Serological analysis of SEREX-defined medullary breast carcinoma-associated antigens

Medullary breast carcinoma (MBC) despite anaplastic features and high grade has a good prognosis that can be related to prominent lymphocytic infiltration. We analyzed the frequency of antibody response toward 41 SEREX (serological recombinant expression cloning)-defined MBC antigens in sera of allogeneic MBC patients using serological plaque-spot assay and examined the mRNA expression profile of some antigens in MBC tissues. This preliminary study allowed us to select 18 autoantigens as potential MBC-associated antigens. Further studies in large cohorts of breast cancer patients will be performed for their evaluation as targets for cancer diagnostics or therapy.     

Paget's disease of the nipple. Immunohistochemical localization of milk fat globule membrane antigens

The authors have used the indirect immunoperoxidase technique to examine the presence and distribution of milk fat globule membrane antigens in 12 cases of mammary Paget's disease using two monoclonal antibodies, HMFG-1 and HMFG-2. These stain breast epithelial cells but do not stain normal epidermis. The Paget's cells showed a similar pattern of cytoplasmic staining to that seen in the underlying intraduct or invasive carcinoma, therefore confirming them to be malignant ductal cells. To support this one of the cases was stained with the antibody LE 61 which is specific for nonepidermal epithelial cytokeratins. The result was strongly positive in Paget's cells in the epidermis but not in squamous cells.     

Immunohistochemical expression and localization of somatostatin receptor subtypes in androgen ablated prostate cancer

Objective

The aim was to examine the expression and localization of the five somatostatin receptors (termed SSTR1 to 5) in radical prostatectomies (RPs) from patients with prostatic adenocarcinoma (PCa) under complete androgen ablation (CAA) before operation.

Material

The five SSTRs were evaluated in the epithelial, smooth muscle and endothelial cells of normal-looking epithelium (Nep), high-grade prostatic intraepithelial neoplasia (HGPIN) and PCa in 20 RPs with clinically detected PCa from patients under CAA. Twenty RPs with clinically detected PCa from hormonally untreated patients were used as control group.

Results

Concerning the secretory cells (i) Membrane staining was seen for SSTR3 and SSTR4; the mean percentages of positive cells, higher in SSTR3 than in SSTR4, decreased sharply in HGPIN and PCa compared with Nep; the mean percentages in the androgen ablated group were 30% to 90% lower than in the untreated; (ii) Cytoplasmic staining was seen for all five SSTRs; the mean percentages of positive cells in Nep, HGPIN and PCa of the untreated group were similar, and in general as high as 80% or more; in the treated group, the Nep values were similar to those in the untreated, whereas the values in HGPIN and PCa were lower for SSTR1, three and five, with a decrease of 30% for SSTR1; (iii) Nuclear staining was seen with SSTR4 and SSTR5, the mean percentages for the former being much lower than for the latter; treatment affected both HGPIN and PCa, whose proportions of stained cells were 30% to 55% lower than in the untreated group. Cytoplasmic staining in the basal cells was seen for all five SSTRs, both in Nep and HGPIN. The values in the treated group were lower than in the other, the difference between the two group being in general comprised between 10% and 40%. Treatment did not affect SSTR staining in the smooth muscle and endothelial cells.

Conclusions

The present study expands our knowledge on the expression and localization of the five SSTRs in the prostate following CAA.     

应用血清蛋白质组分析筛选人胃癌相关抗原

背景与目的:寻找胃癌特异性或相对特异性标志物是胃癌研究亟待解决的课题.本研究利用血清蛋白质组分析方法,建立人胃癌组织蛋白质双向凝胶电泳图谱以及分别与胃癌患者血清和非癌人群血清的免疫印迹图谱,初步筛选人胃癌相关抗原.方法:运用2-DE分离人胃癌组织的总蛋白质;一个样品跑3块凝胶,1块考马斯亮染显色作为平行胶,另2块分别以胃癌患者自身血清和非癌人群血清作一抗进行免疫印迹分析,获得免疫印迹图谱;对图谱进行比较分析,识别差异反应的蛋白质点,根据差异反应蛋白质点位置在平行胶上找出匹配的蛋白质点;对差异反应的蛋白质点应用MALDI-TOF-MS进行质谱分析鉴定,获得相应的肽质指纹图谱,通过数据库搜索鉴定出差异反应的蛋白质.结果:人胃癌组织2-DE分离后分别与胃癌患者自身血清和非癌人群血清进行免疫印迹,获得分辨率较高的反应图谱;图像分析找出差异反应的蛋白质点14个,质谱鉴定获得hnRNP K、hnRNP H1、hnRNP H2、CK18、Fkbp52与Hsp90复合物、DBP、SBP1、eEF-1、ACTB、CKB、PSMC3、DDAH-1、NADH脱氢酶硫-铁蛋白1等13个蛋白质.结论:获得分辨事较高的人胃癌组织蛋白质组与胃癌患者自身血清及非癌人群血清反应的免疫印迹反应图谱,初步筛选了13个人胃癌相关抗原.为进一步筛选可应用于临床早期诊断、治疗、预后评估及疾病检测的人胃癌分子标志物奠定了基础.     

Canine perianal gland carcinoma-associated antigens defined by monoclonal antibodies

This study was conducted to distinguish canine perianal gland carcinomas from adenomas using monoclonal antibodies (MAbs). The adenomas generally retain the lobular architecture, but some may contain focal areas of cellular pleomorphism. These changes may suggest malignant transformation and have led to discordant interpretations. To address this histopathological confusion, two perianal gland carcinoma-associated antigens were defined by mouse MAbs 4A9 and 1A10. These MAbs, generated against a canine mammary carcinoma cell line, reacted strongly with perianal gland carcinoma in preliminary screening and therefore were selected for further investigation. Cellular expression of antigens was examined by indirect immunoperoxidase (IP) assay using MAbs 4A9 and 1A10 against formalin-fixed, paraffin-embedded sections of normal and tumor tissue. Of 25 perianal gland carcinomas, 4A9 antigen was expressed in 100% and 1A10 antigen in 84%. In contrast, perianal gland adenomas were negative for both antigens, and little or no reactivity was detected with normal perianal glands. With eight perianal gland tumors, diagnosis of carcinoma versus adenoma was histologically equivocal, while IP assays consistently revealed focal expression of the 4A9 and 1A10 antigens in these tumors, and the staining coincided with foci of anaplastic cells having a high mitotic index. This group of tumors was designated adenoma/carcinoma in situ. Results suggest that 4A9 and 1A10 antigens are markers of carcinoma and malignant transformation in canine perianal gland tumors, and can be very useful as diagnostic reagents where the identification of carcinoma versus adenoma requires additional clarification beyond routine histopathological examination.     

Human carcinoma-associated precursor antigens of the NM blood group system

Blood group NM specificities occur in healthy, benign and carcinomatous breast glands and those of the gastrointestinal (G.I.) tract, but the precursors in their biosynthesis, T (Thomsen-Friedenreich) and Tn, are found in adenocarcinomata and not in benign or healthy tissues. T-and Tn-antigenic specificities are thus human carcinoma-associated. All humans possess anti-T and anti-Tn antibodies. Patients with breast or G.I. tract carcinoma show statistically significant alteration of anti-T titer levels when compared to patients with benign disease and to healthy controls. Breast carcinoma patients but not healthy people showed cellular immunity to T antigen in vitro and in vivo. Most striking was the delayed-type hypersensitivity reaction, which was positive in over 90% of ductal breast carcinoma patients tested and negative in all presumably healthy individuals. T antigen is readily prepared from healthy human red blood cells in uncontaminated form, and free of HL-A and Au antigens. T antigen and anti-T antibodies may be useful in combating some human adenocarcinomata.     

Biological behavior of human breast carcinoma-associated antigens expressed during cellular proliferation

The cell surface-binding properties of two murine monoclonal antibodies reactive with human mammary tumor cells are described. Fluorescence-activated cell sorter analyses demonstrate that both monoclonal antibodies, B6.2 and B38.1, are reactive with the surface of the majority of human breast tumor cell lines tested but are unreactive with a variety of normal human cell lines, melanomas, sarcomas, and lymphoid tumors. Antibody B6.2 was also reactive with selective carcinomas, while antibody B38.1 showed even broader reactivity. The two monoclonal antibodies were unreactive with the surface of a variety of normal human tissues obtained at biopsy, including lymph nodes, bone marrow, and spleen, but were reactive with mammary tumor cells obtained from four of six pleural effusions. Surface binding to mammary tumor cells by both monoclonal antibodies was shown to decrease during density-dependent arrest; further cell cycle analysis demonstrated differential antibody surface binding during S phase. Prolonged exposure of mammary tumor cells to antibody showed no evidence of antigen capping or internalization. Both monoclonal antibodies were shown to lyse mammary tumor cells in antibody-dependent cell-mediated cytotoxicity.     

'1-8 interferon inducible gene family': putative colon carcinoma-associated antigens

D(b-/-)xbeta2 microglobulin (beta2m) null mice transgenic for a chimeric HLA-A2.1/D(b)-beta2m single chain (HHD mice) are an effective biological tool to evaluate the antitumour cytotoxic T-lymphocyte response of known major histocompatibility-restricted peptide tumour-associated antigens, and to screen for putative unknown novel peptides. We utilised HHD lymphocytes to identify immunodominant epitopes of colon carcinoma overexpressed genes. We screened with HHD-derived lymphocytes over 500 HLA-A2.1-restricted peptides derived from colon carcinoma overexpressed genes. This procedure culminated in the identification of seven immunogenic peptides, three of these were derived from the 'human 1-8D gene from interferon inducible gene' (1-8D). The 1-8D gene was shown to be overexpressed in fresh tumour samples. The three 1-8D peptides were both antigenic and immunogenic in the HHD mice. The peptides induce cytotoxic T lymphocytes that were able to kill a colon carcinoma cell line HCT/HHD, in vitro and retard its growth in vivo. One of the peptides shared by all the 1-8 gene family primed efficiently normal human cytotoxic T lymphocyte precursors. These results highlight the 1-8D gene and its homologues as putative immunodominant tumour-associated antigens of colon carcinoma.     

Monoclonal antibodies against ovarian carcinoma-associated antigens, raised by immunization with cyst fluids

Intrasplenic immunization with ovarian tumour derived cyst fluids was used to generate monoclonal antibodies (Mabs) against human ovarian carcinomas. Seven Mabs were characterized immunocytochemically using indirect immunofluorescence assays (IFA) on frozen sections of gynaecologic tumours (n = 85), non-gynaecologic tumours (n = 50), normal human tissues (n = 66), and on 8 ovarian carcinoma cell lines. The Mabs reacted with 67-96% of 49 ovarian carcinoma tissues tested. Although 6 out of 7 Mabs showed slight cross reactivity with some endometrial, cervical, and oviductal carcinomas, little reactivity with non-gynaecologic tumours or normal tissues was found. Based on immunoelectronmicroscopy, all the antigenic determinants defined by the Mabs appeared to be associated with the tumour cell membrane. Among the seven Mabs, OV-TL 16 (IgG1 subclass) and OV-TL 23 (IgM class) reacted positively with 96% and 82% of the ovarian carcinomas respectively. Staining of 100% of the ovarian carcinomas examined (n = 49) could be achieved by combining OV-TL 16 with 23, as also by other combinations of the Mabs. It is concluded that a panel of these newly developed Mabs may be suitable for diagnostic purposes in histopathology.     

Characterization of human ovarian carcinoma-associated antigens defined by novel monoclonal antibodies with tumor-restricted specificity

Three new monoclonal antibodies (MAbs) (MOv16, MOv18 and MOv19) were raised against human ovarian carcinoma. To obtain more specific reagents than those produced so far, we adopted the following experimental approach which consisted of: the selection of a poorly differentiated ovarian carcinoma which was unreactive with all the MAb previously selected in our laboratory; and the application of a particular immunization protocol. The reactivity of the selected MAbs was studied by solid-phase RIA on live and fixed cells from tumor cell lines and by immunofluorescence on frozen sections from surgical specimens. The MAb MOv16 reacted with 60% of ovarian carcinomas as well as with a high percentage of other carcinomas and with some normal tissues. In contrast, MOv18 and MOv19 appeared to have restricted specificities for ovarian carcinomas and cystadenomas. Reactivity on other carcinomas was only observed in a few cases and no reactivity was found on non-epithelial tumors or normal tissues. Immunoprecipitation experiments indicated that MOv16 recognizes a 48-50-kDA protein, whereas MOv18 and MOv19 both identify a 38-40 kDA glycoprotein band. Cross-competition experiments, together with a double-determinant immunoradiometric assay which uses MOv18 as catcher and MOv19 as tracer, suggested that they recognize different epitopes carried by the same molecule. The affinity constants of MOv18 and MOv19 were estimated to be in the range of 10(8)-10(9) M-1. Taken together, the properties of these antibodies, their restricted ovarian tumor specificities and relative high affinity constants, suggest that they could represent promising tools for in vivo applications.     

Immunohistochemical localization of lysosomal cathepsin D in schwannomas

The immunohistochemical localization of cathepsin D (CD) was demonstrated for the first time in 54 schwannomas (32 intra- and 22 extracranial; 47 benign and 7 malignant) and 5 normal nerve fibers. Granular or vesicular CD-reactive structures were observed in all normal Schwann cells. All tumors contained CD-reactive tumor cells, although the population of CD-reactive tumor cells, the density, intracellular localization, and morphology of CD-reactive structures, and the intensity of CD immunore-activity varied from case to case, portion to portion, and cell to cell, differing variously from those in normal Schwann cells. The variations were greater in malignant than in benign schwannomas. In mildly degenerate tumor cells, CD immunoreactivity was increased, possibly in response to the increased intracellular degenerate proteins, suggesting that the mechanism of induction of lysosomal proteases preserved in normal cells is not affected by the process of neoplastic transformation. In lesions of severe degeneration or necrosis, CD immunoreactivity was lost in most tumor cells but was strong in macrophages invading the lesions and perivascular regions. CD immunoreactivity was observed at various intensities in tumor cells in the Antoni type A area but not in most tumor cells in the Antoni type B area, suggesting that Antoni type B lesions show degenerative changes. The presence of CD-reactive tumor cells in all tumors examined and strong CD immunoreactivity observed at the invasion front of tumors in some cases of benign or malignant schwannoma suggests the possible role of CD in tumor invasion in some cases.     

Expression of transforming growth factor-beta 1 and growth in soft agar differentiate prostate carcinoma-associated fibroblasts from normal prostate fibroblasts

Carcinoma-associated fibroblasts (CAF) promote tumor progression of pre-neoplastic epithelial cells. To investigate the basis of this phenomenon, we compared the properties of fibroblasts cultured from normal human prostate (NHPF) to prostate CAF. NHPF and CAF were assayed for growth potential, cell death and proliferative capacity by measuring population doubling time, cell cycle distribution and capability to form colonies in soft agar. Resistance to genotoxic (UV radiation: 0-50 J/cm2) and chemotoxic (0-200 nM Taxol) agents were compared between CAF and NHPF by measuring cell viability and cell cycle analysis. Transforming growth factor beta1 (TGF-beta1) immunoreactivity was assessed in non-malignant and malignant prostatic tissue. No detectable differences were found when comparing CAF and NHPF with respect to population doubling time, cell cycle distribution and response to genotoxic and chemotoxic agents. The mean number of colonies in soft agar was 120.5 for CAF vs. 18.2 for NHPF (p < 0.05). Because TGF-beta1 and matrix metalloproteinase (MMP)-9 have been associated with growth of fibroblasts in soft agar and tumor promotion, we measured the expression of these factors in NHPF and CAF by ELISA. There was no difference in expression of MMP-9; however, TGF-beta1 was expressed in higher concentrations in CAF than in NHPF (p < 0.0014). Furthermore, TGF-beta1 expression was higher in the carcinoma-associated stroma of prostate cancer tissue than stroma of non-malignant prostatic tissue. Increased capability of CAF as compared to NHPF to form colonies in soft agar may be due to a higher expression of TGF-beta1 and correlates with the ability of CAF to promote malignant progression of prostate epithelial cells.     

Significance of breast carcinoma-associated antigens as a monitor of tumor burden: characterization by monoclonal antibodies

Use of monoclonal antibodies (Mc 3 and Mc 8) prepared against human mammary-epithelial antigens of human mild fat globule membranes has enabled characterization of breast carcinoma (BC) associated antigens (BCAA), antibodies, and circulating immune complexes (CIC). For this study, BC patients were grouped on the basis of measurable tumor burden: Group I patients with no evidence of disease at sampling time; Group II patients with tumor burden less than or equal to 5 g; and Group III patients with known regional or distal metastases. In an in vitro simulation of tumor burden change, selected BC patients' sera were admixed with Mc 3 and Mc 8 at optimal concentration. CIC reduction (dissociation) for Groups I and III and increment (formation) for Group II were noted. Unlike Group I sera, Groups II and III sera required 4- to 16-fold dilution of Mc 3 and 4-fold more concentrated Mc 8 to achieve maximal CIC changes. Serum BCAA isolated by use of both Mc 3 and Mc 8 immunobead procedures showed apparent Mr 33,000 monomer, 66,000 dimer, and 95,000 trimer. When BCAA were added to BC patients' sera, autologous combinations resulted in small (7.7S) CIC for Groups I and III, and medium (9 to 12S) CIC for Group II. Conversely, allogenic combinations resulted in mainly small CIC for Group I, and intermediate CIC for Group II and Group III. Evaluation of circulating BCAA concentration by use of a three-step radioligand technique demonstrated significant discrimination between BC patients' sera (mean = 105 ng/ml) and normal control sera (less than or equal to 20 ng/ml). BCAA were found to be elevated in 31 of 46 (67%) Group I (mean = 70 ng/ml), 41 of 43 (95%) Group II (mean = 197 ng/ml), and 30 of 46 (65%) Group III (mean = 50 ng/ml) patients' sera, as compared to "background" levels in malignant melanoma and normal controls. Benign breast disease sera showed moderate BCAA increases (mean = 48 ng/ml) in 20 of 35 (57%) patients. Furthermore, serial sample determination of BCAA in 36 selected BC patients confirmed the above pattern, indicating that this assay can be used with some restriction to monitor tumor burden. Whereas in early breast carcinoma increase in BCAA concentration was concurrent with or antedated clinical objective evidence of tumor burden increase, significant decreased BCAA concentration was observed with tumor burden reduction. Overall, increased BCAA levels were associated with limited tumor burden (Group II) while decreased BCAA levels were observed with no evidence of disease (Group I) and known regional or distal metastatic advanced disease (Group III) during patients' follow-up.     

Experience of ultrasound-based daily prostate localization

PURPOSE: The NOMOS (Sewickley, PA) B-mode Acquisition and Targeting System (BAT) ultrasound system provides a rapid means of correcting for interfraction prostate positional variation. In this investigation, we report our experience on the clinical issues relevant to the daily use of the BAT system and the analysis of combined setup error and organ motion for 3509 BAT alignment procedures in 147 consecutive patients treated with IMRT for prostate cancer. METHODS AND MATERIALS: After setup to external skin marks, therapists performed the BAT ultrasound alignment procedure before each IMRT treatment. In this study, a single physician (A.C.) reviewed all BAT images and classified image quality and accuracy of image alignment by the therapist. On a scale of 1-3, near-perfect image quality or alignment was given a 1, fair image quality or misalignment > or = 5 mm (likely within the PTV) was given a 2, and unacceptable image quality or misalignment >5 mm (potential to violate the PTV) was given a value of 3. The distribution of shifts made was analyzed in each dimension and for all patients. The time required to perform the BAT alignment was also assessed in 17 patients. RESULTS: Among the 3509 attempted BAT procedures, the image quality was judged to be poor or unacceptable in 5.1% (181). Of the remaining 3328 BAT images, with quality scores of 1-2, alignments were unacceptable (>5 mm misalignment as judged by the reviewing physician) in 3% (100). The mean shift in each direction, averaged over all patients, was 0.5-0.7 mm. Interfraction standard deviation (1 SD) of prostate position based on combined setup error and internal organ motion is 4.9 mm, 4.4 mm, and 2.8 mm in the anteroposterior (AP), superior-inferior (SI), and lateral (RL) dimensions, respectively. The distribution of the shifts was a near-random Gaussian-type in all three major axes, with greater variations in AP and SI directions. The percent of BAT procedures in which the shift was >5 mm was 28.6% in AP, 23% in SI, and 9% in RL directions. The average BAT procedure took extra 5 min out of a 20-min time slot in a typical eight-field IMRT treatment. CONCLUSIONS: The quality of the daily ultrasound images was deemed acceptable in 95%. Major alignment errors by therapists were only 3%. The BAT system is clinically effective and feasible in a matter of 5 min. Although the accuracy of the BAT was not addressed in this investigation, we found a significant percentage of large shifts being made from the initial alignment position.     

Immunohistochemical localization of basic fibroblast growth factor in astrocytomas

Because of the prominent neovascularization observed in the growth of brain tumors, we studied the occurrence of basic fibroblast growth factor (bFGF), a potent angiogenic factor in astrocytomas, the most aggressive of which often have marked vascular hyperplasia. Using immunohistochemical methods, we examined 21 examples of such tumors, 7 glioblastomas multiforme, 7 anaplastic astrocytomas, and 7 low grade astrocytomas. Using polyclonal and affinity-purified rabbit antisera to human bFGF, we detected immunoreactive bFGF in all cases of glioblastoma multiforme. bFGF was present in both endothelial cells and neoplastic astrocytes. In 4 of 7 anaplastic astrocytomas, the tumor astrocytes had bFGF immunoreactivity and, in 5 of 7 cases, endothelial cells were also immunopositive. In glioblastomas multiforme and anaplastic astrocytomas, capillaries adjacent to tumor showed bFGF immunoreactivity, whereas capillaries distant from the tumors were not immunostained. In low grade astrocytomas, astrocytic cells were weakly immunoreactive in 2 of 7 cases, and in only 1 of the 7 cases capillaries were immunostained. In each grade, reactive astroglial cells showed variable bFGF immunoreactivity. The immunostaining was not seen with the flow-through fraction obtained after affinity purification of the bFGF antiserum with pure recombinant bFGF. These results suggest a possible role for bFGF in tumor growth and in angiogenesis in astrocytomas.     

Immunohistochemical localization of glycosaminoglycans in experimental rat glioma models

Changes of glycosaminoglycan distribution in and around C6 glioma and ethylnitrosourea(ENU)-induced glioma in rats were investigated using monoclonal antibodies that specifically recognize epitopes on chondroitin-0-sulfate proteoglycan (C-0-S), chondroitin-4-sulfate proteoglycan (C-4-S), dermatan sulfate proteoglycan (DS), chondroitin-6-sulfate proteoglycan (C-6-S) and keratan sulfate proteoglycan (KS) after chondroitinase ABC digestion. In the normal brain tissues, C-0-S was located on the surface of the neurons. In addition, extracellular staining in the cerebral cortex and axoplasmic staining in the brain stem and the reticular thalamic nucleus were seen. C-0-S was negative, however, both in the C6 and ENU-induced gliomas. C-4-S or DS was detected only in some of the neurons in the normal brain tissues. They were detected in the peripheral part of the ENU-induced gliomas, but not in the C6 gliomas. C-6-S was located on the surface of some neurons and in the white matter of the normal brain, but it was not detected in C6 gliomas. In all ENU-induced gliomas, C-6-S was identified in the adventitia of the vascular structures within the tumor. In some of them, C-6-S appeared in the peripheral part of the tumor. KS was immunostained in the glial cells in the hippocampus, corpus callosum, brain stem, and the floor of the third ventricle. It was also detected in the peritumoral brain tissues both in the C6 and ENU-induced rat gliomas. The significance of glycosaminoglycans in these glioma models was discussed.     

Marker seed migration in prostate localization

PURPOSE: Marker seed location was analyzed to test the hypothesis that there is no intraseed migration within the prostate, a premise fundamental to the technique of marker seed localization of this organ. Despite increasing interest in the use of implanted seeds as fiducial markers for gland location, there are few data available with which to evaluate the validity of this technique, particularly over the entire course of external beam radiation therapy. METHODS AND MATERIALS: Between May 2001 and December 2001, after obtaining fully informed written consent, 9 patients with early stage prostate cancer were enrolled on an institutionally reviewed protocol. Patients had four to five marker seeds implanted into the prostate under transrectal ultrasound guidance before definitive radiotherapy. The porous gold seeds were each 1.2 x 2.0 mm in dimension. Seed locations from orthogonal radiographs based on the initial simulation and weekly orthogonal films were digitized using a CMS Focus planning system, thereby facilitating the determination of intraseed spacing. The digitization of the isocenter from each orthogonal pair of radiographs was used to determine digitizing error for seed localization. Pubic symphysis, bilateral femoral heads, and isocenter were also digitized and will be analyzed at a later date. RESULTS: Overall, the average migration of all the seeds in the patients was 1.2 +/- 0.2 (SD) mm. The greatest average movement of any seed in any patient was 1.9 mm over the entire 7-week course of radiotherapy. The smallest average movement was 0.6 mm. The greatest change in intraseed spacing in any of the patients during the full course of therapy was 6.6 mm. One seed in 1 patient was lost at the start of the third week of therapy and censored from analysis. Digitizing error in seed localization was calculated to be 0.20 +/- 0.03 (SD) mm. CONCLUSIONS: As an aggregate, there is negligible seed migration within the prostate over the entire course of definitive radiotherapy. However, there are small, detectable movements in individual seed locations, perhaps resulting from topographic changes in the gland secondary to seed placement, anatomic changes in bladder and rectum, or treatment itself. With respect to seed migration, prostate marker seeds represent an accurate and reliable surrogate of gland location during a full course of radiotherapy.     

Immunohistochemical localization of NADPH-cytochrome P450 reductase in human tissues

In an attempt to elucidate the mechanism(s) behind the susceptibility of tissues to the toxic effects of chemicals, NADPH-cytochrome P450 reductase IgG has been used to map the distribution and localization of the cytochrome P450 system in hepatic and extrahepatic human tissues. Employing the Western blot procedure this antibody recognized a single band in human liver microsomes which corresponded in mol. wt to the purified reductase. Immunoreactive NADPH-cytochrome P450 reductase staining was detected in all zones of the liver acinus, with maximal staining in hepatocytes adjacent to the terminal hepatic venules (zone 3). Considerable variation in the intensity of reductase staining was observed in different segments of the gastrointestinal tract. Staining was most intense in the enterocytes of the small intestine, with maximal staining at the tips of the villi. Colonic epithelial cells were variably positive while the rectum was negative. Pancreatic ductal cells were positive whereas exocrine cells were negative. In the lung, reductase was detected in bronchiolar and bronchial epithelial cells, Clara cells and alveolar lining cells. In the kidney, the proximal and distal convoluted tubules, the loops of Henle, the collecting ducts in the medulla and the transitional epithelium all stained positively for reductase. The results demonstrate specific cellular localization of NADPH-cytochrome P450 reductase, and hence the cytochrome P450 system, in human tissues. The differential distribution of the reductase within human tissues may lead to a better understanding of the mechanism underlining site-specific carcinogenesis.     

Immunohistochemical localization of parathyroid hormone-related protein in human breast cancer

Parathyroid hormone-related protein (PTHrP) is known to be a causative factor in humoral hypercalcemia of malignancy. A polyclonal rabbit antiserum directed against the amino-terminal region of the protein and immunoperoxidase methods have been used to locate the presence of PTHrP in a series of 102 consecutive invasive breast tumors removed surgically from normocalcemic women. Positive PTHrP staining was detected in 60% of the tumors but not in the accompanying normal breast tissue. Positive staining was related to the progesterone receptor status of the tumor (P = 0.039) and to the prognostic index of the patient (P = 0.046) and not to estrogen receptor status, patient age, tumor size, histological grade, or nodal status.     

Immunohistochemical localization of S-100 protein in granular cell myoblastoma

The presence and distribution of S-100 protein were studied in three cases of granular cell myoblastoma using the peroxidase-anti-peroxidase method. Positive immunostaining was observed in all cases. The densely brown reaction products were found both in the nuclei and in the cytoplasm of myoblastoma cells. The staining of the nuclei was more intense than that of the cytoplasm. In addition, both normal Schwann cells and tumor cells of Schwannomas were also stained with anti-S-100 antibody. This result further supports the concept of the neurogenic origin of the granular cell myoblastoma.