Evaluation of immunoreactivity with monoclonal antibody NCRC 11 in breast carcinoma

Immunocytochemical staining with monoclonal antibody NCRC 11 of formalin fixed paraffin embedded tumour tissue has been studied in 444 cases of primary breast cancer with a minimum follow period of 6 years. The relationship between extent of staining, assessed on a four point scale, and patient survival has been confirmed. There are significant relationships between staining and both histological grade and oestrogen receptor status. No association has been shown between staining and lymph node stage or tumour size. Simplification of staining assessment by modification to two staining groups still allows significant separation of patients into prognostic groups and incorporation into an existing prognostic index.     

常规模拟定位条件下贲门位置判定方法的探讨

目的:探讨一种在常规模拟定位条件下正确判定贲门位置的方法。方法:95例受试者按年龄、性别、体型和胃型分组,通过行上消化道钡餐造影(GI),对贲门口中心位置(M点)随体位改变而发生位移的过程进行了动态观察和摄片。在1:1.1的胶片上,建立了以左侧膈顶为零点的二维直角坐标系,分别测量了各组在仰卧和俯卧位的M点坐标值,并进行了统计学处理。结果:(1)证实因生理解剖学原因,常规模拟定位采取的方法及体位,难以充分暴露M点。(2)M点随体位变化并有一定的规律。而且其坐标值在不同体型组间有显著性差异,但与性别、年龄无关。(3)卧位可借助相应的坐标值推出M点。结论:建立了卧位M点定位参照值,为常规拟定位条件下确定贲门位置提供了一个可靠而实用的判定指标。     

A single-chain antibody fragment against human thyroglobulin: construction and evaluation of immunoreactivity

Smaller recombinant antibody fragments are at the forefront of in vivo diagnosis and therapy. These units possess better distribution and faster clearance than larger molecules. Among these, single chain antibody fragments (scFv) are emerging as credible alternatives. These proteins are shown to have same specificities and affinities for their antigens as the parental monoclonal antibody (MAb). We have attempted to produce scFv against human thyroglobulin (H-Tg) using anti-Tg secreting hybridoma cells and PCR-based cloning approach. Hybridoma secreting anti-Tg MAb B10IV was established. cDNA was prepared from hybridoma cells. The V(H) and V(L) genes were amplified and cloned. The gene sequences were submitted to Genebank database (accession nos. AJ508533 and AM072962, respectively.) V(L) and V(H) genes were then linked together with a linker peptide and successfully cloned in pET28a and expressed as His-tag fusion protein in expression host BL21 (DE3). The scFv protein from IPTG-induced cells was purified under native conditions by immobilized metal affinity chromatography on a Ni-NTA agarose column. The yield expressed in Escherichia coli was approximately 8 mg/L. ScFv could be labeled with (125)I and its immunoreactivity evaluated in radioassays. Although scFv demonstrated specific binding to H-Tg, the immunoreactivity was low (10.3%) compared to the parental MAb B10IV, which showed immunoreactivity of 37.27%. Inhibition radioassays exhibited that scFv and MAb interact with the same epitope on the target antigen, indicating its specificity.     

Immunoenzyme method of determining brain-specific alpha 1-globulin in glial tumors

Specific alpha 1-globulin of the brain was assayed by an immunoenzyme technique in blood sera from cases of various brain pathologies and healthy donors. Relative elevation in brain specific alpha 1-globulin level, as compared with that in healthy subjects, was more frequent in patients with malignant glial tumors of the brain, less frequent in cases of benign glial tumors and in very few patients suffering from other brain diseases. Raised level of specific alpha 1-globulin may be caused by disorders in the blood-brain barrier or lesions in brain or tumor tissues which is peculiar to glial malignancies of the brain. The said test offers much advantage in diagnosis of glial tumors.     

Potentialities of an x-ray method of determining malignant changes in polyps of the large intestine

The study was concerned with evaluation of the clinical and roentgenologic data on 57 cases of malignant polyps of the colon. Malignant polyps reaching 10 mm in length were found. Malignancies were identified using endoscopy and biopsy in 58.6% and X-ray examination--45.8%. Among the most effective methods of X-ray examination of the large bowel for cancer should be: high-voltage roentgenography employing incomplete barium enema, primary double contrast examination, direct blow-up and parietography. X-ray signs of malignant transformation of polyps are: persistent retention of barium, wavy and indistinct contours, irregular pattern, rapid growth of neoplasm, retraction of tumor base and disparity between polyp size and its pedicle length.     

Use of protamine sulfate in a method for determining the hormonal sensitivity of breast cancer

A new procedure for estrogen receptor assay in breast cancer tissue is discussed. According to the method cytosol receptor level is interpreted as the estradiol binding capacity of protein which is precipitated by protamine sulfate from the cytosol fraction of tumor.     

A method for detecting antibody internalization in drug targeting

BCM43/2E5 monoclonal antibody with a specificity for human ovarian carcinoma was evaluated for its binding and internalization behaviour on both fixed and live cells. Internalization of this antibody by the targeted cells was demonstrated by ELISA, flow cytometry and immunoperoxidase. At reduced temperatures or in the presence of electron transfer blockers, internalization of bound antibody was prevented. This method is rapid and reproducible and may be used efficiently for the selection of antibodies suitable for drug targeting in cancer therapy.     

alpha-Neoendorphin-like immunoreactivity in medullary carcinoma of the thyroid

alpha-Neoendorphin-like immunoreactivities (alpha-NE-IR) were demonstrated in tissues from three patients with medullary carcinoma of the thyroid (MCT). A large amount of alpha-NE-IR was detected in the extracts of primary tumors and metastatic lymphatic tissues by a highly sensitive and specific radioimmunoassay (RIA). Gel filtration analyses showed two different molecular-weight forms of the alpha-NE-IR: One eluted at the void fraction and the other at the position of [125I]-alpha-NE on Sephadex (Pharmacia Fine Chemical, Uppsala, Sweden) G-50 chromatography. Immunohistochemical examination revealed the presence of alpha-NE-IR in the C-cell carcinoma. These data presumably reflect that alpha-NE, the opioid peptide derived from preproenkephalin B, is synthesized in the MCT.     

鼠伤寒沙门氏菌回复突变试验中通过吸光度值测定菌液浓度的方法研究

目的：研究鼠伤寒沙门氏菌回复突变试验（Ames试验）中菌液浓度和吸光度值[D（λ）值，λ为波长数值]的关系，开发通过吸光度值测定菌液浓度的方法。方法：选取Ames试验中常用的营养缺陷型鼠伤寒沙门氏菌TA102作为试验菌株，严格控制菌液扩增条件，对比分析540和600 nm波长下菌液的吸光度值的高低以确定最佳测定波长，对比分析菌株扩增0、2、4、6、8、10、12、14 h后的菌液浓度以确定最佳扩增时间。在最佳测定波长和最佳扩增时间下，测定菌液吸光度值，并通过平板菌落计数法测定菌液浓度，获得菌液浓度和吸光度值的回归方程。结果：菌株TA102的最佳测定波长为540 nm，最佳培养条件为37℃、100 r/min、10 h，菌液浓度和D（540）值的直线回归方程为y=4×10⁹x[方程中x为D（540）值；y为菌液浓度，单位为个/mL]。结论：应用上述菌液浓度和D（540）值的直线回归方程，可通过测定D（540）值实时控制菌液达到要求浓度，从而确保试验菌液的质量。     

The value of immunoscintigraphy for the operative retreatment of colorectal cancer. Limitations of a new diagnostic method

In 42 patients with suspected recurrent colorectal cancer, results of conventional diagnostic methods were compared with those of immunoscintigraphy. In 69% of all cases, the intraoperative findings of a second-look operation served for validation, whereas in 31% close follow-up was used. Recurrent tumors were successfully localized in 83% of patients by conventional methods, whereas immunoscintigraphy was expressive in 57% of cases. Immunoscintigraphy was disappointing especially because of its low sensitivity (23%) and low predictive value (positive, 33%; negative, 37%) with regard to successful diagnosis of extrahepatic tumors compared with the results of conventional methods (77% sensitivity; positive, 94%; negative, 79%). The rate of false-positive results was relatively high with immunoscintigraphy (n = 12), 83% of which were related to extrahepatic recurrent tumors. The value of immunoscintigraphy using an immunococktail of 131-J-labeled F(ab')2 fragments of monoclonal antibodies against CEA, with Ca 19-9 as an additional diagnostic tool for early detection of recurrent colorectal cancer, must therefore be viewed critically.     

Monoclonal antibody 83D4 immunoreactivity in human tissues: cellular distribution and microcytophotometric analysis of immunoprecipitates on tissue sections.

Immunocytochemical assays were performed on cell cultures as well as on a wide range of human tissues using a monoclonal antibody, MAb 83D4, produced by a murine hybridoma generated by immunization with a cell suspension from a paraffin block of human breast carcinoma tissue. Frozen breast tissue samples (n = 49) were compared to fixed and paraffin-embedded samples (n = 62). Paraffin sections (n = 194) from a variety of human tissues were compared to breast immunoreactivity. Immunoprecipitates, resulting from positive reactions between 83D4 and Avidin Biotin Peroxidase, were evaluated by computer-assisted microcytophotometry (SAMBA). In some frozen breast samples (n = 27), 83D4 antigen distribution was correlated with tumor cell DNA index, ploidy balance, growth fraction (Ki67), hormone receptor (ER, PR) antigenic sites, NORsAg and oncoprotein pHER-2/neu cell content. MAb 83D4 reacted with 3 breast cancer cells lines (MCF7, T47D and H466B) but not with normal epithelial breast cells in culture. The immunostaining in frozen paraffin sections from breast were similar. Like most normal tissues, normal breast did not react with 83D4. Cellular MAb 83D4 antigen concentration increases with the degree of malignancy but is independent of DNA nuclear content, ER, PR, growth fraction and pHER-2neu in cancers. These results suggested that routine immunohistochemical procedures using MAb 83D4 could facilitate the grading of breast cancer, in particular by allowing detection of microvascular invasions in the breast as well as at a distance and of blood-born micrometastases, especially in bone marrow.