungs and then subjected to differential display with combinations of oligo(dT) and various arbitrary PCR primers. Approximately 50 differentially expressed bands were observed. Several of these DNA bands were isolated, reamplified, and cloned. The cloned DNA fragments were used as probes to perform Northern hybridization on RNA from P. carinii-infected and noninfected rat lungs. One clone was found to react with a 3-kb mRNA from noninfected but not from P. carinii-infected rat lung, suggesting that the gene represented by this clone was down-regulated during P. carinii infection. The nucleotide sequence of this clone was determined and found to be 97% homologous to the mouse GATA-2 transcription factor. In situ hybridization using RNA probes derived from this clone revealed that alveolar macrophages, resident lung monocytes, and bronchial epithelial cells express the GATA-2 gene in the lung.

Abdominal and pelvic needle aspiration biopsies: can we perform them well when using small needles?

BACKGROUND: The aim of this study was to compare abdominal fine-needle aspirations (FNAs) performed with large (>/=20-gauge) or small (

Preclinical in vivo testing of a rotational mechanical thrombolytic device.

PURPOSE: To establish the safety and efficacy of the Arrow Trerotola mechanical percutaneous thrombolytic device (PTD) for restoring patency of thrombosed hemodialysis grafts. MATERIALS AND METHODS: The hindlimb model of dialysis grafts was created in six dogs. Animals had either unilateral (n = 4) or bilateral (n = 2) polytetrafluoroethylene grafts, totaling eight grafts. Grafts were deliberately clotted 48 hours before thrombolysis. Thrombolysis was performed with five different versions of the PTD constructed of stainless steel (n = 12) or nitinol (n = 26) and rotated with use of a hand-held motor drive. After thrombolysis, fistulography was performed. RESULTS: Thirty-eight procedures were performed with the PTD, with 100% success. Thirty-day patency, evaluated in a subset of 15 procedures, was 100%. Complications included a single arterial embolus (2.6%) and eight device breakages (21%, all but two with the stainless steel version); none had any clinical consequences. A final modification of the nitinol device yielded 11 consecutive procedures without further breakage. No residual thrombus occurred in any procedure. Pathologic examination showed no significant injury to the vessels or neointima. CONCLUSION: The PTD is highly effective for mechanical thrombolysis in an animal model of clotted dialysis grafts. Based on this animal model, the device appears safe in its final modified form.

Pulmonary emboli from pulse-spray and mechanical thrombolysis: evaluation with an animal dialysis-graft model.

PURPOSE: To compare pulmonary emboli resulting from pulse-spray pharmacomechanical thrombolysis (PSPMT) and mechanical thrombolysis performed to declot dialysis-access grafts. MATERIALS AND METHODS: Polytetrafluoroethylene arteriovenous shunts were created in eight dogs and were deliberately clotted at monthly intervals. Animals were randomly assigned to treatment with pulse-spray urokinase thrombolysis or a low-speed rotational percutaneous thrombolytic device. Perfusion imaging, pulmonary-artery pressure measurements, and pulmonary arteriography were performed before and after each procedure. RESULTS: A total of 22 procedures were performed (11 PSPMT and 11 mechanical thrombolysis). Declotting was successful in all procedures, with 100% 30-day patency. Segmental defects were seen on perfusion images after 10 (91%) of 11 PSPMT procedures and two (18%) of 11 mechanical thrombolysis procedures (P < .002). Transient increases in pulmonary-artery pressure occurred in the PSPMT group. Complete resolution of emboli and return to baseline pressures were seen in all cases, even after multiple (up to four) procedures in the same animal. There was no histologic evidence of pulmonary infarction in either group. CONCLUSION: The percutaneous thrombolytic device is effective for declotting dialysis grafts in dogs and results in statistically significantly fewer pulmonary emboli compared with PSPMT.

Study of thrombus from thrombosed hemodialysis access grafts.

PURPOSE: To determine the volume and composition of clot within thrombosed hemodialysis access grafts. MATERIALS AND METHODS: Clots were collected in 22 patients at surgical thrombectomy of polytetrafluoroethylene grafts. Histologic analysis was performed in 10 of these clots plus 21 randomly selected clots from the pathology archives. RESULTS: A small, firm piece of whitish thrombus ("arterial plug") was almost always recovered from the arterial limb of the graft. This plug had a concave surface and ranged from 5 mm to 3 cm in length. The remaining clot was soft, red thrombus. The mean weight of all clots was 3.4 g, and mean volume was 3.2 cm3. Average graft length was 42 cm. Histologically, the arterial plug had a characteristic appearance of densely compacted alternating layers of erythrocytes and fibrin. CONCLUSION: Clot volume in thrombosed dialysis grafts is much less (approximately equal to 25%) than would be expected if the graft were completely filled with thrombus, a finding of significance to mechanical thrombolytic techniques. Resistance of the arterial plug to pulse-spray thrombolysis is likely due to compaction.

Aspiration biopsy cytology of smooth muscle tumors. A cytologic approach to the differentiation between leiomyosarcoma and leiomyoma.

From 1970 to 1991, 51 cases of smooth muscle tumors were diagnosed by fine needle aspiration biopsy. All were verified histologically and/or clinically, with no false-positive results. Of the 51 tumors, 41 were leiomyosarcoma and 10 leiomyoma. Among the 41 cases of leiomyosarcoma, 29 were the well-differentiated type; 8, poorly differentiated type; and 4, epithelioid type. Of the 10 cases of leiomyoma, 2 were the epithelioid type. The cytomorphologic features and cytologic patterns of various types of smooth muscle tumors observed in aspirate preparations are presented and compared in order to establish the cytologic criteria for differentiating malignant from benign smooth muscle tumors. Cytomorphologically the various types of smooth muscle tumors were different, and their cytologic features were sufficiently distinctive to distinguish one from the others. It appears possible to differentiate a well-differentiated leiomyosarcoma from a leiomyoma on the basis of cytologic findings observed in aspirate preparations. The recognition of different cytomorphologic features of various types of smooth muscle tumors is important in establishing an accurate cytologic diagnosis, which may be of practical significance to clinical management.

Comparative ultrastructure of needle aspiration biopsy and surgical resection specimens of lung tumors.

Ultrastructural examination affords conclusive evidence for classification of lung tumors. Tissue properly fixed for electron microscopy is not available in many cases, however. Ultrastructural diagnosis of resected specimens obviously follows, rather than directs, the surgical treatment. Fine-needle aspiration (FNA) of lung masses is recommended as a means to obtain lung tumor tissue for electron microscopy. Nevertheless, no comparison has been made between ultrastructural information gained from aspiration specimens and resected specimens. Electron microscopy was performed on transthoracic FNA specimens of 10 lung tumors for which surgical resection was subsequently performed. Glutaraldehyde-fixed specimens from FNA and surgical resection were prepared for electron microscopy according to routine procedures. Fixation of the FNA specimens was equivalent or superior to that of the resected specimens in 9 of the cases. Three of the FNA specimens contained necrotic as well as viable tissue. Features essential for diagnosis such as desmosomes, junctions, neurosecretory granules, intermediate filaments, glycogen, lipid, mucin, and microvilli were identifiable in both FNA and resected specimens. FNA specimens therefore yield a representative sample of the ultrastructural features of lung tumors when adequate cellular material is obtained. Use of a coaxial needle sampling technique with immediate microscopic assessment reduces the likelihood of retrieving only blood or necrotic tissue in the electron microscopy specimens.

Recent developments in hereditary nephritis (Alport's syndrome).

Hereditary nephritis with deafness, or Alport's syndrome, is a familial disorder characterized by progressive renal insufficiency, sensorineural hearing loss and ocular abnormalities. The Alport's nephropathy appears to result from a primary biochemical defect of the glomerular basement membrane, specifically an alteration of type IV collagen. The cardinal clinical manifestation of Alport's is chronic hematuria. End-stage renal disease develops in most affected males, while affected females generally experience a benign renal course. The diagnosis depends on characteristic electron microscopy findings of a variably thickened and thinned glomerular basement membrane with lamellation and basket weaving. Alport's is a genetically heterogeneous disorder with several modes of inheritance. A review of the literature with discussion of the clinical and basic science aspects of hereditary nephritis is presented.

Epiphysiodesis by electrocautery in the rabbit and dog.

Numerous experiments have demonstrated arrest of longitudinal bone growth by traumatizing the epiphyseal growth plate. Recently, promising studies have been performed using less invasive means to produce epiphysiodesis than the conventional surgical technique. The distal femoral physeal plate was cauterized in 20 rabbits and 17 dogs through an inserted needle and a standard operating room electrocautery machine. The result showed a progressive distal femoral length reduction without angular deformities in the experimental limb averaging 3 mm at two weeks to 11.6 mm at 20 weeks after the surgical procedure in the rabbit and 11.5 mm at 12 weeks in the dog. Histologic sections demonstrated thermal injury to the physis followed by a progressive narrowing and eventual resorption of the growth plate.

Unusual presentations of lymphoma of the head and neck in childhood.

Lymphoma of the head and neck in children can pose a significant diagnostic problem, especially when histologic analysis indicates non-Hodgkin's lymphoma and the initial site of involvement is extranodal. This report describes 15 pediatric cases of lymphoma seen from 1981 to 1987 with an initial presentation in the head and neck. Cervical lymph nodes represented the initial site of involvement in 10 of the cases. The other five cases presented with disease in the tonsillar fossa; maxillary sinus and mandible; parotid; pharyngeal wall; trachea and thyroid gland; and ethmoid sinus, sphenoid sinus, and anterior fossa. The histologic type was non-Hodgkin's lymphoma in 12 cases and Hodgkin's lymphoma in 3 cases. Our experience has shown that lymphoma of the head and neck in children presents a confusing clinical picture and was initially confused with inflammatory disease, polymorphic reticulosis, and other neoplasms such as rhabdomyosarcoma. In one patient, Epstein-Barr virus infection and an inherited immunodeficiency state probably played a role in the pathogenesis of the lymphoma.

High-frequency percussive ventilation compared with conventional mechanical ventilation.

In seven patients with severe respiratory distress, conventional mechanical ventilation and PEEP were used initially for respiratory support, which was changed to high-frequency percussive ventilation (HFPV) at the same level of airway pressure and FIO2. During both modes of ventilation, patients could breathe spontaneously via a low-threshold demand valve. With HFPV, PaO2 improved significantly (p less than .01) compared with PaO2 during conventional methods. Cardiac output was unaffected by the change to HFPV.

Suppression of murine immunologic functions by fresh and cultured human tumor extracts.

The role of human tumor-derived immunoregulatory factors (IRF) in the suppression of murine in vitro cell-mediated immune systems was investigated. IRF was extracted from a fresh human colon carcinoma and a liposarcoma using 3 M KCl. These extracts have previously been shown to suppress in vitro human immune responses. Both IRF extracts inhibited PHA-stimulated murine splenocyte [3H]Tdr uptake in a dose-dependent manner while extracts of normal tissue were not inhibitory. To further investigate in vitro immunosuppression a (C57BL/6 X A/J) F1 anti-B10. BR mixed lymphocyte reaction (MLR) was developed. Optimal [3H]Tdr incorporation was on Day 4 with 1 X 10(5) responders and 2 X 10(5) irradiated stimulators. Addition of IRF caused a 56% inhibition of this response but did not alter the kinetics of the MLR response. Induction of cell-mediated cytotoxicity (C57BL/6 X A/J F1 vs B10.D2) was significantly inhibited by addition of IRF during in vitro sensitization. Release of 51Cr from P-815 targets was decreased to spontaneous release levels at an effector:target (E:T) ratio of 20:1 when IRF was present during sensitization. At this E:T ratio, cells sensitized in the presence of a normal muscle 3 M KCl extract or medium caused 71 and 60% 51Cr release, respectively. IRF activity could also reproducibly be extracted from two small cell lung carcinoma tissue culture lines grown under a variety of culture conditions or passaged in nu/nu mice. The biochemical characteristics of the factor inhibiting human and murine lymphoid cell proliferation were identical. Thus, this system provides a convenient model for assessing the activity of human tumor-derived immunoregulatory factors.

Increasing phantom limb pain as a symptom of cancer recurrence.

Previous studies have shown that phantom limb pain following major amputation reaches its greatest severity 2 to 3 weeks following amputation; it then gradually diminishes over subsequent months and years. Transient episodes of severe phantom limb pain are sometimes temporally related to specific activities such as urination, sexual intercourse, or local pressure applied to the amputation stump. Also, neuroma formation may be associated with transient episodes of increased discomfort usually associated with the application of local pressure. Phantom limb pain progressively increasing over an extended period of time has not been previously reported. The authors observed phantom limb pain of increasing severity to be associated with locally recurrent extremity sarcoma in two patients. In both patients increasing phantom limb pain was the first indication of recurrent cancer and led to radiologic studies and biopsy which confirmed the diagnosis of recurrent disease. It is suggested that phantom limb pain of progressive increasing severity may be a symptom of locally recurrent cancer in an amputation stump.

Tumor stasis factor (TSF): a possible mechanism for the regulation of tumor cell proliferation.

A new class of factors that regulates tumor cell division in vitro can be isolated from fresh and cultured tumor cells by 3 M KCl extraction. Tumor stasis factors (TSF) inhibiting cultured tumor cell proliferation were extracted from 8 of 11 fresh human tumors and 2 cultured tumor cell lines. TSF inhibited [3H]Tdr incorporation by allogeneic and autologous cultured tumor cells in a dose-dependent manner. Extracts of normal human tissues and benign tumors did not demonstrate inhibition with the exception of liver. The mechanism of inhibition was cytostatic and not cytotoxic as demonstrated with trypan blue exclusion by tumor cells following TSF treatment, maintenance of intact tumor cell monolayers following addition of TSF, and lack of inhibition of Con A-mediated lymphocyte proliferation by TSF. TSF activity could be reversed by washing for up to 48 hr of incubation and was resistant to heat, pH alterations, reducing agents, proteases, and glycosidases. However, the active moiety bound to lentil lectin and could be purified 80-fold by preparative isoelectric focusing. These factors may represent a novel regulatory mechanism for tumor cell proliferation.

Syngeneic humoral immune responses to tumor-associated antigens expressed by K-1735 UV-induced melanoma and its metastases.

An enzyme-linked immunoassay (ELISA) was developed to study syngeneic humoral immune response to a primary tumor and its metastases in the K-1735 ultraviolet light (UV)-induced C3H murine melanoma system. Binding of sera from syngeneic animals previously immunized with primary tumor or metastatic tumor tissue (M-3, M-4) to corresponding 3 M KCl extracts of tumor was significantly greater than binding of control C3H mouse serum. Antibody binding was not significantly reduced by competitive binding with syngeneic murine muscle or liver extracts, indicating the presence of tumor antigen(s) not shared by normal murine tissue. Antibodies to the tumor-associated antigens were selectively removed by competitive binding with syngeneic K-1735 tumor extracts but not by the unrelated 102 murine sarcoma from C57BL/6. However, EL-4 extracts (C57BL/6) did inhibit antibody binding to the primary and both metastases. Further competitive binding studies demonstrated the presence of a common antigen(s) present on the primary tumor and both metastases. We conclude that the K-1735 UV-induced melanoma primary tumor and its metastases express serologically detectable shared antigenic determinate.