An approach for high sensitivity detection of breast cancer by analysis of changes in structure of the cytoplasmic matrix of lymphocytes specifically induced by a specific breast tumour antigen (MUC-l/SEC).

An alternative procedure for detection breast cancer was examined based on the observation that lymphocytes re-exposed in vitro to antigenic stimulation will change their intracellular structuredness as measured by polarization of fluorescent light emitted by fluorescein labeled cells (SCM test). The specific antigen MUC-l/SEC was used to elicit such response in lymphocytes of patients with and without breast cancer. Eighty-five samples with breast cancer were tested, of which 72 were correctly diagnosed. Of the 41 controls, 35 were correctly identified as healthy subjects. The sensitivity of the test was 85% and the specificity was 81%. These results suggest a possible valuable method for screening and early detection of breast cancer. The clinical importance of this procedure lies in the ability to screen high-risk populations with higher specificity and sensitivity than any combinations of currently available procedures for breast cancer detection.

Early detection of malignant process in benign lesions of breast tumor by measurements of changes in structuredness of cytoplasmic matrix in circulating lymphocytes (SCM test) reinduced in vitro by specific tumor antigen.

Early detection is crucial for successful treatment of all types of cancer. The specificity and sensitivity of the current methods vary from 50% to 80%. The use of specific tumor antigens and cytometric technology has resulted in the development of a new procedure for the early detection of breast tumors. This new method is reported. The test utilizes static cytometry, which records polarization and intensity changes in fluorescent light emitted from each individual lymphocyte obtained from tumor-bearing patients stimulated by the relevant specific tumor antigen. Using MUC-1/SEC as the specific antigen, we detected breast tumors with 85% specificity and 81% sensitivity in 137 breast tumor-bearing women. A significant linear correlation was found between the SCM test and the conventional classification of relative risk for breast cancer in benign lesions, suggesting that this is a precise method that could be used in mass screening for early detection of breast cancer.

Preliminary evaluation of epidural morphine for treatment of heroin withdrawal.

OBJECTIVE: To evaluate the efficacy of epidural morphine in treating heroin withdrawal in patients who failed to detoxify by the other methods. DESIGN: Prospective study. SETTING: Department of Psychiatry of a general hospital. PATIENTS: 8 ASA physical status I patients, aged 26 to 42 years, not having concurrent diseases requiring medication, and who had previously failed other methods of detoxification. INTERVENTIONS: Epidural catheters were inserted at the L(3)-L(4) interspace. Bolus injections of morphine sulfate, 3.0 mg in normal saline, were administered epidurally at 24-hour intervals. Treatment continued for 10 to 12 days. MEASUREMENTS: Withdrawal symptoms, such as mydriasis, insomnia, rhinorrhea, arthralgia, muscular pain, tooth pain, vomiting, diarrhea, dysphoria, and drug craving were monitored. MAIN RESULTS: Withdrawal symptoms ceased within 10 days. Withdrawal symptoms were diminished or entirely abolished by the treatment, and no patient requested to drop out of the program. Discontinuation of the epidural injections did not cause relapse of withdrawal. All patients reported that withdrawal with epidural morphine was considerably easier compared to other methods that they had previously experienced. CONCLUSIONS: A preliminary evaluation of epidural morphine in addicts that failed previous detoxifications showed high effectiveness of this method in reducing withdrawal symptoms.

Comparison of the effects of dopaminergic and serotonergic activity in the CNS on the activity of the immune system.

This work demonstrates that the natural killing function of the innate immune system is affected in psychiatric disorders related primarily to serotonergic pathways in the CNS rather than to psychiatric disorders which involve mainly dopaminergic pathways. Only depressive patients demonstrated low natural killer (NK) cell activity, which is inversely correlated to the intensity of depression and could be reversed by serotonin selective re-uptake inhibitors concomitant with clinical improvement. This phenomenon is absent in Parkinson's and schizophrenic patients, in whom no reduction in NK activity was observed. Also, no effect on NK activity could be demonstrated following the specific respective treatments by dopamine (D2) blockers or agonists.

An approach for high sensitivity detection of prostate cancer by analysis of changes in structuredness of the cytoplasmic matrix of lymphocytes specifically induced by PSA-ACT.

PURPOSE: An alternative procedure for detection of prostate cancer was examined based on the observation that cells reexposed in vitro to antigenic or mitogenic stimulation will change their intracellular structuredness as measured by polarization of fluorescent light emitted by labeled cells (SCM test). MATERIALS AND METHODS: Lymphocytes derived from patients bearing a nonmalignant prostate tumor and healthy individuals were exposed to PSA-ACT, PHA, and MUC-1. RESULTS: Of sixty-five patients with prostate carcinoma (CaP), sixty-two were correctly diagnosed by the test. Of the eighty males in the control group, five were incorrectly diagnosed as having the disease and seventy-five were correctly diagnosed as healthy subjects. The sensitivity of the test was 96.8%. The specificity was 91.1%. The BPH (Benign Prostatic Hyperplasia) control group exhibited a sensitivity of 9.38%, but the specificity was 91.1%. Similar percentages for specificity and sensitivity were observed in the NRT (Non-Relevant Tumor) control group. CONCLUSIONS: The results shown here indicate the possibility of a different use of PSA-ACT for detection of prostate cancer with high specificity and sensitivity.

A rabbit model for evaluation of spinal anesthesia: chronic cannulation of the subarachnoid space.

A rabbit model for evaluation of spinal anesthesia is presented. Chronic cannulation of the subarachnoid space was performed in 44 rabbits using the translumbar approach. An autopsy was performed 24 h after the operation on four of the animals. Intrathecal injections of methylene blue did not reveal any leakage from the spinal space. X-ray examination performed on the second and 30th days after the implantation indicated free spread of the injected solution in the subarachnoid space without any obstruction. Repeated injections of four identical doses of bupivacaine at 3-day intervals showed reproducible pharmacologic effects. Administration of different doses of the anesthetic produced a clear dose-response relationship. The relative activity of the anesthetic agents was found to be identical to that previously obtained in humans. No significant complications after the implantation have been recorded. We suggest the current model as an additional appropriate tool for the investigation of spinal anesthesia.

Combination of a long-acting delivery system for luteinizing hormone-releasing hormone agonist with Novantrone chemotherapy: increased efficacy in the rat prostate cancer model.

The combination of hormonal treatment based on a long-acting delivery system for the agonist [6-D-tryptophan]luteinizing hormone-releasing hormone ([D-Trp6]-LH-RH) with the chemotherapeutic agent Novantrone (mitoxantrone dihydrochloride) was studied in the Dunning R3327H rat prostate cancer model. Microcapsules of [D-Trp6]-LH-RH formulated from poly(DL-lactide-co-glycolide) and calculated to release a controlled dose of 25 micrograms/day were injected intramuscularly once a month. Novantrone (0.25 mg/kg) was injected intravenously once every 3 weeks. Three separate experiments were carried out. When the therapy was started 45 days after transplantation and continued for 70 days, tumor volume in the presence of the microcapsules (966 +/- 219 mm3) or Novantrone (3606 +/- 785 mm3) given alone was significantly decreased compared to controls (14,476 +/- 3045 mm3). However, the combination of microcapsules and Novantrone caused a greater inhibition of tumor growth (189 +/- 31 mm3) than the single agents. Similar effects were seen when the percent increase in tumor volume was examined. Tumor volume increased 10,527 +/- 1803% for the control group. The inhibition of growth caused by the [D-Trp6]LH-RH microcapsules alone (672 +/- 153% increase in volume) was again greater than that caused by Novantrone alone (2722 +/- 421% increase). The combination of the two agents was again the most effective, resulting in an increase in tumor volume of only 105 +/- 29%. Control tumors weighed 30.0 +/- 6.5 g. Tumor weights were much less in the groups treated with either microcapsules (3.28 +/- 0.69 g) or Novantrone (19.53 +/- 3.3 g) alone. The lowest tumor weights after 70 days of treatment were obtained in the group that received the combination of [D-Trp6]LH-RH microcapsules and Novantrone (1.02 +/- 0.2 g). Testes and ventral prostate weights were significantly diminished by the administration of microcapsules of [D-Trp6]LH-RH alone or in combination with Novantrone. In both of these groups, luteinizing hormone and prolactin levels were reduced and serum testosterone was suppressed to undetectable levels. Similar results were obtained in two other experiments in which the duration of treatment was 60 or 105 days. These results suggest that the overall response could reflect the inhibition of proliferation of hormone-independent cancer cells by Novantrone in addition to the suppressive effect of [D-Trp6]LH-RH on the growth of androgen-dependent tumor cells. The administration of Novantrone in combination with microcapsules of [D-Trp6]LH-RH might produce a better clinical response than LH-RH analog alone in patients with advanced prostate carcinoma.

Enrichment of in vitro and in vivo immunologic activity of purified fractions of calf thymic hormone.

Thymus humoral factor (THF), a thymus hormone which participates in the processes leading to acquisition of immunocompetence of lymphoid cells has been isolated in our laboratory by a stepwise gel filtration through various Sephadex columns. THF so isolated appears to be a polypeptide of 3,000 mol wt which contains approximately 30 amino acid residues. Here we have tested the biological activity of THF fractions of successive degrees of purity upon lymphoid cells from both intact and neonatally thymectomized mice. The lymphoid cell populations were treated with the various THF fractions by in vitro incubation for a short time and by repeated injection in vivo. The treated cells evidenced increased ability to react in the graft-versus-host assay in vivo and in mixed lymphocyte cultures in vitro concomitantly with the rise of intracellular cAMP. On the other hand no activity whatsoever was shown by any of the control materials tested. These bioassays permitted isolation of fractions progressively more active than the original crude dialyzate of thymus extract tested. Thus the active peptide component of THF eluted from DEAE Sephadex A-25 column was estimated to be 2 X 10(4)-fold more active than the crude dialyzate of thymus extract which served as a starting material.

Effect of thymic humoral factor on intracellular levels of cyclic AMP in human peripheral blood lymphocytes.

The exposure of human peripheral blood lymphocytes to thymus hormone causes an increase in the intracellular cyclic AMP level in these cells. Studies of the thymus-hormone-induced changes in cellular cyclic AMP levels revealed that cord blood lymphocytes contain significantly more target cells for the hormone than adult peripheral blood lymphocytes. The data obtained also suggest that the interaction between the hormone and its target cell is not readily reversible and that only a certain proportion of target cells in a given population of lymphocytes can be activated by a constant and limited amount of thymus humoral factor.

Enhancement of DNA synthesis and cAMP content of mouse thymocytes by mediator(s) derived from adherent cells.

Supernatants of adherent mouse peritoneal exudate cells or human mononuclear cells were used as the source of lymphocyte activation factor (LAF). LAF was found to potentiate the effect of mitogens such as PHA and Con A on DNA synthesis by mouse thymocytes. However, LAF also was capable of reducing vigorous thymosyte reactions to Con A. Thus, LAF usually enhanced the effect of PHA on DNA synthesis by BALB/c thymocytes to a relatively greater degree than that of Con A. This change in the ratio of Con A to PHA response of thymocytes suggests that LAF can serve as a regulator of thymocyte DNA synthesis. Moreover, in the presence of LAF, allogeneic thymocytes developed the ability to have bidirectional mixed thymocyte reactions. Exposure to LAF not only improved the ability of parental thymocytes to act as responder cells, but, in addition, led to increased stimulatory activity of F1 thymocytes, presumably by promoting the differentiation of stimulator cells. These indications that LAF affected differentiation were investigated further by studying its effect on the cAMP content of thymocytes. LAF stimulated significant immediate but transient elevations of intracellular cAMP and adenylate cyclase activity in thymocyte membranes. In contrast, the mitogens themselves failed to elevate or to influence the effect of LAF on the content of intracellular cAMP of thymocytes. Furthermore, the potentiating effect of LAF on mitogen-induced thymocyte DNA synthesis at times was enhanced by exogenous cGMP, carbachol, or imidazole. These findings suggest that LAF, through its stimulation of cAMP levels in thymocytes may in turn promote thymocytes to differentiate sufficiently to become competent to proliferative in response to mitogens.

The control exerted by thymic hormone (THF) on cellular cAMP levels and immune reactivity of spleen cells in the MLC assay.

We have shown previously that THF induces immunocompetence in lymphoid cells through the control of cellular cAMP levels and DNA synthesis. By the use of a one-way MLC assay it was now shown that the exposure of spleen cells to THF is accompanied by an increase, in the proliferative response of the responding cells. These effects are mediated by regulation of cellular levels of cAMP during the course of the MLC response. Thus, substances which modify cellular levels of cAMP such as DBcAMP, theophylline, and imidazole change the reactivity of cells exposed to THF in the MLC assay. It was also found here that cellular cAMP levels are critical in controlling the reactivity of normal spleen cells in the absence of THF. It is suggested that THF increases the number of competent cells in the spleen cell population capable of responding to antigenic stimulation in the MLC assay.

Intracellular events involved in the induction of immune competence in lymphoid cells by a thymus humoral factor.

A humoral factor extracted from calf thymus (THF) restores the immunocompetence of spleen cells from neonatally thymectomized (NTx) mice to induce an in vitro graft-vs-host (GVH) response. This acquistion of immunocompetence consists of a series of biochemical events, the first of which involves an obligatory rapid increase in adenyl cyclase acitivity and in intracellular levels of cyclic AMP. Protein synthesis occurs as a further step in the events leading to induction of immunocompetence by THF and could be blocked by cycloheximide with the consequent abolishment of the immunocompetence of the lymphoid cells tested. The induction of competence by THF is accompanied by a simultaneous reduction in DNA synthesis resulting from the increase in intracellular cyclic AMP levels. These steps have been studied in the absence of antigenic stimulation which is not required for the induction of competence by THF. Spleen extracts prepared by a similar procedure as THF were found to be devoid of the properties described above.

Hormone-like activity of a thymus humoral factor on the induction of immune competence in lymphoid cells.

Experiments reported here were performed to understand the mechanism by which THF increases the immunocompetence of spleen cells from NTx mice. Dibutyryl cAMP or substances which increase intracellular levels of cAMP in lymphocytes such as Poly(A:U), theophylline, or PGE(2) were shown to mimic the effect of THF and confer reactivity in an in vitro GvH response to spleen cells from NTx mice. Flufenamic acid, an antagonist to PGE(2), was shown to inhibit the induction of competence by this substance. It was found that THF induces competence by activating membranal adenyl cyclase which leads to a rise in intracellular cAMP in thymus-derived cells only. These biochemical changes occur before antigenic stimulation and are unrelated to antigenic challenge. These findings indicate that THF exerts its effect via cAMP and are in agreement with the concepts which permit to classify THF as a thymus hormone.