Early Detection and Monitoring of Cancer with the Anti-Malignin Antibody Test "

ABSTRACT: The serum anti-malignin antibody (AMA) test determines the antibody to malignin. a IO,OOO-Da peptide present in patients with a wide variety of cancers.l~ A total of 3315 double-blind tests demonstrated that AMA is a general transformation antibody, elevated in active no.nterminal cancer, regardless of the site or tissue type, with sensitivity and specificity of95% on the flTst determination and >99% on repeat determinations. - 9 Data have not however been published yet that indicate whether, in daily clinical practice, the AMA test provides accurate prospective and predictive information. Fony-two physicians from II states, who ordered the AMA test, performed blind, report here on their results on 208 determinations in the first consecutive 181 patients and controls. Used in monitoring treatment in 56 patients, the test predicted or agreed 94.1 % overall with the clinical status. Used in early detection in 125 patients and controls, of which 118 now have confirmed diagnoses. AMA was elevated in 21, all of whom were proven to have cancer; AMA was normal in 97, none of whom had cancer. Transient elevated AMA occurred in 3%, followed by normal values. Seven patients with still uncertain diagnosis who have had elevated AMA on repeated tests for I year or longer include six who are symptomatic, and three whose families have a high frequency of cancer. The conditions of these 7 may include undetected cancer because of the 118 with now certain diagnosis the AMA test predicted all correctly. From our experience, the AMA test should be used together with other routine procedures whenever signs and symptoms suggest cancer to facilitate early detection

Picogram per Cell Determination of DNA by Flow Cytofluorometry

Using nuclei isolated from less than 0.2 g tissue or 107 cells, a method is presented for the quantitative determination of amounts of DNA per cell at the picogram level. This technique is based on the enhanced fluorescence of 4′,6-diamidino-2-phenylindole (DAPI) when it binds to DNA. A rapid, one-step nuclear isolation and DNA staining procedure is used to prepare tissue samples for flow cytometric analysis. Frozen tissues give results comparable to those for fresh tissue. Both chicken and trout erythrocyte nuclei were used as reference standards in the determination of amounts of DNA per diploid cell for several mammals and Amazon molly fish. The consistent values obtained for different tissues from the same organism show the accuracy of this method for DNA measurement

Cytophotometric Comparisons of DNA Levels in Neuronal and Glial Cells of the Cerebellum: A Comparative Study

Several cytochemical studies of the DNA content and ploidy status of neuronal cell nuclei in the central nervous system have reported the occurrence of hyperdiploid amounts of DNA in Purkinje cells and suggest the existence of some type of ‘extra’ DNA, the biological significance of which is, as yet, unknown. To explore this phenomenon further, the DNA content of glial and Purkinje cell nuclei was determined in several vertebrate species, using the DNA‐specific fluorochrome 4′,6‐diamidino‐2‐phenylindole (DAPI) to stain isolated cerebellar nuclei for analysis with a single parameter flow cytometer. The Feulgen reaction for DNA was used to stain liver and cerebellar tissue imprints for the measurement of individual nuclei with a Vickers M86 integrating microdensitometer. In both types of analyses, chicken erythrocyte nuclei served as an internal reference standard of 2.5 pg DNA per cell. The mean DNA content of Purkinje cells and glial or granule cells was essentially the same as that found for diploid (2C) non‐neuronal cells, such as hepatocytes, in rainbow trout, Amazon molly fish, salamander (Plethodon), mouse, rat, rabbit, cat, dog, monkey and human. Although Purkinje cell nuclei with 4C DNA levels were found in all of these species, except salamander and rabbit, the frequency of such cells was low (1–7%) and varied with the species. There was a low incidence of Purkinje cell nuclei with interclass DNA amounts in all species examined. Our data show that most neuronal cell nuclei in the cerebellum contain 2C levels of DNA

Cytologically Malignant Squamous‐Cell Carcinoma Arising in a Verrucous Carcinoma of the Penis

Abstract. A case of verrucous carcinoma with a focus of cytologically malignant squamous‐cell carcinoma is presented. This usually occurs following radiation therapy of the verrucous carcinoma, but may rarely occur de novo, as in this case. The potential usefulness of fine‐needle aspiration in detecting focal anaplasia in verrucous carcinoma is discussed. This technique may be especially useful if the lesion is to be destroyed cryosurgically

Modulation of Glucocorticoid Hormone Receptor Levels in Chicken Lymphoid Tissue Following Treatment With Androgens in Vivo

Lymphatic tissues are highly sensitive to androgens and androgens are thought to contribute to sex differences in the immune response. In this study we have examined the effects of androgens on cytosolic glucocorticoid receptor levels in lymphoid tissues. The immature chick was chosen for our experimental model because it allows the separate evaluation of the bursa of Fabricius (primarily B-cells) compared to the thymus (primary T-cells). Treatment with dihydrotestosterone (a potent androgen in chicks) for 3-12 days in vivo reduced the cytosolic glucocorticoid (triamcinolone acetonide-[3H]) receptors in the bursa tissue to ∼ 42% of control levels after 5 days and ≤ 5% of control levels after 7 days of treatment. The chick thymus tissues were still ~ 92% of control triamcinolone acetonide receptor levels after 5 days of androgen treatments. However, the thymus levels had dropped to ≤ 5% of control values after 12 treatment days. Thus a difference in the rate of decrease in the bursa of Fabricius compared to the thymus was indicated. The blastogenesis index (BI), a measurement of the percentage of cells progressing through the cell cycle, was figured using fluorescent DNA staining with diamidino phenylindole followed by flow cytomctry analysis. After 3, 5, or 7 days of androgen treatment, the bursa of Fabricius from dihydrotestosterone treated chicks (2 mg/day/chick) had a mean BI = 11.17 (±3.07 SD) which was significantly lower than the bursa of Fabricius from control chicks which showed a mean BI = 27.33 (±3.42 SD). The thymus from dihydrotestosterone treated chicks had a mean BI = 19.57 (±2.19 SD) which was slightly but not significantly higher than the control thymus BI = 17.38 (±0.89 SD). In summary, androgen treatment in vivo induced a decrease in the cytosolic glucocorticoid hormone receptor levels in both the chick thymus and bursa of Fabricius tissues while decreasing the blastogenesis index in the bursa cells but not in the thymus cells

Flow cytometric enumeration of colonies for in vitro chemotherapeutic drug evaluation

The percentage of 50–100 μ colonies formed by LX‐T cells in medium containing agarose was determined microscopically, and this value was compared with the percentage determined by a flow cytometric method based on the forward and 90° light scatter of the colonies. As assessed by both in vitro methods, LX‐T cells exposed to chemotherapeutic agents formed fewer colonies as the drug concentration increased. However, flow cytometric analysis indicated that a change in the number of colonies formed was a consequence of changing chemotherapeutic drug concentration, whereas microscopic colony counting did not always detect the corresponding change in colony number. These experiments demonstrate that measurement of a drug's chemotherapeutic potential by flow cytometric counting of colonies is an alternative to the enumeration of colonies microscopically

Prognostic Indicators Including DNA Histogram Type, Receptor Content, and Staging Related to Human Breast Cancer Patient Survival

Primary tumors from breast cancer patients were evaluated for the biochemical presence of three steroid cytosolic receptors and by DNA histogram analysis using flow cytometry. These parameters were compared with the histological and staging diagnoses and the patients\u27 survival over a 36-month period. A total of 74 patients with primary breast tumors were evaluated. The breast samples invariably demonstrated a peak population of diploid Gw cells which contained 2C amounts of DNA, as determined by mixing experiments using normal human breast tissues or trout erythrocytes as fixed standards. The tumors were classified into five DNA histogram types based on their DNA index distributions established by flow cytometry. These results showed that 21% of the tumors were diploid and indistinguishable from the diploid population of normal breast cells, 8% were hypodiploid, 11% were hypertetraploid,8% were multiploid, and the remaining 52% were hyperdiploid. The DNA index values varied from 0.78 (hypodiploid) to 2.60 (hypertetraploid). The percentages of S-phase cells were lowest in the diploid and hypertetraploid tumors and highest in the hypodiploid tumors. Among the 24 patients who died during the 36-month follow-up, 92% (22 of 24) were classified in one of the aneuploid groups. Three high-risk groups identified on the basis of survival after 36 months were distinguished: hypodiploid (50% survival); multiploid (43% survival); and hyperdiploid (50% survival). Rates of survival in the diploid and hyperdiploid groups were 87 and 71%, respectively. The hypodiploid group was distinguished by having the lowest mean estrogen cytosolic receptor value [26 ± 13 (S.D.) fmol/mg], progesterone cytosolic receptor value (13 ± 15 fmol/ mg), and androgen cytosolic receptor value (\u3c1 ± 1 fmol/mg). In contrast, the diploid tumors had some of the highest receptor values, with mean estrogen cytosolic receptor value equal to 102 ± 114 fmol/mg, progesterone cytosolic receptor value equal to 74 ± 110 fmol/mg, and androgen cytosolic receptor value equal to 65 ± 80 fmol/mg. The lowest survival rates (17% after 36 months) occurred in patients over 67 years of age who had aneuploid tumors, compared to 100% survival in patients over 67 years of age with diploid tumors. Our results demonstrate the value of using flow cytometry and steroid receptor values as supplements to histopathokxjy for the characterization of subgroups of mammary cancer patients. The ability to identify patients with a good prognosis compared to those at high risk of recurrence and death will be valuable in the design of future prospective treatment studies

The Cancer Recognition (CARE) Antibody Test

The cancer recognition (CARE) antibody (Ab) test is a serologic assay for a specific IgM that is elevated in cancer patients. All tests are measured using an indirect enzyme-linked immunosorbent assay (ELISA) of human serum. The target polypeptide in the CARE Ab test is the IgM binding epitope (LT-11) of the CARE antigen (Ag) consisting of a 16 mer structure that has been produced synthetically. The mean relative concentration (MRC) is determined relative to standard, normalized human plasma. Non-parametric analysis showed median MRC values of healthy volunteers (HVs) with no history of cancer (n=47), family history of cancer (n=126) and a previous cancer history (n=24) to be 26, 34 and 46, respectively. It was determined that there was no significance found among the medians of the three HV groups (P=0.53). The specificity of the HV types was between 87 and 98%. Benign/non-cancer surgical patients (n=27) had a median value of 20 with a specificity of 96%. The cancer patients (n=61) had a median value of 246 with a sensitivity of 89%. There was a significant difference between the HV and cancer patients (P\u3c0.0001) as well as between the benign/surgical non-cancerous group and cancer patients (P\u3c0.0001). The IgM antibody is heat stable at room temperature for two days versus being frozen at -80°C (r2=0.97). Either serum or plasma samples may be used in the CARE Ab test (r2=0.92). The CARE Ab was almost exclusively IgM with no serum conversion to IgG in sequential measurements of patients with cancer over a six-month period. Preliminary data from patients undergoing post-operative cancer treatment showed that decreasing Ab levels revealed patients negative for residual cancer or undergoing remission, while relapsing patients show an increase in Ab levels. A return to a positive Ab level shortly after treatment is a poor prognostic sign while in advanced cancers the Ab levels may be depressed significantly