Dietary protein and chronic toxicity of 1,2-dimethylhydrazine fed to mice.

1,2-Dimethylhydrazine-HCl (DMH-2HCl) is derived from the natural toxin cycasin, and is extensively used to induce cancers in experiments with rodents. We examined the toxicity of DMH-2HCl, incorporated into purified diets varying in protein, to determine concentrations compatible with long-term survival in B6C3H1 mice. Initial studies showed single-dose oral LD50 values (95% confidence intervals) of 26 (18-32) mg DMH-2HCl/kg body weight for males, and 60 (53-65) for females. A 6-wk study was performed with diets containing 10 or 40% soybean protein with doses of 0, 11.25, 22.5, 45, 90, and 180 mg DMH-2HCl/kg diet. All mice fed the highest dose were removed from the study due to severe toxicity. Declines in food consumption and body weight occurred in both sexes, accelerated with increasing log(DMH) dose, and were substantially more severe in groups fed 10% protein. A 5-mo study was subsequently performed with male mice fed 10 or 40% protein diets containing doses of 0, 15, 30, or 45 mg DMH-2HCl/kg diet. In this longer study, dose-related declines of food intake and body weight were also more pronounced with 10% protein. Histopathologic examination of samples from 29 organs/tissues revealed hepatic changes most commonly, and these were more severe at higher DMH levels. Lesions ranged from focal centrilobular hepatocellular necrosis to severe toxic hepatitis, associated with lobular disorganization and hepatocellular hypertrophy. Frequent dose-dependent lesions were also found in kidneys, adrenals, and heart. Renal changes included focal subcapsular fibrosis with atrophy, and hyperplasia of the tubular epithelium. Adrenal cortical hypertrophy was noted at the two highest DMH doses. Focal cardiac myocytolysis was also noted at high DMH doses. Renal damage occurred only rarely in the absence of liver pathology, and adrenal hypertrophy only rarely without renal damage. Cardiac myocytolysis was found in 14% of mice without hepatic, renal, or adrenal damage, but in 62% of those with lesions in each of those organs. No evidence of gastrointestinal toxicity was observed. Hepatic, renal, and adrenal lesions were more frequent and severe in mice fed the low-protein diet. The protective effect of high protein was DMH-dose dependent. The lower doses in these studies could be used to investigate effects of diet, cocarcinogens, or chemopreventative agents on carcinogenesis resulting from chronic, low-level dietary exposure to DMH.

The implementation of a knowledge-based Pathology Hypertext under HyperCard.

A knowledge-based Hypertext of Pathology integrating videodisc-based images and computer-generated graphics with the textual cognitive information of an undergraduate pathology curriculum has been developed. The system described in this paper was implemented under HyperCard during 1988 and 1989. Three earlier versions of the system that were developed on different platforms are contrasted with the present system. Strengths, weaknesses, and future extensions of the system are enumerated. The conceptual basis and organizational principles of the knowledge base are also briefly discussed.

Computers and videodiscs in pathology education: ECLIPS as an example of one approach.

We have enumerated ways in which the evolving computer and videodisc technologies are being used in pathology education and discussed in some detail the particular use with which we are most familiar, text management. While it is probably premature to speculate as to how these technologies will ultimately affect pathology education, one recent trend--the convergence that seems to be developing between those working on expert consulting systems and those working primarily on educational applications--will probably influence this impact substantially. We believe that we are moving, from opposite directions, toward the same end result, namely, the use of machine intelligence to facilitate and augment human learning. We expect that, as the two groups come closer together, very powerful, interesting, and eminently useful educational tools will emerge. While this is occurring, we think that most would agree that one of the very urgent needs is to develop forums in which the academic and practice communities can interact with researchers and developers. With apologies to Clemenceau, computers are rapidly becoming too important to be left exclusively to computer scientists. Such forums would serve to give these communities a chance to learn what the new technologies have to offer and give developers a better idea of where these technologies can make the greatest contributions.

Immunologic cross-reactions between the specifically localizing cells (SLC) generated in response to sheep RBC and the red cells of other species.

Some of the lymphoid cells selectively incorporating radioactive thymidine 3 days after primary immunization with SRBC which are capable of specifically localizing in the lymph nodes of adoptively immunized syngeneic recipients challenged with SRBC (SLC-SRBC)2 are also capable of specifically localizing in lymph nodes challenged with the red blood cells of a varieth of other mammalian species. The one nonmammalian RBC tested, CRBC, failed to cross-react with SRBC by this parameter even qualitatively, although it remains possible that a feeble cross-reaction might be demonstrated with large enough experimental groups. As expected, the magnitude of the observed cross-reactions seems to vary inversely with the phylogenetic distance between the species, with ORBC showing the strongest cross-reaction (41 to 49%), ARBC showing the weakest (8 to 13%), and BRBC and HRBC occupying intermediate positions (13 to 20%). Cross-reaction between secondary anti-SRBC antibodies and the red cells of the other species were weak and inconstant, suggesting that the observed cross-reactions of specific localization may involve T cells more than B cells. During the course of these studies, it was also possible to verify experimentally our impression that with existing methods of selectively labeling SLC it is possible to study the phenomenon of specific localization using only one population of labeled cells, providing certain essential controls are employed.

Selective DNA synthesis by cells specifically localizing in response to xenogeneic erythrocytes.

The present studies have shown that cells capable of specific localization in response to challenge with CRBC or SRBC synthesize DNA very rapidly during the period from 2-5 days (peak 3 days) post primary immunization. This has been done by incubating the antigenically stimulated lymphoid cells with [(3)H] or [(14)C]thymidine in vitro for 45 min before adoptive transfer to syngeneic recipients. Specifically localizing cells (SLC) labeled in this way may ultimately account for up to 50% of the (3)H or (14)C present in a set of specifically challenged lymph nodes 3 days later. The data presented are consistent with the hypothesis that SLC numerically constitute only a very small fraction of the total number of recirculating lymphocytes trapped in antigenically stimulated lymph nodes, and that the demonstration of specific localization therefore depends upon selectively labeling these SLC relative to other recirculating cells. Attempts to selectively label the RNA of SLC with the precursor uridine have to date met with only very limited success.

Mechanism of graft-versus-host-induced lymphadenopathy in mice. Trapping vs. proliferation.

Graft-vs.-host (GVH)-induced lymphadenopathy of the popliteal lymph node has been produced in C57BL/6 x A/J F(1) (BAF(1)) mice by injecting A/J spleen cells into the rear footpads. By giving (51)Cr-labeled BAF(1) lymphoid cells intravenously to the hosts, 24 h before sacrifice, we have demonstrated that a large portion of the GVH-induced lymphadenopathy is due to the trapping of circuating lymphocytes in the challenged lymph nodes. Most of the remaining enlargement can be attributed to proliferation of host cells within the reacting lymph nodes. Conditions have been defined under which the weights and [(14)C]thymidine incorporation of the popliteal nodes can be plotted against the dose of injected A/J spleen cells on a double-log scale to give a linear dose-response. The popliteal lymph node GVH assay is a simple and effective means of quantitating immune reactivity to histocompatibility antigens in mice.

The immunologically specific retention of recirculating long-lived lymphocytes in lymph nodes stimulated by xenogeneic erythrocytes.

The lymph nodes of mice actively or adoptively immunized to sheep RBC and/or chicken RBC selectively retain long-lived lymphocytes after challenge with the appropriate antigen. This retention is demonstrable within 8 hr of the time of stimulation, though it probably begins even before this, and it is essentially complete within the first 24 hr. A similar selective retention is seen in nodes regional to the injection of some nonimmunogenic substances such as turpentine, but not others such as colloidal carbon or syngeneic RBC. In animals adoptively immunized to sheep and chicken RBC simultaneously, there is a preferential accumulation of the labeled long-lived lymphocytes of donors immunized to sheep RBC in lymph nodes challenged with sheep RBC, and a preferential accumulation of lymphocytes (labeled with a different radioisotope) from donors immunized to chicken RBC in lymph nodes challenged with this antigen. This immunologically specific component is demonstrable whether the antigen is given before or after adoptive immunization, suggesting that the only labeled cells capable of specific localization in this system are those cells that normally remain in the recirculating pool. In the present experiments, 31 out of 31 sets of antigenically stimulated lymph nodes have shown radiochemical evidence of immunological specificity in the distribution of donor lymphocytes between them, while corresponding sets of nonstimulated lymph nodes have shown only small random variations in the distribution of donor cells. Two different mechanisms are postulated whereby antigenic stimulation can alter the traffic of recirculating long-lived lymphocytes through stimulated lymph nodes. One affects recirculating cells of a particular immunological specificity, while the other affects recirculating cells without regard to their immunological specificity.