Distinct Cell Borders Research Articles

Effects of intraocular epinephrine on the corneal endothelium of rabbits.

Epinephrine is frequently used in the phacoemulsification to dilate pupils. To determine the effects of different concentration of epinephrine on the corneal endothelial cells, twenty-eight rabbit eyes were equally divided into four groups. Solutions, which contained normal saline, 1:1000 epinephrine, 1:5000 epinephrine and 1:10000 epinephrine respectively, were injected into the anterior chambers of the eyes of four groups of rabbits. In vivo morphological changes of corneal endothelium and changes of thickness were checked with specular microscopy. In vitro morphological evaluation of corneal endothelium was observed in excised corneal buttons stained with alizarin red with trypan blue, and with scanning electron microscopy. Our results showed that there was no significant difference in cell density and corneal thickness among the four groups. Alizarin red with trypan blue stain and SEM exam revealed smooth and distinct cell borders of endothelial cells in each group. Intracameral injection of epinephrine does not produce toxic effect on corneal endothelial cells in rabbits.

The influence of age on the cytology of the liver in healthy dogs.

Liver cytology was evaluated in 28 healthy dogs 1-14 years of age with normal liver structure and function. Smears were stained with May-Grünwald-Giemsa. Hepatocytes had distinct cell borders, and cells did not overlap. Cells with two nuclei and cells with intranuclear crystalloid structures were observed regularly. Cytoplasm contained small numbers of vacuoles characteristic of glycogen and lipid and small amounts of pigment consistent with ceroid or bile. Nuclei were uniform. Small numbers of biliary epithelial cells were seen in most samples. Lymphocytes and neutrophils occurred in small numbers, with lipocytes, mast cells, fibrocytes, mesothelial cells, eosinophils, and Kupffer macrophages seen less frequently. Mean parenchymal cell sizes were significantly greater in older dogs, but no age-related differences were observed in nuclear size. Older dogs also had a significantly increased number of nuclei per cell. There were more neutrophils in young and old dogs than in middle-aged dogs.

Histopathologic changes of thymoma preoperatively treated with corticosteroids.

Preoperative treatment of thymoma in advanced stages with corticosteroids may reduce the size of the tumor, but no precise histologic evaluation has been performed. We examined the histopathologic features of pretreatment biopsy and posttreatment surgical specimens of eleven cases of thymoma with such treatment to see the changes of the histologic subtypes based on Muller-Hermelink classification. All specimens were also assessed immunohistochemically for MIB-1 labeling and apoptotic cells to verify the effectiveness of this pretreatment. Seven tumors clinically diminished in size after the treatment with corticosteroids. Fungal infection occurred in three cases postoperatively. The histology of mixed thymomas (two cases) was converted to that of medullary thymoma. Predominantly cortical thymomas (four cases) and cortical thymomas (three cases) changed to show similar histologic features; both became epithelial-rich thymoma with large polygonal tumor cells having indistinct cell borders. In contrast, two well-differentiated thymic carcinomas showed at surgery more prominent squamoid appearance with distinct cell borders. The apoptotic indices of epithelial cells were increased (P = 0.001), and the MIB-1 indices tended to be decreased with corticosteroid treatment. These results suggest that there may be a histogenetic relationship between medullary and mixed thymomas and also between predominantly cortical and cortical thymomas. Corticosteroids may cause degenerative changes in the epithelial cells and lymphocytes and, in thymomas in advanced stages, corticosteroid pretreatment may be warranted, although attention should be paid to infection after surgery.

Benign Clear Cell Tumor of the Lung

A 49-year-old woman presented with a solitary pulmonary nodule in the right lung. The tumor was well circumscribed and showed a reddish brown cut surface. It showed a diffuse growth pattern of polygonal cells with clear abundant cytoplasmand distinct cell border around thin-walled vascular spaces and sinusoid-type vessels. Based on morphological findings, the patient was diagnosed as having benign clear cell tumor of the lung. Silver impregnation and PAM stains showed fine reticular fibers continuously surrounding the vessels and individual neoplastic clear cells. Strong immunostaining for type IV collagen was observed surrounding all the individual clear cells. A few clear cells were positive for HMB-45 and NCAM 123C3 ( CD56 ). Electron microscopically, clear cells had numerous membrane-bound glycogen granules and a large amount of non—membrane-bound glycogen. The neoplastic cells were surrounded by external lamina and cytoplasmic processes. The neoplastic clear cells showed morphologic features seen in pericytes, melanocytic cells, and neuroendocrine cells.

Three-Dimensional Cytomorphology and Its Relationship with Clinical Stage in Fine Needle Aspiration Biopsy of Papillary Thyroid Carcinoma

To elucidate three-dimensional (3-D) cytomorphology and its relationship with clinical stage in fine needle aspiration biopsy (FNAB) of papillary thyroid carcinoma (PTC). FNAB was performed in tumors from 34 patients with PTC. The aspirate was stained and observed under a light microscope (LM). The aspirate was also fixed, dehydrated, critical point dried, spattered with gold ions and observed with a scanning electron microscope (SEM). Findings under SEM were correlated with the presentation under LM and also with clinical stage. As a control, FNAB was also performed on a normal part of the thyroid in six patients. Under SEM, normal thyroid cells appeared as round, scattered cells or sheets with a uniform honeycomb cellular arrangement. In PTC, cell sheets with uniform nuclear morphology under LM usually showed a uniform cell arrangement and with distinct cell borders under SEM. Cell sheets with nuclear pleomorphism under LM usually showed a more-disorganized arrangement and with indistinct cell borders under SEM. PTC at advanced clinical stages usually presented with a disorganized cell arrangement, indistinct cell borders and loss of microvilli. 3-D cytomorphology in FNAB of PTC was related to clinical stage and might be a prognostic factor for PTC.

Angiomatoid neuroendocrine carcinoma of the thymus: Report of a distinctive morphological variant of neuroendocrine tumor of the thymus resembling a vascular neoplasm

Three cases of primary thymic neuroendocrine tumors characterized by prominent angiomatoid features that resembled a vascular neoplasm are presented. The patients were all men between 52 and 59 years of age who presented with chest pain and shortness of breath attributable to a large anterior mediastinal mass. The lesions ranged in size from 6 cm to 15 cm in greatest diameter, and were grossly soft and well circumscribed, but not encapsulated. The cut surface was remarkable for multiple blood-filled cyst-like spaces admixed with focal solid, hemorrhagic areas. Histologically, the tumors contained multiple cystically dilated spaces filled with blood which imparted the lesion with a striking angiomatoid appearance. The walls of the cysts were lined by a monotonous proliferation of round to oval cells with distinct cell borders, round central nuclei, and abundant eosinophilic cytoplasm. Mitotic activity was present in all cases and varied from 3 to 8 mitoses per 10 high-power fields. Immunohistochemical studies performed in two cases showed positivity of the tumor cells for keratin, Leu 7, and synaptophysin, and focal chromogranin positivity in one. Follow-up information obtained in two patients showed that both had died of tumor 4 and 8 years after initial diagnosis. The present cases show an unusual morphological appearance of thymic neuroendocrine tumors that may be mistaken for a vascular neoplasm. Immunohistochemical stains may be of importance in such instances in arriving at the correct diagnosis.

Oncocytoid artifact of the parotid gland: A newly reported artifact

Electrocautery can induce significant alterations in the connective tissues and epithelium of specimens removed for diagnostic or therapeutic purposes. When electrocautery is used during parotid surgery, it can cause an oncocytoid artifact. The alterations described in this article are enlarged, tightly packed serous acinar cells with coarse to granular eosinophilic cytoplasm, distinct cell borders, and round basal nuclei that on cursory microscopic examination resemble oncocytes with respect to morphology. These changes are seen in conjunction with other, more recognized changes secondary to electrocautery and are believed to occur as a consequence of the electrothermal discharge. On the basis of our findings, this artifact is common in parotid surgical specimens and was misdiagnosed as benign oncocytic lesions in 5 cases.

Detection of t(11;22)(q24;q12) translocation of Ewing's sarcoma in paraffin embedded tissue by nested reverse transcription-polymerase chain reaction.

Ewing's sarcoma is a poorly characterized malignant tumor with a relatively uniform histologic appearance, made up of densely packed small cells with round to oval nuclei, without distinct cell borders and without any structural differentiation. Often the diagnosis has to be made by exclusion. Recently, it has been made possible to identify characteristic chromosomal rearrangements associated with certain solid tumors. More than 85% of Ewing's sarcoma and peripheral neuroectodermal tumor present a specific t(11;22)(q24;q12) balanced translocation, resulting in the production of a novel chimerical EWS/FLI-1 message. Using oligonucleotide primers derived from EWS and FLI-1 complementary DNAs, we were able to use reverse transcription-polymerase chain reaction (RT-PCR) as a diagnostic tool. The described nested RT-PCR method as another supportive diagnostic method enables pathologists to differentiate small blue cell tumors not only to make correct diagnosis but also to investigate retrospective archival tumor samples, using formalin fixed paraffin embedded tissue as a source of RNA.

Renal Cell Carcinoma: Diagnosis Based on Metastatic Manifestations

Renal Cell Carcinoma: Diagnosis Based on Metastatic Manifestations

Occurrence of monocytoid B lymphocytes in lymph nodes of patients treated by chemotherapy

Occurrence of monocytoi B lymphocytes (MBL) in the lymph nodes of patients receiving preoperative chemotherapy for cancer was examined and compared to lymph nodes in controls who had not received chemotherapy. Number of patients receiving and not receiving preoperative chemotherapy were 3 and 10 cases in ovarian cancer, 7 and 11 in testicular cancer, and 22 and 8 in lung cancer, respectively. Chemotherapeutic agents for ovarian, testicular, and lung cancer consisted of cisplatin, Adriamycin, and cyclophosphamide; cisplatin, vinblastine, and bleomycin; and cisplatin, vindesin, and mitomycin, respectively. MBL were defined morphologically as having abundant pale cytoplasm with distinct cell borders and small nucleus. Immunohistochemistry revealed a B-cell nature of these cells, i.e., CD20+ and/or MB-1+ together with negative reactivity for antibodies for T lymphocytes (CD43, CD45RO, OPD4) and macrophages (KP-1, PGM-1). Monocytoid cells in two cases showed a positive reactivity for CD43 together with CD20. The occurrence rate of MBL in patients with ovarian, lung, and testicular cancer receiving and not receiving chemotherapy was 67% (2/3) and 10% (1/10), 59% (13/22) and 75% (6/8), and 43% (3/7) and 9% (1/11), respectively. The occurrence rate in the total patients receiving chemotherapy (56%) was significantly higher than for those not receiving chemotherapy (28%) (P < 0.05). These findings suggest that chemotherapy-induced depressed immune function is causative for the occurrence of MBL in the lymph nodes. MBL might be found more frequently in nodes from patients who have received chemotherapy in certain settings.

A scanning electron-microscopic analysis of the morphology of equine lower lip sinus hair.

Sinus hairs, which are tactile organs of most mammals, are differentiated into cavernous and sinus types. The horse has the cavernous type. Horse lower lips were perfused in diluted Karnovsky's fixative and sinus hair processed for scanning electron microscopy. Anastomosing trabeculae of different thickness and shape originate from the internal connective tissue layer and extend to the external layer, thus forming a dense meshwork suspending the hair root like a net. However, many of the distal trabeculae do not reach the external wall of the blood sinus but end in a finger-like knob within the sinus, thus sharing some features of the sinus type. The sinus wall and trabeculae are completely covered by polygonal endothelial cells showing characteristically protruding nuclei, sparse microvilli and distinct cell borders. Endothelial cells on the trabeculae are more elongated. Openings of small blood vessels were found exclusively on the internal wall, indicating that blood exchange occurs via the internal wall only and that turnover of blood may be minimal. The supposed significance and function of the finger-like trabeculae in the distal part of the blood sinus lying close to the majority of nerve terminals is discussed.

Incidence of latent infection of Epstein–Barr virus in lung cancers—an analysis of EBER1 expression in lung cancers by in situ hybridization

To evaluate the presence of Epstein-Barr virus (EBV) in lung cancers of Japanese patients, 81 lung cancers were examined using a highly sensitive in situ hybridization (ISH) method, employing an antisense oligonucleotide probe for EBV-encoded small nuclear RNA-1 (EBER). EBER1 expression was demonstrated in one poorly differentiated squamous cell carcinoma associated with marked lymphoid stroma (PDSCC-LS), two well differentiated adenocarcinomas, and two moderately differentiated squamous cell carcinomas, but was not detectable in other lung cancers, including small cell carcinomas. Unlike lymphoepithelioma-like undifferentiated carcinoma (LELC) of the lung, the PDSCC-LS consisted of poorly differentiated cells with distinct cell borders and nuclei with a coarse chromatin pattern and some prominent nucleoli. Most of the cancer cells expressed intense EBER1 signals. Although small to moderate numbers of cells positive for EBER1 were present in two adenocarcinomas and two squamous cell carcinomas, EBER1 signals varied in intensity and number in these four cases. Although polymerase chain reaction (PCR) and Southern blot hybridization with a 32P-labelled probe internal to the primers were conducted to detect the EBV genome in 24 lung cancers, including five EBER1-positive cases, the genome was found to be positive in the five cases with EBER1-positive staining, including the PDSCC-LS, two adenocarcinomas and two squamous cell carcinomas, but not in the other cases. This study indicates that the morphological features of EBV-associated lung cancers are not restricted to the typical LELC type.

Monocytoid B-cell lymphoma arising in extranodal organs

Six cases of monocytoid B-cell lymphoma (MBCL) developing in extranodal sites (thyroid, three; stomach, two; rectum, one) are described. Patients were all women aged 46 to 65 years (median, 53 years). Three patients with thyroid lymphoma presented with an increasing goiter, two with positive serum antithyroid antibodies, and one patient had chronic lymphocytic thyroiditis (CLTH). The histologic type of surgically resected specimens was a diffuse proliferation of atypical lymphoid cells with monocytoid appearance, i.e., abundant pale cytoplasm with distinct cell border and small reniform nucleus. These cells were CD20+, 22+, 24-, 9- showing their B-cell origin. The monoclonal nature of the proliferating cells was confirmed by restricted expression of immunoglobulin (Ig) light chain and/or gene rearrangement study in three cases. Two cases of thyroid lymphoma in which the monoclonality could not be confirmed had histologic appearances characteristic of malignant lymphoma. All three patients with gastric or rectal lymphoma had reactive lymphoid hyperplasia (RLH) near the tumors. These findings showed presence of MBCL in the extranodal sites with invariable coexistence with lymphoid follicles formed by CLTH in thyroid or RLH in stomach and rectum.

Fine‐Needle Aspiration Biopsy Diagnosis of Endodermal Sinus Tumor: Histologic and Ultrastructural Correlations

Fine-needle aspiration biopsies from five patients with endodermal sinus tumors (ESTs) were reviewed, and the findings were correlated with histologic and ultrastructural appearances. In the aspiration smears, two types of tumor cells were seen, forming clusters of variable sizes. Type A cells had distinct cell borders, and their cytoplasm contained only occasional vacuoles. Type B cells had ill-defined cell borders and formed syncytial clusters; their cytoplasm was characterized by large numbers of rounded vacuoles. The background contained patches of mucoid material and macrophages with foamy cytoplasm. Eosinophilic hyaline cytoplasmic bodies and irregular deposits of intercellular basement membrane-like material were recognized easily in aspiration smears. These features correlated well with histologic and ultrastructural appearances. The significance of these findings in the fine-needle aspiration biopsy diagnosis of EST and its distinction from other germ-cell and non-germ-cell tumors is discussed.

Isolation and characterization of rabbit ovarian surface epithelium, granulosa cells, and peritoneal mesothelium in primary culture

Mammalian ovarian surface epithelial (OSE) cells and peritoneal mesothelial (PM) cells have a common embryologic origin, yet certain morphologic and histochemical characteristics are different in the adult. In this study, a two-step culture method was developed to examine the characteristics of these two cell types in vitro. OSE, PM, and ovarian granulosa (GC) cells were isolated from estrous rabbits and cultured for 6 d in 5% serum-supplemented D-valine medium (to inhibit fibroblast growth), then incubated for a further 2 d in serum-free McCoy's 5A medium. This study showed that rabbit OSE and PM cells in vitro maintained certain in vivo morphologic characteristics; OSE cells exhibited distinct cell borders and abundant microvilli of homogeneous size and shape, whereas PM cells were characterized by obscure cell borders and abundant microvilli of heterogeneous form. GC in vitro exhibited overlapping cell borders and sparse microvilli of homogeneous structure. This study showed for the first time that cultured rabbit OSE and PM cells, but not GC, contain distinct filaments of cytokeratin 18. In addition, rabbit OSE cells and GC, but not PM cells, contained 17 beta-hydroxysteroid dehydrogenase. However, only GC contained delta 5-3 beta hydroxysteroid dehydrogenase. OSE, PM, and GC maintained their ultrastructural and histochemical characteristics in serum-free medium. These results suggest that rabbit OSE cells in vitro could be distinguished from PM cells by histochemical and ultrastructural differences. Furthermore, because these characteristics were not altered in serum-free medium, the two-step culture method will be valuable in further hormonal studies of these cells in vitro.

Stromal macrophage-histiocytes in Hodgkin's disease. Their relation to fever.

Morphologic variations of Concanavalin A-binding histiocytes were studied in biopsy specimens of 140 untreated patients with Hodgkin's disease (72 asymptomatic, and 68 with constitutional symptoms). Fever was the most common symptom, present in 57 of the 68 patients. Three morphologic types of stromal histiocytes were recognized: medium-sized cells similar to those seen in reactive follicles, characterized by distinct cytoplasm and cell borders, and uniform nuclei (Type A); damaged-appearing Type A cells marked by rarefied or ragged cytoplasm, disrupted or indistinct cell borders, and varying sized nuclei (Type B); and large spindling or stellate cells (Type C). Type A cells were predominant in 52 patients; Type B cells in 51; Type C cells in seven; and Type A cells were mixed with Type B in 30. Fever was present in one of 52 patients (1.9%) with Type A predominance; 43 of 51 (84.3%) with Type B cell predominance; none of seven (0%) with Type C predominance; and 13 of 30 (43.3%) with mixed Type A and B cells. Logistic regression analysis of the data showed that the association of fever with Type B cell predominance was highly significant, and was not attributable to the known association of fever with other variables. Morphologic evidence suggests that fever in Hodgkin's disease may be a clinical manifestation of damaged macrophage-histiocytes rather than an acute-phase response of inflammatory or immune reaction.

Short-term estrogen test for cytodiagnosis in postmenopausal women.

A short-term estrogen test was used to obtain a correct cytologic diagnosis in 73 patients with an equivocal atrophic cellular pattern. Overestimation in cytodiagnosis was markedly eliminated, and the correct cytologic diagnosis was made in 81% of the cell samples after the estrogen test. The purpose of this study was not only to confirm the usefulness of the short-term estrogen test, but also to observe the cytomorphologic changes before and after the test. Thick cytoplasm, vacuoles in the cytoplasm, and a distinct cell border increased, while the amorphous chromatin pattern was eliminated after the estrogen test. Maturation of atypical cells in cases of dysplasia, carcinoma in situ, and invasive squamous-cell carcinoma was investigated before and after the estrogen test. Although malignant cells were not influenced by estrogen, maturation of dysplastic cells was induced after the test.

A Case Combined Germ Cell Tumor in Testic

Primary germ cell tumor of the testis is, rare, which occupies 1 to 2% of all reported malignant male neoplasms. Combined primary germ ceil tumor of the testis composed of embryonal carcinoma and seminoma is more rare tumor. The authors experienced a case of 50-year old male who presented with painless enlargement of right testicle. He has had radical orchiectomy and diagnosed by pathologically as combined germ cell tumor of testis. The testicle is measuring 180 gm in weight and cm in dimension, and almostly replaced by tumor mass. Grossly the tumor is rubbery solid smooth tumor mass, with variegated cut surface with geographically outlined diffuse necrotic area. Histologically the tumor is composed of two components of tumor, which are solid growth pattern of large round to polyhedral cells with clear or granular cytoplasm and distinct cell border, and anastomosing glandular and papillary arrangement of anaplastic epithelial cells. The former corresponds to seminoma, and the latter to embryonal carcinoma. Each tumor lobule is separated by abundant fibrous stroma.

Quantitative SEM analysis of injury to the endothelium of rabbit aorta and carotid artery during experimental atherosclerosis

The luminal surface of the aorta and the carotid artery in normal and cholesterol-fed rabbits (3 weeks, 6 weeks, and 8 months of alimentary hypercholesterolemia) was studied by scanning electron microscopy (SEM). To study endothelial injury the vessels were perfused and stained under physiological pressure. The frequency of large and small endothelial defects was determined per surface unit of endothelium in the normal and experimental groups of rabbits. Loss of endothelial cells was regarded as a large defect, argyrophilic cells, craters, and stomata were regarded as small ones. It was found that the percentage of regions without endothelial cells was similar in both control rabbits and in rabbits with experimental atherosclerosis (0.005-0.04% of the total surface examined). The frequency of small endothelial defects increased in rabbits after 3 weeks of hypercholesterolemia but decreased to the control level after 6 weeks of hypercholesterolemia. In rabbits with 8 months of hypercholesterolemia the frequency and area of defects outside plaques did not differ from the control group. In the group with hypercholesterolemia for 8 months 39.2% of the plaque surface contained endothelial cells in which there were no distinct silver-stained cell borders. Kevex X-ray spectrometric data of silver topography indicated that the plaque surface without distinct cell borders was not an area devoid of cells. The data obtained do not support the assumption that morphological endothelial injury is the structural precursor of plaque formation.

Effect of carcinogenic polycyclic aromatic hydrocarbons on mouse embryonic cells in culture: Induction of spindle-shaped cells

In cultured mouse embroyonic cells (MECs) treated with benzo[ a]pyrene(B[ a]P), there appeared unusual type of fibroblasts, spindle-shaped cells (SP cells), which were characterized by their narrow bipolar shape, long cellular processes and optically distinct cell borders. Appearance of SP cells was massive and irreversible. The amount of SP cells increased with increased with increasing concentrations of B[ a]P, while early cytotoxicity did not. In various polycyclic aromatic hydrocarbons (PAHs) tested, only potent carcinogens {7,12-dimethylabenz[ a]anthracene (DMBA), 3-methylcholanthrene (MCA), B[ a]P and dibenz[ a, e]pyrene (DB[ a, e]P)} induced SP cells. Among them, PAH having higher Iball's index induced SP cells at lower concentration and at an earlier time. Weak or non-carcinogenic PAHs including 3-hydroxybenzo-[ a]-pyrene(3-H-B[ a]P) did not induce SP cells. α-Napthoflavon (αNF) suppressed the induction of SP cell by carcinogenic PAH. SP cells did not appear spontaneously under various abnormal culture conditions. These results indicate that carcinogenic PAHs induce the appearance of a specific type of fibroblast, SP cells in MEC cultures in accordance with their carcinogenicity.