A surgical pathology reporting and encoding system using industry-standard hardware and software.

I describe an efficient, inexpensive, industry-standard computer system that is designed for word processing of surgical pathology reports as well as encoding in Systematized Nomenclature of Medicine (SNOMED) and data storage. This system is capable of maintaining a large database of patient information with coded diagnoses. It is designed to be utilized with readily available stock computer hardware and software without significant customization or any additional programming. The hardware is easily purchased from and can be maintained by a retail, business-oriented computer store. The system described incorporates WordPerfect for word processing and the CAP/SNOMED system for diagnosis encoding and data storage and retrieval.

Cellular effects of human leukocyte hydrolases III: inflammatory exudate and synovial fibroblasts.

Experiments were undertaken to determine whether lysosomal enzymes obtained from human polymorphonuclear leukocytes (PMN) might adversely affect the viability of human synovial fibroblasts. The effects of the PMN granule enzymes were additionally determined in the presence of an inflammatory exudate. These in vitro results indicated that as a result of these experimental conditions, the lysosomal enzymes, although present in relatively high concentration, were incapable of cell destruction and could only release cells from their growth surfaces; as could other proteases. However, even this effect was not expressed in the presence of naturally occurring inhibitors widely distributed in body fluids, such as serum; and most importantly which are also present in inflammatory exudates. This was in spite of the relatively high dilution of serum and inflammatory exudate used. So that, in effect, the relatively dilute inhibitors present in both serum and inflammatory exudates prevented the relatively concentrated lysosomal enzymes from exerting any discernible effects on either the cells, or the intercellular substance under these experimental conditions. This possibly suggests that the role of PMN granule enzymes as mediators of the cellular destruction observed in many inflammatory diseases, needs further elucidation.

Cellular effects of human leukocyte hydrolases. I. Human gingival fibroblasts.

Experiments were carried out to determine whether lysosomal enzymes had the ability to adversely affect the viability of human gingival fibroblasts. These studies were accomplished in vitro using lysosomal enzymes derived from human polymorphonuclear leukocytes. The results strongly indicated that under the conditions of these experiments, lysosomal enzymes were incapable of mediating cell destruction. In fact, the only measurable effect was to release the cells from their growth surface in a manner similar to other known proteases. Even this capability was not expressed in the presence of naturally occurring inhibitors which are widely distributed in many body fluids, such as serum and inflammatory exudates. These results suggest that PMN leukocyte granule enzymes may not contribute significantly as mediators of the cellular destruction observed in inflammatory diseases in general, and inflammatory periodontal disease in particular.

Cellular effects of human leukocyte hydrolases. II. Ultrastructural studies on fibroblasts.

Morphologic ultrastructural changes induced in human gingival fibroblasts in vitro by treatment with human neutrophil granule enzymes were studied. Surface mucopolysaccharide of the fibroblasts was examined by ruthenium red staining. The only alteration that could be identified was the removal of surface mucopolysaccharide after treatment with granule enzymes, but not when soybean trypsin inhibitor was present. When the cells were released from their growth surface they assumed the characteristics of floating cells rather than spreading fibroblasts, but were otherwise unremarkable.

Detachment of endothelial cells by polymorphonuclear leukocytes in vitro: potentiation by antibody coating and prevention by inflammatory exudate.

A widely advocated hypothesis suggests that polymorphonuclear leukocytes release enzymes at sites of inflammation which in turn cause tissue damage, especially vasculitis. To test this we isolated and grew human endothelial cells and labelled them with chromium-51. Polymorphonuclear leukocytes were separated from whole blood and added to the labelled endothelial cells. Upon incubation with the polymorphonuclear leukocytes the endothelial cells rapidly detached but were not lysed. Detachment was prevented by the addition of small amounts of inflammatory exudate. Prior coating of endothelial cells with a specific antibody accelerated detachment of the cells by polymorphonuclear leukocytes but this was still prevented by addition of inflammatory exudate in low concentration. In a separate experiment extracellular basement membrane synthesized in vitro by endothelial cells was labelled by incubating the cells with 14C-proline. Polymorphonuclear leukocytes caused release of 14C but this was again prevented by addition of either inflammatory exudate or serum. We conclude that polymorphonuclear leukocytes alone could cause vascular intimal damage in vivo if inhibitors, which are present in serum and inflammatory exudate, are excluded or overwhelmed.

Manganese inhibition of sarcoma induction by benzo[a]pyrene in rats.

The effects of Mn and Cr dusts upon carcinogenesis by polycyclic aromatic hydrocarbons were tested in male Fischer 344 rats. In Experiment I, rats were given i.m. injections of benzo[a]pyrene (BP, 1.2 mg), Mn dust (4.4 mg), and Cr dust (4.4 mg), alone, and in various combinations. By 100 weeks, sarcomas occurred at the injection site in 17/20 rats that received BP alone, versus 10/19 rats that received BP plus Mn dust (P < 0.05). The sarcoma incidences were 0/20 in 3 control groups that received the vehicle, Mn dust alone, or Cr dust alone, and 20/20 in rats that received BP plus Cr dust. In Experiment II, rats were given i.m. injections of BP (0.6 mg), 7,12-dimethylbenz[a]anthracene (DMBA, 0.6 mg), Mn dust (4.4 mg), and Cr dust (4.4 mg), alone, and in various combinations. Local sarcomas occurred in 17/20 rats that received BP alone, versus 5/17 rats that received BP plus Mn dust (P < 0.001). Sarcoma induction was not inhibited when BP was injected in the right thigh and Mn dust in the left thigh (sarcoma incidence = 16/20). The sarcoma incidences were 0/19 in a vehicle control group; 0/18 in 2 control groups that received Mn dust or Cr dust alone; 18/20 in rats that received BP plus Cr dust; 14/20 in rats that received DMBA alone; 17/19 in rats that received DMBA plus Mn dust; and 13/18 in rats that received DMBA plus Cr dust. These experiments show that Mn dust inhibits the carcinogenicity of BP when the Mn dust and BP are administered to rats by a single i.m. injection. Under identical conditions, Mn dust does not affect the carcinogenicity of DMBA; Cr dust does not affect the carcinogenicity of BP or DMBA.

Screening tests for cell-mediated immunodeficiency diseases.

As our knowledge of immunology has become more sophisticated we have had to alter our ideas of the etiology of many immune deficiency diseases. Indeed, current concepts now prevalent have led to reclassification of a number of disease entities. In order to keep our diagnostic efforts abreast of the information being generated by the extensive immunology research programs now in progress, the clinical laboratory has been required to offer a new array of sophisticated tests on a relatively routine basis. This article is intended to serve as a brief review of immunobiology and immunodeficiency diseases with an indepth coverage of specialized tests generally available at the large centers. With an understanding of the principles, procedures, and pitfalls of the tests carried out the laboratory scientist is in a better position to assist the clinician in reaching the correct diagnosis. The detailed review is concerned with methods available to separate, classify, and subclassify lymphocytes and thereby allow a categorization of immune deficiency diseases. Toward that end there is a discussion of surface markers, rosetting, mitogenic and antigenic responsiveness as well as lymphokine production. With a view to present day research tests that might eventually find their way into the armamentarium of the clinical laboratory in the future, there is brief discussion of the methods presently used to classify T-cells as helper, suppressor, or effector cells, assays of some of the lymphokines, and measurement of antibody synthesis in cell culture.

Interaction of C1s and C4. A binding phenomenon.

Addition of enzymatically active 125-I-labeled C1s (the esterase which is part of the activated complex protein of serum designated as the first component of complement or C1) to purified C4 (the naturally occurring fourth component of human serum complement) results in binding of a portion of the C1s to C4 as indicated by sucrose density gradient ultracentrifugation. Demonstration of binding requires hemolytically active C4, but not enzymatically active C1s. The latter was demonstrated by using DFP inactivated C1s as well as fragments of C1s produced by prior protease treatment of the C1s. While treatment of C1s with proteases (human leukocyte lysosomal enzymes, trypsin or plasmin) resulted in progressive inactivation of the enzymatic activity, the decline in esteratic activity occurred at a much slower rate than the decline in functional activity (inactivation of C4 in free solution). The data lead to the probable conclusion that C1s contains an enzymatic (or esteratic) site in addition to a binding site. The latter might be important for positioning a large molecule, such as C4, in order to effect proteolytic cleavage at the proper bond and hence prepare C4 to participate in the complement sequence.

Cell-detaching activity mediated by an enzyme(s) obtained from human leukocyte granules.

Granule proteins have been isolated from human peripheral blood leukocytes and their effects on intact tissue culture cells have been determined. Three hours after the addition of the mixture of granule hydrolases, both HeLa cells and human newborn fibroblasts were detached from one another and from their growth surface, but they seemed to remain viable, since cell-bound -51Cr was retained. Further studies with HeLa cells indicated that after as long as 24 hours in the presence of these enzymes the cells remained viable as judged by several independent criteria. The enzyme activity responsible for cell detachment was subject to inhibition by various protease inhibitors. Its molecular size, as determined by gel filtration, was approximately 20,000 daltons. In addition, the naturally occurring serum inhibitor of this enzyme activity was characterized as either alpha(1)-antitrypsin or another protein with similar properties.

Inability of Ni++ and Co++ to release histamine from rat peritoneal mast cells.

Ni++ and Co++ concentrations from 10 minus 3M to 10 minus 6M were added to rat peritoneal mast cells. These metal ions, at the concentrations indicated, did not cause histamine release from the mast cells, and did not inhibit the histamine release mediated by compound 48/80. On the basis of these studies, anaphylactoid edema of the rat following injection of i++ or Co++ is on a basis other than a direct effect of the m-tal ion on mast cells.