Fine structure of the human gallbladder with cholesterosis with special reference to the mechanism of lipid accumulation

Eight excised gallbladders with cholesterosis were studied by light and electron microscopy. Lipid droplets were found not only in the submucosa, but also in the infranuclear cytoplasm of epithelial cells. These contained well developed mitochondria and agranular reticulum. Lipid droplets were seen to form in the agranular reticulum. Macrophages were often present between the epithelial cells and the submucosa and contained numerous processes and well developed cell organelles, abundant lysosomes and lipid droplets. With the advance of lipid deposition, macrophages were filled with lipid droplets and became foam cells. It is suggested that these may become too large and rigid to pass through the endothelium of lymph vessels or that the lumen of the lymph vessels may be obstructed by large foam cells resulting in the destruction of these vessels and the accumulation of foam cells in the submucosa.     

Fine structure of the human gallbladder with cholesterosis with special reference to the mechanism of lipid accumulation.

Eight excised gallbladders with cholesterosis were studied by light and electron microscopy. Lipid droplets were found not only in the submucosa, but also in the infranuclear cytoplasm of epithelial cells. These contained well developed mitochondria and agranular reticulum. Lipid droplets were seen to form in the agranular reticulum. Macrophages were often present between the epithelial cells and the submucosa and contained numerous processes and well developed cell organelles, abundant lysosomes and lipid droplets. With the advance of lipid deposition, macrophages were filled with lipid droplets and became foam cells. It is suggested that these may become too large and rigid to pass through the endothelium of lymph vessels or that the lumen of the lymph vessels may be obstructed by large foam cells resulting in the destruction of these vessels and the accumulation of foam cells in the submucosa.     

Studies on fine structure and location of lipids in quick-freeze replicas of atherosclerotic aorta of WHHL rabbits

Summary The fine structure of intracellular and extracellular lipids in the atherosclerotic aorta of Watanabe-heritable hyperlipidemic (WHHL) rabbits was demonstrated by a quick-freeze etching technique. Many lipid droplets, with and without a membrane, were observed in the foam cells. Membrane-free droplets were observed as onionlike structure with a concentric lamellar structure surrounded by 10 nm filaments. Droplets surrounded by a limited membrane probably correspond to lipid-laden lysosomes.In the extracellular connective tissue space, marked accumulation of lipids with a vesicular structure was seen among collagen fibers. The appearance of these lipids was similar to that of lipids in lysosomes of foam cells.     

小鼠类巨噬细胞系超微结构的观察

本文报道了小鼠类巨噬细胞系(MMC-1)的超微结构。在透射电镜下,可见暗细胞、亮细胞及空细胞。这三种细胞有一定的比例。在其他培养细胞中也见到了类似的三种细胞。暗细胞电子致密度最高,呈圆形、椭圆形或梭形。表面有许多突起,有的呈丘状。胞质可分内、外两部分。外胞质无细胞器;内胞质中有丰富的线粒体、粗面内质网和聚核蛋白体。溶酶体易见,高尔基复合器发达,有成堆的微纤维。胞核不规则,多呈肾形,位于细胞的一侧。核膜清楚,常染色质较多。有的胞核中可见核仁。亮细胞突起少,线粒体肿胀,溶酶体与脂滴易见,核肿胀。空细胞的突起更少,内质网明显减少或消失,溶酶体多,次级溶酶体易见胞核大,染色质稀疏。本文根据此细胞的超微结构特点,证实其为巨噬细胞,并根据其来源称之为类巨噬细胞。     

Repopulation of lymph nodes and spleen in thymus chimeras after lethal irradiation and bone marrow transplantation: dependence on the age of the thymus

CAP and Lewis rats were thymectomized and received a syngeneic thymus graft followed by lethal irradiation and syngeneic bone marrow transplantation. In three groups (A: recipient 15 months old, thymus graft 3 months old; B: recipient 3 months old, thymus graft 15 months old; C: recipient and thymus graft both 3 months old), we performed an immunohistologic analysis of the splenic white and red pulp and the paracortical zone of the lymph nodes. The repopulation of these regions was demonstrated with monoclonal antibodies that react with Thy-1 positive cells, peripheral T cells, T helper cells, and T non-helper cells. In the splenic red pulp, more Thy-1 positive lymphocytes were found in group B than in group C. The proportion of T lymphocytes and T helper lymphocytes in the region of the periarteriolar lymphocyte sheath of the splenic white pulp was higher when a young thymus was transplanted (groups A and C) than when an old one was (group B). In contrast, in the splenic red pulp, more T lymphocytes were found in group A than in groups B and C. In the paracortical zone of the lymph nodes, this was demonstrable only for group C versus group B. The proportion of T non-helper lymphocytes in the region of the splenic red pulp was higher in group B than in group C. These results indicate that the repopulation of lymph nodes and spleen after transplantation of an old thymus is delayed, quantitatively reduced, and qualitatively different (more T non-helper lymphocytes).     

Age-related changes in localization of injected radiolabelled lymphocytes in the lymph nodes of antigen-stimulated mice.

The localization of i.v. injected syngeneic lymph node cells, radiolabelled with 51Cr or 75Se-L-selenomethionine, was studied in male CBA/H mice aged between 3 and 30 months. The following results were obtained. (1) Localization of cells from young adult donors was greater in the s.c. lymph nodes of old than of young recipients, the main increase being between 15 and 17 months of age. Increases in lymph node weight and DNA-synthesis were also seen at this time; but the rise in cell localization was significant even when calculated per unit of tissue weight. Splenic localization either declined slightly with age or, like the liver, showed no significant change. (2) Local antigenic stimulation by a single injection of sheep erythrocytes into one front footpad, 24 hr before lymph node cell injection, resulted in increased localization in the regional lymph nodes of 3-17 month old, but rarely of 24-30 month old mice. (3) No consistent differences in localization were observed between lymph node cells from 4-month and 25-month old donors. Both age-related and antigen-related increases in cell localization were at least partly attributable to an enhanced rate of entry of lymphocytes from the blood to the lymph nodes. Although the changes underlying the decline in antigen-related localization of cells in old recipients have still to be clarified, it is probable that the defective immune responses of old mice result partly from this decline.     

Some observations on the fine structure of human granulosa cell tumors

Summary The ultrastructural features of three granulosa cell tumors are presented. The neoplastic non-luteinized granulosa cells are characterized at sub-microscopical level by severely indented nuclei with prominent nucleoli, sparse to moderately developed predominantly granular endoplasmic reticulum, scanty lipids and lysosomes, small mitochondria with lamellar cristae and abundant intracytoplasmic filamentous material. The luteinized cells display a strongly developed tubular agranular endoplasmic reticulum and mitochondria with tubular cristae.These findings are compared with those of previous reports and discussed in relation to the well-known hormonal activity of these tumors.     

T and B lymphocytes in New Zealand Black mice: an analysis of the theta, TL and MBLA markers

The lymphocytes of thymus, blood, spleen, lymph nodes and bone marrow were studied in NZB mice between the ages of 1 and 14 months, and compared with lymphocytes of A and CBA strain mice of the same age. By standard cytotoxicity techniques, the proportion of cells possessing the θ, TL and MBLA markers was found to be similar in NZB in mice and controls. In 14-month old NZBs, whose spleen was largely replaced by reticulum cell sarcoma, and in younger recipients of the passaged tumour, there was a reduction in the percentage of θ and MBLA-positive cells in the spleen. In a few mice at 4 and 9 months, small numbers of MBLA-positive cells were present in the thymus and there was a corresponding decrease in θ-positive cells. TL-positive cells were not present outside the thymus, and θ-positive cells were not present in the bone marrow in unusual numbers. NZB peripheral lymphocytes appeared to have the same surface concentration of θ as those of A or CBA mice, as judged by anti-θ titration curves. The reticulum cell sarcoma was shown to be θ-negative and MBLA-negative, while an NZB thymoma was θ-positive, TL-positive and MBLA-negative. It was concluded that the peripheral lymphoid organs contain a large population of T lymphocytes of abnormal character.     

In-vitro effects of bromocriptine on isolated pituitary adenoma cells. Ultrastructural and morphometrical studies

3 pituitary adenomas in hyperprolactinemia and 3 GH and prolactin producing tumours were analysed. The adenoma cells were prepared and held in suspension so that they could be treated with bromocriptine (10 ng and 100 ng). At different times after treatment (0.5, 60 and 90 minutes), the cells were fixed and prepared for conventional electronmicroscopy. Electron microscopic photographs were quantitatively analysed by the point counting method. The results were compared to those of an untreated control group. After bromocriptine influence, there was a decrease of the hormone secretion into the supernatant (2 of 3 prolactin producing adenomas). The prolactin secretion was unchanged in all 3 adenomas which produced prolactin and GH, but there was a decrease in the GH production in 1 of these cases. Ultrastructural morphometry revealed the following results: In prolactin producing adenomas, there was a decrease in the number of exocytoses, an increase in the volume density of lysosomes (2 cases) and an increase of the rough endoplasmic reticulum (1 case). The decrease of the "unorganized" cytoplasm was observed in all 3 cases, but was significant only in 1 case. There was a significant increase in secretory granules (1 case). In adenomas which produced prolactin and GH displayed a significant increase of the rough endoplasmic reticulum and of the granules. The outlines of the cellular membranes seemed smoother (1 case). The heterogeneous results may be interpreted as an expression of the reduced hormone secretion (secretory granules, lysosomes), some data are in accordance with the beginning of necrobiotic phenomena (rough endoplasmic reticulum). The decrease of the "unorganized" cytoplasm may be due to a shrinking process.     

Reversible phenotypic modulation induced by deprivation of exogenous essential fatty acids

Essential fatty acid deficiency, produced by deprivation of omega-6 and omega-3 fatty acids, is a condition characterized by renal disease, dermatitis, and infertility. Although many of the biochemical aspects of this disorder have been investigated, little is known about the ultrastructural changes induced by essential fatty acid deficiency. Using a unique fatty acid-deficient cell line (EFD-1), which demonstrates the in vivo fatty acid changes of essential fatty acid deficiency, and the prostaglandin E2-producing mouse fibrosarcoma line from which it was derived (HSDM1C1), we correlated ultrastructural and biochemical changes induced by prolonged deprivation of all exogenous lipids and subsequent repletion of selected essential fatty acids. We found that in cells deprived of all exogenous lipids, there was dilation of rough endoplasmic reticulum and an associated defect in protein secretion; these changes were specifically reversed by arachidonate. There was also an accumulation of secondary lysosomes containing degraded membranes in these cells with an associated increase in phospholipids relative to parent HSDM1C1 cells. Cytoplasmic lipid bodies present in parent cells disappeared, with an associated decrease in triacylglycerol. After just 2 days in lipid-free medium, all these changes were apparent, and prostaglandin E2 production was markedly impaired despite normal amounts of cellular arachidonate. Incubation of EFD-1 cells with arachidonate, the major prostaglandin precursor fatty acid, induced a reversion to the HSDM1C1 phenotype, whereas other fatty acids were totally ineffective. These results indicate changes in fatty acid metabolism in essential fatty acid deficiency are associated with marked alterations in ultrastructure and secretion of protein from cells.     

Salivary duct carcinoma: A clinicopathologic study of three cases with a review of the literature

Three cases of salivary duct carcinoma are presented. They occurred in a 60 year old man, a 66 year old man and a 57 year old woman. All of the lesions were located In the parotid gland. The tumor size ranged from 3 to 5 cm across the largest diameter. Facial paralysis was observed in two cases. Histologically, intreductal and invasive adenocarcinoma showing papillary, cribriform, and solid patterns with comede like necrosis was observed. lmmunohistochemically, the tumor cells were positive for keratin and epithelial membrane antigen. No myoepithelial cells were demonstrated within the tumor by staining for S-100 protein, α-smooth muscle actin or muscle specific actin. Ultrastncturally, intracytoplasmic lumina with microvilli, a moderate number of mltochrondria, lysosomes, and tight junctions were found. Regional lymph node metastasis was observed in one case, and distant metastasis developed in two cases. All of me patients were treated with adjuvant postoperative irradiation. One patient died of disease at 11 months after the initial diagnosis, another was alive with disease at 8 months, and the third patient was alive without disease at 2 years and 3 months. Salivary duct carcinoma should be ditterentiated from low-grade salivary gland carcinomas using morphologic and clinical criteria because of its poor prognosis even with aggressive therapy.     

TUMOR GROWTH OF THE RETICULOENDOTHELIAL SYSTEM

From the author's hitherto studies on the reticuloendothelial system (RES) it was concluded that the RES is not a single cell system of identical origin, morphology and function but is a group of several types of cells of different origin. From this point of view the heterogeneity of tumors of the RES was studied to reveal the following results. Tumors of the "Reticulo-endothel" (Aschoff) reveal pictures of endothelioma, while tumors of histiocytes in connective tissue show findings of fibrohistiocytoma. Histiocytes and reticulum cells of lymph nodes are respectively independent cells, and reticulum cells do not partake in the development of histiocytosis or histiocytic sarcoma. Follicular lymphoma is a neoplastic growth of reticulum cells having desmosomes in lymph follicles, and tumor cells of the majority of reticulum cell sarcoma are similar to the cells forming the lymph node anlage in the early fetal stage (lymphoreticular cell). Ewing's sarcoma is considered to be a kind of angiopericytoma.     

Tumor growth of the reticuloendothelial system.

From the author's hitherto studies on the reticuloendothelial system (RES) it was concluded that the RES is not a single cell system of identical origin, morphology and function but is a group of several types of cells of different origin. From this point of view the heterogeneity of tumors of the RES was studied to reveal the following results. Tumors of the "Reticuloendothel" (Aschoff) reveal pictures of endothelioma, while tumors of histiocytes in connective tissue show findings of fibrohistiocytoma. Histiocytes and reticulum cells of lymph nodes are respectively independent cells, and reticulum cells do not partake in the in the development of histiocytosis or histiocytic sarcoma. Follicular lymphoma is a neoplastic growth of reticulum cells having desmosomes in lymph follicles, and tumor cells of the majority of reticulum cell sarcoma are similar to the cells forming the lymph node anlage in the early fetal stage (lymphoreticular cell). Ewing's sarcoma is considered to be a kind of angiopericytoma.     

SANFILIPPO B SYNDROME — A CASE REPORT —

An autopsy case of a 9 years and 5 months old gargoyle girl diagnosed as Sanfilippo B syndrome by the biochemical demonstration of a large amount of heparan sulfate in urine and some organs and of deficiency of α-N-acetyl-D-glucosaminidase in the liver and brain was reported. The morphological changes characterized by cytoplasmic swelling and vacuolization were more generalized than those which had been described in previously reported cases. Histochemically, accumulation of variable amounts of acidic glycosaminoglycans and compound lipids, presumably gangliosides and phospholipids, was substantiated in the vacuolated cells of various visceral organs and in the ballooned neuronal cells. Ultrastructurally, numerous inclusions found in these cells were largely divided Into two types; flocculent reticulogranular and osmiophilic, mostly laminated materials, many of which were bound by a single unit membrane. Enzyme cytochemistry proved acid phosphatase activity in the majority of the inclusions in fibroblasts and nbrocytes biopsied from the skin. Rough endoplasmic reticulum in these cells was markedly dilated with reticulogranular materials. The morphological changes of the present case and their pathogenesis were discussed.     

The subcellular localization of immunoglobulin in mouse plasma cells,as studied with immunoferritin cytochemistry on ultrathin frozen sections

Cells of mineral oil-induced plasmacytomas (MOPC) and normal plasma cells from mouse lymph nodes were used to study the intracellular localization of IgG by means of immunoferritin cytochemistry on ultrathin frozen sections. IgG was demonstrated in the rough endoplasmic reticulum, in virus-containing smooth ER of the tumor cells, in peripheral elements at the cis-side of the Golgi complex, and in all Golgi cisternae. It is suggested that the peripheral elements transfer IgG molecules from the RER to the Golgi complex. Vacuoles showing a strong immunoreaction occurred at the trans-side of the Golgi complex. These vacuoles were normal in lymph node plasma cells and were occasionally seen in the tumor plasma cells. It is proposed that these vacuoles carry IgG from the Golgi complex to the cell membrane and, hence, can be considered as secretory vacuoles.     

Isolation of large mononuclear Ia-positive veiled cells from the mouse thoracic duct

Veiled cells (VC), are not normally present in the mouse thoracic duct lymph (TDL). However, TDL from mesenteric lymph-adenectomized (MLNX) mice contained about 1% VC. After exposure to whole body irradiation of 350-500 rads., or 800 rads., 1.5% and 7% VC respectively were present in the TDL. These cells could be enriched further by density gradient centrifugation on 14.5% metrizamide, about 60-70% VC being present in the cells from the interface. The VC had long sweeping cytoplasmic extensions or veils which were continuously extended and retracted when the VC were incubated at 37 degrees C. Electron microscopy revealed that the VC had a lobulated nucleus, and that the cytoplasm contained many mitochondria, a well-developed Golgi zone, rough endoplasmic reticulum and a few lysosomes. The cells were strongly Ia positive and over 70% showed adenosine triphosphatase activity. These features of the mouse VC from the TDL are similar to those described for VC isolated from different sources in other experimental animals and from the afferent lymph in man.     

Fine structural studies of a human thyroid adenoma, with special reference to psammoma bodies.

The fine structural morphologic features of a microfollicular thyroid adenoma from a 28 year old female were examined. Although the patient had been laking exogenous thyroxine therapy for 14 months, the morphology of the adenoma was characterized by numerous small to medium sized follicles composed of metabolically active, well differentiated columnar cells with numerous colloid droplets, dilated granular endoplasmic reticulum, large numbers of coated vesicles and lysosomes, large colloid containing "lakes," microtubules, microfilaments, and prominent apical microvillous projections. Of special inetrest were small spherical psammomatous calcospherites ecountered in histiocytes and the interstitium. Also noteworthy were ropelike configurations observed in most of the follicular lumina. Structural-functional correlations and potential origins of psammoma bodies and calcospherites are discussed.     

Histological development of bovine abomasum

The histological development of the bovine abomasum during fetal and neonatal periods was studied. The abomasum in a fetus of 2.3 cm in length (estimated to be 1 month old) was a separated compartment situated to be caudo-ventral to the primordium on the median plane. On the later stages, the primitive stomach became distinctly separated into rumen, reticulum, omasum and abomasum. The epithelium of abomasum was pseudostratified histologically, consisting of cells low differentiation. The stomach in a fetus of 13-14 cm in length (estimated to be 3 months old) displayed the morphological feature with nearly same proportion as an adult stomach. In this stage, abomasal epithelium had a shape of simple column and also formed gastric pits. Pyloric gland cells could be recognized earliest of all the gastric exocrine cells in fetuses of 16-18 cm in length (estimated to be 3-4 months old). Mucous neck cells could be found in fetuses 43-45 cm in length (estimated to be 5-6 months old). Pyloric gland cell and mucous neck cell contained neutral and sialo- or sulfo mucosubstances in neonates. Chief cells, could be noticed in fetuses 58-65 cm in length (estimated to be 6-7 months old), and were devoid of demonstrable mucosubstance as well as parietal cells. Main abomasal gland cells began to develop to increase rapidly in number in the latter half period of gestation. All the types of gastric cells became to be present and mature in form at birth.     

An ultrastructural and cytochemical investigation of the development of inclusions in gastric chief cells and parietal cells of mice with the Chediak-Higashi syndrome

Gastric epithelium of the beige mouse, with a mutation thought to be analogous to that in Chediak-Higashi syndrome of man, has been examined by ultrastructural morphologic and cytochemical methods. The gastric chief cell in beige mice at 2 months of age or older disclosed two types of abnormal inclusion bodies each having distinctive morphologic and cytochemical features and a different distribution pattern and relationship to other organelles. On the basis of these findings, the first type of inclusion was thought to originate from zymogen granules, in a process of crinophagy, and the second type was interpreted as arising from the maturing face of the Golgi lamellae by the route for genesis of secondary lysosomes or lipofuscins. Each type of inclusion showed evidence both for participating in autophagic processes and for fusing with each other to produce giant inclusions. Additional observations in this study provided evidence for a role of Golgi endoplasmic reticulum lysosome in genesis of secretory granules and of the mature face of the Golgi complex in development of secondary lysosomes in chief cells. The findings also afforded evidence of migration of chief cells toward the bottom of the gland in the course of their maturation. The gastric parietal cell of control black mice disclosed secondary lysosomes, thought to arise from fusion between multivesicular bodies and mitochondria. These autophagic secondary lysosomes were enlarged in beige mice.     

The effects in the excess administration of prostaglandin E1 on the indicator enzymes of organella membrane in gastric mucosa

Administration of excess vitamin A to rats causes gastric ulceration. In this study the effects on the gastric mucosa of excess vitamin A and excess prostaglandin E1, alone and in combination, were studied. Prostaglandin E1 protected against ulceration by vitamin A. Vitamin A labilized marker enzymes from four different membrane systems, namely those of the lysosomes, mitochondria, endoplasmic reticulum and plasma membrane, whereas only the effect on lysosomes was prevented by prostaglandin E1. Indeed, the prostaglandin alone labilized the enzymes from plasma membrane and endoplasmic reticulum and also damaged mitochondrial membranes. Both vitamin A and prostaglandin E1 caused a reduction in the total number and an increase in irregularly-shaped mitochondria in the parietal cells and produced dilation of the endoplasmic reticulum in both parietal and chief cells. It is noteworthy that prostaglandin E1 effectively prevents ulceration by vitamin A despite the extent to which it damages these membrane systems. These findings lend support to the hypothesis that vitamin A ulceration of the gastric mucosa is mediated via release of lysosomal enzymes, following damage to the lysosomal membranes.