Scanning electron microscopic and immunohistochemical studies of pelvic endometriosis

The pathogenesis of pelvic endometriosis has been studied by using scanning electron and light microscopy, observing the surface structure of bluish lesions obtained from 26 patients during laparotomy. Paraffin sections included another 17 tissue samples of endometriosis, based on immunohistochemical responses to epithelial membrane antigen, keratin and vimentin. Ultrastructurally, the surface epithelial cells could not be detected in 13 out of 17 pelvic peritoneal endometriosis samples. In one case in which the surface peritoneal cells were seen histologically to dip into the subperitoneal stroma, many surface peritoneal infoldings were observed, and ciliated cells were detected at the edge of these infoldings. Ovarian endometriosis was composed of three types of cells, none of which had any cilia. These findings were observed in continuity with adjacent normal mesothelial cells. No characteristic structure of the endometrial surface was observed for the bluish lesion, but the gland surface of endometriosis located in the subperitoneal stroma initially had ciliated cells. The immunoreactions in both the columnar mesothelial cells with surface peritoneal infoldings and the glands of endometriotic tissues were similar to those of normal endometrial glands, but different from those of normal mesothelial cells. Pelvic endometriosis might originate by a process of metaplasia from the pelvic peritoneum.     

Sulphatide antigen in islets of Langerhans and in diabetic glomeruli, and anti-sulphatide antibodies in type 1 diabetes mellitus

Clinical coincidence between diabetes and neurological disorders, and sharing of antigen determinants between islets of Langerhans and neural tissue, has been suggested. Sulphatide is a neural epitope which can be visualized with a monoclonal antibody Sulph I. Different tissues were examined by immunohistological methods. Sulphatide and anti-sulphatide antibodies were determined by thin-layer chromatographic techniques. IgG was isolated using protein A columns. A specific staining by Sulph I was found of rat islets, assigned to the secretory granules of both alpha and beta cells. No labelling of the exocrine tissue or other body tissues was seen, except for nerve and kidney structures. The latter showed staining of the distal tubules and, in addition, but only in the diabetic kidney, of glomeruli located in the subendothelial area in the capillary loops and the mesangial space. Sera from 38% of 40 spontaneously diabetic BB rats displayed anti-sulphatide antibodies, mainly IgG, whereas all 30 control Lewis rats were negative. Most recently we have demonstrated anti-sulphatide antibodies in 88% of 57 patients with newly diagnosed Type 1 diabetes (titres of > 1:400); all 135 healthy control persons were negative. The sulphatide antibody reactivity was present in the IgG fractions of the patients' sera. Thus, sulphatide is demonstrated in islets of Langerhans and in kidney related to the diabetic lesion, and, furthermore, anti-sulphatide antibodies exist in Type 1 diabetes mellitus.     

Idiopathic membranous glomerulonephritis associated with diabetes mellitus: light, immunofluorescence and electron microscopic study

The present study described 3 patients with idiopathic membranous glomerulonephritis associated with diabetes mellitus. Clinical characteristics of the 3 patients contrasted with diabetic glomerulosclerosis in the following manner: absence of diabetic retinopathy and neuropathy, and presence of nephrotic syndrome associated with relatively short duration of diabetes mellitus. Renal histology showed the characteristic changes of membranous glomerulonephritis along with those of diabetic glomerulosclerosis. Immunofluorescent studies demonstrated a granular pattern of IgG and C3 deposits along the glomerular capillary wall. Electron microscopic study also demonstrated thickening of glomerular basement membrane and increase of mesangial matrix as well as the presence of electron-dense deposits primarily in the subepithelial and mesangial areas.     

Transplantation of islets of Langerhans in patients with insulin-requiring diabetes mellitus undergoing orthotopic liver transplantation--the Miami experience

Most patients with cirrhosis of the liver have detectable insulin resistance. In 60-80% of patients with cirrhosis, impaired glucose tolerance can be uncovered; approximately 20% of these patients eventually develop overt diabetes. Theoretically, insulin resistance and glucose intolerance could be improved or reversed by orthotopic liver transplantation alone or in association with a simultaneous transplant of pancreatic islet cells from the same donor. To investigate these possibilities we initiated a pilot study of simultaneous liver and pancreatic islet cell transplantation in seven patients with diabetes and liver cirrhosis. Donor bone marrow cells were also infused to enhance the acceptance of the grafts. Seven patients who received only orthotopic liver transplantation and donor bone marrow cells were used as historical controls. The preliminary results of this pilot trial suggest that islet cell transplantation in conjunction with orthotopic liver transplantation improves glucose metabolism in patients with liver cirrhosis in association with reduced insulin requirements and HbA1c levels. These results were evident in spite of pre- and post-transplant basal C-peptide levels that were unchanged. Further evaluation of the effects of orthotopic liver transplantation with or without islet cell transplantation will require a randomized prospective trial including accurate metabolic evaluation with the euglycemic insulin clamp technique.     

Regional differences in the pineal gland of the cotton rat, Sigmodon hispidus: light microscopic, electron microscopic, and immunohistochemical observations

Light microscopic, electron microscopic and immunohistochemical observations of the various portions of the pineal gland of the cotton rat (Sigmodon hispidus) were made. The volume of the proximal half occupied about 30% of the whole organ, and pinealocytes were slightly smaller in size in the proximal portion than elsewhere. The distal and intermediate portions contained few interstitial cells and numerous astrocytes, but the proximal portion lacked interstitial cells and had more abundant astrocytes than elsewhere. Astrocytes, which were immunoreactive for glial fibrillary acidic protein, mainly lined the pericapillary spaces in the distal and intermediate portions, but in the proximal portion these cells often surrounded isolated or groups of pinealocytes. In the distal and intermediate portions, abundant sympathetic fibers and less numerous non-sympathetic, peptidergic fibers were mainly localized in the pericapillary spaces; these fibers were sparsely distributed in the parenchyma close to interstitial cells or astrocytes. In the proximal portion, non-sympathetic fibers were scarce and sympathetic fibers were distributed abundantly and almost exclusively in the parenchyma. Most of the sympathetic fibers were adjacent to astrocytes and, occasionally, made specialized contact with them. Fenestrae in the capillary endothelium were numerous in the distal portion but absent in the proximal portion. Thus, marked differences in structure existed between the distal and proximal portions of the pineal gland of the cotton rat suggesting that both portions are functionally dissimilar. In addition, the present study indicates that the proximal portion of the cotton rat was well developed and showed morphological features similar to the deeply situated pineal glands of other mammals.     

Identification of orbital lymphatics: enzyme histochemical light microscopic and electron microscopic studies

The presence of orbital lymphatics in the primate model is demonstrated using light and electron microscopic enzyme histochemistry. In addition, strictly morphological definitions of lymphatics, such as discontinuous basal lamina, thin and irregular walls, anchoring filaments, and attenuated endothelial cell cytoplasm, were applied. This study confirmed the presence of conjunctival lymphatics reported by others. It also clearly demonstrated the presence of orbital arachnoid and lacrimal gland lymphatics that have not been previously described. A few areas of the extraocular muscles and connective tissue at the orbital apex also showed evidence of the presence of lymphatic vessels. Additional work is needed to define the nature and extent of orbital lymphatics as well as their connection to the extraorbital lymphatic system.     

Secretory meningioma: clinical, histologic, and immunohistochemical findings in 31 cases

BACKGROUND: Secretory meningioma is a rare histologic variant characterized by a unique epithelial differentiation of meningothelial cells resulting in the production of hyaline inclusions. Most previous reports have presented single case observations. The authors selected 31 cases for a clinicopathologic study to characterize this type of tumor further. METHODS: Clinical data were compiled and the extent of peritumoral edema was assessed from preoperative computed tomography or magnetic resonance imaging scans. Preparations of surgical specimens of all tumors were studied after both conventional histologic and immunohistochemical preparations were made. Immunostaining was performed by either the avidin-biotin complex method or the alkaline phosphatase-antialkaline phosphatase method using 22 primary antibodies. RESULTS: In the tumor collection used in this study, secretory meningiomas represented 3% of meningiomas. The female-to-male ratio was 9:1. Most tumors were located at the sphenoid ridge or at the frontal convexity, and recurrences were not observed. Eighty-four percent of tumors presented with slight to marked peritumoral edema. The MIB-1 staining index showed a mean of 3.8%. Inclusions and surrounding cells consistently expressed epithelial membrane antigen, cytokeratins, carcinoembryonic antigen, and carbohydrate antigen 19-9. In decreasing frequency, they also contained alpha1-antitrypsin, immunoglobulin (Ig)A, alpha1-antichymotrypsin, IgM, and IgG. Cells positive for vimentin and S-100 did not contain inclusions. All tumors were positive for progesterone receptors. Macrophages were stained with antibodies to factor XIIIa, human leukocyte antigen-DR, and alpha1-antitrypsin. In 64% of cases, tumor vessels lacked expression of glucose transporter protein 1. CONCLUSIONS: The classification of secretory meningioma as a distinct variant has been justified on clinical, histologic, and immunohistochemical grounds. The unique epithelial features call attention to the broad spectrum of differentiation properties found in meningiomas.     

Vibrio cholerae adherence and colonization in experimental cholera: electron microscopic studies

Colonization of the intestinal epithelium by Vibrio cholerae was examined in two model systems, in ligated ileal loops of adult rabbits and in the patent gut of infant rabbits, using both scanning and transmission electron microscopy. Time studies in the adult model showed a lag period of up to 1 h before the attachment of significant numbers of the vibrios. The bacteria appeared initially in small patches on the sides of the villi, predominantly along the transverse furrows. The number of adherent bacteria steadily increased, reaching a maximum between 4 and 7 h, when a dense mat of bacteria several layers thick covered much of the villi. After this time there was a rapid decline in the number of V. cholerae bound. By 12 to 16 h only a few bacteria could be seen on the surface of the villi, which had a rough, patchy appearance at these later times. Globular protrusions, with vibrios attached, may play a role in the clearance of bacteria. Colonization and clearance in the patent intestine of the infant rabbit occurred much as in the adult model. However, the bacteria adhered more uniformly and there was no lag in attachment. In both models the majority of bacteria were aligned horizontally with the epithelial surface, but some were attached in an end-on manner, with their flagella extending into the lumen. The bacteria adhered via their surface coats directly to the tips of the microvilli, except for a few vibrios that were partly embedded into the brush border. Some changes in the microvilli occurred as a consequence of the bacterial attachment.     

Rhabdomyoma of the tunica vaginalis of the testis: a histologic, immunohistochemical, and ultrastructural study

A case of an unusual tumor of skeletal muscle origin is described. The tumor was located in the tunica vaginalis of the testis in a 19-year-old man. Histologic examination showed a proliferation of elongated or round cells, with clearly discernible cross striations, surrounded by abundant mature connective tissue, consistent with genital rhabdomyoma. Immunohistochemical and electron microscopic features supported this diagnosis. Rhabdomyoma must be considered in the differential diagnosis of paratesticular tumors.     

Localization of cholecystokinin-like peptide in neuroendocrine cells of mammalian lungs: a light and electron microscopic immunohistochemical study

Mixing of starved cells of complementary mating types of Tetrahymena thermophila induces shortening of their longitudinal length within 10 min of mixing. This early morphogenetic transformation in preconjugant sexual interaction (costimulation period) was named "costimulation-induced rounding" (CIR). CIR is the earliest morphological change that has ever been found in the costimulation period and differs from "synchronous rounding" in the vegetative cell cycle, because CIR cells are still able to form food vacuoles, while cells in synchronous rounding do not have this ability. When sexual cell-to-cell collisions between two mating types were hampered by unidirectional stirring for 20 min after mixing of the two mating types, both CIR and conjugation were delayed by 20 min. When secreted materials needed for the onset of costimulation were removed by washing the cells with 10 mM Tris-HCl, pH 7.4, before mixing the two mating types, both CIR and conjugation were delayed by about 30 min. CIR-like rounding was not induced by cell-free medium either from the opposite mating type or from mixed costimulated cells. These results indicated that CIR is induced when cells are activated to form conjugating pairs by cell-to-cell collisions between complementary mating types in the presence of secreted molecules.     

Rhabdomyosarcoma mimicking acute leukemia in an adult: report of a case with histologic, flow cytometric, cytogenetic, immunohistochemical, and ultrastructural studies

OBJECTIVES: To measure ground reaction force variables during lameness resulting from impaired tibial nerve function and to determine whether these variables changed significantly as recovery progressed. ANIMALS: 11 healthy Greyhounds of either sex, weighing between 22 and 39 kg. PROCEDURE: On 3 consecutive days before surgery, ground reaction forces were measured by force platform gait analysis at the trot. In dogs under general anesthesia, the left tibial nerve was crushed proximal to innervation of the gastrocnemius muscle. Gait analyses were repeated on days 8 to 10, 28 to 30, 43 to 45, 58 to 60, and 90 to 92 after surgery. Ground reaction force variables and stance time were compared among the 3-day clusters. RESULTS: 10 days after surgery, all dogs had weight-bearing lameness attributed to paralysis of muscles in the caudal compartment of the crus. Nerve regeneration resulted in functional recovery within 3 months. Decreases in vertical force were significant 10 days after surgery; thereafter, changes reflected gradual return of load bearing, with the most marked improvement between 10 and 45 days. At 90 days, vertical force variables were within 3% of presurgical values. Stance time for the left hind limb was significantly longer at 10, 30, and 45 days after surgery and was seen in all dogs, but returned to within 1 % of preoperative stance time at 90 days. The effect of tibial nerve dysfunction on braking or propulsive force was not significant. CONCLUSIONS AND CLINICAL RELEVANCE: The significant changes in vertical ground reaction forces in hind limbs of dogs during lameness that resulted from impaired tibial nerve function are detectable, as is response during recovery, by use of force platform analysis.     

Transplantation of islets of Langerhans in type 1 diabetes. Three cases treated in Sweden to date

The article consists in a report of the first three cases of human islet transplantation to have been carried out in Sweden. Cadaveric pancreatic glands were harvested and flown to the islet transplantation laboratory at the University of Giessen in Germany. After isolation, the islets were returned to Huddinge Hospital in Stockholm. The recipients were diabetic renal transplant patients, who received 5,000-8,000 islets by intraportal injection. Initially blood glucose levels were stabilised and HbA1c levels normalised, but no patient became insulin independent. After a few months serum C-peptide levels diminished, and after 6-10 months were undetectable. Islet function loss is probably to be explained by rejection and cytomegalvirus infection. In future cases new improved immunosuppressive protocols will be implemented.     

Multihormone-producing islet cell tumor of the pancreas associated with somatostatin-immunoreactive amyloid: immunohistochemical and immunoelectron microscopic studies

OBJECTIVE: To assess the association between high but nondiabetic blood glucose levels and the risk of death from all causes, coronary heart disease (CHD), cardiovascular disease, and neoplasms. RESEARCH DESIGN AND METHODS: We studied the 20-year mortality of non-diabetic, working men, age 44-55 years, in three European cohorts known as the Whitehall Study (n = 10,025), the Paris Prospective Study (n = 6,629), and the Helsinki Policeman Study (n = 631). These men were identified by their 2-h glucose levels following an oral glucose tolerance test and by the absence of a prior diagnosis of diabetes. As the protocol for the oral glucose tolerance test and methods for measuring glucose differed between studies, mortality was analyzed according to the percentiles of the 2-h and fasting glucose distributions, using the Cox's proportional hazards model. RESULTS: Men in the upper 20% of the 2-h glucose distributions and those in the upper 2.5% for fasting glucose had a significantly higher risk of all-cause mortality in comparison with men in the lower 80% of these distributions, with age-adjusted hazard ratios of 1.6 (95% CI 1.4-1.9) and 2.0 (1.6-2.6) for the upper 2.5%. For death from cardiovascular and CHD, men in the upper 2.5% of the 2-h and fasting glucose distributions were at higher risk, with age-adjusted hazard ratios for CHD of 1.8 (1.4-2.4) and 2.7 (1.7-4.4), respectively. CONCLUSIONS: If early intervention aimed at lowering blood glucose concentrations can be shown to reduce mortality, it may be justified to lower the levels of both 2-h and fasting glucose, which define diabetes.     

Isolation of the pig islets of Langerhans: evaluation of in vitro and in vivo function

Pig islets are considered the best alternative to human islets in the treatment of insulin-dependent diabetes. Pigs could represent a potential islet donor for xenotransplantation in humans because of the close similarity between human and porcine insulin and the theoretically unlimited availability of porcine pancreas. From November 1991 to January 1997 we performed 221 pig islet isolations from 3 pig sources: group 1: minipigs (age 9-18 months) and white pigs (3-8 months), group 2: large white pigs (5-8 months), group 3: large white pigs (12-24 months). Islets were isolated according to a semi-automated method using enzymatic digestion and purification through discontinuous Euro-Ficoll gradients. The pancreases were surgically removed in our laboratory for group 1, while pancreases from groups 2 and 3 were removed at the slaughterhouse with an average warm ischemia time of 15 minutes. In vitro islet function was assessed by static incubations and perifusions, and in vivo islet function by transplantation under the kidney capsule of nude diabetic mice. The results were as follows: [table: see text] Insulin secretion increased twofold after in vitro glucose stimulation. We obtained restoration of euglycemia in diabetic mice which survived > 3 months after the graft and returned to diabetes after nephrectomy. This study shows that our isolated pig islets are viable and functional in vitro and in vivo after transplantation.     

Scanning and transmission electron microscopic studies of lesional epidermis in herpes zoster

The epidermal skin lesions of herpes zoster were studied by scanning (SEM) and transmission electron microscopy (TEM). When erythematous lesions were observed by TEM, many of the infected keratinocytes showed evidence of cell degeneration, being characterized by swollen nuclei, disappearance of desmosomes, and widening of intercellular spaces. Macrophages and/or lymphocytes migrated through the intercellular spaces between degenerated keratinocytes. In the vesicular lesions, SEM and TEM showed some infiltrating neutrophils, directly adhering to the virus-infected keratinocytes, with swollen nuclei and irregularly clumped chromatin. In some specimens, balloon-degenerated keratinocytes were observed in the cavity. In the pustular stage, ruptured keratinocytes and numerous neutrophils were observed in the reticular-degenerated epidermal tissue. These results suggest that, in herpes zoster, the epidermal damage may be due, at least in part, to cell-mediated host immunity as well as to the cytopathic effect of varicella-zoster virus.     

Subependymal giant cell astrocytoma: clinical, histologic and immunohistochemical characteristic of 3 cases

The role of 5-hydroxytryptamine (5-HT) in neural reflexes regulating secretion was examined in muscle-stripped segments of guinea-pig colon set up in modified flux chambers. A 15-microL pulse of 5-HT (100 microM) to the mucosal bath (1.5 mL), which was continuously perfused, evoked an increase in short-circuit current (Isc). The 5-HT-induced increase in Isc was inhibited by tetrodotoxin, N-acetyl-5-hydroxytryptophyl-5-hydroxytryptophan amide (5-HTP-DP), GR82334 and atropine, but not by tropisetron. 5-HTP-DP reduced the response to a 5-HT pulse over the concentration range of 1 nM to 1 microM. The Isc response to a 5-HT pulse was unaffected by the cyclooxygenase inhibitor, piroxicam. This contrasted with a reduction in the Isc response to mucosal stroking with a brush by piroxicam. The results suggest that a 5-HT pulse, like mucosal stroking, activates a secretory reflex that includes tachykinin and cholinergic neurons but, unlike mucosal stroking, does not release prostaglandins.     

Quantitative autoradiographic light- and electron microscopic studies on the retinohypothalamic connections in the rat

Light microscopic autoradiography performed subsequent to intraocular injection of 3H-leucine revealed silver grains (SG) above axons of the optic tract which could be followed into the ventral and caudal portion of the suprachiasmatic nuclei (SCN) and above the contralateral anterior hypothalamic nucleus (AHN). By high resolution photometric measurement and computer processing the labelled areas were analysed, thus yielding statistical data of the relative grain distribution. The highest SG density was found in the ventrolateral part of both SCN (SCvl), confirming earlier reports concerning retinohypothalamic connections. That area exhibiting a cytoarchitecture different from the remaining nucleus was traversed, however, by numerous labelled axons. In the caudal part of both SCN a specific projection field of retinal fibres could be located. Here, almost no traversing fibres contribute to the rather circumscribed marked area. In the ventral part of the contralateral AHN, diffuse labelling well above background levels could be observed. Distinction between bypassing and terminating fibres within the SCvl could not be made using light microscopy. Analysis of SG distribution of the SCvl with electron microscopic autoradiography revealed a specific localization of SG within presynaptic terminals containing clear vesicles and pale mitochondria.