Excised Tumor Tissue Research Articles

A comparative evaluation of a RARE-based single-shot pulse sequence for diffusion-weighted MRI of musculoskeletal soft-tissue tumors

The purpose of this study was to evaluate the feasibility of a centric-reordered modified rapid acquisition with relaxation enhancement (mRARE) sequence for single-shot diffusion-weighted magnetic resonance imaging (DWI) of soft-tissue tumors in the musculoskeletal system. In the evaluation of this sequence, DWI was performed in a liquid phantom, in excised human tumor samples embedded in bovine muscle, and in nine patients suffering from different types of soft-tissue tumors. The measurements were compared to DWI using a spin-echo sequence and a single-shot echo planar imaging (EPI) sequence. The phantom measurements in water and dimethyl sulfoxide showed a difference of less than 5% when comparing the apparent diffusion coefficients (ADCs) determined by the mRARE sequence and the two other techniques. Comparing mRARE and EPI, the differences in the ADCs were about 10% in the excised tumor tissue and, typically, about 15% in vivo. ADCs between 0.8 x 10(-3) mm2/s and 1.4 x 10(-3) mm2/s, depending on the tumor type, were found in solid tumor tissue; in cystic tumor areas, ADCs greater than 2.0 x 10(-3) mm2/s were determined with the mRARE and the EPI sequences. Diffusion-weighted images of the mRARE sequence were less distorted than those acquired with the single-shot EPI sequence, and provided more anatomic information, since the muscle and fat signals were considerably higher.

Vitamin D3 administration induces nuclear p27 accumulation, restores differentiation, and reduces tumor burden in a mouse model of metastatic follicular thyroid cancer.

We have previously demonstrated in vitro that 1alpha,25-dihydroxyvitamin D3 (calcitriol) treatment increases p27 expression and decreases cell proliferation in cultured thyroid carcinoma cell lines. We hypothesized that in vivo treatment with calcitriol would have a beneficial effect on thyroid carcinoma growth and progression. Five x 10(6) WRO (human thyroid follicular carcinoma derived) cells were implanted in the neck in 4- to 5-wk-old female SCID mice in an orthotopic xenograft model. Animals (n = 15) were treated i.p. three times a week for 21 d with 0.75 microg/kg calcitriol or vehicle. Mice were killed 21 d after tumor implantation, tumor volume was measured, and excised tumor tissue was examined by light microscopy and immunohistochemistry for p27 and thyroglobulin reactivity. Average tumor volume in control mice after 21 d of vehicle treatment was 2002 +/- 207 mm3 compared with a mean tumor volume of 1241 +/- 115 mm3 in animals receiving calcitriol, reflecting a 38% reduction in tumor volume size (P < 0.003). Tumors from vehicle-treated animals demonstrated morphological features of epithelial malignancies with characteristics of insular carcinoma and multiple metastases to the lungs. Tumors excised from calcitriol-treated animals demonstrated signs of differentiation with restoration of thyroglobulin staining. This was associated with a marked accumulation of p27 immunoreactivity in the nuclear compartment. These studies demonstrate that in vivo calcitriol administration can effectively restore p27 accumulation in thyroid carcinoma cells, an effect associated with appreciably enhanced cellular differentiation, reduction in tumor burden, and prevention of metastatic growth.

Simultaneous acquisition of multiple orders of intermolecular multiple-quantum coherence images in vivo

Until recently, NMR imaging with intermolecular multiple-quantum coherences (iMQCs) has been based on the acquisition of a single echo. In vivo studies of iMQC image contrast would greatly benefit from a method that could acquire several orders of quantum coherence during the same acquisition. This would enable comparison of the image contrast for various orders and eliminate image coregistration problems between scans. It has previously been demonstrated that multiple orders of iMQC images can be simultaneously acquired of a simple phantom. Here, we examine the technique and its effect on biological tissue, both in vivo and in vitro. First, we establish the effectiveness of the iMQC sequence in vivo using earthworms as specimens. We then further show that the multi-CRAZED sequence enhances detection of next generation (nanoparticle) contrast agents on excised tumor tissue.

Enhancement of antitumor effects of 5-fluorouracil combined with ultrasound on Ehrlich ascites tumor in vivo

To evaluate the role of ultrasonic waves in enhancing the effect of 5-fluorouracil (5-FU) as anticancer drug, tumor growth, cell ultrastructure and temperature rises of ascites Ehrlich tumor implanted in mice were investigated at a frequency of 0.8 MHz. Different conditions of ultrasonic intensity (1, 2 and 3 W/cm 2), sonocation time (1, 3 and 5 min) and waveform (continuous and pulsed) were studied. A total of 10 mg/kg body weight of 5-FU was IV injected into the mice bearing Ehrlich tumors on days of 1, 3, 5, 8, 10 and 12 of therapy. After 24 h of each injection with 5-FU, tumor sites were sonocated at room temperature of 22°C. Tumor volumes were monitored by using ultrasonic imaging system during treatment just before each 5-FU injection, and on the 15th day of treatment. Density and ultrasonic attenuation of excised tumor tissues were measured in vitro and used to estimate the temperature rises due to ultrasonic absorption. Density and attenuation coefficients of excised tumor tissues were found to be dependent on the treatment regimen. The estimated rate of temperature rise and equilibrium temperature, and the characteristic time to reach equilibrium are given for each group. Results obtained indicate that tumor growth decreases with increasing of ultrasonic intensity and sonocation time. Tumor growth was delayed 4 to 6 days by combined treatment of 5- fluorouracil and ultrasound (US). Ultrastructure investigations of tumor cells showed severe damage in cytoplasmic organelles and cytoplasmic vacuoles that increased with increasing ultrasonic intensity and sonocation time. This damage appears as prominent crowded vacuoles among swollen ruptured organelles, chromatin fragments and severe increase in numbers of pyknotic and apoptotic cells. (E-mail: moustafamm@yahoo.com)

Enhanced cell death in NR-S1 tumor by photodynamic therapy: possible involvement of Fas and Fas ligand system.

To investigate the effect of photodynamic therapy (PDT) on cell death in malignant tumor tissue, the frequency of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive cells and the possible involvement of Fas and Fas ligand system were evaluated. NR-S1 tumor-bearing C3H/HeNCrj mice were treated by PDT with Photofrin(R) (12 mg/kg body weight) and Nd:YAG dye laser (630 nm, 10 Hz, 150 J/cm(2)). Paraffin-embedded tissue sections from the excised tumor tissues at 6, 12, 24, 48 hours after PDT were analyzed by TUNEL for the occurrence of apoptosis and by immunohistochemistry for Fas and Fas ligand (FasL) expression. TUNEL-positive cells as well as Fas- or FasL-positive cells were counted and expressed as a percentage of positive cells per total cells. Based on the percent area of tumor necrotic foci, the most effective conditions for PDT were first determined. Under these conditions, PDT increased the number of TUNEL-positive tumor cells at 12 hours after irradiation. In parallel with the increase in TUNEL-positive cells, Fas-positive tumor cells were also found in the same area where many TUNEL-positive tumor cells were found. The expression of Fas ligand was found in the tumor cells surrounding TUNEL-positive cells on serial sections. A significant increase in FasL-positive lymphocytes was observed at 12 hours, whereas the infiltration of such lymphocytes into the area where TUNEL-positive tumor cells were observed was rare. The possible role of Fas/FasL system in the cell death induced by PDT with Photofrin(R) and Nd:YAG dye laser was suggested. Moreover, the role of infiltrated lymphocytes seemed not to be so much in this model.

マウスへん平上皮癌に対するｉｎｔｅｒｌｅｕｋｉｎ‐２，ｉｎｔｅｒｆｅｒｏｎ‐βと抗癌剤併用投与による抗腫よう効果に関する研究

We studied the in vivo antitumor effects of interleukin-2 (IL-2) and interferon-β(IFN-β) in combination with chemotherapy on murine syngeneic transplantable squamous cell carcinoma, NR-S 1.Tumors were inoculated subcutaneously by injecting NR-S 1 cells into the back of 5-weekold female C 3 H/He mice. The tumor-bearing mice were divided into eight groups, and the following agents and anticancer drugs were administered:(1) control (saline), (2) IL-2+IFN-β, (3) bleomycin (BLM), (4) cyclophosphamide (CY), (5) cisplatinum (CDDP), (6) BLM+IL-2+IFN-β, (7) CY+IL-2+IFN-β, and (8) CDDP+IL-2+IFN-β.Intratumoral injections of saline or IL-2 +IFN-β and intraperitoneal injections of anticancer drugs were continued daily for 14 days. Treatment effects were estimated by measuring the tumor diameter and calculating the survival rate. Fourteen days after treatment, cytotoxic activities (against YAC-1, P815 and NR-S 1 cells) and their splenocyte phenotypes (Thy1.2, L 3 T 4, Lyt-2, and asialo-GM 1) were determined. The excised tumor tissues were also examined histologically.Although tumor size decreased slightly in the IL-2+IFN-β group, the reductions in size were significantly greater in each of the chemotherapy groups (p<0.05). Furthermore, the combination of chemotherapy and IL-2+IFN-β was much more effective in tumor reduction than the corresponding chemotherapy alone. Survival rate was significantly longer in all experimental groups except for the IL-2+IFN-β group. Tumors disappeared completely in two mice in the BLM+IL-2+IFN-β treatment group, which survived for more than 100 days. NK activity was significantly enhanced in mice treated with CDDP, BLM, or CDDP+IL-2+IFN-β. Although low LAK activity and cytotoxic activity against syngeneic NR-S 1 tumor cells were observed in the IL-2+IFN-β grove or the chemotherapy alone group, both of these activities were enhanced when these treatments were combined. Histological examinations disclosed remarkable infiltration of mononuclear cells and tumor necroses in the chemotherapy +IL-2+IFN-β groups. The infiltrated mononuclear cells predominantly expressed Mac-1 or asialo-GM 1 phenotype.These results suggest that the combination of systemic chemotherapy with local administration of IL-2+IFN-β is feasible for cancer therapy. Furthermore, NK cells may play an important role in both antitumor effect and the prolongation of survival.

Adenomyoepithelioma of the Breast. A Case Report

Adenomyoepithelioma is a proliferative disorder of both epithelial and myoepithelial cells. This lesion may be found in salivary glands, skin appendages and, very rarely, in the mammary gland. Adenomyoepithelioma was first described in 1970 and very few cases have so far been reviewed in the literature. This paper reports the clinical, histological and immunohistochemical characteristics of an adenomyoepithelioma in a 24 year old woman; to our knowledge this is the first published case in such a young patient. The clinical feature suggested a fibroadenoma. A more complete study of the excised tumor tissue by immunohistochemical and ultrastructural analysis proved that the correct diagnosis was adenomyoepithelioma. Whether adenomyoepithelioma is a benign or a low-grade malignant lesion is still controversial and, therefore, the therapeutic approach is not well defined.

腺様嚢胞癌の増殖様式

Eleven cases of adenoid cystic carcinomas arising from the minor salivary gland were surgically excised at our department between 1978 and 1987. The histologic examination revealed a tumor negative margin in 5 cases and a positive margin in 6 cases.The 5 tumors of negative margin were located in the tongue (T1aN0M0), palate (T1a N0N/10), oral floor (T2bN2bN.10), upper alveolar bone (TlaN0M0), and cheek (T3bN0N10). This study was attempted to determine the extent of tumor invasion by using serial sections prepared from the excised tumor tissues of negative margin. The results showed extensive and insidious infiltration of tumor cells into adjacent soft tissues, especially involving the submucocutaneous connective tissues and sheaths of muscle, fat and nerves. The degree of peripheral infiltration appeared to elevate with the increased clinical tumor size. The spread of tumor juged by microscopy was 1. 5 to 3. 5 times of the macroscopical size.These findings suggest that more wide excision and combination with radio-and/or chemotherapy are required for a management of this tumor.

Immunohistochemical and immunoelectron microscopic study of an amylase-producing, CA19-9 positive ovarian mucinous cystadenocarcinoma

A 75-year-old woman with a right ovarian tumor revealed high levels of serum amylase and CA19-9 which decreased to within normal limits after the operation. A histopathological study of surgically excised tumor tissue revealed a mucinous cystadenocarcinoma. The tumor was composed of three elements: adenoma, adenoma with low potential malignancy, and adenocarcinoma. Using the light microscopic indirect immunoperoxidase technique for amylase and CEA, and the Avidin-Biotin affinity technique for CA19-9 and CA12-5, the amylase and CA19-9 were stained in the cytoplasm of the adenoma and adenocarcinoma although CEA was stained only in the cytoplasm of the adenocarcinoma. An ultrastructural study using the immunoperoxidase method revealed that CA19-9 was positive in the apical portion of the tumor cells and amylase was positive in the entire secretory vesicles of the tumor cells. Furthermore, ciliated tumor cells derived from fallopian tube epithelium were not observed in the light and electron microscopic specimens.

Chain-fluorinated polyamines as tumour markers. III. Determination of geminal difluoropolyamines and their precursor 2,2-difluoroputrescine in normal tissues and experimental tumours by in vitro and in vivo 19F NMR spectroscopy.

Treatment of laboratory animals bearing transplantable tumours with alpha-difluoromethylornithine, an inhibitor of ornithine decarboxylase, followed by 2,2-difluoroputrescine allows the partial replacement of the endogenous polyamines spermidine and spermine by their fluorine-containing analogs 6,6-difluorospermidine and 6,6-difluorospermine. Quantitative determination of the chain-fluorinated polyamines in excised tumor tissue by in vitro 19F NMR spectroscopy at 11.7 T gave results similar to those obtained by extraction and HPLC analysis. The NMR methods allowed the observation of several as yet unidentified geminal difluoropolyamine metabolites with a detection threshold as low as 1 nmole/g. Using a surface coil at 2.4 T, it was possible to detect in 10-20 min fluorinated polyamines in tumour, liver and bladder (urine) of mice in vivo at concentrations of the order of 50-100 nmol/g, i.e., when 2-5% of the natural polyamines have been replaced by their fluoro analogs. Since 6,6-difluorospermidine and 6,6-difluorospermine preferentially accumulate in tumours and specific tissues with enhanced polyamine metabolism, it is expected that these compounds may serve as probes for the application of 19F NMR methods for tumour diagnosis and therapy control. Since significant quantities of additional fluorine-containing polyamine metabolites were detected in some tissues, 19F NMR spectroscopy will contribute to our knowledge of polyamine metabolism in normal and tumour tissue.

Use of a monoclonal antibody (B72.3) as a novel immunohistochemical adjunct for the diagnosis of carcinomas in fine needle aspiration biopsy specimens

Monoclonal antibody B72.3 has been shown to be reactive with a high-molecular-weight glycoprotein complex termed TAG(tumor-associated glycoprotein)-72. By the avidin-biotin immunoperoxidase method, fine needle aspirates and corresponding surgically excised tumor tissues from both malignant and benign tissues were analyzed for TAG-72 expression. Staining (range, 1 to 100 per cent of tumor cells) with monoclonal antibody B72.3 was observed in needle aspirates from 18 of 18 adenocarcinomas and adenosquamous carcinomas of the lung, 17 of 21 adenocarcinomas of the breast, and six of six adenocarcinomas of the colon, as well as adenocarcinomas from other body sites. In contrast, small cell carcinomas of the lung, malignant melanomas, lymphomas, and sarcomas did not stain with the antibody. Benign lesions from the breast, lung, pancreas, parotid, and thyroid also failed to stain. In 66 patients, tumor-bearing tissue had also been resected and was available for comparative examination with monoclonal antibody B72.3. In 62 of these 66 patients, the staining patterns in the aspirates were found to be predictive of the patterns of antibody reactivity in the comparable surgically resected tissues. From these studies it is concluded that monoclonal antibody B72.3 defines a tumor-associated antigen that is expressed in neoplastic cells but not in benign cells and is most selectively expressed in adenocarcinomas. This monoclonal antibody may be used as a novel adjunct for the diagnosis of carcinoma in fine needle aspiration biopsy specimens.

Intracranial Embryonal Carcinoma mixed with Endodermal Sinus Tumor and Teratomatous Elements in the Anterior Third Ventricle

A 16-year-old male patient was diagnosed as having an intracranial germ cell tumor in the anterior third ventricle. Surgery revealed that the tumor was composed of embryonal carcinoma, endodermal sinus tumor, and teratomatous elements, such as cartilage, myogenic tissue, and keratinizing squamous epithelium. Alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG) were elevated in the serum preoperatively and demonstrated also in the excised tumor tissue by the immunohistochemical method. After postoperative radiation therapy, the serum AFP and HCG returned to normal limits. About 1 year after the first operation, a recurrence of the tumor, with marked elevation of serum AFP, was detected. A second operation was performed, and the histological examination showed mostly endodermal sinus tumor with irradiation effect and teratomatous elements. The embryonal carcinoma portion had almost disappeared as a result of the earlier radiation therapy. Changes in the tumor composition correlated with changes in serum tumor markers. The patient is being followed closely.

Clinical study on pituitary adenoma with hyperprolactinemia (author's transl)

Clinical symptoms, endocrinological findings, neuroradiological findings, and the results of treatments were studied in 24 patients with pituitary adenoma with hyperprolactinemia (over 30 ng/ml). The excised tumor tissues were examined light-microscopically and electron-microscopically, and hormones in the tumor culture medium were measured. The subjects in this study were 19 female and 5 male patients. All of the female patients had had amenorrhea and 14 of them had shown galactorrhea. Nine of the 24 patients had had visual disturbance and 2 had had diplopia or blephaloptosis. These 24 patients were classified into 3 groups according to the prolactin (PRL) values and the neuroradiological findings. The first group consisted of 4 patients who were rather aged (mean age, 44.5 years) and had low PRL values ranging from 30 to 112 ng/ml. The tumor showed suprasellar extension, and all the 4 patients had had visual disturbance. Electron microscopy and immunological staining did not yield confirmation of PRL-productive pituitary adenoma. The second group consisted of 12 patients who had high PRL values ranging from 116 to 1, 040 ng/ml. The tumor was intrasellarly localized. PRL value returned to normal after surgery in 18% of the patients. Menstruation started again in 27% of them. After administration of CB154 the PRL value was restored to normal in 75% of them, and menstruation started again in all of the 12 patients. The third group consisted of the remaining 8 patients where the tumor showed parasellar extension. Sufficient effects could not be obtained by surgery alone. Even after administration of CB154, the PRL value was restored to normal in only 2 of the 8 patients, and menstruation and pregnancy were observed in only 1 of the 4 female patients. Thus, patients who belong to the second and third groups with PRL-productive pituitary adenoma should be treated as follows: In patients with PRL-productive intrasellarly localized pituitary adenoma, the tumor should be totally excised. In patients with PRL-productive pituitary adenoma which invades the cavernous sinus and so cannot be totally excised, CB154 should be administered after surgery.

Feline sarcoma virus induced in vitro progression from premalignant to neoplastic transformation of human diploid cells

Human diploid cells morphologically transformed by feline sarcoma virus were serially propagated under selective cell culture conditions. When injected into nude mice prior to passage in soft agar (0.35%), morphologically transformed cells did not produce tumors. However, when propagated under selective cell culture conditions, transformed cells grew in soft agar and, when injected subcutaneously into the subcapsular region of the n mu/n mu mice, produced neoplastic nodules histopathologically interpreted as fibromas. Karyological examination of cell populations grown out from the tumors confirmed that the tumors were composed of human cells. Examination of electron micrographs of the excised tumor tissue revealed the presence of budding virus particles. Tumor cells isolated from nude mice and morphologically transformed cells both contained the feline oncornavirus-associatied cell membrane antigen. It was concluded that expression of feline oncornavirus-associated cell membrane antigen is associated with an early stage of feline retrovirus-induced carcinogenesis, namely focus formation. In addition, it was shown that FeLV-FeSV can induce morphological transformation in human cells in vitro and that there is a requirement for the cells to passage through soft agar before subsequent tumor formation (neoplastic transformation) can be demonstrated.

Estrogen Receptors in Human Cancers

In the seven years since the first clinical correlations were reported from our laboratory, the utility of estrogen receptor assays on excised tumor tissue has become well established as an aid in deciding which patients with advanced breast cancer should be treated by endocrine ablation (oophorectomy, adrenalectomy, hypophysectomy) or by hormone administration, and which should be placed directly on chemotherapy. The experience of many investigators 1 has confirmed the original conclusion that patients whose tumors contain no detectable estrogen receptor protein (estrophilin) have little chance of benefit from endocrine therapy, whereas most, but not all, women with receptor-containing breast cancers will respond favorably to endocrine manipulation. Recent evidence, 2 though still limited, indicates that estrophilin analyses of stage I and stage II primary tumors can be used to predict response to endocrine therapy when disseminated disease appears at a later time. These observations, if confirmed, suggest the value of routinely

Diffuse Malignant Mesothelioma

Thirteen cases of diffuse malignant mesothelioma, 12 pleural and one peritoneal, were examined and reviewed from the clinical and morphologic points of view. The most frequent symptom and sign were pain and effusion. Periods of survival varied, with one patient surviving 11 years despite morphologic features of the pleural tumor. The diagnosis can be made only on the basis of excised tumor tissue. The best control of the tumor is attained by repeated surgical removal. Neither radiation nor chemotherapy altered significantly the growth of the tumors.

Local Excision of Choroidal Malignant Melanoma

Histopathological analysis of a malignant melanoma locally excised by fullthickness eye wall resection revealed the margins and scleral emissaria to be completely free of tumor. This surgical technique, if applied to tumors that are of appropriate size and location, appears promising as a means of retaining vision and cosmesis in the involved eye. Furthermore, histopathologic examination of the excised tumor tissue is possible. This latter feature is an important advantage of this technique over other forms of local therapy of choroidal malignant melanoma.

Rapid method for the isolation of plasma membranes from a murine fibrosarcoma using an aqueous two-phase polymer system.

Abstract An aqueous two-phase polymer system was used to isolate plasma membranes from a palpable mouse fibrosarcoma. The excised tumor tissue was washed with sterile saline and pushed through nylon screens of decreasing mesh size. This cell suspension was placed in Tris-buffered, isotonic sucrose plus MgSo4 and homogenized by nitrogen cavitation. A pellet was collected from the homogenate by low-speed centrifugation and was added to the aqueous two-phase polymer system. After several brief, low-speed centrifugations, the interfacial material between the polymer phases was collected. Data from enzyme and biochemical assays demonstrated that this fraction was plasma membrane. This method provided a high yield of the surface membrane in less than three hours.

Filamentous particles in human alveolar cell carcinomas: electron microscopy studies of six cases (preliminary report).

Over a 10-month period we observed 6 cases of human alveolar cell carcinoma, a relatively uncommon human tumor. Electron microscopy study of freshly excised tumor tissue revealed intracytoplasmic vacuoles with dense osmophilic inclusions containing filaments in 4 cases. The filamentous structures were about 90 nm in diameter, with a central core about 15 nm in diameter. Spherical bodies about 90–130 nm in diameter in the vacuolar space had a double membrane, and electrondense particles randomly dispersed in their matrices.

Glomus Jugulare Tumors: I. Historical Background

ON APRIL 18, 1942, at The Mount Sinai Hospital, in New York city, I operated upon a patient with a bleeding mass which arose in the middle ear and protruded from his left ear canal. The excised tumor tissue was identical in every characteristic with proven benign carotid body tumors^1,2removed from the typical site in the neck. The presence of a carotid body-like tumor in the middle ear and mastoid bone without any clinically demonstrable tumor in the neck had never hitherto been described, hence my reluctance to report such a finding. When I found Dr. Stacy Guild's³brief communication in the Anatomical Record, describing "A Hitherto Unrecognized Structure, The Glomus Jugulare In Man," I felt that one could postulate that the tumor I had removed could have developed from the glomus jugulare structure described by Guild. It was three years later, in 1945, when my original