Immunocytochemical localization of the 130K and 180K proteins (putative replicase components) of tobacco mosaic virus

We have revealed the cellular localization of the putative replicase components of tobacco mosaic virus (TMV), 130K and 180K proteins, in TMV-infected tobacco leaves by the immunogold technique with antisera which specifically react with these two proteins. When sections of TMV-infected tobacco leaves were treated with anti-130K protein antiserum and then with protein A-gold complex, most of the gold label was strongly localized on granular inclusion bodies which were found specifically in the cytoplasm of TMV-infected cells. Very small amounts of label present in other regions, including the nuclei, chloroplasts, and mitochondria, seemed to be nonspecific. Gold-labeled 180K protein was also dispersed over the granular inclusion bodies. The granular inclusion bodies appeared to be oval-shaped structures with various diameters ranging from 0.2 to 2.8 microm. TMV particles were usually observed near the granular inclusion bodies as aggregates but not inside them. Considering the involvement of the 130K and 180K proteins in replication, the granular inclusion bodies may be the site for replication of TMV RNA.     

Pulmonary blue bodies

Pulmonary blue bodies are intra-alveolar laminated basophilic concretions of uncertain etiology. Blue bodies were studied in lung biopsy speciments from 10 patients. The patients ranged in age from 47 to 69 years and were predominantly men. Three had a history of overt exposure to environmental dusts such as sawdust and asbestos, and two showed occasional ferruginous bodies in the lung, raising the possibility of pneumoconiosis. In eight cases there was intersitial pneumonitis, which resembled desquamative interstitial pneumonia by light microscopy but which was often seen to be patchy and asymmetrically distributed in the lung by chest x-ray examination. Of two other patients, one had xanthogranulomatous inflammation and the other, necrotizing granulomatous inflammation.Light and electron microscopic, histochemical, microchemical, and x-ray diffraction studies of blue bodies were also performed. Calcium carbonate is a major component of blue bodies and is responsible for their birefringence in unstained sections and ready solubility in acid solutions. Blue bodies also contain a mucopolysaccharide matrix and iron. We offer the hypothesis that blue bodies (calcium carbonate) are a product of histiocytic catabolism.     

A stable proportion of Lewy body bearing neurons in the substantia nigra suggests a model in which the Lewy body causes neuronal death

Lewy bodies in Parkinson disease could be innocent bystanders or active agents responsible for neuronal death. Eighteen elderly patients with a Parkinson syndrome were studied prospectively and selected postmortem on the presence of Lewy bodies (14 cases with Parkinson disease, four with dementia with Lewy bodies). Information on disease duration was available in 17 cases. While akinesia and rigidity were linked with the neuronal loss, the percentages of Lewy body bearing neurons and of alpha-synuclein immunoreactive neurons in the substantia nigra were not correlated with the symptoms or the disease duration, and appeared stable, involving 3.6% of the neurons on average. Such stability indicated that, during the whole course of the disease, the destruction of the Lewy bodies was equal to their production. In the model that is proposed here, the Lewy bodies are eliminated when the neurons that bear them die. With the hypothesis that neuronal death is directly related to Lewy bodies, it is possible to estimate their life span, which was calculated to be 6.2 months (15.9 months for any type of alpha-synuclein inclusion).     

The mesolimbic dopamine system: Evidence for a high amine turnover and for a heterogeneity of the dopamine neuron population

By means of a new photographic method to quantitate catecholamine fluorescence in tissue sections it has been possible to demonstrate a high dopamine (DA) turnover within the DA cell bodies of the midbrain. The medially located small-sized DA cell bodies of the A10 DA cell group has a very high DA turnover which was significantly different from that found in the laterally located medium-sized DA cell bodies of the A10 cell group. Thus, the mesolimbic DA pathways may be divided into two systems, the medial system having a very high DA turnover in their cell bodies which may reflect a high functional activity in this system.     

Lamellar bodies are markers of cholinergic neurons in ferret nucleus basalis

Summary Lamellar bodies are composed of stacks of closely-packed, ribosome-free cisterns which are in continuity with the rough endoplasmic reticulum. In the ferret nucleus basalis stained for choline acetyltransferase it was shown, by correlating light with electron microscopy, that only the cholinergic cells there possess lamellar bodies. The significance of lamellar bodies in the cholinergic neurons of the nucleus basalis is not known, but these structures may reflect a peculiar aspect of the functioning of the cholinergic cells which will need to be investigated further.     

The fate of the bodies of executed persons in the Anatomical Institute of Halle between 1933 and 1945

In the period from 1933 to 1945 the Anatomical Institute in Halle (Saale) received bodies of persons, among them politically persecuted women and men, who had been sentenced to death and executed. In this article, we attempt to answer two important questions: (1) What happened to the bodies of those executed; i.e. which anatomical “purposes” did they serve? (2) Were anatomical specimens from these bodies added to the institute’s anatomical collection and are they still present today? If so, can they be traced back to the bodies of politically persecuted people? So far we have discovered that between 1933 and 1936 the institute received 30 bodies, among them the bodies of two politically motivated death sentences. From 1937 until the end of 1942, only a few bodies arrived at the institute, and from November 1942 until the end of the war in 1945 the institute documented the transfer of 64 bodies of executed people. The death sentences pronounced during those early years were usually based on severe criminal acts (e.g. murder). During the war, special courts sentenced people to death mostly because of theft, looting, etc. The bodies of those executed were used in anatomical education, anatomical research, and in preparations of anatomical specimens to be added to the anatomical collection. There are eight macroscopic preparations which can definitely be associated with the bodies of people executed during the Nazi regime. Trial by jury sentenced those people to the maximum penalty because of the severity of their criminal acts. Up to now we have found no evidence that specimens of the anatomical collection were removed from bodies of victims whose execution was politically motivated.     

In Vivo incorporation of choline-3H,leucine-3H and galactose-3H in alveolar type II pneumocytes in relation to surfactant synthesis. A quantitative radioautographic study in mouse by electron microscopy

A quantitative time study of the incorporation of choline-³H, leucine-³H and galactose-³H in lung epithelial cells, was undertaken in vivo with electron microscopic autoradiography. Type II pneumocytes were selectively labeled with choline-³H, a specific precursor of phosphatidylcholine, which is the main component of pulmonary surfactant. Choline was initially localized in endoplasmic reticulum, then was rapidly transferred through the Golgi complex and stored in lamellar bodies. Previously undescribed small lamellar bodies are suggested as phospholipid carriers between Golgi complex and lamellar bodies. After initial incorporation in the endoplasmic reticulum, the leucine label migrated through the Golgi complex into lamellar bodies by fusion of multivesicular bodies, which are the carriers between the two structures. Galactose was modestly incorporated into lamellar bodies via the Golgi complex. Intra-alveolar myelin figures, recognized as excreted surfactant, were labeled 120 minutes after injection with the three precursors. These findings indicate that the synthesis of a secretory product by type II pneumocytes involves phospholipid, protein and polysaccharide precursors; the secretory product is segregated as lamellar bodies which are destined to be excreted into the alveolar cavity to become part of the lining layer.     

The Variable Prevalence of Junctional-Complex-Associated Bodies in the Human Sweat Gland

Sweat gland biopsies have been examined for their content of junctional-complex-associated (JCA) bodies. These bodies have been observed at the lumen of the gland as well as the intercellular canaliculi. A JCA body was encountered in each of two dark cells with uncondensed secretory granules; one of these cells was from a specimen in which no other bodies could be found after an extensive search. All other JCA bodies occurred in either clear or undifferentiated cells. Junctional-complex-associated bodies occupied pairs of adjacent cells rather frequently, but no cell revealed more than one body. The prevalence of JCA bodies varied widely among specimens. Glands lacking morphologic abnormalities, and others with some atypical features, disclosed few or no such bodies. Some glands that contained JCA bodies also contained coil cells with extensive vacuolization, which occurred as a manifestation of Hurler's disease or on an unexplained basis. Glands from a patient with cystic fibrosis exhibited the highest prevalence of JCA bodies. Some of the coil profiles in glands near a scar in one specimen were composed of undifferentiated cells with rare mitotic figures and no identifiable clear or dark cells. Numerous JCA bodies, particularly at the lumen, were found in sweat glands in this and another specimen containing frequent undifferentiated cells.     

Comparison of the reflex responses elicited by stimulation of the separately perfused carotid and aortic body chemoreceptors in the dog

1. In dogs under chloralose and urethane anaesthesia, the carotid and aortic bodies were isolated from the circulation and separately perfused with blood, the composition of which could be controlled at will. The remainder of the systemic circulation was perfused at constant blood flow, thereby enabling the reflex vascular responses to be determined. The systemic venous blood was oxygenated in the isolated perfused lungs of a second dog and the P(O2) and P(CO2) of the systemic arterial blood was maintained constant.2. Using hypoxic hypercapnic blood to stimulate the arterial chemoreceptors, carotid body excitation in spontaneously breathing animals caused an increase in respiratory minute volume approximately seven times larger than that evoked by stimulation of the aortic bodies. Whereas the hyperpnoea of carotid body origin is due to an increase in rate and depth of breathing, that from the aortic bodies is due predominantly to an increase in respiratory frequency.3. Stimulation of the carotid bodies in spontaneously breathing animals caused small variable changes in systemic vascular resistance, whereas stimulation of the aortic bodies invariably increased the vascular resistance.4. When pulmonary ventilation was maintained constant, the vascular response to stimulation of the carotid bodies was considerably modified in that constriction invariably occurred; that from the aortic bodies, however, was little affected. There was now no significant difference in the size of responses from the two groups of chemoreceptors. These constrictor responses represent the primary vascular effects.5. A similar modification of the carotid body vascular response occurred in the spontaneously breathing animal after denervation of the lungs, and is due to abolition of a lung-inflation vasodilator reflex.6. The size of the primary vasoconstrictor responses from the carotid and aortic bodies is reduced by lowering the arterial blood P(CO2).7. The results indicate that there is a fundamental difference in the functions of the carotid and aortic bodies. They exert a quantitatively similar primary control of the ;vasomotor centre' which is in striking contrast to the relatively more powerful influence on respiration by the carotid bodies. In the spontaneously breathing animal, however, the primary vasoconstrictor response from the carotid bodies is offset to a varying degree by the lung-inflation vasodilator reflex initiated by the concomitant hyperpnoea. This is not evident with the aortic bodies because of the relatively weaker respiratory response they evoke.     

Evidence for the presence of substance P in cat nasal receptor afferents

The afferents of the nasal receptors responsible for many upper airway protective reflexes are carried in the ethmoidal branch of the ophthalmic division of the trigeminal nerve. Previous electrophysiological studies indicate that a significant number of ethmoidal afferents respond to noxious stimuli applied to the nose. The objective of the present study was to identify ethmoidal nerve cell bodies within the trigeminal ganglion which demonstrated the presence of the neurotransmitter substance P (SP). SP is believed to be involved in the relay of nociceptive information. A double-labelling technique was employed and involved tracing the afferents to their cell bodies using horseradish peroxidase (HRP) and subsequent identification of SP-immunoreactivity within HRP-filled cells using monoclonal antibody immunohistochemistry. SP-immunoreactive cell bodies constituted 43 per cent-50 per cent of the total number of labelled ethmoidal cell bodies within the trigeminal ganglion. Although ethmoidal cell bodies were much smaller than the overall population of trigeminal ganglion cells, the size of SP-immunoreactive ethmoidal cell bodies was not significantly different from that of ethmoidal cell bodies not exhibiting SP-immunoreactivity.     

Expression of osteopontin messenger RNA by macrophages in ovarian serous papillary cystadenocarcinoma: a possible association with calcification of psammoma bodies

Calcified psammoma bodies often appear in human ovarian serous papillary cystadenocarcinoma. Osteocalcin (OC), osteonectin (ON) and osteopontin (OPN) are three members of non-collagenous bone-related proteins known to be related with mineralization of bone. To clarify possible involvement of these bone matrix proteins in the calcification of the psammoma bodies, the expression of OC, ON and OPN was analyzed by immunohistochemical and in situ hybridization studies using 15 surgical specimens. OPN protein was detected in the calcified area of the psammoma bodies which was positively stained by von Kóssa's staining, while OC and ON proteins were not. OPN protein was not detected in any cells in tissues, but OPN messenger ribonucleic acid (mRNA) was detected in CD68-positive macrophages, indicating that OPN was produced and promptly secreted by macrophages. These results suggest that OPN produced and promptly secreted by macrophages and subsequently translocated to psammoma bodies may be causally related with the calcium phosphate deposition in the psammoma bodies of the ovarian serous papillary cystadenocarcinomas.     

Ultrastructural study of the alveolar lining and the bronchial mucus layer by block staining with oolong tea extract: the role of various surfactant materials

To investigate the roles of various surfactant materials in the lung, we examined rat lung fixed with a mixture of 0.2% oolong tea extract (which contains various polyphenols) and 2.5% glutaraldehyde by electron microscopy. We also measured the surface tension of various isolated surfactant fractions, with a Wilhelm balance. A fraction containing lamellar bodies and a fraction containing lattice-like structures were obtained by discontinuous sucrose gradient centrifugation. From the 15 000 g supernatant, a fraction containing electron-dense amorphous materials was obtained as the 105 g precipitate. The fraction containing lamellar bodies and the fraction containing electron-dense amorphous materials displayed surfactant activity, but the fraction containing lattice-like structures did not. The lamellar bodies were found to be gradually secreted from type II epithelial cells while self-decomposition occurred. The alveolar lining layer had the form of a thin film consisting of electron-dense amorphous materials. These electron-dense amorphous materials may be precursors of the phospholipid film, which exhibits surfactant activity, on the alveolar surface. Lattice-like structures and lamellar bodies were found to be located in the interalveolar pores. The interalveolar pores were filled with surfactant, and this indicated that they do not play a role in the collateral ventilation of the alveoli. It may be that the lattice-like structures serve to connect lung epithelial cells in the interalveolar pores, as well as serving as the basis for the formation of the alveolar ducts. The bronchial mucus layer, which consisted of fibrillar mucous materials, was not divided into an epiphase and a hypophase. A surfactant, in the form of an osmiophilic surface film and some trilaminar materials, was found to cover the mucus layer. Thus, it is possible that the lamellar bodies could be transformed into various surfactant materials, which then serve bronchial or alveolar mechanical and physiological functions.     

Adenovirus and ileocecal intussusception.

Intranuclear inclusion bodies were found by light microscopy in epithelial cells in more than one-third of the specimens from children operated on for ileocecal intussusception. Electron microscopic examination done on hematoxylin and eosin-stained slides showed the intranuclear inclusion bodies to be composed of viral particles in large and small crystalline arrays. Adenovirus of serotypes 2, 3, and 5 were isolated from the five cases with inclusions in which isolation was attempted. These findings strongly suggest a pathogenetic role for adenovirus in those cases of intussusception in which intranuclear inclusion bodies are found in the epithelial cells of the appendix or the terminal ileum.     

Nature and function of endocytosis in Sertoli cells of the rat

The endocytic activity taking place at the apex of Sertoli cells was analyzed by using various tracers to demonstrate fluid-phase and adsorptive endocytosis. Native ferritin and protein-gold complexes, used to demonstrate fluid-phase endocytosis, were internalized by Sertoli cells at all stages of the cycle of the seminiferous epithelium. At 5 min after injection, the tracers were found in the large spherical or C-shaped vesicles seen in the apical processes; at 15 and 30 min, the tracers accumulated in light multivesicular bodies, and at 60 min in dense multivesicular bodies and lysosomes. At later time intervals, an increasing number of lysosomes contained the tracers. Following injection of cationic ferritin or concanavalin A-ferritin, these tracers, used to demonstrate adsorptive endocytosis, were found to be found to the plasma membrane of Sertoli-cell apical processes but were not collected by the large cytoplasmic apical vesicles, multivesicular bodies, or lysosomes. As an exception, a few special multivesicular bodies seen in the large apical processes which encapsulate the heads of late spermatids at stage VII of the cycle contained these tracers. During stage VIII of the cycle, the residual bodies which detach from the mature spermatids are phagocytosed by Sertoli cells. Such residual bodies do not contain their own hydrolytic lysosomal enzymes. However, it was observed that the lysosomes which form as a result of the fluid-phase endocytic activity of the Sertoli cells fuse with the phagocytosed residual bodies and by means of their hydrolytic enzymes contribute to the rapid disintegration of these bodies. Thus, during stage VIII of the cycle, the Sertoli cells integrate two distinct processes, i.e., fluid-phase endocytosis and phagocytosis.     

幽门螺杆菌L型微结构的研究

扫描电镜发现幽门螺杆菌的Ｌ型与其它细菌的Ｌ型一样形态不规则，呈多形性，有圆球体、巨形体、短杆状、串珠状和腊肠样，大小不等。超薄切片可见细胞壁部分或完全缺失，菌体内部结构稀疏，电子密度降低，部分菌体含有大量原生小体，有的正从细胞内释放出来，是细菌Ｌ型繁殖的方式之一。     

Intranuclear Helioid Inclusions in Mammary Intraductal Hyperplasias

Intranuclear helioid bodies were identified by light microscopy in eight cases of mammary intraductal hyperplasia, only one of which was atypical. These structures appear as round, intranuclear eosinophilic bodies on light microscopic examination. When prominent, they may be and have been mistaken for viral inclusions. Ultrastructural analysis of these inclusions shows a single-membrane-bound structure containing a laminated or homogeneously electron-dense core with a corona of radiating filaments. In the first and only previous report of this structure in the breast, helioid bodies were identified incidentally during an ultrastructural analysis of a breast adenoma, but the light microscopic counterpart of this structure was not described or illustrated. In view of the similarity of the laminated inclusions to psammoma bodies, the possibility that helioid bodies serve as a nidus for development of microcalcification in the breast was considered. Actual microcalcification was not evident in the vicinity of any of these inclusions, however, and there was no evidence of calcification in these structures on von Kossa's stain for calcium.     

Immunohistochemical and morphometric analysis of immunoglobulin light-chain immunoreactive amyloid in psammoma bodies of the human choroid plexus

The aim of this research was to establish the presence of amyloid and to quantify immunohistochemical reactions of kappa and lambda light chains of psammoma bodies of the choroid plexus. Choroid plexus tissue obtained from 14 right lateral ventricles postmortem was processed histologically and stained with Congo red, thioflavin T, and monoclonal antibodies for kappa and lambda light chains. Morphological analysis was performed with a light microscope at lens magnifications of 4×, 10×, 20×, 25×, and 40×. The morphometric characteristics of psammoma bodies that were kappa and lambda positive and negative were analyzed with ImageJ. Histological analysis showed that the psammoma bodies, stromal blood vessel walls, and some epithelial cells reacted positively with Congo red and thioflavin T. Psammoma bodies were predominantly positive for lambda light chains. Lambda positivity was detected inside some stromal blood vessels, which pointed to a probable systemic origin for these light chains. Morphometric analysis showed that the mean optical densities of lambda- and kappa-positive psammoma bodies were significantly higher than those that gave a negative reaction. The percentage of lambda-positive psammoma bodies was significantly higher than the percentage of lambda-negative psammoma bodies in 80 % of the cases, while the reaction with kappa light chains was negative in the majority of the cases. Linear regression analysis showed a significant increase in the percentage of lambda-positive psammoma bodies and their mean optical density with age. Finally, it can be concluded that the positive reaction of psammoma bodies in the choroid plexus with respect to amyloid and lambda light chains may point to the presence of light-chain amyloid in their structures.     

Amylase producing lung cancer

Summary A case of the lung cancer associated with marked elevation of serum (7820 IU/l) and of urinary amylase (2225 IU/l) was autopsied. Material was examined histopathologically, electronmicroscopically and biochemically. The pulmonary tumor tissues showed histological pattern of papillary adenocarcinoma. In addition to the small round bodies which were very similar to secretory glanules, many large round bodies were noticed, diffusely distributed in the tumor cell. These large round bodies ranged from 0.2 to 0.7 m in diameter and showed a lamellar or annular pattern. The soluble phase of the homogenized pulmonary tumor tissues had an amylase level of 11,300 IU/g of protein, which consisted of S-type isoamylase with minor components. Cholesterol, triglyceride and phospholipid were also present at greater concentration in the tumor tissue than the normal pulmonary tissue. The large round bodies appeared too, to be amylase including bodies from the electronmicroscopical and biochemical findings.     

Neutralization of Chlamydia trachomatis infectivity with antibodies to the major outer membrane protein.

Rabbit immunoglobulin G (IgG) antibodies raised against the major outer membrane protein of the Chlamydia trachomatis lymphogranuloma venereum strain 434 neutralized the infectivity of the parasite for HeLa 229 cells. The mechanism by which anti-major outer membrane protein IgG prevented C. trachomatis from establishing infection was studied by using intrinsically 14C-radiolabeled elementary bodies. Neutralized elementary bodies were filterable through a polycarbonate filter (pore diameter, 600 nm), demonstrating that reduction in infectivity was not due to the aggregation of elementary bodies by cross-linking IgG. Antibody-neutralized elementary bodies attached to and penetrated HeLa cells at rats nearly identical to those for infectious organisms exposed to nonneutralizing control IgG. These results suggest that antibody interferes with the infectious process of the parasite after its internalization. Anti-major outer membrane protein Fab fragments could not be substituted for neutralizing IgG antibodies. The requirement for intact IgG implies that cross-linking of antibodies to the major outer membrane protein on the surfaces of the organisms may be instrumental in neutralization.     

Glycogen accumulation in the renal tubular cells of spontaneously occurring diabetic WBN/Kob rats

Cytoplasmic and nuclear accumulation of glycogen granules in the kidney cells of 72 male WBN/Kob rats with a long-term diabetic condition was studied histologically and by electron microscopy. The incidence and degree of the accumulation showed good correlation with the blood glucose concentration. In the kidneys, there was evidence of two types of lesion, cytoplasmic glycogen accumulation in the distal convoluted tubules and nuclear accumulation in the ascending thick segment of Henle's loops. Electron microscopically, the cytoplasmic glycogen accumulation was often associated with an increased number of lysosomal bodies containing lamellar bodies. Glycogen bodies, the halo of which was thought to be identical with that of nuclear bodies, were frequently observed in the nuclei containing the glycogen granules. These morphological and topographical differences between the two types of lesion were considered suggestive of different pathogenetic mechanisms for glycogen accumulation in the kidney cells.