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PubMed1988

Ultrastructural changes in dermal pig skin after exposure to heat and electric energy and acid and basic solutions.

In order to describe the ultrastructure of the histopathological changes in dermis after exposure to electrical energy, heat energy and acid and basic solutions the skin of fully anaesthetized Danish Landrace pigs were exposed to direct current, heat (80 degrees C and 450 degrees C) and acid and basic solutions. Biopsies were obtained immediately after the exposure from all types of injury. Biopsies from the cathode areas biopsies were also taken on day 1 and day 2.5 in order to describe the initial calcium deposits. Homogeneous collagen fibres without any birefringence from heat exposed areas were ultrastructurally composed of filamentous materials. Collagen fibres with fine densely spaced cross-striation from cathode areas and areas exposed to basic solutions were shown ultrastructurally to consist of parallelly arranged collagen fibrils with regular waves. It is concluded that the cross-striation of the collagen fibres observed in polarized light are due to a periodic change in the orientation of the fibres seen as waves of the fibres. The ultrastructure of dermal cells were similar to that of epidermal cells following the different types of influence. Characteristically the nuclei were condensed following heat and more electron-lucent following direct current (d.c.) and acid and basic solutions. In cathode areas and areas influenced by basic solutions the electron-lucent nuclei contained fine fibrils. The ultrastructural study supports the suggestion from light microscopic studies that the morphology of anode and cathode lesions shows resemblance to acid induced and basic induced lesions, respectively. Apatite crystals were observed on day 2.5 at the periphery of the collagen fibrils and in the matrix of elastic fibres.

Animals↗
PubMed2004

Ultrastructural changes in tracheobronchial epithelia following experimental traumatic brain injury in rats: protective effect of erythropoietin.

BACKGROUND: We aimed to demonstrate the time dependent ultrastructural changes in tracheobronchial epithelia after traumatic brain injury. And also, protective effect of erythropoietin was demonstrated. METHODS: We used 56 Wistar-Albino female rats weighing 170 to 200 g. The rats were allocated into 7 groups. First group was the control. The second underwent craniotomy without trauma. The third, fourth, and fifth groups were respectively 2-, 8-, and 24-hour trauma groups. The sixth and seventh groups were respectively treatment (erythropoietin, 1,000 IU/kg) and vehicle (0, 4 ml/rat) groups. Weight-drop method was used for achieving head trauma. Samples were obtained from both trachea and main bronchi. Modified electron microscopic scoring model was used to reveal the ultrastructural changes in both trauma and treatment groups. RESULTS: There was no statistical difference between control and sham groups (p >0.05). Scores of all trauma groups were significantly different from the controls (p <0.05). Trauma produced obvious gradual damage on ultrastructure of the tracheobronchial epithelia. Erythropoietin decreased tracheobronchial scores after traumatic brain injury in significant levels. Erythropoietin attenuated ultrastructural scores for each organelle in significant levels (p <0.05 for each organelle). CONCLUSIONS: The data suggested that ultrastructural damage is obvious at 2 hours deteriorating with time. Erythropoietin protects epithelia against damage after traumatic brain injury. Pharmaceutical lung preservation may help gaining efficacious donor lungs in brain death. But, further time dependent experiments are needed to determine the liability of the donor lung after traumatic brain injury. This fact is to be known for achieving higher graft survival rates.

Animals↗
PubMed2004

Ultrastructural evidence of increased tolerance of hibernating myocardium to cardioplegic ischemia-reperfusion injury.

OBJECTIVES: The goal of this study was to investigate the effects of ischemia-reperfusion on myocardial ultrastructure in patients with and without hibernating myocardium. BACKGROUND: It is generally accepted that chronically dysfunctional, hibernating myocardium may remain nonetheless viable for a long time. It has been postulated that hibernating myocytes may survive, despite being subtended by a severe coronary artery stenosis, as they might be less susceptible to ischemic insults. However, whether hibernating myocardium is indeed more resistant to ischemia has never been investigated. METHODS: Myocardial biopsies were taken before cardiac arrest and after reperfusion from the anterior wall of the left ventricle in patients undergoing coronary artery bypass surgery, divided according to presence (n = 7) or absence (n = 7) of hibernating myocardium. Ultrastructural changes were studied by electron microscopy. Because ischemia-reperfusion injury is related to oxidative stress, we also evaluated coronary sinus concentration of the antioxidants alpha-tocopherol, beta-carotene, and ubiquinol, and of lipid peroxidation products pre-ischemia and after reperfusion. RESULTS: Both groups were similar with respect to length of ischemia and changes in the various indexes of oxidative stress. In normally contracting myocardium, ischemia/reperfusion induced moderate overall ultrastructural changes, and marked alterations at the mitochondrial level. In contrast, post-reperfusion biopsies of hibernating myocardium displayed only minor overall ultrastructural changes, and scored significantly better on mitochondrial damage. CONCLUSIONS: Despite similar severity of ischemia/reperfusion, hibernating myocardium showed significantly less ultrastructural evidence of cell injury compared with normally contracting myocardium. These data indicate that human hibernating myocardium is intrinsically more resistant to ischemia/reperfusion injury.

Aged↗
PubMed1986

Protective effect of diltiazem on ultrastructural alterations induced by coronary occlusion and reperfusion in dog hearts.

This study was designed to examine whether diltiazem, a calcium channel-blocker, inhibits the cardiac ultrastructural alterations induced by coronary occlusion with or without reperfusion, in dogs anesthetized with pentobarbital. The left anterior descending coronary artery (LAD) was completely occluded for 60 min with or without reperfusion (induced by release of occlusion) for 20 min. Coronary occlusion increased ST segment in the ischemic area, and also produced typical ultrastructural alterations including decreased glycogen granules, destruction of mitochondria, and margination of the nuclear chromatin, especially in the subendocardium. Reperfusion of the ischemic area resulted in more severe alterations of the myocardial ultrastructure, including many myofibrillar contraction bands. Diltiazem was injected intravenously at the dose of 200 micrograms/kg (bolus injection) 20 min before LAD occlusion, and was then infused intravenously at the rate of 80 micrograms/kg/min for 10 min starting at the beginning of LAD occlusion, the total dose being 1 mg/kg. Diltiazem decreased heart rate and diastolic blood pressure, inhibited the increase in ST segment, and also inhibited the ultrastructural alterations induced by coronary occlusion, regardless of reperfusion. A bolus injection of diltiazem alone (200 micrograms/kg), however, did not inhibit markedly the ultrastructural alterations induced by coronary occlusion, regardless of reperfusion. It is concluded that the large dose of diltiazem (1 mg/kg) protects the myocardium from ischemic injury.

Animals↗
PubMed2001

Sun and shade leaves? Cuticle ultrastructure of Jurassic Komlopteris nordenskioeldii (Nathorst) Barbacka.

An ultrastructural transmission electron microscope (TEM) study of fossil leaf cuticles from the Jurassic pteridosperm Komlopteris nordenskioeldii (Nathorst) Barbacka from the Mecsek Mountains (South Hungary) was conducted. Remnants of cuticles of leaves originating from so-called "sun and shade" environments were sectioned with a diamond knife, transversally as well as longitudinally. Although the present study showed a simple type of cuticle in this pteridosperm, differences were observed in the occurrence of its components, such as electron lucent amorphous material and various densities of granules, which give rise to different zones. The included fibrilous elements appeared to be made of aggregated and aligned granules, equivalent in size and electron density to nearby non-fibrilous granular regions. The combinations of these ultrastructural features allow distinctions between four types of cuticle: sun upper, sun lower, shade upper and shade lower. Considering the distinction made earlier in two types of cuticle and supposed to be related to sun and shade on the basis of macroscopical and microscopical features, four types only on the basis of differences in thickness, the present study reinforces the distinctions with ultrastructural microcharacteristics. As this study shows the variations in ultrastructure of cuticle among the four types, the differences observed may reveal the great sensitivity of some plants to environment. At the same time, it points out the importance, in ultrastructural studies of cuticles, of studying a number of samples for one taxon.

Journal Article↗
PubMed1990

Ultrastructural heterogeneity of lung carcinomas: representativity of samples for electron microscopy in tumor classification.

Histologic heterogeneity of tumors is a well-known phenomenon, which has been repeated studied at the light microscopic level. Electron microscopy has been advocated as an adjunct in classification of tumors which pose difficulties on light microscopic classification. However, in view of tumor heterogeneity, it might be anticipated that the problem of sample error could detract from the usefulness of electron microscopy in tumor typing. Ultrastructural heterogeneity of tumors has thus far not been systematically investigated. We performed an ultrastructural randomized and blinded study of superficial and deep samples of 44 resected lung carcinomas, tumors which are notorious for their histologic heterogeneity. Neuroendocrine and squamous differentiation, as well as adenodifferentiation, were assessed separately and semiquantitatively in each sample. Twenty-six tumors showed more than one type of differentiation in at least one sample. However, in only two cases did the main type of differentiation differ between the two samples. A further nine cases showed one predominant differentiation type in both samples, but a similarly pronounced second differentiation type in one of the samples. Thus, in terms of ultrastructural diagnosis, the two samples showed a major discrepancy in two of 44 cases, and a minor discrepancy in nine of 44 cases. We conclude that ultrastructural heterogeneity of lung tumors is a common occurrence, but that it only rarely leads to totally different ultrastructural diagnoses.

Carcinoma↗
PubMed1982

The ultrastructure of poorly differentiated rhabdomyosarcomas: a case report and literature review.

The ultrastructure of a case of rhabdomyosarcoma is reported, and the world literature on the ultrastructure of rhabdomyosarcomas is reviewed. The reported case was first observed as an undifferentiated malignancy originating in the maxillary antrum of a 46-year-old white woman and later showed differentiated cells in metastatic sites. A specific feature of the sarcomere within a representative malignant cell is necessary and sufficient for diagnosis of rhabdomyosarcoma by ultrastructure. Examples of specific features include thick and thin filaments in a hexagonal array, an unambiguous Z line, or A bands with H and M bands. The presence of thick and thin filaments without additional features of the sarcomere indicate a myogenous origin, either leiomyosarcoma or rhabdomyosarcoma. Other ultrastructural features observed in rhabdomyosarcomas, for which the diagnosis is confirmed by the presence of diagnostic ultrastructural features or by the presence of cross-striations observed by light microscopy, include large irregularly shaped nuclei with projections and invaginations, free ribosomes closely associated with filaments, rough endoplasmic reticulum that is often dilated, glycogen, and external lamina, prominent nucleoli, and pleomorphic mitochondria. When this spectrum of features is present, the possibility of rhabdomyosarcoma cannot be excluded, and a diligent search for the diagnostic components of the sarcomere should be made. Small intercellular junctions, pinocytotic vesicles, and paranuclear cilia occurred in the poorly differentiated metastasis of this case; the presence of these nonspecific features permits neither diagnosis nor exclusion of rhabdomyosarcoma.

Breast Neoplasms↗
PubMed2002

Ultrastructural differences in bovine morulae classified as high and low qualities by morphological evaluation.

Bovine morulae collected from superovulated cows were classified into four groups (excellent, good, fair, poor) using morphological evaluation under a phase contrast microscope. The number of blastomeres in morulae classified as excellent and good (high quality) was significantly higher than the number in fair and poor (low quality) morulae. The ultrastructure of morulae in each group was compared. In morulae classified as excellent and good, two characteristic cell types could be identified by the electron density of their cytoplasm and by their ultrastructural features. One type (presumptive trophoblast cells) was located in the peripheral layer of the embryo mass, was generally characterized by low electron density of cytoplasm, and possessed numerous cellular organelles such as mitochondria and Golgi apparatus and had numerous microvilli projecting into the perivitelline space. The other type (presumptive embryonic cells) was located in the interior of the embryo mass and distinguished by cytoplasm that stained more densely than that of the lighter-appearing cells. The darker-appearing cells generally possessed fewer organelles than the lighter cells, but many lysosome-like structures were present in the cytoplasm. All high quality morulae contained well-developed mature nucleoli. Many embryos classified as fair contained both types of cells, while cells in some fair and most poor quality morulae could not be distinguished by the electron density of their cytoplasm and ultrastructural features. The morulae classified as fair and poor showed less well-developed junctional complexes between cells, less well-developed apical microvilli on the blastomeres, and contained large numbers of lipid droplets, vesicles, immature mitochondria, and nucleoli with low transcriptional activity. This study demonstrates conspicuous differences in the ultrastructural features between the high quality and the low quality morulae, suggesting that these ultrastructural features may reflect the various physiological anomalies observed in previous studies.

Animals↗
PubMed1999

Ultrastructure of vestibular commissural neurons related to velocity storage in the monkey.

The angular vestibulo-ocular reflex maintains gaze during head movements. It is thought to be mediated by two components: direct and velocity storage pathways. The direct angular vestibulo-ocular reflex is conveyed by a three neuron chain from the labyrinth to the ocular motoneurons. The indirect pathway involves a more complex neural network that utilizes a portion of the vestibular commissure. The purpose of the present study was to identify the ultrastructural characteristics of commissural neurons in the medial vestibular nucleus that are related to the velocity storage component of the angular vestibulo-ocular reflex. Ultrastructural studies of degenerating medial vestibular nucleus neurons were conducted in monkeys following midline section of rostral medullary commissural fibers with subsequent behavioral testing. After this lesion, oculomotor and vestibular functions attributable to velocity storage were abolished, whereas the direct angular vestibulo-ocular reflex pathway remained intact. Since this damage was functionally discrete, degenerating neurons were interpreted as potential participants in the velocity storage network. Ultrastructural observations indicate that commissural neurons related to velocity storage are small and medium sized cells having large nuclei with deep indentations and relatively little cytoplasm, which are located in the lateral crescents of rostral medial vestibular nucleus. The morphology of degenerating dendritic profiles varied. Some contained numerous round or tubular mitochondria in a pale cytoplasmic matrix with few other organelles, while others had few mitochondria but many cisterns and vacuoles in dense granular cytoplasm. The commissural nature of these cells was further suggested by the presence of two different types of degenerating axon terminals in the rostral medial vestibular nucleus: those with a moderate density of large spherical synaptic vesicles, and those with pleomorphic, primarily ellipsoid synaptic vesicles. The recognition of two types of degenerating terminals further supports our interpretation that at least two morphological types of commissural neurons participate in the velocity storage network. The degenerating boutons formed contacts with a variety of postsynaptic partners. In particular, synapses were observed between degenerating boutons and non-degenerating dendrites, and between intact terminals and degenerating dendrites. However, degenerating pre- and postsynaptic elements were rarely observed in direct contact, suggesting that additional neurons are interposed in the indirect pathway commissural system. On the basis of these ultrastructural observations, it is concluded that vestibular commissural neurons involved in the mediation of velocity storage have distinguishing ultrastructural features and synaptology, that are different from those of direct pathway neurons.

Animals↗
PubMed1998

Ultrastructural assessment of the myocardium receiving intermittent antegrade warm blood cardioplegia.

Ultrastructural changes in the myocardium during intermittent warm blood cardioplegia were studied in 12 dogs. During cardiopulmonary bypass, dogs received three 5-minute periods of warm blood cardioplegia, interrupted by three 10-minute episodes of ischemia in group A or three 20-minute episodes of ischemia in group B. Biopsy specimens were taken from the right ventricle before each cardioplegic infusion and at the end of the aortic cross-clamp period to analyze ultrastructural changes. Functional recovery was measured after cardiopulmonary bypass. Mild and principally reversible ultrastructural changes were observed in group A. In group B, moderate ultrastructural changes were observed, and these changes suggested cumulative ischemic injury. Functional recovery was good in group A, but significantly deteriorated in group B. We concluded that three 10-minute interruptions of warm blood cardioplegia cause mild ultrastructural changes and seem to be safe, but three 20-minute interruptions cause cumulative ischemic injury in normal dog hearts.

Animals↗
PubMed2001

The effect of acute oxidative stress on the ultrastructure of the perfused rat liver.

Ageing and liver disease are associated with ultrastructural changes in the hepatic sinusoid. Because of the possibility that reactive oxygen species could mediate these processes, we examined the effect of acute oxidative stress on the ultrastructure of the intact liver. Rat livers were perfused ex vivo, in situ with hydrogen peroxide via the portal vein. The livers were then fixed and the ultrastructure of the liver tissue examined with transmission and scanning electron microscopy. The effects of hydrogen peroxide were largely confined to the perisinusoidal areas. The sinusoidal endothelial cells became swollen and more porous, with large gaps replacing sieve plates. The space of Disse showed an increase in volume and the density of hepatocyte projections decreased. Kupffer cell activation was noted. Little or no ultrastructural change was observed within the hepatocytes. Oxidative stress delivered via the portal vein dramatically alters the ultrastructure of the perisinusoidal regions of the liver. This process may contribute to the pathogenesis of disease and age-related changes in the liver.

Animals↗
PubMed2006

Ultrastructural features of lymphocyte suppression induced by anthrax lethal toxin and treated with chloroquine.

Antibacterial therapy does not fully protect against anthrax because of severe systemic intoxication. Lysosomal processing of anthrax lethal toxin (LTX) is a key event in the disease pathogenesis, and agents interfering with this process, like chloroquine (CQ), may have practical applications. Although LTX is known to induce T-cell suppression, precise mechanisms of this phenomenon are not completely characterized. In the present study, we investigated alterations of lymphocyte ultrastructure caused by LTX and associated with favorable effect of CQ on the LTX-related dysfunction. Peripheral blood lymphocytes were activated via CD3 crosslinking in the presence or absence of LTX and CQ, and examined by transmission electron microscopy, flow cytometry and immunoblotting. Crosslinking of CD3 induced ultrastructural signs of lymphocyte activation, mostly disappeared after LTX treatment. The cell ultrastructure was well preserved in LTX-treated cells, despite dose- and time-dependent inhibition of T-cell function associated with impaired activation of mitogen-activated protein kinase. Regardless of intracellular signaling abnormalities, LTX did not decrease T-cell viability. CQ restored expression of CD69 (P<0.001) and improved phosphorylation of p38 (P=0.022) in LTX-exposed T lymphocytes. The exposure of cells to CQ, with or without LTX, led to appearance of many phagolysosomes with heterogeneous content, possibly representing unprocessed internalized material. In conclusion, LTX suppressed T-cell functions, but did not affect the viability and caused no ultrastructural damage. Ultrastructural observations indicated that CQ reduced harmful effects of LTX, possibly by interfering with lysosomal activity.

Analysis of Variance↗
PubMed2001

Ultrastructural features resembling those of harlequin ichthyosis in patients with severe congenital ichthyosiform erythroderma.

Congenital ichthyoses are a group of heterogeneous disorders of cornification. Autosomal recessive congenital ichthyosis (ARCI) can be clinically subdivided into congenital ichthyosiform erythroderma and lamellar ichthyosis. Ultrastructurally, ARCI is classified into four groups: ichthyosis congenita (IC) types I-IV. The genetic background of the ARCI disorders is heterogeneous, but only one disease gene, transglutaminase 1, has been detected so far. We describe six patients with severe congenital ichthyosis from six different Scandinavian families. They could not be classified ultrastructurally into the four IC groups because of atypical findings of electron microscopy. These included abnormal lamellar bodies, alterations in keratohyalin, remnant organelles and lipid inclusions in the upper epidermal cells, which resembled the ultrastructural findings of harlequin ichthyosis (HI), although the HI phenotype was not present at birth. Some clinical features, such as thick scales, erythroderma, alopecia and ectropion were common to all patients. Ichthyosis was usually accentuated in the scalp and four patients had clumped fingers and toes. None of the patients carried the transglutaminase 1 mutation. We conclude that ultrastructural findings resembling those detected in previous HI cases (type 1 and 2) can also be found in patients who do not have classic clinical features of that rare ichthyosis. This may be due to lack of specificity of ultrastructural markers for HI or to its clinical heterogeneity.

Adolescent↗
PubMed1999

Ultrastructural features of nerve fascicles and basal lamina abnormalities in Hirschsprung's disease.

Although the pathogenesis of Hirschsprung's disease (HD) is not completely resolved, both the absence of nerve cells and the hypertrophy of nerve fascicles within the aganglionic colonic segment have been attributed to an abnormal intestinal microenvironment. Studies on animal models for HD revealed an altered ultrastructure of ingrowing nerve fascicles and abnormalities of basal laminae (BL). Therefore, the purpose of this study was to examine the ultrastructure of hypertrophied nerve fascicles in human HD with special reference to structural abnormalities of BL. Colonic specimens were obtained from patients with HD (n = 10) and controls (n = 5) and processed for electron-microscopical examination. Hypertrophied nerve fascicles were characterized by a prominent perineural sheath surrounded by large amounts of collagen bundles, a collagen-filled endoneurium, vasa nervorum and abundant glial cells of extraenteric ultrastructure, which were arranged in mono- or oligoaxonal units and frequently displayed different stages of myelination. As these ultrastructural characteristics resembled typical features of extrinsic nerves and were similar to those observed in subserosal nerves, the prominent intramural nerve fascicles were considered to be of extraenteric origin. Most likely their overabundance contributes to the functional obstruction of the terminal colon. Morphological abnormalities of BL encountered in the aganglionic colonic segment consisted of an extensive multilamination of BL surrounding glial processes and an irregular thickening of BL surrounding perineurocytes and smooth muscle cells of the muscularis mucosae. Similar alterations in BL have also been described in inherited peripheral and diabetic autonomic neuropathies and attributed to reactivated schwann cells. Thus, the overproduction of BL material within the hypertrophied nerve fascicles in HD may reflect an increased activity of proliferating glial cells. Since the smooth muscle cells of the muscularis mucosae showed abnormalities of BL similar to those observed in murine models for HD, it is suggestive that also in human HD the aganglionic colon is affected by a disturbed intestinal micro-environment impairing the neuronal colonisation and promoting the ingrowth of extrinsic nerves. The ultrastructurally observed alterations in BL of both neuronal and non-neuronal cells, as well as the increased amount of perineural and endoneural collagen provide further evidence that extracellular matrix components are abnormally distributed and overproduced within the bowel wall of patients affected by HD.

Animals↗
PubMedDate not supplied

Pituitary carcinoma: an ultrastructural study of eleven cases.

Pituitary carcinomas are very rare. Defined as adenohypophysial tumors that undergo craniospinal and/or systemic metastasis, most are PRL- or ACTH-producing. Their ultrastructural features, particularly relative to benign adenomas of similar functional type, have not been sufficiently explored. Eleven cases of immunohistochemically characterized pituitary carcinoma with documented cerebrospinal and/or systemic metastases were collected from various institutions and studied by transmission electron microscopy. The tumors were surgically removed from 7 women and 4 men ranging in age between 28 and 74 years (mean, 50 years). All were endocrinologically functioning. Six tumors secreted PRL; three were ACTH-producing; one each was GH/PRL- and TSH-producing. The patients with the ACTH-producing tumors had all presented with Cushing's disease and two of them had undergone adrenalectomy (Nelson syndrome). In most cases significant cellular atypia and mitotic activity were observed. In terms of morphologic features of functional differentiation, electron microscopy revealed that in 9 cases the tumor cells maintained at least some ultrastructural markers of their basic phenotype. A unique feature in 2 ACTH carcinomas was the variable admixture of smooth endoplasmic reticulum with intermediate (cytokeratin) filaments. In 2 cases, both PRL-producing carcinomas, the cell type comprising the tumor could not be identified on an ultrastructural basis alone. Ultrastructural investigation of pituitary carcinomas confirms their endocrine nature and, in most but not all cases, reveals their functional differentiation. Despite the diagnostic utility of electron microscopy in the assessment of these rare tumors, the distinction of pituitary carcinoma from pituitary adenoma cannot be firmly made on ultrastructural grounds alone.

Adenoma↗
PubMedDate not supplied

Pediatric and adult hepatic embryonal sarcoma: a comparative ultrastructural study with morphologic correlations.

Hepatic embryonal (undifferentiated) sarcoma (ES) is a rare pediatric tumor occurring predominantly in the first decade of life, but a few examples of adult ES have also been described. Isolated ultrastructural reports describe contradictory lines of differentiation in these tumors. Four pediatric and 3 adult ES cases were studied ultrastructurally and features were correlated with morphology. Morphologically, tumors were composed of mixture of plump spindle cells and bizarre giant cells, showing abundant cytoplasmic eosinophilic globules. Ultrastructurally, the hallmark features in all cases included dilated RERs and secondary lysosomes with dense precipitates. Dilated mitochondria and mitochondrial-RER complexes were often seen. Other features included intracytoplasmic fat droplets, scant actin microfilaments, and focal glycogen pools. In summary, pediatric and adult ES show similar morphologic and ultrastructural features. Ultrastructurally, hepatic ES have distinctive findings, including dilated RER and electron-dense lysosomal precipitates, which correlate with the eosinophilic hyaline bodies seen microscopically. These findings suggest that ES are composed of fibroblastic, fibrohistiocytic, and undifferentiated cells. Other lines of differentiation were not identified.

Adult↗
PubMed1969

Ion-induced ultrastructural transformations in isolated mitochondria. The energized uptake of calcium.

The energized uptake of low levels of Ca(2+) in the presence and absence of phosphate by isolated rat liver mitochondria, and the perturbation effected by this activity on ultrastructural and metabolic parameters of mitochondria have been investigated. In the presence of phosphate, low levels of Ca(2+) are taken up by mitochondria and result in various degrees of ultrastructural expansion of the inner mitochondrial compartment. This indicates that low levels of Ca(2+) in the presence of phosphate, are accumulated in an osmotically active form into the water phase of the inner compartment. The first clearly observable quantitative increase in the volume of the inner compartment occurs after the accumulation of 100 nmoles Ca(2+)/mg protein. An accumulation of 150-200 nmoles Ca(2+)/mg protein, which is equivalent to the osmolar concentration of endogenous K(+), is required to effect a doubling of the volume of the inner compartment. This degree of osmotic perturbation occurs as mitochondria transform from a condensed to an orthodox conformation. The osmotically induced orthodox conformation differs from the mechanochemically induced orthodox conformation previously described, in that its development is concomitant with a marked decrease in acceptor control and oxidative phosphorylation efficiency and it fails to transform to a condensed conformation in response to addition of ADP. In the absence of added phosphate, a maximum of 190 nmoles Ca(2+)/mg protein was found to be taken up by mitochondria (state 6). Ca(2+) is apparently bound under state 6 conditions since the uptake does not effect an ultrastructural expansion of the inner compartment. Phosphate added after state 6 Ca(2+) binding, however, results in an immediate ultrastructural expansion of the inner compartment. The addition of phosphate to mitochondria in the absence of exogenous Ca(2-) fails to effect an osmotic ultrastructural transformation. Under state 6 conditions, the binding of between 40 and 190 nmoles Ca(2+)/mg protein results in the formation of dense matrix inclusions which appear to be composed of tightly packed, concentrically oriented membranes. Under conditions in which the bound Ca(2+) is subsequently released, there is a concomitant loss in the density of these matrix inclusions, leaving behind morphologically distinct membrane whorls in the mitochondrial matrix.

Adenine Nucleotides↗
PubMedDate not supplied

Ultrastructure and viability of isolated bovine preantral follicles.

Techniques for the isolation of ovarian follicles and maturation of oocytes in vitro have enormous reproductive potential. Preservation of normal tissue function is vital. This study emphasizes the ultrastructure and viability of mechanically isolated bovine small (diameter 40-100 microm) preantral and large (140-450 microm) preantral/early antral follicles. Viability studies were performed for small preantral follicles. The presence of esterase activity, active mitochondria and dead cells served as parameters of oocyte and granulosa cell viability. After 1 day of culture, all follicles had a viable granulosa, displaying active mitochondria and/or esterase activity in all their cells, although a few (generally <5) dead granulosa cells were present in 17% of the follicles. Of the oocytes, 35 and 80% had esterase activity and active mitochondria respectively, whereas 8% appeared dead. The percentages of oocytes showing esterase activity and active mitochondria decreased during culture, whereas the percentage of follicles with dead oocytes or dead granulosa cells strongly increased. More than 90% of the isolated small follicles showed a poor ultrastructure, especially of their oocyte, which points to a negative selective isolation of poor follicles in the present study and/or an isolation procedure-induced damage of follicles. With respect to large preantral follicles, 42% of those distributed in the cortex and 64% of those isolated and cultured for 1 day had a poor ultrastructure. In contrast with the small ones, the percentage of ultrastructurally poor large preantral follicles had decreased to 27% after 5 days of culture, possibly due to better isolation and culture conditions. It is recommended to use ultrastructural and/or viability cell markers for in-vitro grown follicles to evaluate their quality, and particularly that of their oocytes.

Animals↗