Expression of heat shock proteins in myocardium of patients with atrial fibrillation

Atrial fibrillation (AF) is the most common sustained arrhythmia. Because heat shock proteins (Hsp) can protect cells from stress, we compared the levels of Hsp60, Hsp72, Hsc73, and Hsp27 in atrial myocardium from 17 patients with AF (8 paroxysmal and 9 persistent) and 7 controls in sinus rhythm (SR). Hsp60, Hsp72, and Hsc73 levels were not significantly different among the 3 groups. Hsp27 expression was slightly higher in paroxysmal AF than in SR and in persistent AF, and a borderline significant difference (P = 0.064) was seen between the paroxysmal and persistent AF subgroups. Hsp60 levels in the moderate, severe, and profound myolysis groups were significantly lower than the light myolysis group, but no differences were found in other Hsps. In summary, the data indicate that expression of Hsp27 and Hsc73 may be associated with different stages of AF and that Hsp60 also may be associated with the degree of atrial myolysis.     

Type-II alveolocytes in the treatment of the pulmonary form of oxygen poisoning (electron microscopic and morphometric research)

The effect of cytochrome C, thymalin and their combination has been studied concerning morphofunctional state of alveolocytes of the II type (A II) in the lungs of 33 non-inbred white rats at the pulmonary form of oxygen poisoning. The phenomenon develops, when the animals are in pure oxygen under pressure of 0.25 MPa for 10 h. The ultrastructural stereological analysis demonstrates that after exposure of the mice in the barochamber, immediately after decompression and during the 1st and the 3d day in the animals not given the pharmacological preparations in the A II diffuse and local edema develops in hyaloplasm, certain changes develop in mitochondria and endoplasmic reticulum. Comparing to intact animals, a relative volume of the lamellar bodies decreases nearly two times, the volume of mitochondria increases by 1.5 times, amount of A II drops. While treating the pulmonary form of oxygen poisoning with cytochrome C or thymalin, in 3 days after beginning to administer the preparations, the relative volume of the lamellar bodies increase by 1.5 times in comparison with those in the group of untreated animals, and at the combined administration-by 2.5 times. This demonstrates stimulation of the pulmonary surfactant synthesis. When the preparations are applied together, by the 3d day the relative volume of mitochondria and amount of A II do not differ from the corresponding indices in intact animals.     

Studies on Habu Snake Venom

It has been found that there are two factors responsible for myolysis in Habu snake venom; heat-labile and heat-stable myolytic factors. As reported previously, the former is a proteinase contained in Habu venom. In this article purification and chemical properties of the heat-stable myolytic factor are presented. Using the method of fractionation with ammonium sulfate and with column chromatography, a heat-stable myolytic factor was obtained ultracentrifugally in a homogeneous state. This factor caused severe myolysis with the aid of both Mg⁺⁺ ion and phospholipase A. It has been found that heat-stable myolytic factor has no activities such as those of proteinase, esterase or ribonuclease. The relationship between the heat-stable myolytic factor and phospholipase A in myolytic activity is discussed.     

Phosphorothioate Antisense Oligonucleotides Induce the Formation of Nuclear Bodies

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20–30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150–300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides.     

Cell differentiation in microsporangia of Pinus sylvestris. V. Diakinesis to tetrad formation

Following the diffuse stage, the progression of meiosis in Pinus sylvestris unlike that earlier in meiosis, was conspicuously asynchronous. During the diffuse stage of meiosis tapetal cells dedifferentiated. Plasmodesmata were formed, the cells developed a uniform, meristematic appearance and the nuclei underwent mitosis. Throughout the stages covered by this report tapetal cells redifferentiated, again becoming hypersecretory cells, and Ubisch bodies (orbicules) formed. In angio-sperms Ubisch bodies apparently form only once whereas in Pinus they are produced several or many times with a different and characteristic form each time. The future Ubisch bodies are filled from connections with cisternae of the endoplasmic reticu-lum, then coated by plasma membrane and its glycocalyx. The plasma membrane and glycocalyx coating are likely to be responsible for the specific exine form of Ubisch bodies. Cytokinesis after meiosis was typical of plant cells, but no cell wall formed. Thus deep invasions of callose between microspores give an appearance of furrowing, as was often suggested in classical literature.     

The major immediate-early proteins IE1 and IE2 of human cytomegalovirus colocalize with and disrupt PML-associated nuclear bodies at very early times in infected permissive cells.

The major immediate-early (MIE) gene products of human cytomegalovirus (HCMV) are nuclear phosphoproteins that are thought to play key roles in initiating lytic cycle gene regulation pathways. We have examined the intranuclear localization pattern of both the IE1 and IE2 proteins in virus-infected and DNA-transfected cells. When HCMV-infected human diploid fibroblast (HF) cells were stained with specific monoclonal antibodies, IE1 localized as a mixture of nuclear diffuse and punctate patterns at very early times (2 h) but changed to an exclusively nuclear diffuse pattern at later times. In contrast, IE2 was distributed predominantly in nuclear punctate structures continuously from 2 to at least 12 h after infection. These punctate structures resembled the preexisting PML-associated nuclear bodies (ND10 or PML oncogenic domains [PODs]) that are disrupted and dispersed by the IE110 protein as a very early event in herpes simplex virus (HSV) infection. However, HCMV differed from HSV by leading instead to a change in both the PML and SP100 protein distribution from punctate bodies to uniform diffuse patterns, a process that was complete in 50% of the cells at 2 h and in 90% of the cells by 4 h after infection. Confocal double-label indirect immunofluorescence assay analysis confirmed that both IE1 and IE2 colocalized transiently with PML in punctate bodies at very early times after infection. In transient expression assays, introduction of IE1-encoding plasmid DNA alone into Vero or HF cells produced the typical total redistribution of PML into a uniform nuclear diffuse pattern together with the IE1 protein, whereas introduction of IE2-encoding plasmid DNA alone resulted in stable colocalization of the IE2 protein with PML in the PODs. A truncated mutant form of IE1 gave large nuclear aggregates and failed to redistribute PML, and similarly a deleted mutant form of IE2 failed to colocalize with the punctate PML bodies, confirming the specificity of these effects. Furthermore, both Vero and U373 cell lines constitutively expressing IE1 also showed total PML relocalization together with the IE1 protein into a nuclear diffuse pattern, although a very small percentage of the cells which failed to express IE1 reverted to a punctate PML pattern. Finally, the PML redistribution activity of IE1 and the direct association of IE2 with PML punctate bodies were both confirmed by infection with E1A-negative recombinant adenovirus vectors expressing either IE1 or IE2 alone. These results confirm that transient colocalization with and disruption of PML-associated nuclear bodies by IE1 and continuous targeting to PML-associated nuclear bodies by IE2 are intrinsic properties of these two MIE regulatory proteins, which we suggest may represent critical initial events for efficient lytic cycle infection by HCMV.     

Glutamate-like immunoreactivity in chick cerebellum and optic tectum

Glutamate was coupled via glutaraldehyde to bovine serum albumin. The conjugate was used for raising specific anti-glutamate antibodies. The purified antibody was used for immunostaining of chick cerebellum and optic tectum. Staining was intense in the molecular layer and in cell bodies of the granule cell layer. In the optic tectum a diffuse staining was detected in the superficial layers of stratum griseum fibrosum superficiale and in cell bodies especially in the layers a and e. Large cell bodies located in the stratum griseum centrale were also stained.     

Uterine artery embolization for the treatment of uterine fibroids

Uterine artery embolization can be regarded as a less invasive procedure for the treatment of fibroids compared with myomectomy, hysterectomy, and laparoscopic myolysis. The aim of this study was the evaluation of safety and efficacy of uterine artery embolization and of womens' opinion about this treatment. After gynecological examination sixty-nine premenopausal women underwent uterine artery embolization. All procedures but four were technically successful; three women underwent unilateral embolization because of vascular malformation and one of them had an allergic reaction to contrast medium. Of the 69 patients: 58 went home the day after embolization, and 11 within first week. The follow-up examinations after 3, 6 and 12 month showed a significant reduction of uterine and fibroid volume with significant improvement of bleeding. Therefore, according to this report, uterine artery embolization is a successful, minimal invasive treatment of myoma that preserves the uterus and requires shorter hospitalization and recovery times than surgery.     

TRIM5 alpha cytoplasmic bodies are highly dynamic structures

Tripartite motif (TRIM)5 alpha has recently been identified as a host restriction factor that has the ability to block infection by certain retroviruses in a species-dependent manner. One interesting feature of this protein is that it is localized in distinct cytoplasmic clusters designated as cytoplasmic bodies. The potential role of these cytoplasmic bodies in TRIM5 alpha function remains to be defined. By using fluorescent fusion proteins and live cell microscopy, we studied the localization and dynamics of TRIM5 alpha cytoplasmic bodies. This analysis reveals that cytoplasmic bodies are highly mobile, exhibiting both short saltatory movements and unidirectional long-distance movements along the microtubule network. The morphology of the cytoplasmic bodies is also dynamic. Finally, photobleaching and photoactivation analysis reveals that the TRIM5 alpha protein present in the cytoplasmic bodies is very dynamic, rapidly exchanging between cytoplasmic bodies and a more diffuse cytoplasmic population. Therefore, TRIM5 alpha cytoplasmic bodies are dynamic structures more consistent with a role in function or regulation rather than protein aggregates or inclusion bodies that represent dead-end static structures.     

Differences in the volume distributions of human lung mast cell granules and lipid bodies: evidence that the size of these organelles is regulated by distinct mechanisms

We analyzed transmission electron micrographs of human lung mast cells by digitized planimetry and point counting to determine the cross-sectional areas of two distinct cytoplasmic organelles: specific granules and lipid bodies. Specific granules have a limiting membrane and often contain one or more cylindrical scroll-like inclusions. By contrast, lipid bodies are on average much larger than granules and lack both limiting membranes and inclusions. The measured cross-sectional areas of lipid bodies and scroll-containing granules were converted to equivalent volumes, and the noise in the frequency distribution of these volumes was smoothed using a moving bin technique. This analysis revealed (a) a periodic, multimodal distribution of granule equivalent volumes in which the modes fell at volumes that were integral multiples of the volume defined by the first mode (the "unit volume"), and (b) a modal granule equivalent volume frequency that occurred at a magnitude equal to four "unit volumes." Thus, specific granules appear to be composed of units of a narrowly fixed volume. Furthermore, the mean volume of intragranule inclusions was 0.0061 mu3, a value very similar to that calculated for the "unit volume" (0.0071 mu3). This result suggests that each "unit volume" comprising the individual scroll-type granules contains (or is capable of generating or accommodating) a single scroll-like inclusion. In contrast to the specific granules, mast cell lipid bodies lack a periodic, multimodal volume distribution. Taken together, these findings suggest that the volumes of human lung mast cell granules and lipid bodies are regulated by distinct mechanisms.     

Comparsion of staining characteristics of Toto bodies

Toto bodies are eosinophilic structures that resemble the cells of the superficial cell layer of the oral epithelium. Toto bodies commonly are associated with inflammatory gingival and other mucosal lesions including pyogenic granuloma, irritational fibroma, epulis fissuratum, peripheral giant cell granuloma and inflammatory hyperplastic gingivitis. We evaluated staining characteristics of Toto bodies to establish their origin and to identify their significance in lesions. We investigated pyogenic granuloma, fibroma and leukoplakia with epithelium that exhibited Toto bodies after hematoxylin and eosin (staining. Sections were stained with Alcian blue, periodic acid-Schiff and Ayoub-Shklar stains to evaluate staining intensity and distribution. More Toto bodies were found in pyogenic granuloma than in fibroma and leukoplakia. PAS and Alcian blue staining exhibited mild intensity and did not establish the origin of Toto bodies. High staining intensity and diffuse distribution of stain was observed using Ayoub-Shklar staining, which indicated that Toto bodies originate from keratin.     

Effect of starvation on nuclear bodies and rough endoplasmic reticulum in the bovine hepatocyte

The frequency of simple nuclear bodies and the relative volume of rough endoplasmic reticulum was determined in hepatocytes of normal lactating cows and of lactating cows starved for 6 days. Starvation resulted in a highly significant decrease in frequency of simple nuclear bodies and in the relative volume of rough endoplasmic reticulum. It is suggested that simple nuclear bodies are normal nuclear organelles which can respond to physiological changes.     

Fine structure of the macronucleus during the cell division cycle of Euplotes eurystomus,a Ciliate Protozoon

This paper reports new observations obtained from a study of macronuclear fine structure throughout various stages of the cell division cycle of Euplotes. Study of the ultrastructural organization of the macronuclear chromatin indicates that much of the chromatin is organized into continuous masses, portions of which appear to be attached to the nuclear envelope. The macronuclear envelope appears unchanged in the region of a replication band, and apparent attachments of the chromatin to the inner membrane of the nuclear envelope are maintained in the reticular and diffuse zones. Intranuclear helices were never observed in the diffuse zone. During macronuclear division, linear elements (fibrils or microtubules) were observed in close association with both chromatin bodies and nucleoli. The ultrastructural data suggest that the intranuclear linear fibrils have two functions: elongation of the dividing nucleus, and attachment of chromatin bodies and nucleoli to the envelope. The significance of these observations for macronuclear division and chromatin segregation is considered.     

Further morphological and histochemical studies on the yolk nucleus and associated cell components in the developing oocyte of the Indian wall lizard

In March through April when the oocyte growth in the ovaries of the wall lizard (Hemidactylus) is very rapid, the yolk nucleus continues to persist through various stages of previtellogenesis. This persisting yolk nucleus and associated cell components have been studied with histochemical techniques. The spherical and dense yolk nucleus stains for protein, lipoprotein and RNA. It does not form any close morphological association with the other cell components such as the mitochondria, lipid bodies (L₂), spaces or canals, diffuse sudanophilic substance and dense bodies, which are arranged into three zones round the yolk nucleus proper. The mitochondria stain for lipoprotein; the L₂ bodies consist of phospholipid; the spaces do not contain any material demonstrable with histochemical techniques; and the ooplasm containing the diffuse sudanophilic substance and dense bodies shows lipoprotein, protein and RNA. Eventually, the yolk nucleus disintegrates, and its substance as well as the other cell components are distributed in the cortical ooplasm of oocytes which are ready to form the yolk bodies. Concepts of the origin, morphology, cytochemistry and function of the yolk nucleus in the oocytes of invertebrates and vertebrates, which have come about recently through the application of cytochemical and submicroscopical techniques, are discussed.     

Calreticulin mRNA and protein are localized to protein bodies in storage maize callus cells

Maize callus cells possess numerous protein bodies which develop as sub-compartments of the endoplasmic reticulum. We localized maize calreticulin mRNAs and protein in maize callus cells using in situ hybridization and immunocytochemistry. Calreticulin mRNAs were selectively targeted to the endoplasmic reticulum (ER) subdomains surrounding protein bodies. Profilin mRNAs, used as a positive control for in situ hybridization experiments, showed distinct and rather diffuse localization pattern. Using both, immunofluorescence and immunogold electron microscopy localization techniques, calreticulin was found to be enriched around and within protein bodies in maize callus storage cells. As a positive control for reticuloplasmins, HDEL antibody revealed labelling of protein bodies and of the nuclear envelope. The identity of protein bodies was confirmed by specific binding of an α zein antibody. These data suggest that calreticulin mRNA is targeted towards protein body forming subdomains of the ER, and that calreticulin is localized and enriched in these protein bodies. The possibility that calreticulin plays an important role in zein retention within the ER and/or its assembly and packaging into protein bodies during protein body biogenesis in maize callus is discussed.     

Twenty-four-hour variations in subcellular structures of rat type II alveolar epithelial cells

Summary Subcellular structures of type II alveolar epithelial cells in the rat lung were analyzed at six evenly spaced times over 24 h (light period: 06.00 h–18.00 h), using a morphometric technique. The cell volumes were maximal at 16.00 h and minimal at 08.00 h. The volume and surface densities of rough endoplasmic reticulum and mitochondria were low during the light period, and high during the dark period. Morphometric parameters of multivesicular bodies did not significantly fluctuate over 24 h, but they increased from 04.00 h to 08.00 h. The volume densities of lamellar bodies increased from 16.00 h to 20.00 h, and decreased from 00.00 h to 08.00 h. The change in numerical densities of lamellar bodies was inversely correlated to that in the volume densities. As shown by electron microscopy, small lamellar bodies predominated at 08.00 h, larger lamellar bodies increasing at 16.00h. Composite bodies often appeared at 08.00 h and 12.00 h. Type II cells thus appear to fluctuate, showing three phases over 24 h: formation, accumulation and secretion of lamellar bodies. In particular, it is noteworthy that the accumulation stage occurs during the resting phase of the rat, whereas the secretion stage occurs during its body-active phase.     

Cell cycle-dependent alteration in NAC1 nuclear body dynamics and morphology

NAC1, a BTB/POZ family member, has been suggested to participate in maintaining the stemness of embryonic stem cells and has been implicated in the pathogenesis of human cancer. In ovarian cancer, NAC1 upregulation is associated with disease aggressiveness and with the development of chemoresistance. Like other BTB/POZ proteins, NAC1 forms discrete nuclear bodies in non-dividing cells. To investigate the biological role of NAC1 nuclear bodies, we characterized the expression dynamics of NAC1 nuclear bodies during different phases of the cell cycle. Fluorescence recovery after photobleaching assays revealed that NAC1 was rapidly exchanged between the nucleoplasm and NAC1 nuclear bodies in interphase cells. The number of NAC1 bodies significantly increased and their size decreased in the S phase as compared to the G?/G? and G? phases. NAC1 nuclear bodies disappeared and NAC1 became diffuse during mitosis. NAC1 nuclear bodies reappeared immediately after completion of mitosis. These results indicate that a cell cycle-dependent regulatory mechanism controls NAC1 body formation in the nucleus and suggest that NAC1 body dynamics are associated with mitosis or cytokinesis.     

甘蓝型油菜油体数量及面积之和与含油量的相关性

利用荧光染料尼罗红染色和激光扫描共聚焦显微观察技术,建立了油菜油体观察或生物体内中性脂类物质定性鉴定的研究体系。对高油品种宁油14号、宁油18号、ZH-088和低油品种ZL-366、NjY008、Westar共6个甘蓝型油菜品种子叶贮藏细胞内的油体进行了观察。研究发现：油菜种子成熟过程中,油体从着色不明显的小颗粒,逐渐发育形成着色清晰的球状大油体。种子成熟干燥后,油体间很少发生聚合。在成熟干燥的种子中,油体集中分布于子叶贮藏细胞中央,呈椭圆形或不规则形状,较少为圆形。通过研究种子内油体与含油量的关系,发现高油品种组与低油品种组之间在单个子叶贮藏细胞内油体数量和截面积之和存在明显差异,而在高油品种组内或低油品种组内的差异不明显。结果显示,油菜种子细胞中油体的数量和总面积与含油量之间存在正相关,可作为高油分材料的选择依据。