Fritz Herz

Alkaline phosphatase in HT-29, a human colon cancer cell line: influence of sodium butyrate and hyperosmolality.

Abstract HT-29, a cell line derived from a human colon carcinoma, exhibits very low alkaline phosphatase activity. The enzyme is thermolabile and is of the intestinal type. Hyperosmolality and/or sodium butyrate induce increased levels of activity. The increase is most pronounced with HT-29 cells growing in hyperosmolar medium containing sodium butyrate. Under these conditions specific activity rises over 1000-fold. The effect of hyperosmolality is blocked by cycloheximide and that of sodium butyrate by thymidine, cordycepin, and cycloheximide. By contrast to other human cancer cell lines, the enzyme of HT-29 is not influenced by cell density and glucocorticoid hormones. 5-Bromo-2′-deoxyuridine and inhibitors of DNA synthesis cause a slight increase in specific activity.

Flow cytometric analysis of the DNA distribution in human brain tumors

SummaryFlow cytofluorometric analysis was used to determine the distribution of the DNA content in cells from selected areas of normal human brain and in benign and malignant brain tumors. Propidium iodide was employed as DNA fluorochrome and the analysis was carried out on a suspension of single cells. Normal, nonstimulated human lymphocytes were used as diploid controls. With nonneoplastic tissue an average of 91% of the cells were diploid (presumably in G0 or G1 stage of the cell cycle). The cells of most benign tumors were mainly diploid (77–98%), nine specimens of pituitary adenomas had large numbers of aneuploid cells. In glioblastoma multiforme the proportion of diploid cells was significantly diminished and polyploid cells were frequently seen. Similar results were obtained in other malignant tumors, with metastatic tumors showing the greatest ploidy variation, which included triploid, tetraploid, and hypertetraploid cells. The analytical method used provides valuable information of significant clinical importance on the DNA distribution in brain tumor cells.

Comparative immunohistochemical study on the expression of αB crystallin, ubiquitin and stress-response protein 27 in ballooned neurons in various disorders

This Report deals with a comparative study on the expression of αB crystallin, ubiquitin, stress‐response protein 27 (srp 27), srp 72 and phosphorylated neurofilament protein (pNFP) by ballooned neurons in Picks disease, Creutzfeldt‐Jakob disease (CJD), amyotrophic lateral sclerosis (ALS), leptomeningeal carcinomatosis, anterior spinal artery syndrome and pellagra. Immunohistochemical techniques were used. αB Crystallin was expressed by the majority of ballooned neurons of Picks disease and CJD, but not by those of the other disorders. Ubiquitin and srp 27 expression was also restricted to abnormal neurons of Picks disease and CJD, but the proportion of stained cells was less than that expressing aB‐crystallin. There was no evidence of ballooned neurons expressing srp 72. Except for those of pellagra patients, phosphorylated neurofilament protein (pNFP) was detected in most abnormal neurons. Our results suggest that the mechanisms involved in formation and maintenance of swollen neurons in Picks disease and CJD may be different than those of ballooned neurons in the other entities studied.

Ultrastructural and immunohistochemical studies on ballooned cortical neurons in Creutzfeldt-Jakob disease: expression of αB-crystallin, ubiquitin and stress-response protein 27

SummaryThis report concerns ultrastructural and immunohistochemical studies on ballooned neurons of ten patients with Creutzfeldt-Jakob disease (CJD). While abundant ballooned neurons and severe white matter degeneration was seen in six Japanese cases, only occasional ballooned neurons and no white matter degeneration was observed in four cases from the files of Montefiore Medical Center. Ultrastructurally, the ballooned neurons contained granule-coated fibrils of 25 to 40 nm in width and 10-nm neurofilaments. The immunohistochemical studies revealed that most ballooned neurons expressed αB-crystallin, with deposits of reaction products observed in the cytoplasm. A similar intracellular staining pattern was also seen with the antibody to phosphorylated neurofilament proteins (pNFP). Although the proportion of stained ballooned neurons was less, a positive reaction was also observed with antibodies against ubiquitin, stress-response protein 27 (srp 27) and synptophysin, but not with an antibody to srp 72. Our findings suggest that expression of pNFP and synaptophysin by ballooned neurons may reflect axonal impairment and that the presence of αB-crystallin, srp 27 and ubiquitin may be related to the degenerative processes that neurons undergo in CJD.

Predictive value of DNA measurements in bladder washings. Comparison of flow cytometry, image cytophotometry, and cytology in patients with a past history of urothelial tumors

Comparative DNA ploidy measurements were carried out by flow cytometry and by image analysis on cells in 71 bladder washing specimens from 50 patients with past histories of bladder tumors. Among the specimens classified as diploid or questionable by flow cytometry, 14 showed the presence of aneuploid DNA values documented by image analysis. In 18 of the 50 patients, recurrent tumors were observed during a relatively brief period of follow‐up. In 15 of them the DNA pattern was aneuploid and in three it was questionable. In nine of the 15 patients, both methods of DNA analysis disclosed aneuploidy, but in six patients aneuploidy was detected by image analysis only. A combination of DNA aneuploidy, whether observed by flow cytometry, image analysis, or both, and of positive or suspicious urine cytology is highly predictive of recurrence of high grade bladder tumors. Image analysis of DNA content in bladder washings adds information of clinical value above and beyond that obtained by flow cytometry.

Chromosome abnormalities in meningiomas

Cytogenetic analyses of eight meningiomas grown in culture for 1 week are reported. Normal karyotypes were found in three cases and hypodiploidy in the remaining five. In the five hypodiploid meningiomas, one chromosome #22 was missing in four cases, and one case exhibited a 22q- deletion. In four of these five cases, chromosome #14 was either lost or altered. Chromosome #1 was lost or altered in three, and chromosome #6 in two. These findings lend further support for the association of total or partial loss of chromosome #22 in meningiomas and suggest the involvement of other chromosomes in the clonal evolution of these tumors.

Cell cycle-dependent reactivity with the monoclonal antibody Ki-67 during myeloid cell differentiation☆

The specificity and sensitivity of the monoclonal antibody Ki-67 in identifying proliferating cell compartments was tested with the human promyelocytic leukemia cell line HL-60 using multi-parameter flow cytometry. While correlated measurements of DNA content and Ki-67 immunofluorescence indicated that the antigen was present in all phases of the cell cycle, reactivity with the antibody was highest in proliferating S and G2+M cells. The analysis of the BrdU content of cells sorted on the basis of reactivity with Ki-67 showed a correlation between Ki-67 reactivity and BrdU uptake. In HL-60 cells induced to differentiate with dimethyl sulfoxide (DMSO), the loss of reactivity with Ki-67 paralleled the exit of cells from the cell cycle. This was not observed in DMSO-resistant HL-60 cells. These results validate the usefulness of the Ki-67 antibody for determining the proliferative stage of mammalian cells in culture.

Expression of stress-response (heat-shock) protein 27 in human brain tumors: an immunohistochemical study

SummaryThis report concerns the expression of the low molecular weight stress-response (heat-shock) protein 27 (srp 27) in a variety of human brain tumors. Immunohistochemical techniques were used; cells of the breast cancer line MCF7 served as positive controls. The reaction product was found exclusively in the cytoplasm. Srp 27 was detected in 5/5 breast tumor metastases to the brain and in 5/21 meningiomas. The protein was also detected in 5/11 glioblastomas and 2/5 pituitary adenomas. By comparison, positive staining was observed in only 1/15 astrocytomas and 1/7 medulloblastoma and no reaction was seen with the oligodendrogliomas, schwannomas and gangliogliomas tested. These observations demonstrate that srp 27 is expressed by certain primary intracranial tumors.

Comparative study on the expression of stress-response protein (srp) 72, srp27, αB-crystallin and ubiquitin in brain tumours. An immunohistochemical investigation

This immunohistochemical study compares the expression of stress‐response (heat‐shock) protein (srp) 72, srp 27, αB‐crystallin and ubiquitin in 86 primary human brain tumours and 21 carcinoma metastases to the central nervous system. Normal brain tissues were included for control purposes. Serial sections of formalin‐fixed, paraffin‐embedded tissues were used. Most meningiomas (17/23), glioblastomas (11/12) and breast carcinoma metastases (9/10) and some astrocytomas (7/13), pituitary tumours (4/9) and lung cancer metastases (5/11) had tumour cells that reacted with one or more of the antibodies used. Around 43% of the meningiomas and 25% of the glioblastomas expressed srp 72 only. Sole expression of srp 2 7, αB‐crystallin or ubiquitin was seen in several tumours. Some meningiomas (3/23) and breast cancer metastases (4/10) co‐expressed srp 72 and srp 27, and 1/3 of the glioblastomas co‐expressed srp 27 and αB‐crystallin. We conclude that primary and metastatic tumours of the brain produce stress‐related proteins and that certain tumours concurrently express two or more srps.

Flow-through cytometry of meningiomas and cultured meningioma cells.

SummaryFlow cytometric techniques were used to compare the DNA content, size and viability of meningioma cells obtained directly from surgical specimens with the same cells after a period of culture. Cells isolated from the original meningiomas and cells in primary culture were similar with regard to size and DNA content, regardless of the histologic subclassification of tumor. The cell populations were essentially diploid with a small proportion of tetraploid cells. Viable cells were smaller and more uniform in size than the nonviable cells. An increase in the number of cells having an elevated DNA content was seen with cultures repeatedly transferred. The latter results suggest that any transfer of information from long-term cultured meningioma cells to the in vivo situation must be done with caution.