Intranuclear inclusions in Bence Jones lambda plasma cell myeloma

A patient with plasma cell myeloma producing only Bence Jones lambda protein was found to have pale intranuclear inclusions in the majority of the bone marrow plasma cells. These inclusions, previously undescribed in myeloma patients producing only Bence Jones protein, contained Bence Jones lambda protein, were non-electron dense, bound by a single membrane, and contained no cytoplasmic structures. Intracytoplasmic inclusions were not present, and the perinuclear cistern was not dilated. Thus, the inclusions may represent intranuclear protein synthesis with anomalous release in the abnormal cells.     

Interactions between live Bartonella bacilliformis and endothelial cells.

Bartonella bacilliformis, a gram-negative, flagellated, motile bacterium, is the etiologic agent of verruca peruana. It is found within the verruca, where it can form large cytoplasmic (Rocha-Lima) inclusions in endothelial cells. Previously, an activity has been described in homogenates of B. bacilliformis that in vitro increases the proliferation of endothelial cells and their production of tissue-type plasminogen activator (t-PA) and in vivo is angiogenic. The aim of the present study was to determine if live B. bacilliformis similarly stimulated endothelial cells and produced the Rocha-Lima inclusion. By measuring proliferation of cells and the production of t-PA in vitro, it was found that the live bacteria increased both parameters in a fashion similar to the homogenates of B. bacilliformis. Interaction between the bacteria and endothelial cells appeared to be necessary for proliferation. On electron microscopy, bacteria penetrated the endothelial cell within 1 h, forming a small membrane-bound inclusion. By 12 h, a large membrane-bound inclusion, similar to the Rocha-Lima inclusion, containing numerous bacteria was present. These data provide further evidence that B. bacilliformis has an angiogenic activity and that the bacteria are at least in part responsible for the vascular proliferation of the verruca.     

Development of the smooth muscle foam cell: uptake of macrophage lipid inclusions.

A possible mechanism for the formation of smooth muscle foam cells in the atherosclerotic lesion was explored. Cultured macrophages (J774 cell line) were induced to form cytoplasmic cholesteryl ester inclusions by exposure to acetylated low density lipoprotein in the presence of cholesterol-rich phospholipid dispersions. The macrophages were disrupted by brief sonication, and the inclusions were isolated by flotation. When these inclusions were placed in direct contact with cultured smooth muscle cells, cellular uptake of the inclusions in a time- and dose-dependent manner was observed. Light and electron microscopy indicated the presence of lipid inclusions throughout the cytoplasm of the cells. Uptake of inclusion lipid by the smooth muscle cells was inhibited by several metabolic inhibitors, indicating that the process is dependent on metabolic activity. A modest but significant hydrolysis of the cholesteryl ester was observed, showing that the stored cholesteryl esters are metabolically available.     

Specific cytoplasmic crystalline inclusions in multiple myeloma with amyloidosis

It is widely known that multiple myeloma is sometimes followed by amyloidosis. It is also not particularly rare that inclusions exist in myeloma cells. However, there has been no previous report of a case of myeloma with both inclusions and amyloidosis. A 60-year-old female initially complained of a tendency to bleed, which was caused by fibrinolysis. Amyloid deposition in bone marrow stroma and the gastric submucosa was recognized, in addition to crystalline inclusion in the cytoplasm of myeloma cells. An immunoelectron microscopic study demonstrated the amyloid fibrils and the crystals to react positively to anti-lambda serum. No crystals were found in macrophages, and no relationship was recognized between lysosomes and crystalline inclusions in the cytoplasm of myeloma cells. This case had disturbed transportation or secretion of the lambda type L chain and it was considered that crystals derived from the lambda type L chain were formed in the cytoplasm of myeloma cells.     

急型克山病患者心肌线粒体内含物电镜X射线显微分析

电子显微镜观察发现,急型克山病人心肌线粒体内出现两种电子密度内含物,一种是无定形的,另一种是针状高电子密度的,这两种内含物的出现是心肌细胞不可逆损伤的重要标志,经电子染色,X 射线显微分析,以及次氯酸钾消化三种方法鉴定结果证明,中等电子密度无定形内含物是线粒体蛋白质的变性凝聚,针状高电子密度内含物是含钙物质。本文对这两类内含物出现的条件作了分析和讨论。     

Ubiquitin is associated with abnormal cytoplasmic filaments characteristic of neurodegenerative diseases.

Several degenerative diseases of the nervous system are characterized by the presence of neuronal inclusions. Most of these inclusions are made of abnormal filaments and share epitopes with cytoskeletal proteins. One of these inclusions, the neurofibrillary tangle of Alzheimer disease, has recently been shown to contain ubiquitin, a regulatory protein thought to play a role in the degradation of abnormal proteins. We carried out light and electron microscopic immunocytochemistry with several polyclonal and monoclonal antibodies to investigate the presence of ubiquitin in neuronal inclusions of neurodegenerative diseases. Ubiquitin was present not only in paired helical filaments that form the neurofibrillary tangle of Alzheimer disease, but also in the filamentous components of the inclusion characteristic of Parkinson disease, Pick disease, and progressive supranuclear palsy. In contrast, ubiquitin was not detected in other neuronal inclusions often found in aging and in Alzheimer disease, such as Hirano bodies and granulovacuolar degeneration. Reactivity with monoclonal antibodies suggests differences in the ubiquitin-acceptor proteins present in the inclusions studied. It is concluded that ubiquitin is selectively present in neuronal inclusions of degenerative diseases.     

Ultrastructure of abnormal membrane inclusions in nuclei of human myocardial cells.

The ultrastructure of the nuclear envelope was studied in cardiac muscle cells of 12 patients undergoing cardiopulmonary bypass surgery. Two main types of membrane inclusions, globular and tubular, were observed in the nuclei. The globular type was found in 6 patients. The globular inclusions were about 0-5mu to 1-5mu in diameter and lined by two unit membranes equal in structure to that of the nuclear envelope. The lining was probably always in continuity with the nuclear membranes. Such inclusions contained a granular matrix in which no cytoplasmic organelles could be seen, with the exception of a limited number of ribosomes. The tubular type of nuclear inclusion was observed in the cardiac tissue of 3 patients. These tubules ran in straight or slightly bent segments of about 0-2 mu to 1-6 mu length, interrupted by bendings at obtuse or right angles. The diameter of their lumina measured about 300 A to 800 A. Such tubules were nearly always found in areas of the nucleoplasm rich in heterochromatin. Nuclear pore-like structures were occasionally found in the tubular wall. The tubular wall consisted of a single unit membrane shown to be in continuity with the inner membrane of the nuclear envelope. Since these nuclear aberrances have been observed at all of the stages examined, the possibility is considered that they may represent a specific nuclear response to the process of cardiac hypertrophy.     

Ultrastructure of abnormal membrane inclusions in nuclei of human myocardial cells.

The ultrastructure of the nuclear envelope was studied in cardiac muscle cells of 12 patients undergoing cardiopulmonary bypass surgery. Two main types of membrane inclusions, globular and tubular, were observed in the nuclei. The globular type was found in 6 patients. The globular inclusions were about 0-5mu to 1-5mu in diameter and lined by two unit membranes equal in structure to that of the nuclear envelope. The lining was probably always in continuity with the nuclear membranes. Such inclusions contained a granular matrix in which no cytoplasmic organelles could be seen, with the exception of a limited number of ribosomes. The tubular type of nuclear inclusion was observed in the cardiac tissue of 3 patients. These tubules ran in straight or slightly bent segments of about 0-2 mu to 1-6 mu length, interrupted by bendings at obtuse or right angles. The diameter of their lumina measured about 300 A to 800 A. Such tubules were nearly always found in areas of the nucleoplasm rich in heterochromatin. Nuclear pore-like structures were occasionally found in the tubular wall. The tubular wall consisted of a single unit membrane shown to be in continuity with the inner membrane of the nuclear envelope. Since these nuclear aberrances have been observed at all of the stages examined, the possibility is considered that they may represent a specific nuclear response to the process of cardiac hypertrophy.     

The fine structure of adult Onchocerca volvulus IV. The hypodermal chords of the female worm

The morphology of the hypodermal chords in the anterior, midbody and posterior region and in worms of different ages are described and the amphids, the cephalic papillae and the nerve ring are demonstrated. At the anterior end, the hypodermal cap comprises the sensory organs. Close behind, the hypodermal chords are differentiated, and several cell margins are found in all of them. Four sublateral chords are differentiated, and several cell margins are found in all of them. Four sublateral chords are also found in the anterior region. A row of median cells is found between the dorsal and ventral syncytia further behind and at the posterior end. In the midbody region, the lateral chords are very large but without central cells. Various particles and inclusions such as bacteria, dense bodies and fibrils are found in the plasm of the hypodermal chords. A large multilayered inclusion was found in the degenerating chord of a female worm at least five years old.     

Target tissues associated with genital infection of female guinea pigs by the chlamydial agent of guinea pig inclusion conjunctivitis.

Female guinea pigs were experimentally infected in the genital tract with the strain of Chlamydia psittaci that causes guinea pig inclusion conjunctivitis. Chlamydiae were found frequently in superficial squamous epithelial cells of the exocervix, along with heavy involvement at the squamocolumnar junction. In this zone chlamydiae were observed in columnar cells, but inflammation and chlamydial inclusions were not observed in true endocervical epithelium. These observations were supported by results of immunofluorescence microscopy. Electron micrographs revealed typical chlamydiae within surface epithelial cells at the squamocolumnar junction; various stages of the growth cycle (elementary, reticulate, and intermediate bodies) were also present. Cytopathologic features were not remarkable, but peripheral distribution of organelles, rarefaction of the cytoplasm, and loss of microvilli were observed.     

Auer rod-like intracytoplasmic inclusions in multiple myeloma. A case report and review of the literature

Several intracytoplasmic morphological changes in the plasma cells of multiple myeloma have been described previously. However, Auer rod-like inclusions are rarely found in these types of cells. Here, we report a case of multiple myeloma with Auer rod-like, needle-shaped intracytoplasmic inclusions in plasma cells. A review of all published cases revealed that this phenomenon is exclusively found in myeloma with kappa-type paraprotein. The nature of these intracellular inclusions and the relationship to prognostic implications and concomitant illnesses are discussed.     

Simulation of Inclusion Particle Motion Behavior under Interfacial Tension in Continuous Casting Mold

Inclusions entrapped by the solidifying front during continuous casting adversely affect the properties of the final steel products. In this study, we investigated the effect of the interfacial tension due to surfactant concentration, particularly sulfur, on alumina inclusion motion behavior during molten steel solidification in a continuous casting mold. A two-dimensional numerical model was developed in Ansys Fluent software to simulate the inclusion motion in a continuous casting mold. Further, the impacts of different values of the alumina inclusion diameter, sulfur concentration, and melt temperature were studied to understand the inclusion motion behavior. The inclusion diameter affected the inclusion distribution throughout the domain. The alumina inclusion entrapment percentage varied in the case of sulfur mixing (using an empirical relationship for modeling). It was found that the removal percentage varied according to the sulfur concentration. The addition of sulfur at concentrations from 10 ppm to 70 ppm resulted in a 4% increase in the removal of alumina inclusions (trapped in the solidifying shell), except for the 100-ppm case. Smaller-sized inclusion particles had a 25% higher chance of entrapment at the top level of the mold. Under the effect of a higher surface tension gradient between inclusions and the melt, the predicted findings show that inclusions were vulnerable to engulfment by the solidification front.     

Chlamydia psittaci: growth characteristics and enumeration of serotypes 1 and 2 in cultured cells.

The growth characteristics of Chlamydia psittaci serotypes 1 and 2 (ovine and bovine origin) were studed in mouse L cells. Formation of inclusions and yield of infectious progeny for serotype 1 were maximal when host cells were treated with cycloheximide and the pH in the cell culture medium was 7.2-7.4. The number of cells that contained inclusions and the infectivity yield for serotype 2 were maximal when the pH was 6.6-7.0. Treatments with diethylaminoethyl dextran and cycloheximide increased inclusion formation in serotype 2 but decreased the yield per infected cell. For both serotypes, centrifugation of the chlamydiae onto monolayers infected 50-1,200 times the number of cells infected with use of stationary adsorption. An enumeration method for L-cell infectious units was developed that gave higher titers than the chicken embryo 50% lethal dose after yolk sac inoculation.     

Effects of heat shock, heat shock protein 40 (HDJ-2), and proteasome inhibition on protein aggregation in cellular models of Huntington's disease

Huntington's disease (HD), spinocerebellar ataxias types 1 and 3 (SCA1, SCA3), and spinobulbar muscular atrophy (SBMA) are caused by CAG/polyglutamine expansion mutations. A feature of these diseases is ubiquitinated intraneuronal inclusions derived from the mutant proteins, which colocalize with heat shock proteins (HSPs) in SCA1 and SBMA and proteasomal components in SCA1, SCA3, and SBMA. Previous studies suggested that HSPs might protect against inclusion formation, because overexpression of HDJ-2/HSDJ (a human HSP40 homologue) reduced ataxin-1 (SCA1) and androgen receptor (SBMA) aggregate formation in HeLa cells. We investigated these phenomena by transiently transfecting part of huntingtin exon 1 in COS-7, PC12, and SH-SY5Y cells. Inclusion formation was not seen with constructs expressing 23 glutamines but was repeat length and time dependent for mutant constructs with 43-74 repeats. HSP70, HSP40, the 20S proteasome and ubiquitin colocalized with inclusions. Treatment with heat shock and lactacystin, a proteasome inhibitor, increased the proportion of mutant huntingtin exon 1-expressing cells with inclusions. Thus, inclusion formation may be enhanced in polyglutamine diseases, if the pathological process results in proteasome inhibition or a heat-shock response. Overexpression of HDJ-2/HSDJ did not modify inclusion formation in PC12 and SH-SY5Y cells but increased inclusion formation in COS-7 cells. To our knowledge, this is the first report of an HSP increasing aggregation of an abnormally folded protein in mammalian cells and expands the current understanding of the roles of HDJ-2/HSDJ in protein folding.     

The Hermansky-Pudlak Syndrome: Inclusions in Circulating Leucocytes

S ummary . Electron microscopy of circulating leucocytes from a patient with the Hermansky-Pudlak syndrome revealed two different types of inclusions in a small percentage of monocytic cells. One of the inclusions closely resembled the large masses of lipopigment which fill the bone marrow macrophages in this disorder. The second type was unlike any inclusion previously reported in human leucocytes.     

Intramitochondrial inclusions in the myocardial cells of human hearts with coronary disease

Moderately electron-dense intramitochondrial inclusions occurred in the myocytes present in biopsies from akinetic or dyskinetic left ventricle from 8 of 10 patients with old myocardial infarction, and in biopsies from 1 of 2 patients having apparently normal left ventricles but presenting with unstable angina. These inclusions were classified according to their presence in necrotic or viable cells into two categoris: (1) The necrotic cell type inclusion occurred in aneurysmal ventricular wall. The inclusion material occupied enlarged matrix spaces, masked cristal membrane profiles and often invaded intracristal spaces of disrupted mitochondria. Cytochrome oxidase activity was shown to be absent in these mitochondria. (2) The viable cell type inclusion occurred in the presence of various focal ultrastructural changes, but myofibrils, glycogen, nuclei and sarcolemma did not show any of the changes described by others for the early irreversible phase of cell injury in animal models of myocardial ischemia. This type of inclusion, 100 to 600 mμ in diameter, developed in the matrix of about 15% of total mitochondrial profiles. Cytochrome oxidase was demonstrated to be active in most of these mitochondria, including those which contained the inclusions. In right ventricular biopsy controls from five of eight patients with old myocardial infarction and from one of two patients with unstable angina, viable cell type inclusions were only rarely seen (less than 1%). The two types of inclusions we found in human hearts are compared with the three distinct types of intramitochondrial densities described by others in animal models of acute myocardial ischemia.     

Inclusions of Hemoglobin in Erythroblasts and Erythrocytes of Thalassemia

Large, usually single inclusions, having the staining properties of Heinz-bodies and occurring spontaneously in thalassemic cells, are described. Theirfrequency is greatest in the normoblasts, either circulating ones or those foundin the bone marrow, and decreases sharply in more mature red cells. Splenectomy unmasks this inclusion phenomenon to a large extent. Some stainingand other properties of these inclusion bodies are described.

It is concluded that these inclusions represent precipitated hemoglobin,very likely uncombined -chains. The findings are discussed and related topresent-day knowledge on hemoglobin synthesis and erythrocyte turnover inthalassemia.

Submitted on July 23, 1962     

Effect of estradiol-17 beta on the somatrotroph cells of the adenohypophysis in the newt (author's transl)]

The somatotroph cells of the female newt's adenohypophysis are characterized by PAS positive, acidophils, electron dense large inclusions (4-6 micrometer). The apparition of these inclusions has been studied in females during the puberty. A correlation between the ovary stages and apparition, number and ultrastructural aspects (paracrystalline arrangement) of the inclusions is established. Same inclusion genesis and aspects has been observed after treatment of young newt males by oestradiol-17 beta (5 mg/liter diluted in the aquarium water). It is concluded that endocrine activity of ovaries may be responsible of the inclusions occurence in the females.     

Cholesteryl ester cycle in cultured hepatoma cells

The existence of a cholesteryl ester cycle in cultured Fu5AH hepatoma cells was documented and factors affecting the rate of turnover of the cholesteryl ester cycle in this cell line were explored. The influence of the physical state of the lipid inclusion in which the cholesteryl esters are stored could be addressed in this cell line because these cells can be induced to store cholesteryl esters in anisotropic (liquid-crystalline) cytoplasmic inclusions by exposure to free cholesterol-rich phospholipid dispersions or in isotropic (liquid) inclusions by addition of oleic acid to the phospholipid dispersions. To examine the relative rates of turnover of the cholesteryl ester cycle in the cells with the two types of inclusions, the fraction of cholesteryl linolenate, a cholesteryl ester present in low amounts in these inclusions, was examined after cells were exposed to medium containing linolenate. After 12 h, cells with anisotropic inclusions contained 17.5% cholesteryl linolenate and cells with isotropic inclusions contained 29.8% cholesteryl linolenate, suggesting an approximately 2-fold difference in turnover of the cholesteryl ester pool. To determine whether this difference was due to a differential rate of cholesteryl ester hydrolysis, the acyl CoA: cholesterol acyl transferase arm of the cholesteryl ester cycle was blocked using a specific inhibitor, Sandoz 58-035. In the presence of this compound, cholesteryl ester was hydrolysed twice as fast in cells with isotropic inclusions as compared to that in cells with anisotropic inclusions. The difference in rate of turnover of the cholesteryl ester cycle was shown to be related to the rate of hydrolysis of cholesteryl ester which, in turn, is related to the physical state of the stored cholesteryl ester.     

Familial visceral neuropathy with neuronal intranuclear inclusions: diagnosis by rectal biopsy.

A family with a visceral neuropathy manifested as chronic idiopathic intestinal pseudo-obstruction is reported. Diagnoses were made histologically by simple rectal biopsy. Discrete eosinophilic intranuclear inclusions, diagnostic of a disease known as neuronal intranuclear inclusion disease, were found in the submucosal ganglion cells. Abnormalities of the autonomic nervous system were identified by pupillary examination and electroretinography. In this family, three of four siblings were affected by the disease, which is apparently transmitted from the paternal side. This pedigree was unique for several reasons: (a) diagnosis in multiple members of two generations indicates that this familial visceral neuropathy was expressed in an autosomal dominant manner, (b) central autonomic nervous system abnormalities were detected by eye examination, and (c) the definitive pathological diagnosis was established antemortem by rectal biopsy in all cases.