Endometriosis: Scanning electron microscopic and immunohistochemical studies of pelvic endometriosis

The pathogenesis of pelvic endometriosis has been studied byusing scanning electron and light microscopy, observing thesurface structure of bluish lesions obtained from 26 patientsduring laparotomy. Paraffin sections included another 17 tissuesamples of endometriosis, based on immunohisto-chemical responsesto epithelial membrane antigen, keratin and vimentin. Ultrastructurally,the surface epithelial cells could not be detected in 13 outof 17 pelvic peritoneal endometriosis samples. In one case inwhich the surface peritoneal cells were seen histologicallyto dip into the subperitoneal stroma, many surface peritonealinfoldings were observed, and ciliated cells were detected atthe edge of these infoldings. Ovarian endometriosis was composedof three types of cells, none of which had any cilia. Thesefindings were observed in continuity with adjacent normal mesothelialcells. No characteristic structure of the endometrial surfacewas observed for the bluish lesion, but the gland surface ofendometriosis located in the subperitoneal stroma initiallyhad ciliated cells. The immunoreactions in both the columnarmesothelial cells with surface peritoneal infoldings and theglands of endometriotic tissues were similar to those of normalendometrial glands, but different from those of normal mesothelialcells. Pelvic endometriosis might originate by a process ofmetaplasia from the pelvic peritoneum.     

Electron microscopic and immunohistochemical studies of oral malignant melanoma

A rare variety of malignant melanoma in the maxilla was studied by electron microscopic observation and an immunohistochemical investigation of the transferrin receptor (TfR). A number of low-stage melanosomes were seen, and the TfR was found in both the cell membrane and the cytoplasm.     

Hepatoblastoma cells producing alpha-fetoprotein: morphometric, immunohistochemical and electron microscopic studies

Morphometric, immunohistochemical, and electron-microscopic studies were undertaken in an attempt to identify the types of hepatoblastoma cellular elements responsible for the synthesis of alpha-fetoprotein (AFP), and to see how they may relate to serum AFP levels and the metastatic spread and prognosis of the hepatoblastoma. Morphometric studies of 21 hepatoblastomas with statistical treatment of the results revealed a moderately strong reliable correlation of the AFP serum titer with the volume ratio of embryonal tumor cells and with that of those tumor elements of endodermal hepatic diverticulum which are similar to the latter cells with regard to degree of differentiation. Also, a consistent, reliable negative correlation was demonstrated between serum AFP titer and the volume of fetal hepatoblastoma cells. The volume ratio of stromal elements was found to be subject to chance variations and not to correlate with serum AFP level. Immunohistochemical and electron-microscopic studies confirmed the morphometric findings and showed AFP synthesis to be effected by poorly differentiated hepatoblastoma cells--by endodermal hepatic diverticulum elements at first and by embryonal and intermediate tumor cells later--and to decrease as the liver tumor cells differentiate further. It is concluded that a high serum AFP level is, generally, an indication that the hepatoblastoma is an extensive one and consists of poorly differentiated cells so that the prognosis is unfavorable.     

Cell death in retinoblastoma: electron microscopic, immunohistochemical, and DNA fragmentation studies

Apparent cell loss by apoptosis occurs in carcinomatous tissue. To investigate cell death in retinoblastoma (Rb), ultrastructural examination, ApopTag staining, electrophoresis to detect apoptotic DNA fragmentation, and flow cytometric studies were performed. Immunostaining for the oncogenic products bcl-2 and p53 was also carried out. Relationships between the proliferation fraction (PF), apoptotic index (AI), and the distribution of bcl-2 and p53 were investigated according to the degree of histologic differentiation of Rb. Ultrastructurally, two patterns of cell death were seen. Necrotic cells exhibited vacuolation of cytoplasmic organelles with a marked lytic change in the karyoplasm and cytoplasm. In contrast, apoptotic cells were characterized by crescentic margination of chromatin, condensation of karyoplasm and cytoplasm, and fragmentation of the nucleus. Differentiated Rb had a low AI value (< 1%), whereas undifferentiated Rb had a high AI value (> 8%). The PF of undifferentiated RB (31%) was significantly higher than that of differentiated RB (14%). Analysis of DNA fragmentation using 3'-end labeling with terminal transferase indicated that undifferentiated Rb has increased DNA cleavage. The distribution of apoptotic bodies within Rb was inversely correlated with the expression of bcl-2. A majority of tumor cells of differentiated Rb were negative for p53, whereas 20-40% of tumor cells of undifferentiated Rb showed a positive reaction for p53. These findings suggest that the degree of susceptibility to apoptosis is closely related to PF, is inversely related to the degree of differentiation of Rb, and is protected by oncogene bcl-2.     

Distribution of neuropeptides in rat pterygopalatine ganglion: light and electron microscopic immunohistochemical studies.

The distribution and quantity of neuropeptides in the rat pterygopalatine ganglion were studied by using complete serial paraffin sections of the ganglion immunostained with antiserum against several neuropeptides. The pterygopalatine ganglion, composed of 4932 +/- 291 (mean +/- SD) neurons, was triangular in shape with a tapering caudal tail. The most commonly found peptide in neurons was vasoactive intestinal polypeptide (VIP) (99.0%), followed by neuropeptide Y (NPY) (54.1%) and enkephalin (10.5%). The rostro-ventromedial and caudal parts of the ganglion where intensely VIP-immunoreactive neurons predominate project to the nasal mucosa, while the rostro-dorsolateral part of the ganglion where NPY-immunoreactive neurons predominate projects to the Harderian gland. The coexistence of VIP/NPY (47.4%), VIP/NPY/enkephalin (6.6%) or VIP/enkephalin (3.9%) in the ganglionic neurons was recognized. Calcitonin gene-related peptide (CGRP)- and substance P-immunoreactive varicosities formed synaptic contacts with the somatic spine or soma, which confirmed that the reflex arch, composed of axon collaterals of trigeminal ganglionic neurons and parasympathetic ganglionic neurons, operates through direct synapses. Enkephalin-immunoreactive varicosities, which were probably derived from parasympathetic preganglionic neurons, also made synaptic contact with the somatic spine.     

Liver cancer: immunohistochemical and electron microscopic study

Primary liver tumors, including 7 low-grade hepatocellular carcinomas (HC), 12 average-grade HC (including 2 mixed tumors and 2 cholangiocellular carcinomas (ChC)), obtained from 23 patients, were histologically, immunohistochemically, and electron microscopically. Certain markers were immunohistochemically studied to identify HC and ChC and differentiate liver carcinoma from metastatic tumors of the same organ.     

Quantitative and electron microscopic studies of sensory ganglion cells of the Sprawling mouse

Summary The L4–6 sensory root ganglia of young and adult Sprawling (Swl) and normal mice were studied. Cell counts showed a great reduction in the total number of ganglion cells inSwl. Cell degeneration was observed in youngSwl animals but not in normal littermates. Most of the remaining ganglion cells showed morphological abnormalities very similar to those seen in chromatolytic neurons — enlarged nucleolus, eccentric nucleus with an infolded nuclear membrane, loss of juxtanuclear Nissl bodies and an increase in neurofilaments, Golgi membranes, autophagic vacuoles, and dense bodies. In contrast to the classical changes of chromatolysis the abnormalities inSwl neurons persisted throughout the lifespan of the animal. Reconstructions from serial sections showed that ganglion cells inSwl were highly irregular in shape.     

Repeated light- and electron microscopic studies of the small-bowel mucosa in Whipple's disease.

The small-bowel mucosa of a patient with Whipple's disease in the florid state and one year after starting antibiotic treatment, in complete clinical remission was examined by the author. As a result of treatment, light-microscopic changes led to total regression. Electronmicroscopically, in the florid state, destruction of small-bowel epithelial cells was observed, mostly in the Lieberkühn's crypts. The changes of epithelial cells regressed after one year of treatment. In the lamina propria, a large amount of macrophages containing characteristic inclusions were found which persisted even after treatment. The composition of inclusions has, however, changed. The macrophages were cells poor in organelles and of a primitive differentiation. Presumably, this morphological picture is associated with the digestion incapability of cells and this may lead to the development of "acquired storage disease".     

Hyaline globules in yolk sac tumor. Histochemical, immunohistochemical and electron microscopic studies

Four cases of yolk sac tumor of the testis and ovary were examined by histochemistry, immunohistochemistry, and electron microscopy, and particular attention was directed toward the close relationship between hyaline globules and alpha fetoprotein (AFP). The hyaline globules were easily identified as eosinophilic globular structures located in the cytoplasm of the tumor cell as well as in the extracellular space. They were considered to be of glycoprotein in nature by their histochemical staining property. By an indirect immunoperoxidase method, the majority of intracytoplasmic hyaline globules were positive for AFP. Electronmicroscopically, the hyaline globules appeared as electron dense bodies with no limiting membranes. They were divided into two types by their electron density and inner structure: one is an electron high dense body consisting of homogeneous or fine granular materials and the other is a moderately electron dense body composed of fine filamentous structures similar to the basement membrane materials. It is suggested that the former corresponds with the AFP positive hyaline globules produces by the tumor cells of visceral type, while the latter represents the AFP negative globules formed by the cells of parietal type in non-neoplastic counterparts.     

Age-related changes in capillaries of rat oral mucosa. A quantitative electron microscopic study

Ultrastructural age changes in capillaries of the buccal mucosa were examined in montages of cross sections made from electron micrographs at 37,500 x. Six rats aged 6 months and 6 aged 30 months were perfused with glutaraldehyde and conventional thin sections obtained. Two capillaries located within a connective tissue papilla were studied from each rat. Capillaries of the old group differed from those of the young group by statistically significant increases in several parameters. The endothelial cell was increased in thickness, especially in the vicinity of junctions. The frequency of pinocytotic vesicles/unit length of the cell circumference was nearly doubled. Junctions were of nearly double length and took a more oblique course. All parts of the basement membranes were thickened, though perhaps less than seems true in skin. In striking contrast to epidermis, the epithelium of oral mucosa undergoes no appreciable thinning with age. We suggest that the observed age increases in frequency of pinocytotic vesicles and in the length of junctions may facilitate blood/tissue exchange, thus compensating for impaired exchange due to the thickened basement membranes. These compensatory changes in conjunction with the unchanged size of the mucosal capillary bed in the aged rat (demonstrated previously) could explain the unchanged thickness of the oral epithelium.     

Two cases of acquired toxoplasmic lymphadenitis. Light and electron microscopic and immunohistochemical studies

We report two cases of acquired toxoplasmic lymphadenitis, one with toxoplasmic cysts and the organisms of Toxoplasma gondii and the other with the organisms only. These cysts and organisms were observed in paraffin-embedded sections, touch smears and ultrathin sections for electron microscopy. Touch smears were especially valuable for the quick and accurate diagnosis of toxoplasmic lymphadenitis. We also studied immature sinus histiocytosis (ISH) in these cases. The predominant cells of ISH were confirmed to be B lymphocytes immunohistochemically, the majority being positive for polyclonal surface IgM. ISH was observed in the perifollicular and paracortical areas surrounding post-capillary venules (PCV), whereas the sinuses were only partially involved.     

Mixed epithelial and stromal tumor of the kidney: preliminary immunohistochemical and electron microscopic studies of the epithelial component

Mixed epithelial and stromal tumor of the kidney is a rare biphasic tumor composed of cysts and tubules embedded in the spindle cell stroma. Although the histogenesis of this tumor is unknown, it has been proposed that both components of the tumor, i.e., stromal and epithelial, are neoplastic. The authors report preliminary immunohistochemical and electron microscopic studies of the epithelial component from one case of a typical, benign, mixed epithelial, and stromal tumor of the kidney. In this study, some tubules showed positivity for proximal, while others showed positivity for distal, nephron immunomarkers. By electron microscopy, some tubules had features of proximal tubular epithelium, while other tubules had features of the loop of Henle (thin segments). The authors believe that in a benign tumor such morphologic heterogeneity is inconsistent with neoplastic proliferation. Therefore, they postulate that in mixed epithelial and stromal tumor of the kidney the tubules are entrapped rather than neoplastic. Additional studies are needed to address this issue and electron microscopy should play a significant role in this process.     

The electron microscopic localization of alkaline phosphatase in Pacinian corpuscles of the cat

The ultrastructural localization of alkaline phosphatase has been studied in Pacinian corpuscles of the cat mesentery by the method of Mayahara et al. (1967) with 3 substrates. As control studies, specimens were incubated in the medium containing L-cysteine (10 mmol) or EDTA (5 mmol). The electron opaque final reaction product was observed on plasmic membranes and in cytoplasm and pinocytotic vesicles of the inner core cells. The precipitate was present also in rough endoplasmic reticulum, multivesicular bodies, and cytoplasmic vacuoles of the inner core lamellae. The axon revealed the positive enzymatic activity in the axolemma and the scattered precipitate was found in axoplasm. The pinocytotic vesicles in the capillary endothelium entering Pacinian corpuscles contained the reaction product, too. The capsule lamellae were devoid of precipitate. Localization of alkaline phosphatase in pinocytotic vesicles of the inner core lamellae and capillary wall support the opinion that this enzyme plays the significant role in the phenomenon of the transport of molecules through inner core lamellae from capillaries to the axon in Pacinian corpuscles.     

Scanning electron microscopic studies on the subepithelial tissue of the gastrointestinal mucosa of the rat

The three-dimensional architecture of the subepithelial tissue of the gastric, the small and the large intestinal mucosa of the rat was observed by scanning electron microscopy (SEM) after removal of cellular elements through prolonged osmication followed by ultrasonication (the method by Highison and Low, 1982), or by cell-maceration with a low temperature NaOH solution (the method by Ohtani, 1987). The basal lamina was exposed by the former method, and the collagenous fibrous sheet immediately under the basal lamina was disclosed by the latter. The surface of the subepithelial tissue is grossly smooth in the pyloric gland and crypts of the small and the large intestine. However, in the fundic glands of the stomach, the surface structure of the subepithelial tissue greatly differs according to glandular location. In the pit of the fundic glands, the surface of both the basal lamina and the sub-basal laminar fibrous sheet is smooth. In the neck region, however, shallow round depressions are seen. The most striking feature of the subepithelial tissue of the fundic glands is the presence of numerous hemispherical concavities in the middle and basal regions of the glands which harbor parietal cells. On the surface of the small intestinal villi, the basal lamina is elevated by the underlying capillary network and forms a meshwork of ridges that surround shallow basins in which numerous round fenestrations 1-5 microns in diameter are seen. The fibrous sheath of the marginal arteriole is observed as a cord-like protuberance on the apical margin of the villi; this suggests its role in the maintenance of the structural integrity of the villi. In the large intestine, well defined round fenestrations are clearly seen, mainly distributed on the upper third of the crypts, and continuing to the lamina propria.     

腹腔内纤维瘤病临床病理、免疫组化及超微结构研究

目的；探讨腹内纤维瘤病临床病理、免疫组化和超微结构特点,讨论其鉴别诊断。方法；用免疫且化及电镜的方法，结合HE形态学观察，对17例腹内纤维瘤病进行研究。结果：男7例，女10例，年龄19－78岁，平均42．9岁，中位年龄42岁。4例有手术史，2例有Gardner综合征。15例单结节，2例2枚结节。肿块直径5－25cm，平均13．5cm，多数境界清楚、切面灰白质韧、实性、半透明状。光镜下主要有纤细梭长和星芒状两种形态的(肌纤维母细胞，间质大量胶原。超微结构显示增生细胞周围大量胶原原纤维，质膜清楚，无基板，胞质内有丰富的粗面内质网。免疫表型：Vim弥漫强阳性；46．7％（7／15）；α－SMA局灶阳性；40％（6／15）MSA局灶阳性；CD117、CD34、S－100蛋白、PGP9．5、desmin阴性。行完整肿块及部分肠段切除者15 例。7例无复发（2－13年，平均8年），3例失访，5例随访中。姑息术者2例，1例死于肠梗阻，1例再手术后失访。结论：腹内纤维瘤病是少见且易误诊的疾病，部分与手术史以有Gardner综合征有关，可能的机制为胶原合成和降解失衡，免疫组化显示部分肌纤维母细胞分化的特点，手术完整切除者治疗效果好。     

Oral squamous cell carcinoma: electron microscopic and immunohistochemical characteristics

Oral squamous cell carcinoma is the most common oral malignancy, and we performed electron microscopic and immunohistochemical investigation of the tumor. In patients with cervical metastasis, microvilli were developed and a small number of desmosomes were found, regardless of the width of the intercellular spaces. In patients without the metastasis, few microvilli were found in relatively wide intercellular spaces, or numerous microvilli were found in narrow intercellular spaces, and a large number of desmosomes were shown. However, these findings were different from those of tumors that had received radiotherapy, in which numerous microvilli and a small number of desmosomes were found in the nonmetastatic cases. Transferrin receptor, which is a marker of cell proliferation, was localized on the cell membrane, especially in microvilli. Ultrastructural similarity between the primary tumor and the metastatic tumor was recognized, however, the features of microvilli, desmosomes, and the intercellular spaces differed between them in most cases. It is suggested that microvilli might be related to the metastasis of oral squamous cell carcinoma cells. Immunohistochemically, the protein expression of p53 and pRb2/p130 was related to the clinical course of the patients with oral squamous cell carcinoma; the mechanism of the synthesis of these proteins should be investigated in order to understand the biological behavior of the tumor.