Immunohistochemical study of characteristics of bile duct dysplasia evaluated on the basis of expression of metastasis/invasion-related factors and p53

Background/Purpose Bile duct carcinoma still continues to have an unfavorable prognosis, despite an improved rate of curative resection and the development of surgical techniques. We evaluated the expression of matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and intercellular adhesive factors (E-cadherin, -catenin, -catenin) in cancer, dysplastic lesions, and normal (nonatypical) epithelia (glandulous ducts) in patients with bile duct carcinoma. Positivity rates for these factors were compared among the three histological types to examine the characteristics of bile duct dysplasia.Methods Among 89 patients with bile duct carcinoma resected at our hospital during the past 10 years, we studied 18 patients who concurrently had a cancerous lesion, dysplastic lesion, and normal (nonatypical) epithelia adjoining each other in excised specimens. The immunohistochemical expressions of MMPs, TIMPs, E-cadherin, -catenin, and -catenin were investigated.Results Positivity rates for MMP-9, TIMP-1, TIMP-2, membrane type (MT) 1-MMP, MT2-MMP, and E-cadherin were significantly higher in cancerous than in normal epithelium. Only the positivity rate for MT1-MMP was significantly higher in dysplasia than in normal epithelium. Positivity for MMP-related factors correlated with the degree of atypia of the bile duct epithelium. Differences between cancer and dysplasia were slightly greater than those between dysplasia and normal epithelium. Likelihood ratios between cancer and dysplasia, cancer and normal epithelium, and dysplasia and normal epithelium were higher than 5 for all metastasis-related factors and higher than 10 for most factors. This finding suggests that a normal-dysplasia-carcinoma sequence underlies the development of bile duct cancer accompanied by dysplasia.Conclusions The phenotypic characteristics of dysplasia are closer to those of normal epithelium than to those of cancerous epithelium. A normal-dysplasia-carcinoma sequence is apparently involved in the development of bile duct cancer accompanied by dysplastic cells.     

A study of dysplasia associated with bladder cancer--histopathological findings of bladder giant sections and related urinary cytology

Giant sections were prepared from 31 totally removed bladders because of transitional cell carcinoma and examined for accompanied urothelial dysplasia. Preoperatory urinary cytology related to dysplasia was also investigated. Dysplasia of the urinary bladder was morphologically defined as the urothelium with intermediate atypism between normal transitional epithelium and carcinoma in situ (CIS) of grade 3 anaplasia. Dysplasia was divided into four groups from two points of view: slight and severe dysplasia on the basis of cellular anaplasia, epithelial stratification and cellular polarity, and clear cell and compact cell dysplasia on the basis of features of dysplastic cell cytoplasm. Compact cell slight dysplasia was recognized in all specimens examined. Compact cell severe dysplasia as well as CIS was frequently accompanied with high grade papillary and non-papillary cancers, but not with low grade papillary cancer. Both slight and severe clear cell dysplasia tended to be frequently associated with high grade cancer. Because of no distinctiveness in nuclear atypia from CIS, severe dysplasia could be regarded as so called CIS of grade 2 anaplasia. From the current results, it appears likely that severe dysplasia is related to the occurrence of high grade bladder cancer. In addition, dysplastic cells exfoliated in to urine retained features of each type of original dysplasia, which were identified with a more than 70% accuracy. The result indicates that urinary cytology offers histologic aspects of coexisting dysplasia as well as bladder cancer.     

Increased c-Met tyrosine kinase expression in segmental renal dysplasia

Renal dysplasia (RD) is a disorganized development of renal parenchyma that results in a deficit of functional renal tissue. It has been suggested in the animal model that increased expression of HGF receptor, c-Met tyrosine kinase in the epithelial cells during kidney development may induce a growth of dysplastic epithelia and result in RD. The aim of this study was to investigate the immunoreactivity of c-Met tyrosine kinase in the dysplastic kidney in order to further understand the pathogenesis of RD. Specimens of dysplastic upper pole kidney were obtained from 19 patients during upper pole partial nephrectomy for non-functioning upper moiety of duplex kidney. In the dysplastic kidney, there was strong c-Met immunoreactivity in the epithelium of primitive tubules. In contrast, c-Met immunoreactivity was barely detectable in the normal kidney. Markedly increased expression of HGF receptor, c-Met tyrosine kinase in renal dysplasia suggests that HGF may be involved in the development of renal dysplasia.     

Treatment of dysplasia of the gastric mucosal epithelium]

Examined were 53 patients with dysplasia of the gastric mucosal epithelium revealed at endoscopic and histologic study. In detection of moderately pronounced dysplasia, the conservative treatment of the underlying disease with endoscopic and histologic control every 6 mos is indicated; in severe dysplasia--conservative treatment with dynamic control every 3 mos; in absence of positive dynamics of the dysplastic process within a year, or in suspected malignant transformation--the operative intervention, the volume of which is determined by spreading of the foci of dysplasia. The patients operated on for severe dysplasia of the gastric mucosal epithelium should be under continuous supervision.     

Ureteral structure and ultrastructure. Part V. The dysplastic ureter

The ureters in neonates and infants can be anatomically and functionally so severely compromised to warrant the term "dysplasia". The clinicopathologic criteria for this diagnosis are: 1) a variable degree of dilatation on the excretory urogram and/or cystogram. 2) poor or absent ureteral motility as observed on cine-fluoroscopy or at operation, 3) poor muscularization and decreased muscle/connective tissue ratio under light microscopy and 4) small and deformed muscle cells with nexuses that are markedly decreased or absent and intracellular organelles that are poorly defined under electron microscopy. Excessive collagen and ground substance are present throughout the ureter. Although these qualitative and quantitative structural abnormalities are non-specific and can occur as a result of prenatal obstruction and postnatal infection, their severity in early postnatal life and their frequent association with dysplastic kidneys suggest congenital dysplasia as their etiology.     

Characteristics of separated epithelial and stromal subfractions of prostate: II. Human prostate

Surgical samples of human benign prostatic hyperplasia tissue (BPH) were fractionated into epithelial clumps and stromal fractions, using the "optimal" tissue dissociation procedure developed for rat prostate described in the preceding report. The separated cellular fractions were compared to control unfractionated tissue (wherein extracellular secretory products had been removed) with respect to the concentrations of androgen receptor and enzyme markers on a DNA basis; cell damage was also evaluated by light and electron microscopy (EM). EM revealed extensive cell damage in epithelial clumps and stromal fractions, which had appeared normal when examined by light microscopy. Damage to the ultrastructure of individual epithelial cells present in clump fractions was very variable, involving vacuolization of the cytoplasm and condensation of nuclear chromatin in some cells, vacuolization of just the cytoplasm in other cells; only a small fraction of the cells in clumps had normal ultrastructure. Ultrastructural damage to stromal cells was much greater in fibroblasts than in muscle fibers. The cell damage observed in both subfractions of human prostate was associated with a marked degree of receptor loss. The mean decreases in the number of androgen receptors per unit DNA relative to control unfractionated tissue was 68.5 and 62.5% recovered in epithelial and stromal fractions, respectively. Measurement of various enzymes as "markers" revealed that acid phosphatase activity (per unit DNA) was associated exclusively with the epithelial clump fraction. Prolyl hydroxylase and myosin ATPase activities (per unit DNA) were restricted to the stromal fraction. The limitations of using mechanically separated subfractions of human prostate tissue for evaluation of the cellular distribution or the initial concentration of steroid receptors in human prostate tissue are discussed.     

Tubular dysplasia and carcinoma in situ: precursors of renal cell carcinoma

OBJECTIVES: To identify the dysplastic changes in tubules adjacent to or remote from renal cell carcinoma (RCC) and to assess proliferating cell nuclear antigen (PCNA) expression of normal tubule and carcinoma cells. METHODS: The study analyzed 62 kidneys with RCC that were removed by radical nephrectomy. Pathologic sections were stained with hematoxylin-eosin and evaluated for the presence of dysplasia. Sections that contained dysplasia were then stained by the avidin-biotin immunoperoxidase technique after epitope retrieval for PCNA. RESULTS: Dysplastic changes in normal kidney were identified in 14 cases (23%). Dysplastic changes were adjacent to the tumor in 10 cases. Dysplasia was adjacent to the tumor and diffuse in 6 cases (3 clear cell [CRCC], 2 chromophobe [ChRCC], 1 sarcomatoid RCC [SRCC]), adjacent to the tumor and focal in 4 cases (2 CRCC, 1 papillary RCC, 1 SRCC), remote and focal in 3 cases (1 granular RCC, 1 ChRCC, 1 SRCC), and remote and diffuse in 1 case (CRCC). The lesions represented a focus that could be defined as carcinoma in situ in 3 cases. PCNA immunostaining in dysplastic epithelia was more intense than that in normal tubules and was as intense or even more intense than that in carcinoma cells. CONCLUSIONS: Dysplasia of tubular epithelium is probably a biologic precursor of at least some RCC. Tubular dysplasia warrants further study as an important phase that will provide new insights into the pathogenesis, biologic behavior, and natural history of RCC. Its impact on the surgical management of small unilateral RCC needs to be investigated.     

Suprabasal expression of Ki-67 antigen as a marker for the presence and severity of oral epithelial dysplasia

BACKGROUND: Suprabasal expression of Ki-67 is assessed as a marker for oral dysplasia. The study involved non-neoplastic epithelium adjacent to 74 oral squamous cell carcinomas. METHODS: An immunohistochemical technique was carried out (peroxidase-antiperoxidase) with the monoclonal antibody MIB-1. Epithelial expression of Ki-67 was classified as being absent, basal, and suprabasal. The epithelium was normal in 19 cases, hyperplastic in 38 cases, and dysplastic in 37 cases. The dysplasia was slight in 20 cases, moderate in 12 cases, and severe in 5 cases. RESULTS: The results of the expression of Ki-67 were in normal epithelium, basal expression 9 cases, absent 10 cases; in hyperplastic epithelium, basal expression 18 cases, absent 20 cases; in dysplastic epithelium, basal and suprabasal expression (always jointly) 27 cases, absent 10 cases; all the severe and moderate dysplasia cases expressed suprabasal Ki-67. A significant association was observed between the presence (p <.0001) and severity (p <.007) of the dysplasia and the suprabasal expression of Ki-67.     

Paneth cell-like change of the prostate gland. A histological, immunohistochemical, and electron microscopic study.

Paneth cell-like change (PCLC) of the prostatic glandular epithelium was focally observed in one case of normal glandular epithelium, two cases of glandular and stromal hyperplasia, one case of prostatic intraepithelial neoplasia, and four cases of prostatic adenocarcinoma. The distinctive cells were characterized by bright, eosinophilic cytoplasmic granules on routine hematoxylin and eosin-stained material. The cytoplasmic granules in the benign prostatic epithelium were periodate-Schiff's procedure (PAS)-positive and diastase resistant and immunohistochemically negative for lysozyme, neuron-specific enolase, chromogranin, and serotonin. The eosinophilic granules in the prostatic intraepithelial neoplasia and adenocarcinoma cases were immunohistochemically positive for chromogranin, serotonin, and neuron-specific enolase, and negative for lysozyme. By electron microscopy the eosinophilic granules represented exocrine-like or lysosomal-like vesicles in the benign epithelium and neuro-endocrine granules in the malignant epithelium. The lesion represents a prostatic epithelial PCLC rather than a Paneth cell metaplasia. PCLC is the common histological manifestation of two different phenomena: (a) a PAS-positive and diastase-resistant eosinophilic cytoplasmic granular change in benign prostatic epithelium, and (b) endocrine differentiation with neuroendocrine granules in dysplastic and malignant prostatic epithelia. The importance of recognizing PCLC lies in its differentiation from other possible prostatic cytoplasmic inclusions.     

Renal prognosis in women with hereditary nephritis

Renal prognosis cannot be easily predicted in females with hereditary nephritis (HN). Thirty-six women with persistent urinary abnormalities, belonging to 24 families with progressive HN, were studied. Renal biopsy specimens were available in 23 patients and were studied by light and electron microscopy (EM). Nine women (group I) progressed to early renal failure, at 35 yrs of age or less. Five women (group II) progressed to late renal failure at 45 yrs of age or more. In contrast, 22 patients have so far normal renal function, and 14 of these (group IV) range from 31 to 62 yrs of age. Diffuse glomerular basement membrane (GBM) thickening was found by EM in 6 of 7 cases of group I, whereas it was found in no patient of group IV. The following features are suggestive of progressive nephritis in females: gross hematuria in childhood, nephrotic syndrome, and diffuse GBM thickening by EM. In contrast, family history of HN without nerve deafness, normal or nearly normal kidney on repeat biopsy, and thin or normal GBM by EM are suggestive of less or nonprogressive renal disease. Further follow-up is needed to assess the prognostic significance of these features.     

Clinicopathological characteristics of renal light chain deposition disease coexisted with cast nephropathy

Objective To investigate the clinicopathological characteristics of renal light chain deposition disease coexisted with cast nephropathy (LCDD&LCN). Methods Patients with LCDD&LCN (n=10), isolated LCDD (I-LCDD, n=21) and isolated LCN(I-LCN, n=17) diagnosed by renal biopsy in Peking University First Hospital from January 1, 2000 to March 31, 2018 were enrolled, and all cases were examined by light microscopy, immunofluorescence (IF) (including light chain) and electron microscopy (EM). The semi-quantitative evaluation of the main features of renal pathology was performed. The clinical manifestations and pathological features were reviewed and compared. Results LCDD&LCN was more prevalent in middle-aged males. Nine patients showed acute renal insufficiency with small molecular proteinuria (97.1%) and microscopic hematuria. The hematologic diseases included 9 patients of multiple myeloma. The type of monoclonal light chain in serum and urine by immunofixation electrophoresis showed λ dominant (5/8). By light microscopy, glomerular lesions presented with mild mesangial proliferation in most patients, and only one of them displayed mesangial nodular sclerosis. At the same time, acute tubular injury with light chain casts was the prominent feature, and the clinical manifestations and histological features of LCDD&LCN were similar to that of I-LCN. IF revealed linear staining of monoclonal light chain along the glomerular basement membrane (GBM), tubular basement membrane (TBM) and Bowman's capsule, and also positive in tubular casts. By electron microscopy, diffuse powder-like or granular electron-dense deposits located in the inner side of the GBM, the outer layer of the TBM, renal interstitium and arteriolar walls were observed. Conclusions Patients with LCDD&LCN manifest as acute renal insufficiency, and the majority have multiple myeloma. The pathology of LCDD&LCN possesses the features of both I-LCDD and I-LCN. The IF stain of light chains(κ, λ) and ultrastructural examination by electron microscopy are the inevitable methods for the diagnosis of LCDD&LCN.     

The neurotoxicity of epidural hyaluronic acid in rabbits: a light and electron microscopic examination

Because hyaluronic acid (HA) has an antiinflammatory effect and prevents and/or reduces tissue adhesion, we believed it possible that epidurally-administered HA during epidural adhesiolysis procedures could alleviate pain in patients with chronic lower back pain. Therefore, we performed this pre-clinical trial evaluation of epidurally-administered HA neurotoxicity by light microscopy (LM) and electron microscopy (EM) in rabbits. Twenty rabbits were randomly divided into two groups, a normal saline (NS) group (n = 10) and a HA group (n = 10). Saline (0.2 mL/kg of 0.9% solution) and the same volume of HA were injected into the epidural space. No rabbits showed any sensory-motor or behavior change during the 3-wk period, except for one rabbit in the NS group that showed decreased appetite, activity, and weight loss. By LM, two rabbits in the NS group showed abnormal findings considered to be the result of trauma and infection associated with epidural catheterization. EM findings showed no significant neurotoxic findings in either group. In conclusion, epidurally-administered HA did not cause neurotoxicity in rabbits. IMPLICATIONS: We performed a pre-clinical trial evaluation on the neurotoxicity of hyaluronic acid administered epidurally by light microscopy and electron microscopy in rabbits. Epidurally-administered hyaluronic acid did not produce any sign of neurotoxicity in rabbits.     

Quantitative analysis of optical coherence tomography and histopathology images of normal and dysplastic oral mucosal tissues

Selecting the most representative site for biopsy is crucial in establishing a definitive diagnosis of oral epithelial dysplasia. The current process involves clinical examination that can be subjective and prone to sampling errors. The aim of this study was therefore to investigate the use of optical coherence tomography (OCT) for differentiation of normal and dysplastic oral epithelial samples, with a view to developing an objective and reproducible approach for biopsy site selection. Biopsy samples from patients with fibro-epithelial polyps (n = 13), mild dysplasia (n = 2), and moderate/severe dysplasia (n = 4) were scanned at 5-μm intervals using an OCT microscope and subsequently processed and stained with hematoxylin and eosin (H&E). Epithelial differentiation was measured from the rate of change (gradient) of the backscattered light intensity in the OCT signal as a function of depth. This parameter is directly related to the density of optical scattering from the cell nuclei. OCT images of normal oral epithelium showed a clear delineation of the mucosal layers observed in the matching histology. However, OCT images of oral dysplasia did not clearly identify the individual mucosal layers because of the increased density of abnormal cell nuclei, which impeded light penetration. Quantitative analysis on 2D-OCT and histology images differentiated dysplasia from normal control samples. Similar analysis on 3D-OCT datasets resulted in the reclassification of biopsy samples into the normal/mild and moderate/severe groups. Quantitative differentiation of normal and dysplastic lesions using OCT offers a non-invasive objective approach for localizing the most representative site to biopsy, particularly in oral lesions with similar clinical features.     

Intra-epithelial mast cells in human airway epithelium: evidence for smoking-induced changes in their frequency.

A morphological and quantitative study was performed on the respiratory epithelium of human distal airways in 20 lungs removed at operation for the treatment of carcinoma. Using 1 micron araldite sections stained with Toluidine Blue for light microscopy, with further electron microscopic study where required, we have described mast cells in the epithelium of bronchioles, terminal bronchioles, and respiratory bronchioles. Examination of these cells at both microscopic levels demonstrates many of the typical features of mast cells--namely, metachromatic granules on staining with toluidine blue at light microscopic level, surface microvillous processes, and numerous dense intracytoplasmic granules with a whorled substructure at electron microscopy. Furthermore, we have identified cells devoid of granular material which may be recognised as mast cells by virtue of their nuclear and cell surface characteristics--the occurrence of a spectrum of change between those cells devoid of granular material and those with full granule content; and a few cells in which we found both typical dense, whorled granules and large lucent vacuoles devoid of such material. Quantitation of all respiratory epithelial cells by light microscopy shows that mast cells comprise 0-2% of the total population. A higher proportion of mast cells was found in the epithelium of smokers than in non-smokers. Because of the presence of many epithelial mast cells devoid of granule content, we suggest that the use of traditional 4 microns paraffin sections stained with thiazine dyes to study mast cells would lead to a marked underestimation of the epithelial population.     

Effects of dilation with a balloon catheter on the endothelium of the internal thoracic artery.

Controlled dilation of the internal thoracic artery with a balloon catheter has been reported to effectively treat intraoperative arterial spasm. It has been shown in laboratory animals that dilation of the internal thoracic artery at prescribed shear force levels will not cause intimal damage. Using scanning electron and light microscopy, we have examined the effects of calibrated balloon dilation on the endothelium of the human internal thoracic artery. In 10 patients with bilateral internal thoracic artery grafting, the artery was dilated with a Fogarty IMAG balloon catheter (Baxter Healthcare Corporation, Edwards Division, Santa Ana, Calif.) that was withdrawn at tensions of 20 or 30 gm. Arterial segments and nondilated control specimens were prepared for scanning electron microscopy. The intimal surface of each internal thoracic artery was evaluated by assigning a score (from 0 to 3) to 10 examined scanning electron microscopy fields; subsequently the arterial tissue was viewed by light microscopy with paraffin-embedded sections stained for elastic tissue. Arteries were obtained from three additional patients so that the microscopic appearance of the arteries could be observed after rough manipulation or removal of the balloon without shearing. The results of this study are as follows: (1) By scanning electron microscopy, dilated internal thoracic arteries yielded consistently higher scores than the control arteries, reflecting severe, tension-dependent alterations of the endothelium, which included marked desquamation of endothelial cells, with extensive areas of complete denudation and pronounced attachment of platelets to these areas; (2) endothelial injury occurred by inflation alone, without shearing by the inflated balloon; (3) by light microscopy, the internal thoracic arteries showed (a) fenestrations of the internal elastic lamina with occasional transmigration of smooth muscle cells through these gaps and (b) foci of intimal thickening without overt atherosclerotic lesions. We conclude that the endothelium of human internal thoracic arteries is highly vulnerable to balloon dilation, which can severely injure the intimal surface. For this reason we prefer not to include this procedure in our protocol for preparing the internal thoracic artery.     

Immunocytochemical demonstration of prostatic acid phosphatase: different secretion kinetics between normal, hyperplastic and neoplastic prostates

The ultrastructural localization and distribution of prostatic specific acid phosphatase in normal, hyperplastic and neoplastic human prostates was studied by immunocytochemical methods. In normal or hyperplastic prostates, the localization of prostatic specific acid phosphatase was uniformly observed at the apical portion of the glandular epithelium of apical cells under the light microscope. Electron microscopy revealed prostatic specific acid phosphatase localized in the microvilli lining prostatic and vesicular bodies of apical cells. Occasionally the limiting membrane of the blebs and vesicles extruded into the glandular lumen and were stained positively. Light microscopic examination of neoplastic prostates revealed a more intense and uniform staining of tumor cells and the glandular epithelium of well differentiated adenocarcinomas, whereas less intense and more variable staining was seen in neoplastic cells of moderately or poorly differentiated adenocarcinomas. Furthermore, under electron microscopic study, prostatic specific acid phosphatase granules were uniformly and intensely condensed in intracytoplasmic vacuoles in well differentiated adenocarcinomas, whereas in moderately or poorly differentiated adenocarcinomas 2 types of staining were observed. Among neoplastic cells, positive granules with less intensity were found between collagen fibers as well as adjacent to the endothelium of the stromal capillaries in anaplastic tissue.     

Fatty acid synthase expression is increased in neoplastic lesions of the oral tongue.

BACKGROUND: Fatty acid synthase (FASE) is required for fatty acid synthesis. Elevated levels of FASE have been observed in a variety of malignancies. METHODS: We examined the expression of FASE in 56 primary squamous cell carcinomas (SCC) of the tongue using immunohistochemistry (IHC) with a monoclonal antibody to FASE. RESULTS: Immunoreactivity was low in histologically normal epithelium (0.42 +/- .07, n = 43), moderate in mildly dysplastic epithelium (1.41 +/- 11, n = 40), and strong in SCC of the tongue (1.64 +/- 10, n = 50). Both mild dysplasia and SCC stained more strongly than histologically normal epithelium (p<0.00001). Well-differentiated tumors showed increased immunoreactivity when compared to less well-differentiated tumors (p=0.044). Decreased overall survival was observed among patients with tumors with low immunoreactivity (p = 0.04). CONCLUSIONS: Increased expression of FASE in dysplasia and squamous carcinomas of the oral tongue may be an indicator of both differentiation and early neoplastic change. FASE expression may be useful diagnostically, prognostically, and as a potential target for therapy.