Telepathology between Richmond and Beckley Veterans Affairs Hospitals: report on the first 1000 cases.

OBJECTIVE: A telepathology connection between Richmond VAMC and Beckley VAMC using dynamic robotic telepathology to provide pathology services remotely was established. MATERIALS AND METHODS: This study reports a 14-month experience using telepathology to diagnose surgical specimens obtained from patients at the Beckley VA Medical Center and viewed in Richmond 250 miles away. Over 14 months, 2325 slides representing 1000 cases were viewed. RESULTS: Discrepancies were observed in 20 of 2325 slides, or 0.86% of the total. None of the patients, where a discrepancy was found, were adversely affected by the preliminary report given. CONCLUSIONS: This study demonstrates that telepathology is a reliable and cost-effective alternative to on-site pathology services and reviews advantages and disadvantages of the system.

Prognostic histomorphometry in colorectal adenocarcinoma.

In order to examine the use of morphometry in the prognosis of colorectal adenocarcinoma, histologic material from 30 patients was analyzed. The volume-weighted nuclear volume (Vvnuclei), mean nucleolar diameters (dnucleoli) and their standard deviation (snucleoli) were determined in each case. These variables were compared between patients who died of the disease and those who did not. Patients with disease-free survival had larger Vvnuclei than did those who died of their disease (P < .02). These data suggest that a neoplastic specimen from colorectal adenocarcinoma may contain quantifiable nuclear characteristics that may be prognostically useful.

Identification of Charcot-Leyden crystals by electron microscopy.

Observations of the ultrastructure of Charcot-Leyden crystals are sporadic in the literature. These crystals appear occasionally in clinical materials, however, and may pose diagnostic dilemmas if not correctly identified. Two cases in which unusual crystallike structures were seen on electron micrographs of specimens were evaluated for diagnostic purposes. These structures were tentatively identified as Charcot-Leyden crystals and subsequently were confirmed as such by immunoperoxidase labeling. The cases are reported together with a review of the ultrastructure, histology, immunology, and natural history of Charcot-Leyden crystals.

Specific attachment of mesenteric IgA lymphoblasts to specialized endothelium of intestinal mucosa lamina propria capillaries.

The bulk of IgA secreted in the gut is mostly contributed by locally dwelling plasma cells derived from B cells originating in the gut-associated lymphoid tissues (GALT). These IgA cells originate in Peyer's patches and recirculate, returning to the gut upon maturity. The precise mechanism of homing to secretory mucosae is to date not fully understood. It has been demonstrated, however, that specialized endothelium of small vascular spaces in peripheral nodes (PN) and endothelia of mucosal vessels are the site of receptor recognition for B and T cells. In their sojourn, IgA blasts have been shown to stop momentarily in mesenteric nodes (MN) before proceeding to their final destination, the lamina propria (LP) of the gut mucosa. They then develop into IgA-secreting plasma cells. In the present work, we show that IgA MN lymphoblasts, when compared to PN lymphoblasts, attach preferentially to LP venule and capillary endothelium, The B-cell maturation in the mesenteric lymph nodes, where IgA is the sole membrane-bound immunoglobulin, allows attachment of most of these cells. Our work suggests that the site of exit of IgA cells from the circulation are these specialized lamina propria venules and capillaries.

Nucleolar prominence as a diagnostic variable in prostatic carcinoma.

The diagnosis of well-differentiated adenocarcinoma of the prostate can be difficult on needle biopsy specimens. Nucleolar prominence has proven to be a useful diagnostic variable, but its objective evaluation has had limited study. To find nucleolar criteria that might differentiate benign from malignant conditions, we examined 41 open prostatectomy specimens, 25 of which were removed for well-differentiated adenocarcinoma and 16 of which were removed for benign prostatic hypertrophy (BPH). Four acini of carcinoma, prostatic intraepithelial neoplasia (PIN), atypical adenomatous hyperplasia (AAH), nodular hyperplasia, and normal tissue were examined. The total number of nuclei with nucleoli 3 microns or greater (N'), the fraction of nuclei with nucleoli 3 microns or greater (N'/N), and the average diameters of nucleoli 3 microns or greater (AVG) were recorded. Hyperplastic and normal areas, when compared with carcinoma, had significantly smaller N',N'/N, and AVG values (P less than 0.005). The N' and N'/N values were significantly higher in hyperplasia when compared with normal acini (P less than 0.005). In addition, N' and N'/N values in PIN were significantly greater than those in AAH (P less than 0.0001). In comparing prostates with and without carcinoma, N' and N'/N were significantly different for hyperplastic areas. In only two cancer areas and one PIN area was the N'/N ratio less than 0.31, which was the highest value for either hyperplastic or normal areas. Although AVG also were significantly different, they did not improve discrimination between the groups. We conclude that N'/N ratios are useful in diagnosing well-differentiated prostatic adenocarcinoma on small tissue samples.

A rat histone H4 gene closely associated with the testis-specific H1t gene.

A rat histone H4 gene closely associated with the testis-specific H1t gene was isolated by screening the Sargent-Bonner rat genomic library using cloned human histone genes as probes. Both the H4 gene and the H1t gene are located on a 7-kb EcoRI genomic DNA fragment. Although the deduced amino acid sequence of the rat H4 histone is identical to that of the sequence of human histone H4, the nucleotide sequence of the coding region differs significantly from the coding region of the human H4 gene. Moreover, the relative spacing between the 5'-consensus sequence elements is unique for an H4 gene. S1-nuclease protection analyses reveal that both the H4 and H1t mRNA species are present in a fraction of rat testis cells highly enriched in pachytene spermatocytes, while only the H4 mRNA species is present in a rat myeloma cell line (Y3-Ag1.2.3). During a 1-h hydroxyurea treatment of the Y3 cells, which produces a 99% inhibition of DNA synthesis, the level of this H4 mRNA drops by only 50%, indicating that the stability of this mRNA is only partially coupled with DNA synthesis.

Gut-associated lymphoid tissue as source of an IgA immune response in respiratory tissues after oral immunization and intrabronchial challenge.

Most IgA plasma cells in the digestive tract are thought to derive from gut-associated lymphoid tissue, whereas IgA plasma cells in the respiratory mucosa are thought to originate largely in bronchus-associated lymphoid tissue. However, previous work has also shown that IgA antibodies to gut antigens can be detected in immunocytes of the bronchial mucosa and in bronchial secretions after appropriate stimulation via the gut. To analyze the cellular origin of such respiratory antibodies, mice were orally immunized with ferritin for 40 days and then segregated for intrabronchial challenge as follows: one group was given saline, another group Formalin-fixed Escherichia coli as a nonspecific challenge, and a third group ferritin. Lungs and intestines from these animals were then examined by immunofluorescence for the presence of plasma cells containing particular isotypes of antibody to ferritin. Animals fed ferritin and given saline or E. coli intrabronchially showed a greater than 6-fold increment in IgA antiferritin plasma cells in the bronchial mucosa, compared to animals which had not received ferritin, whereas orally immunized animals challenged intrabronchially with ferritin showed a greater than 15-fold increase. In other experiments, ferritin-naive animals transfused with mesenteric node cells that were obtained from donors that had been orally immunized with ferritin and were already committed to IgA production showed a 4-fold or greater increase in IgA antiferritin plasma cells in respiratory mucosa after intrabronchial challenge with ferritin when compared to recipients of peripheral node cells from the same donors or to recipients of mesenteric node cells that had not been specifically boosted intrabronchially. These results suggest that immunologically specific IgA immunocytes from gut-associated lymphoid tissue can migrate to the respiratory mucosa after oral immunization, and that migration and/or local cell division are enhanced by subsequent intrabronchial challenge. In providing further evidence for interrelations between gut-associated and bronchus-associated lymphoid tissue, the findings lend added support to the overall concept of a generalized secretory immune system.

Binding and uptake of immunoglobulins by mouse mammary gland epithelial cells in hormone-treated cultures.

To study binding and uptake of immunoglobulins by mammary gland epithelial cells in culture, mouse mammary glands were taken from lactating animals. The exocrine epithelial cells were dispersed and cultured as monolayers in the presence of insulin. Insulin, progesterone, estrogen and prolactin in combination were then used during the final period of subculture. Such four-hormone treatment induced the production of differentiated products such as casein and the structural changes seen in fully developed lactating mammary gland epithelial cells. Induced cells exposed to mouse IgM, IgG, monomeric or dimeric serum-type IgA, or secretory IgA exhibited significant binding and internalization mainly of dimeric serum IgA as assessed by immunofluorescence and enzyme immunoassay.

Organ and isotype distribution of plasma cells producing specific antibody after oral immunization: evidence for a generalized secretory immune system.

Mice were induced to produce IgA antibodies against ferritin after oral immunization. Such antibodies were detected by immunofluorescence in plasma cells in the intestinal mucosa as well as in secretory sites located elsewhere, such as the lactating mammary gland, salivary gland, and respiratory tract. The observation suggested that cells immunized locally via the gut could home to distant secretory sites. To confirm this hypothesis, lymphocyte transfer studies were done with mesenteric node (MN) versus peripheral node (PN) cells from orally immunized donors into nonimmunized recipients. IgA anti-ferritin cells from MN homed to exocrine targets, whereas IgM and IgG anti-ferritin cells homed to PN. The findings overall support the concept of a generalized and interrelated secretory immune system.

Membrane-bound antibodies to bloodstream Trypanosoma cruzi in mice: strain differences in susceptibility to complement-mediated lysis.

The Y, CL and other strains of Trypanosoma cruzi display different morphological and immunological characteristics. Such observations are here extended to the interaction of bloodstream forms of different strains of T. cruzi with components of the complement system. We demonstrate that the bloodstream forms of the Y and B strains, but not those of the CL strain, are lysed by normal human serum. Lysis is mediated by combined activities of the alternative and classical complement pathways. These activities are triggered by antibodies on the surface of the parasites as shown by: (a) binding of fluorescein or radiolabelled anti-mouse immunoglobulin to the parasite's membrane and (b) the finding that bloodstream forms from lethally irradiated mice can be sensitized and rendered susceptible to complement-mediated lysis by incubation with sera from acutely infected animals. Bloodstream forms of the CL strain also bear surface immunoglobulin and sensitizing antibodies are present in the sera of mice infected with this strain. However, CL trypomastigotes from acutely infected mice fail to be lysed by human or mouse complement unless the parasites are pre-incubated with sera from chronically infected animals. The basis of the different interactions between CL and Y trypomastigotes with antibodies and the complement system, and their biological significance are discussed.

Hormonal induction of the secretory immune system in the mammary gland.

The secretory immune system of the mammary gland is undeveloped in virgin mice but becomes active at term and during lactation. This change appears to depend on migration to the mammary gland of precursors of IgA-secreting cells derived from the gut-associated lymphoid tissue, an origin which explains the specificity of milk IgA antibodies for enteric organisms. Because development of the epithelial components of the mammary gland is clearly under hormonal control, we examined the effect of mammotropic hormones on differentiation of the immune elements. Under a combined regimen of progesterone, estrogen, and prolactin, development of the glandular epithelium occurs with concomitant increments in the number of IgA-secreting plasma cells and amount of intraepithelial IgA. These increases appear to be due to enhanced capacity of the gland to attract or retain precursors of IgA plasma cells derived from gut-associated lymphoid tissue. Testosterone, which antagonizes lactation, also antagonizes development of the secretory immune system and decreases cellular trapping in the lactating gland. The ability of the gland to trap IgA immunoblasts is probably contingent upon a hormone-induced increase in receptors.

Development of the IgA system in the mammary gland.

1) Lymphoblasts in gut-associated lymphoid tissue, committed to the production of IgA, can home to the mammary glands of syngeneic mice and differentiate there into IgA-containing plasmablasts. The phenomenon is limited to near term and lactating recipients. 2) The ability of lymphocytes originating in gut-associated lymphoid tissue and sensitized to intestinal antigens to migrate to the mammary gland can account for the specificity of milk IgA toward intestinal microorganisms and the consequent passive protection offered to suckling infants. 3) The secretory immune system of the mammary gland is apparently under hormonal control since mammotropic hormones given to virgin females can induce morphological and functional characteristics seen naturally only during pregnancy and lactation. Examples are increased numbers of IgA plasma cells and the ability to trap their circulating precursors taken from mesenteric lymph nodes.

Origin of IgA-secreting plasma cells in the mammary gland.

Lymphoblasts from the mesenteric lymph nodes (MN) of mice home to the mammary glands of syngeneic recipients late in pregnancy and during lactation, and within hours of transfer most can be shown to contain IgA. Homing does not occur in virgins, in early pregnancy, or after weaning. Homing MN lymphoblasts are sensitive to antiserum to IgA plus complement, but not to other class-specific antisera. Thus, lymphoblasts in MN with the potential to home to the mammary gland are already committed to IgA synthesis and bear surface IgA before reaching their destination. These results explain observations, made by others, of specific IgA antibodies and IgA plasma cells in milk and colostrum after oral immunization. Under natural conditions it is likely that IgA precursor cells, after stimulation in the gut-associated lymphoid tissue by intestinal antigens, migrate to the mammary gland where they secrete antibodies which constitute an important defense mechanism of the newborn. In the absence of lactation, these cells probably form part of the normal traffic to the lamina propria of the small intestine.

Plasma cells and epithelial immunoglobulins in the mouse mammary gland during pregnancy and lactation.

Mammary glands of virgin, pregnant, lactating, and post-weaning CAF1 mice were studied by immunofluorescence for epithelial immunoglobulins and stromal plasma cells. In normal virgins, both females and males, only occasional plasma cells and but scanty intraepithelial immunoglobulins were present. During pregnancy and the early days of lactation, an increase in the number of plasma cells occurs, and by 1 week of lactation, there is a marked increase in the number, and most are synthesizing IgA. This increment parallels the development and proliferation of the glandular epithelium, in anatomical relation to which the plasma cells are observed. The intraepithelial content of IgA is also maximal when the glandular epithelium is most developed. These findings are consistent with a local production of the IgA in milk. Weaning, or deliberate interruption of suckling for more than 10 days results in a sheep decrease in the number of IgA plasma cells and an involution of the epithelium, whose content of immunoglobulin also declines. The changes in the secretory IgA system which occur locally during lactation are thought to be hormone-dependent.

Extragenital mixed heterologous tumor of Müllerian type arising in the cecal peritoneum: report of a case.

A mixed heterologous tumor of müllerian type arising in the cecal serosa of an elderly woman is reported. There was no history of endometriosis, and no evidence of the latter was found in the surgical specimen or at autopsy. Evidence of mesothelial origin was found microscopically in the adjacent cecal peritoneum. A review of medical literature on mixed heterologous tumors of müllerian type reveals that heretofore they have originated only in the genital system.

Secretory immunoglobulins in colonic neoplasms.

Secretory immunoglobulins are found in nongoblet columnar cells of normal intestinal epithelium. These molecules consist of a secretory component portion, which is synthesized in the columnar cells, and an immunoglobulin portion which enters the columnar cells from plasma cells in the adjacent lamina propria. In the present work, the synthesis and transport of these various subunits have been studied by immunofluorescence in benign polyps and cancers of the colon. In both the epithelium and plasma cells of benign and malignant tumors, as well as in normal tissue, IgA is the principal immunoglobulin, followed by IgM. However, when compared to normal tissue, neoplastic epithelium contains less immunoglobulin and also less secretory component; the decrement usually inversely parallels the degree of differentiation. Thus, benign polyps closely resemble normal colonic mucosa in so far as the secretory immunoglobulin system is concerned. In contrast, atypical areas of benign polyps and carcinomas exhibit greatly decreased or absent synthesis and transport of secretory IgA. Plasma cells tend to be markedly decreased in the stroma of carcinomas, suggestive of an alteration in the normal mechanism for attracting the circulating precursors of local IgA plasma cells. Whenever neoplastic epithelium contained IgA, plasma cells with IgA could be observed in the vicinity; this is in keeping with the concept of local synthesis of secretory IgA. In some instances in which local plasma cells were plentiful, neoplastic cells were deficient in secretory component and IgA, which suggested impairment of the mechanisms for transporting IgA across epithelium. The possible role of secretory component in such transport and in attracting lymphoblasts to mucous membranes is dicussed.