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Experimental vasospasm in cultured arterial smooth-muscle cells. Part 1: Contractile and ultrastructural changes caused by oxyhemoglobin.

Smooth-muscle cells were cultured from rat aortic media, then oxyhemoglobin and other agents including serotonin, norepinephrine, and angiotensin II were added separately to the medium. Contractile and ultrastructural changes of the cells were examined with electron microscopy during the first 2 weeks of incubation. Oxyhemoglobin not only produced progressive contraction of the arterial smooth-muscle cells, but it also caused ultrastructural changes that resembled myonecrosis. In contrast, there was no evidence of progressive contraction or ultrastructural changes either in control cultures or in cultures with the other vasoactive agents. Although washout of oxyhemoglobin 3 hours after administration prevented continued contraction of the cells, washout 24 hours or longer after administration had no preventive effect. Judging from these results and from the fact that the culture medium was changed every 2 days, it is unlikely that accumulation of exogenous vasoactive agents caused these changes. The contraction and suggestive myonecrosis of the arterial smooth-muscle cells are probably caused by some intrinsic process initiated by oxyhemoglobin. The culture of cerebral arterial smooth-muscle cells requires further technical improvement; nevertheless, these results obtained with the smooth-muscle cells of rat aortic media indicate that arterial smooth-muscle cells in culture provide a promising new experimental model for chronic in vitro study of cerebral arterial spasm. It is suggested from these results that cerebral arteries are particularly prone to vasospasm because of structural differences as compared to noncerebral arteries.

Animals↗

Ultrastructural characteristics of hemorrhagic, nonhemorrhagic, and recurrent cavernous malformations.

OBJECT: Ultrastructural characteristics of hemorrhagic, nonhemorrhagic, primary, and recurrent central nervous system cavernous malformations (CMs) were examined in an attempt to clarify their pathological mechanisms. METHODS: Thirteen specimens (nine from samples of CMs and four from healthy control tissue) were processed for ultrastructural study immediately after surgical or postmortem removal, by fixation in glutaraldehyde/formalin and postfixation in OsO4. Transmission electron microscopy was used to examine the vascular walls, endothelium, subendothelium, and cytoplasmic organelles. The vascular walls in CMs demonstrated abnormal ultrastructure with no basement membranes and astrocytic foot processes. Pericytes were rarely seen. Single-layer lining endothelial cells showed fenestrated luminal surfaces. Large gaps were observed at intercellular junctions between endothelial cells, and large vesicles with extremely thin plasma membranes bordering the lumen were common in the lesions that had previously hemorrhaged. Endothelial cells of recurrent CMs had more Weibel-Palade bodies, filopodia, cytoplasmic processes, micropinocytotic vesicles, and filaments than those in primary lesions and normal control tissues. CONCLUSIONS: The absence of the blood-brain barrier, normal supporting wall structure, and large vesicles bordering the lumen of CM vessels may explain leakage of red blood cells into surrounding brain in the absence of major hemorrhage. Proliferation of residual abnormal endothelial cells may contribute to the recurrence of surgically removed CMs.

Adolescent↗

Effect of mexiletine on lipid peroxidation and early ultrastructural findings in experimental spinal cord injury.

OBJECT: The purpose of this study was to investigate the effect of mexiletine on lipid peroxidation and on ultrastructural findings after induced spinal cord injury (SCI). The authors also compared the activity of mexiletine to that of the well-known antioxidant, methylprednisolone sodium succinate (MPSS). METHODS: Wistar rats were divided into seven groups, (Groups 1-7). Those in Groups 1 and 2 were control animals that underwent laminectomy only, after which nontraumatized spinal cord samples were obtained immediately (Group 1) and 2 hours postsurgery (Group 2). Spinal cord injury was induced in all other groups, and cord samples were obtained at 2 hours postsurgery. The rats in Group 3 underwent SCI alone; those in Group 4 received 30 mg/kg of MPSS intraperitoneally immediately after trauma was induced; and those in Groups 5, 6, and 7 received 1, 10, and 50 mg/kg of mexiletine, respectively, by intraperitoneal injection immediately after trauma was induced. Compared with the levels in control animals, lipid peroxidation was significantly elevated in rats in Groups 3 and 5, but there were no statistical differences among those in Groups 1, 2, 4, 6 and 7 in this regard. Compared with the findings in rats in Group 3, ultrastructural damage post-SCI was minor in rats in Groups 4 and 5, and there was even less damage evident in rats in Group 7. CONCLUSIONS: Analysis of these findings showed that administration of 50 mg/kg mexiletine significantly decreased the level of lipid peroxidation and protected spinal cord ultrastructure following SCI.

Analysis of Variance↗

Ouabain-induced mechanical toxicity: aberrations in left ventricular function, calcium concentration, and ultrastructure.

The isolated rabbit heart was employed to determine if the myocardial mechanical dysfunction (mechanical toxicity) produced by digitalis was accompanied by ultrastructural lesions and an excessive accumulation of calcium by the cardiac cell. Ouabain (1.2 or 2.4 microM) produced a transient increase and then a continuous decrease in the systolic pressure development by the isovolumic left ventricle. The end-diastolic pressure increased progressively during the time that the systolic pressure was diminishing. The ultrastructural abnormalities that were observed in the ouabain-treated hearts included swollen mitochondria, hypercontracted sarcomeres and, occasionally, myofibrillar disruption. The left ventricular cellular calcium concentration was increased (P less than 0.05) in these mechanically toxic hearts and this increase correlated significantly (r = 0.65, P less than 0.05) with the magnitude of the increase in end-diastolic pressure. Furthermore, a reduction in the perfusate calcium concentration did not attenuate the ouabain-stimulated uptake of calcium. The data suggest that the impaired mechanical performance and the ultrastructural aberrations that may develop in the digitalis-treated heart are related to an excessive uptake of calcium by the left ventricle.

Animals↗

Ultrastructural characterization of SARS coronavirus.

Severe acute respiratory syndrome (SARS) was first described during a 2002-2003 global outbreak of severe pneumonia associated with human deaths and person-to-person disease transmission. The etiologic agent was initially identified as a coronavirus by thin-section electron microscopic examination of a virus isolate. Virions were spherical, 78 nm in mean diameter, and composed of a helical nucleocapsid within an envelope with surface projections. We show that infection with the SARS-associated coronavirus resulted in distinct ultrastructural features: double-membrane vesicles, nucleocapsid inclusions, and large granular areas of cytoplasm. These three structures and the coronavirus particles were shown to be positive for viral proteins and RNA by using ultrastructural immunogold and in situ hybridization assays. In addition, ultrastructural examination of a bronchiolar lavage specimen from a SARS patient showed numerous coronavirus-infected cells with features similar to those in infected culture cells. Electron microscopic studies were critical in identifying the etiologic agent of the SARS outbreak and in guiding subsequent laboratory and epidemiologic investigations.

Animals↗

[Ultrastructure of the integument of adult Clonorchis sinensis]

Ultrastructural studies on the integument of digenetic trematodes were investigated by several workers (Senft et al., 1961; Bjorkman and Thorsell, 1964; Burton, l966; Erasmus, l968; Morris and Threadgold, 1967, l968). Clonorchis sinensis is a common parasite of human beings in the Far East. But as far little has been written on ultrastructure of the integument of this parasite. This study was carried out systematically to elucidate the ultrastructure of the integument of adult C. sinensis, and the results are summarized as follows: 1)The integument of C. sinensis was organized in two parts; the epidermis and the muscular layer. 2)The apical surface and basal surface of the epidermis were covered with typical plasma membrane and irregularly invaginated from the surface. Plasma membrane at perisuckers was extremely folded and appeared as if the epidermis might be divided with the layer. 3)The syncytial epidermis contained mitochondria and numerous secretory bodies, and which was connected by cytoplasmic processes to nucleated epidermal cells lying in the parenchyma. 4)The mitochondriae, the secretory bodies, the granular endoplasmic reticulum, the Golgi bodies and the free ribosomes were present in the perinuclear cytoplasm of epidermal cells, but no organelle was observed in processes of epidermal cell except some secretory bodies. 5)The fibrous layer consists of argentophile fiber and collagenous fiber, in which the circular muscle bundles and longitudinal muscle bundles were embedded. Bundles of muscle fiber are well developed at the perisucker, and mitochondriae in this portion contain many cristae as compared with any other parts of C. sinensis.

Journal Article↗

Surface ultrastructure of Metagonimus miyatai metacercariae and adults.

A scanning electron microscopic study was performed to observe surface ultrastructures of excysted metacercariae and adults of Metagonimus miyatai. Metacercariae were collected from the scale of the pale chub (Zacco platypus), and adult flukes were harvested 1-4 weeks after infection to rats. In excysted metacercariae, the oral sucker was devoid of tegumental spines and had type I and type II sensory papillae. Anteriorly to the ventral sucker, spines were dense and digitated into 5-7 points, whereas near the posterior end of the body spines were sparse and digitated into 2-3 points. In one-week adults, 7 type II sensory papillae were arranged around the lip of the oral sucker, and at inner side of the lip one pair of small and two pairs of large type 1 sensory papillae were seen on each side. The distribution of tegumental spines was similar to that of metacercariae, but they were more differentiated with 9-11 pointed tips. In two- to four-week old adults, the surface ultrastructure was nearly the same as in one-week old adults, however, sperms were frequently seen entering into the Laurer's canal. Conclusively, the surface ultrastructure of M. miyatai was generally similar to that of M. yokogawai, however, differentiation of tegumental spines and distribution of sensory papillae around the oral sucker were different between the two species, which may be of taxonomic significance.

Animals↗

Experimental hypercholesterolemia induces ultrastructural changes in the elastic laminae of rabbit aortic valve.

Atherosclerosis is the most severe problem in the high-pressure systemic circulation and similar changes also occur in the high-pressure loading valve. This study was designed to test the hypothesis that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. However, it is not known whether this process is also taking place in the cardiac valve at the early stage of atherosclerosis. Animals were fed either a high cholesterol diet (n = 5) or a control diet (n = 5) for 10-12 weeks. Histologic analysis demonstrated that subendothelial thickening and foam-cell infiltration were evident in the arterialis of aortic valves. Confocal microscopy revealed an altered pattern characterized by fragmentation and disorganization of the arterialis elastic laminae of hypercholesterolemic valves. Computerized digital analysis of the images obtained by confocal scanning microscopy demonstrated that compared to normal valves, the arterialis elastic laminae of hypercholesterolemic valves decreased in percentage of their elastin content (29.03 +/- 1.10% vs. 42.94 +/- 1.35%, p = 0.023). Immunohistochemical staining for matrix metalloproteinase-3 (MMP-3) revealed MMP-3 immunoreactivity was increased in hypercholesterolemic valves, predominantly in the arterialis. This study demonstrated that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. The arterialis endothelium of the aortic valve may be a more atherosclerosis-prone area compared with the ventricularis. The presence of ultrastructural defect in the elastic laminae may play a role in chronic degenerative change and a resultant valvular dysfunction.

Animals↗

Effect of monensin on ultrastructure and cellular invasion by the turkey coccidia Eimeria adenoeides and Eimeria meleagrimitis.

Freshly excysted sporozoites (SZ) of the turkey coccidia Eimeria meleagrimitis and Eimeria adenoeides were incubated at 41 C in concentrations of monensin from .01 to 1.0 microgram/mL, washed free of the drug, and either processed for phase, fluorescence, and transmission electron microscopy or inoculated into cultures of turkey kidney cells. Phase microscopy indicated that after 1.5 h incubation in 1.0 micrograms/mL monensin, about 60% of the SZ of E. meleagrimitis had become notably rounded or displayed localized protrusions. These alterations were accompanied by ultrastructural abnormalities (in 90% of the SZ) including vacuoles in the cytoplasm, bulging and separation of plasma membrane layers, and dense bands in the refractile bodies that extended toward the periphery of the refractile body. Similar morphological and ultrastructural changes were observed in over half of the E. adenoeides SZ after 2 h incubation in 1.0 micrograms/mL monensin. Additionally, some specimens contained a pycnotic nucleus that was usually surrounded by a large vacuole. After 4 h incubation, almost all of the SZ displayed some degree of ultrastructural damage. Indirect fluorescent antibody labeling with parasite-specific monoclonal antibodies demonstrated clouds of antigen surrounding the monensin-treated but not the untreated SZ, suggesting an increase in permeability with incubation in monensin. With both E. meleagrimitis and E. adenoeides, the structural changes were reflected in a significant inhibition of cellular invasion. The inhibitory activity of monensin was concentration- and time-dependent in that the greatest inhibition of invasion was observed in SZ incubated for 4 h in 1.0 micrograms/mL of monensin; shorter incubation times or lower concentrations of monensin having less effect.

Animals↗

Ultrastructural changes of chondrocytes of growth plates of young broiler chickens predisposed to tibial dyschondroplasia.

Growth plates from the proximal tibiotarsal bones of growing broiler chickens from lines exhibiting high and low incidences of tibial dyschondroplasia (TD) were compared for ultrastructural differences. Samples of growth plates were collected from five chickens of each line at 2, 7, 14, and 21 d of age. Each sample was prepared for ultrastructural evaluation after initial fixation in 2.5% glutaraldehyde and secondary fixation in 1% osmium tetroxide, each containing .7% ruthenium hexamine trichloride. No gross TD lesions were observed. The ultrastructure of growth plates of birds at 2, 7, and 14 d of age was similar between the groups. However, at 21 d of age, chondrocytes in the prehypertrophic zone of the high incidence group contained large lipid inclusions and vesiculated and disarranged stacks of rough endoplasmic reticulum with greatly dilated cisternae. Apoptotic and necrotic chondrocytes were present in the hypertrophic zone. Nuclei of apoptotic chondrocytes contained crescentic caps of condensed chromatin. The cytoplasm contained short segments of dilated granular endoplasmic reticulum, lipid droplets, and vacuoles. Necrotic chondrocytes appeared as amorphous masses with pyknotic and karyorrhectic nuclei. These changes were not observed in chondrocytes of the low TD incidence group and have not been previously reported in nonthickened growth plate cartilage.

Animals↗

[Effects of herb therapy for benefiting qi and removing blood stasis on ultrastructure of vascular endothelial cells and vascular smooth muscle cells of uterine endometrium in rabbits with copper intrauterine device].

OBJECTIVE: To observe the effects of herb therapy for benefiting qi and removing blood stasis on ultrastructure of vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs) of uterine endometrium in rabbits with copper intrauterine device. METHODS: Fifty-six rabbits were randomly divided into seven groups, which were normal control group, sham-operation group, untreated group, indomethacin-treated group and low-, medium- and high-dose Gonghuan Zhixueling Recipe (GHZXLR)-treated groups. The rabbits in the last five groups were operated with copper IUD insertion and then orally given distilled water, indomethacin and low-, medium- and high-dose GHZXLR respectively for at least one week. Rabbits in the normal control group and sham-operation group were given distilled water orally. The ultrastructure of VECs and VSMCs of uterine endometrium in rabbits was observed by transmission electron microscopy. RESULTS: The morphological changes of VSMCs revealed intracellular edema, organelle disintegration and decrease of organelle amount, or cell atrophy and vacuolar degeneration of mitochondria in the untreated group, and the amount of collagen fibers also increased outside the VSMCs. Local interstitial edema in subendothelial substance and vacuolar degeneration of mitochondria in VECs were both observed. The ultrastructural damages to the mitochondria, Golgi bodies and myofilament of VECs and VSMCs and the intercellular substance in GHZXLR-treated groups were slighter than those in the untreated group, while these damages had no significant differences as compared with those in the indomethacin-treated group. CONCLUSION: The Chinese herb therapy for benefiting qi for removing blood stasis has the protective effect on VECs of uterine endometrium in the rabbits with copper intrauterine device. It appears to be a good treatment for menorrhagia induced by copper IUD insertion.

Animals↗

Dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress in rats.

AIM: To investigate the dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress (WRS) in rats. METHODS: WRS model of Sprague-Dawley (SD) rats was established. Fifty-six male SD rats were randomly divided into control group, stress group and post-stress group. The stress group was divided into 1, 2 and 4 h stress subgroups. The post-stress group was divided into 24, 48 and 72 h subgroups. The pH value of gastric juice, ulcer index (UI) of gastric mucosa and H(+), K(+)-ATPase activity of gastric parietal cells were measured. Ultrastructural change of parietal cells was observed under transmission electron microscope (TEM). RESULTS: The pH value of gastric juice decreased time-dependently in stress group and increased in post-stress group. The H(+), K(+)-ATPase activity of gastric parietal cells and the UI of gastric mucosa increased time-dependently in stress group and decreased in post-stress group. Compared to control group, the pH value decreased remarkably (P = 0.0001), the UI and H(+), K(+)-ATPase activity increased significantly (P = 0.0001, P = 0.0174) in 4 h stress subgroup. UI was positively related with stress time (r = 0.9876, P < 0.01) but negatively with pH value (r = -0.8724, P < 0.05). The parietal cells became active in stress group, especially in 4 h stress subgroup, in which plenty of intracellular canalicular and mitochondria were observed under TEM. In post-stress group, the parietal cells recovered to resting state. CONCLUSION: The acid secretion of parietal cells is consistent with their ultrastructural changes during the development and healing of stress ulcer induced by WRS and the degree of gastric mucosal lesions, suggesting gastric acid play an important role in the development of stress ulcer and is closely related with the recovery of gastric mucosal lesions induced by WRS.

Animals↗

Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823.

AIM:To investigate the morphological and ultrastructural changes in the human gastric carcinoma cell line BGC-823 after being treated with tachyplesin.METHODS:Tachyplesin was isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes. BGC-823 cells and the cells treated with 2.0mg/L tachyplesin were examined respectively under light microscope, scanning and transmission electron microscope.RESULTS: BGC-823 cells had undergone the restorational alteration in morphology and ultrastructure after tachyplesin treatment. The changes were as follows: the shape of cells was unanimous, the volume enlarged and cells turned to be flat and spread, the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular, the number of nucleolus reduced and its volume lessened,heter-chromatin decreased while euchromatin increased in nucleus. In the cytoplasm, mitochondria grew in number with consistent structure relatively, Golgi complex turned to be typical and well-developed,rough endoplasmic reticulum increased and polyribosome decreased. The microvilli at cellular surface were rare and the filopodia reduced while lamellipodia increased at the cell edge.CONCLUSION:Tachyplesin could alter the malignant morphological and ultrastructural charact-eristics of human gastric carcinoma cells effectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.

Journal Article↗

Ultrastructure and molecular biological changes of chronic gastritis, gastric cancer and gastric precancerous lesions: a comparative study.

AIM: To carry out a comparative study on ultrastructure and molecular biological changes of chronic gastritis (CG), gastric cancer (GC) aand gastric precancerous lesions. METHODS: By the use of histochemical staining, SEM with EDAX, TEM with EDAX, image analysis technique, RIA and chemiluminescence method, gastric mucosa of 168 patients were synchronously analyzed in morphology, trace elements, DNA, cAMP, SOD, (3)H-TdR LCT and serum LPO were also done. RESULTS: The incidence of epithelial nucleoplasmic ratio >1, lobulated nuclei, inter-chromatin aggregation of granules, nucleolar hypertrophy, and the content of DNA, Zn, Cu in nuclei and serum LPO of each group were showed as belows: normal control group (0.0, 0.0, 6.7, 0.0, 12.6+/-2.7, 7.6+/-0.4, 58.4+/-0.3, 2.6+/-0.6), CSG group (5.7, 2.9, 7.4, 2.9, 15.2+/-3.1, 8.1+/-0.5, 58.9+/-0.5, 4.2+/-0.7), CAG group (31.3, 29.7, 45.3, 42.2, 16.5+/-3.1, 8.6+/-0.4, 59.3+/-0.5, 4.5+/-0.6), CA group (100.0, 100.0, 72.2, 50.0, 30.7+/-8.2, 8.8+/-0.3, 59.5+/-0.4, 6.8+/-1.6), ATP(++) group (61.5, 38.5, 23.1, 38.5, 23.5+/-8.9, 8.3+/-0.4, 59.1+/-0.4, 5.1+/-1.2), IM(++)+ATP(++)group (77.8, 55.5, 33.3, 44.4, 25.1+/-7.2, 8.4+/-0.5, 59.5+/-0.4, 6.5+/-1.1), IM(+++)+ATP(++) group (100.0, 100.0, 75.0, 62.5, 28.5+/-9.1, 8.9+/-0.5, 59.7+/-0.4, 7.6+/-0.7), IMII(b) group (100.0, 62.5, 75.0, 50.0, 27.3+/-10.3, 8.6+/-0.3, 59.5+/-0.4, 6.1+/-0.9); whereas the content of Zn, Cu in mitochondria and cAMP, SOD in gastric mucosa, and (3)H-TdR LCT of each group were showen as belows: normal control group (9.2+/-0.5, 58.3+/-0.3, 15.9+/-1.5, 170.5+/-6.1, 1079.7+/-227.4), CSG group (8.6+/-0.5, 57.8+/-0.3, 14.6+/-1.8, 163.3+/-5.6, 867.3+/-240.5), CAG group (8.3+/-0.4, 57.5+/-0.3, 13.4+/-1.8, 161.2+/-4.3, 800.9+/-221.8), CA group (8.9+/-0.4, 57.1+/-0.3, 10.2+/-3.9, 152.2+/-3.8, 325.7+/-186.8), ATP(++) group (9.1+/-0.4, 57.0+/-0.3, 12.4+/-1.8, 161.5+/-3.8, 642.9+/-174.3), IM(++)+ATP(++) group (8.6+/-0.4, 56.9+/-0.3, 12.0+/-2.3, 152.2+/-2.5, 326.3+/-160.3), IM(+++)+ATP(++) group (8.5+/-0.3, 56.8+/-0.2, 10.4+/-0.9, 147.4+/-2.6, 316.1+/-170.7), IMII(b) group (8.6+/-0.3, 56.9+/-0.3, 11.9+/-1.9, 150.0+/-2.8, 318.9+/-145.8), there were significant differences between groups (P<0.05-0.01). CONCLUSION: There was a significant difference between CG and GC in their ultrastructure and molecular biology. Only on the condition of changes of internal environment in combination with the harmful effect of external environment, chronic atrophic gastritis can then develop into gastric cancer. Hence it might have similar epithelial cell ultrastructure and molecular biological changes in ATP(++), IMII(b) and cancer, hence there were similar patterns of occurrence, development and transformation. Recognition of this trend might help to explore problems of prevention and cure.

Atrophy↗

Ultrastructural and immunocytochemical characterization of interstitial cells in pre- and postnatal developing sheep pineal gland.

Pineal gland interstitial cells from 32 sheep embryos (from day 54 of gestation until birth) and 18 sheep (from 1 month to >2 years) were analysed using ultrastructural and immunohistochemical techniques. From day 98 of gestation and throughout postnatal development, a second cell type was observed in addition to pinealocytes; these cells displayed uniform ultrastructural features similar to those of CNS astrocytes. Ultrastructural homogeneity was not matched by the results of histochemical and immunohistochemical analysis. Expression of phosphotungstic acid hematoxylin, glial fibrillary acidic protein and vimentin indicates that the second cell population in the developing ovine pineal gland is, in fact, a combination of glial-astrocyte cells at varying stages of maturity. Pineal interstitial cells started to show signs of functional activity evident in vascular tropism; such activity, evident from around day 98 of gestation, appeared to relate to the exchange of substances between the pineal parenchyma and blood vessels and, though it continued throughout postnatal development, was most evident in animals slaughtered between 9 months and 2 years of age (group II). Morphologically, functional activity in interstitial cells in this age-group was apparent in: 1, formation of specific contact sites between interstitial cells and nerve fibres in the perivascular space; and 2, the presence of numerous gap junctions between the bulbous endings of cytoplasmic processes.

Animals↗

Ultrastructural characteristics of ostrich eggshell: outer shell membrane and the calcified layers.

The ultrastructure of the eggshell of the domestic hen has been well researched and structural studies of other avian species, such as the ostrich, often base their interpretation of egg shell structure on that of the chicken. In the ostrich, lowered hatchability and hatching trauma may be due to shell ultrastructural abnormalities. In the present study the ultrastructure of the calcified portion, and the outer shell membrane (OSM), of domesticated ostrich eggshells was investigated using standard electron microscopic techniques. Transmission and scanning electron microscopy studies demonstrated intimate contact between cup-shaped structures present on the OSM and the mammillary layer of the calcified portion of the shell. The initial calcium carbonate growth of the calcified shell was of a dendritic nature with nucleation sites on the surface of the cup's contents. The dendritic growth gave way to a more randomly-orientated, smaller crystallite growth structure, which changed in form as it neared the vertical crystal layer (VCL). The VCL is described as being both amorphous and 'crumbly' depending on the plane of fracture. These observations suggest that firstly, initial calcification is contained within the cups and is then directed outwards to form the shell and that secondly, the VCL may contain an evolutionary, calcified cuticular layer. These observations serve as a baseline for studies investigating the effect of shell structure and strength on hatchling trauma and the influence of maternal diet.

Animals↗

Ultrastructural features of the follicular wall in developing follicles of the sexually immature ostrich (Struthio camelus).

The ultrastructure of the follicular wall in primordial, previtellogenic and vitellogenic follicles of the sexually immature ostrich is described in the present study. The follicular wall consists of a zona radiata, granulosa cell layer, basal lamina and thecal layer. Cytoplasmic processes from the plasma membranes of the granulosa cell layer and the ovocyte form the zona radiata in previtellogenic and vitellogenic follicles. The granulosa cell layer transforms from simple cuboidal epithelium in primordial follicles to simple columnar or pseudostratified columnar epithelium in previtellogenic and vitellogenic follicles. Transosomes were observed along the apical and lateral plasma membranes of granulosa cells. The thecal layer in previtellogenic and vitellogenic follicles consists of interna and externa components. The fibroblasts in the theca externa contain microfilaments, which are thought to be actin filaments. The study revealed ultrastructural features, which are associated with the transportation of yolk precursors and nutrients into the ovoplasm. In addition, the study indicates that, although the cells in the theca externa contain microfilaments, they do not possess the ultrastructural characteristics of smooth muscle cells.

Animals↗

Mixed mucus-secreting and oncocytic carcinoma of the thyroid: pathologic, histochemical, immunohistochemical, and ultrastructural study of a case.

We report a carcinoma that is, to the best of our knowledge, the first case of a mixed mucus-secreting and oncocytic carcinoma of the thyroid. We also describe the histochemical, immunohistochemical, and ultrastructural features of this tumor. A 59-year-old man complaining of severe bone pain and weight loss underwent clinical and radiologic investigations. The studies revealed a nodule in the left thyroid lobe that was "cold" by (131)I scintiscan and multiple lytic lesions of the skeleton that showed increased uptake by (99m)Tc-Sestamibi scintiscan. Left hemithyroidectomy was performed and the surgical specimen contained a well-circumscribed nodule of 3 cm in the greatest diameter. Light microscopy showed an oncocytic carcinoma with an area of glandular and papillary proliferation of mucin-producing cells. A double histochemical approach (Alcian blue-periodic acid-Schiff and Alcian blue-high-iron diamine) combined with ultrastructural investigation confirmed the presence of true mucus, ruling out the presence of breakdown products of thyroglobulin. Ultrastructural and immunohistochemical studies, together with clinical findings, excluded a possible metastatic origin of the mucin-producing component.

Adenocarcinoma, Mucinous↗