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Rabbit forebrain cholinergic system: morphological characterization of nuclei and distribution of cholinergic terminals in the cerebral cortex and hippocampus.

Although the rabbit brain, in particular the basal forebrain cholinergic system, has become a common model for neuropathological changes associated with Alzheimer's disease, detailed neuroanatomical studies on the morphological organization of basal forebrain cholinergic nuclei and on their output pathways are still awaited. Therefore, we performed quantitative choline acetyltransferase (ChAT) immunocytochemistry to localize major cholinergic nuclei and to determine the number of respective cholinergic neurons in the rabbit forebrain. The density of ChAT-immunoreactive terminals in layer V of distinct neocortical territories and in hippocampal subfields was also measured. Another cholinergic marker, the low-affinity neurotrophin receptor (p75(NTR)), was also employed to identify subsets of cholinergic neurons. Double-immunofluorescence labeling of ChAT and p75(NTR), calbindin D-28k (CB), parvalbumin, calretinin, neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase, or substance P was used to elucidate the neuroanatomical borders of cholinergic nuclei and to analyze the neurochemical complexity of cholinergic cell populations. Cholinergic projection neurons with heterogeneous densities were found in the medial septum, vertical and horizontal diagonal bands of Broca, ventral pallidum, and magnocellular nucleus basalis (MBN)/substantia innominata (SI) complex; cholinergic interneurons were observed in the caudate nucleus, putamen, accumbens nucleus, and olfactory tubercule, whereas the globus pallidus was devoid of cholinergic nerve cells. Cholinergic interneurons were frequently present in the hippocampus and to a lesser extent in cerebral cortex. Cholinergic projection neurons, except those localized in SI, abundantly expressed p75(NTR), and a subset of cholinergic neurons in posterior MBN was immunoreactive for CB and nNOS. A strict laminar distribution pattern of cholinergic terminals was recorded both in the cerebral cortex and in CA1-CA3 and dentate gyrus of the hippocampus. In summary, the structural organization and chemoarchitecture of rabbit basal forebrain may be considered as a transition between that of rodents and that of primates.

Acetylcholine↗

Physiological and morphological characterization of honeybee olfactory neurons combining electrophysiology, calcium imaging and confocal microscopy.

The insect antennal lobe is the first brain structure to process olfactory information. Like the vertebrate olfactory bulb the antennal lobe is substructured in olfactory glomeruli. In insects, glomeruli can be morphologically identified, and have characteristic olfactory response profiles. Local neurons interconnect glomeruli, and output (projection) neurons project to higher-order brain centres. The relationship between their elaborate morphology and their physiology is not understood. We recorded electrophysiologically from antennal lobe neurons, and iontophoretically injected a calcium-sensitive dye. We then measured their spatio-temporal calcium responses to a variety of odours. Finally, we confocally reconstructed the neurons, and identified the innervated glomeruli. An increase or decrease in spiking frequency corresponded to an intracellular calcium increase or decrease in the cell. While intracellular recordings generally lasted between 10 and 30 min, calcium imaging was stable for up to 2 h, allowing a more detailed physiological analysis. The responses indicate that heterogeneous local neurons get input in the glomerulus in which they branch most strongly. In many cases, the physiological response properties of the cells corresponded to the known response profile of the innervated glomerulus. In other words, the large variety of response profiles generally found when comparing antennal lobe neurons is reduced to a more predictable response profile when the innervated glomerulus is known.

Action Potentials↗

Further morphological characterization and structural proteins of infectious bursal disease virus.

An outer layer surrounding the capsid of infectious bursal disease virus was evident from electron micrographs of intact virus particles having diameters of 62 to 63 nm. The capsid was found to be composed of large morphological units or capsomeres, measuring about 12 nm in diameter. The architecture of the capsid appears to be that of T = 3 symmetry, with a probable 32 morphological units by rotational enhancement of image detail. Structural proteins of infectious bursal disease virus consist of seven species, two major and five minor polypeptides. These are P1 to P7, with molecular weights of 133 x 10(3), 124 x 10(3), 98 x 10(3), 51 x 10(3), 33 x 10(3), 26 x 10(3), and 23 x 10(3), respectively.

Capsid↗

Morphological characterization of renal cell lines (BGM and VERO) exposed to low doses of lead nitrate.

The response to lead nitrate has been assessed in two cell lines of renal origin. The range of toxic concentrations was determined by Neutral Red assay after 24-h of exposure. Morphological changes in the Buffalo Green Monkey (BGM) and VERO cell lines after exposure to subcytotoxic doses (1.38 mM and 1.04 mM, respectively) equivalent to EC10 (effective concentrations 10%) of lead nitrate were evaluated at the ultrastructural level by transmission microscopy. The most notable finding in treated cells was the presence of inclusion bodies in the form of irregular granules of varying size in both cytoplasm and lysosomes. Cell membrane integrity was not affected. The number of phagolysosomes and myeline figures associated to the inclusion bodies was higher than in the control cultures. We conclude that the phagolysosomic mechanism fails to digest this metal ion and the BGM and VERO renal cell lines can be considered as useful tools for toxicological studies involving lead nitrate.

Animals↗

[Morphological characterization of the myelopoietic stem cell reserves in the human].

According to microkinomatographic observations made on single cells up to ten days in vitro, there are the following laws of growth in haematopoiesis: Small cells will increase in growth up to five times in size, with their morphologic and kinetic properties being preserved, the blood lymphocyte will grow to immunoblasts, the small pluripotent stem cell to bone-marrow histiocytes. When growing, the myelopoietic stem cell may be gradually deviated into all myelopoietic cell lines. Instead of bone-marrow histiocytes it may differentiate to promyelocytes, promonocytes or proerythroblasts, all having an equal nucleus size, when it is induced by serum factors. Apart from histiocytes these large cells are capable of differentiating to a clon of blood cells, such as granulocytes, monocytes or reticulocytes, by several successive divisions of maturity. Contrary to the stimulated lymphocyte, symmetric mitoses will frequently occur, when small pluripotent stem cells are growing to bone-marrow histiocytes to be no further differentiated. Occasionally, asymmetric divisions may also be observed, i.e. one of the daughter cells will differentiate into one of the myelopoietic lines, whereas the other one will remain a progenitor cell. Moreover, there are various pathological mitoses in all progenitor cell sizes, such as endomitosis, cytoplasm fusion after mitosis, nucleus fusion after cytoplasm conjunction or amitotic nucleus division without cytokinesis. They produce megacaryoblasts further differentiating to megacaryocytes by corresponding pathological mitoses. According to our vital observations the pluripotency of the haematopoetic stem cell is being lost step by step.

Bone Marrow Cells↗

Flow cytometric and morphological characterization of platelet-rich plasma gel.

UNLABELLED: BACKGROUND OF PROBLEMS: Platelet-rich plasma (PRP) gel is derived from an autogenous preparation of concentrated platelets and is widely used in implant dentistry as a vector for cell growth factors. However, limited data are available on its structure and composition. The present study was aimed at providing a flow cytometric and ultrastructural characterization of PRP gel. MATERIALS AND METHODS: Twenty PRP gel samples were obtained from healthy volunteers. These PRP gel specimens were prepared for transmission (TEM) and scanning electron microscopy (SEM) examination of their morphological ultrastructure. Flow cytometry with CD41-PE monoclonal antibody was used to detect platelet cells, as this antibody recognizes human-platelet-specific antigen CD41. RESULTS: Both SEM and TEM showed that PRP gel contains two components: a fibrillar material with striated band similar to fibrin filaments, and a cellular component that contains human platelet cells. Both techniques indicated that no morphological elements were bound between the cellular component and the fibrillar material. The cells were confirmed as platelet cells by flow cytometric study after incubation with specific monoclonal antibody CD41-PE. CONCLUSION: PRP gel contains a fibrillar and a cellular (largely human platelet cell) component. This unique structure may be capable of acting as a vehicle for carrying of cells that are essential for soft/hard tissue regeneration.

Animals↗

Phenotypic and morphological characterization of neuroblastoma cells constitutively expressing B-myb.

B-myb gene is expressed in neuroblastoma cells and down-regulated during differentiation. We used B-myb-transfected LAN-5 cells, which constitutively express high level of B-myb, to detect changes at phenotypic and morphological levels in basal and differentiation conditions. Our results demonstrate that the overexpression of B-myb markedly affects the cytoskeletal composition, the pattern of neurotransmitter enzymes and the extracellular matrix expression. In general, B-myb transfected neuroblastoma cells show a broad potentiality without a direction toward a specific neuroectodermal differentiation pathway. On the other hand, we confirm inhibition of the neuronal differentiation upon retinoic acid (RA) treatment of B-myb transfected cells. Furthermore, the ultrastructural analyses are supportive of a change in the metabolism in B-myb transfected cell treated with RA. Our data suggest that B-myb expression is compatible with an early phase of differentiation of neuroectodermal cells, but must be down-regulated for the completion of the differentiative programme.

Cell Cycle Proteins↗

Morphological characterization of dorsal horn spinal neurons in rats with unilateral constriction nerve injury: a preliminary study.

A new animal model of neuropathic pain utilizing loose ligation of a peripheral nerve has been previously reported. In addition to displaying abnormal pain symptoms such as allodynia and hyperalgesia, physiologic and morphologic changes are seen in spinal cord dorsal horn neurons. Two weeks after ligation of the right common sciatic nerve, rat dorsal horn spinal cord neurons with signs of transsynaptic changes (dark neurons) were found on the side ipsilateral to the nerve injury. A few dark neurons were also found in the contralateral dorsal horn. The distribution of dark neurons in lumbar dorsal horn was limited to the superficial laminae (I-III). The following changes which suggest altered cellular activity were seen under the electron microscope. The nuclear envelope appeared ruffled while the mitochondria appeared normal. In addition, the dense cytoplasm was filled with rosettes of ribosomes as well as extensively developed rough endoplasmic reticulum and distended Golgi apparatus cisternae. While dark neurons had normal appearing somatic synapses, a few appeared atypical. The altered activity of these neurons may lead to abnormal sensory experiences and may be a consequence of central changes in response to persistent peripheral nerve injury. The purpose of the present study was to assess morphologic, hence functional, changes in spinal cord neurons in response to peripheral nerve constriction injury which evokes chronic pain-related behaviour.

Animals↗

Signet ring stomach cancer: morphological characterization and antigenic profile of a newly established cell line (Mz-Sto-1).

A human gastric signet ring cancer cell line (Mz-Sto-1) was established in tissue culture from the ascites fluid of a 54-year-old patient. The tumor cells growing in tissue culture exhibit the morphological characteristics of signet ring cells in phase contrast and transmission electron microscopy. Mz-Sto-1 cells grow as monolayer with a population doubling time of 28-36 hr during exponential growth phase and show a chromosome number between 72 and 74. In the cellular DNA of Mz-Sto-1 cells no amplification of 19 oncogenes studied is observed, c-myc included. Mz-Sto-1 cells secrete 150-250 ng CEA per 10(7) cell in 3 days, but no AFP. In addition Mz-Sto-1 cells and 2 already established gastric cancer cell lines MKN-28 and MKN-45 express HLA- and blood group related antigens (A, Lewis). HLA-DR antigens, which are regularly detected on normal stomach epithelium, are not found on any of the 3 cultured gastric cell lines. Mz-Sto-1 cells represent the first human gastric cancer cell characterized ultrastructurally as signet ring cells. This line will be a valuable tool to study the biology and genetics of gastric carcinoma, to test cytostatic drugs and to define new antigenic markers for stomach cancer.

Adenocarcinoma, Mucinous↗

Morphological characterization and subtyping of silent somatotroph adenomas.

GH-producing adenomas clinically are endocrine-active tumors accompanied with acromegaly in most instances. However, GH-producing adenomas apparently unassociated with acromegaly, or so-called silent somatotroph adenomas (SSA), have recently been reported but rarely. The reported cases are characterized by normal or slightly elevated serum levels of GH but without acromegaly. Tumor cells contain moderate, trace or no GH immunoreactivity. We experienced 7 cases of SSA which were not always similar in morphology and pathogenetic mechanism. They could be further divided into the following 3 subtypes. Subtype 1 (N = 2): a moderate number of cells were immunopositive for GH, and GH mRNA was also expressed in moderate or numerous cells. Densely granulated cells were noted. It is assumed that inhibition of hormone release into circulation. Subtype 2 (N = 3): a small number of cells were immunopositive for GH, while GH mRNA was expressed in numerous tumor cells. They were sparsely granulated cells containing fibrous bodies. These findings suggest that posttranslational processing of the gene product may be defective. Subtype 3 (N = 2): Only a scattered number of cells were immunopositive for GH and GH mRNA was co-localized in immunopositive cells. They were sparsely granulated cells containing poorly developed organelles that did not resemble those of typical sparsely granulated GH cells. The findings indicate that adenoma cells are largely immature with minimal GH lineage differentiation.

Acromegaly↗

Morphological characterization of chicken anaemia agent (CAA).

Chicken anaemia agent (CAA) was characterized as a virion with 25 nm in diameter, with a buoyant density in CsCl of 1.36-1.37 g/cm3, and containing a circular, single-stranded DNA genome. The virus is composed of 32 hollow morphological units representing a regular T = 3 icosahedron.

Anemia↗

Morphological characterization of stationary reticulum cells of the stromal in the mesenteric lymph node of the guinea pig.

The large mesenteric lymph node taken from guinea pigs in a period of time ranging from the 10th day prepartum till the 26th day postpartum has been examined in order to study: the morphological features of the stromal stationary reticulum cells with particular regard to recognize their stages of development; the possible ontogenetic relationship between these cells during the maturation of the lymphoid tissue. Our data support the hypothesis that from local mesenchymal cells originates a pool of poorly differentiated reticulum cells that can give rise to stromal stationary reticulum cells (myofibroblast-like cells, fibroblast-like cells, pericyte-like cells and dendritic cells). These elements have a characteristic distribution pattern likely related to different local functional requirements.

Aging↗

Morphologic characterization of proliferative cells and virus particles in turkeys with lymphoproliferative disease.

The tumors found in turkeys having lymphoproliferative disease (LPD) are histologically characterized by a pleomorphic population of cells of the lymphoid series. Electron microscopy has shown that, despite marked differences in shape and size, the proliferating cells share basic ultrastructural features, indicating their lymphoid origin. Virus particles morphologically and morphogenetically characteristic of type C oncorna-viruses of Retraviridae were found in different organs and plasma samples of diseased or infected turkeys with LPD. This LPD type C virus resembled members of the reticuloendotheliosis virus group but not members of the avian sarcoma virus group.

Animals↗

Morphological characterization of hemangiomatous tumors derived from a novel murine vascular endothelial cell line (F-2).

Hemangiomatous tumors were induced in Balb/c nude mice by inoculating F-2 cells (5 x 10(6)) from a novel tumorigenic murine endothelial cell line which had been established and maintained in our laboratories. These tumors were morphologically investigated in the course of development. Subcutaneous hemorrhage was observed at the inoculation site within 12 hours after the injection of F-2 cells, followed by development of skin tumors of various sizes at the same sites. They were dome-shaped, glossy surfaced, black, soft tumors. Mice finally died of massive blood loss due to internal and/or external hemorrhage (on day 10-77). Light microscopically, F-2 cells formed aggregates immediately after inoculation and then branched into a network of channels and cysts containing erythrocytes. Thereafter, spongiform structures composed of various sizes of cysts appeared with subsequent formation of a single large blood-filled cyst lined by one or two layers of thin cells. Under an electron microscope, F-2 cells, possessing large amounts of cytoplasm, formed narrow spaces which were occasionally incomplete with ambiguous basal lamina at the early stages. However, later, in large cysts, they became attenuated, tightly connected, and produced complete lumen surrounded by a basal lamina. Immunohistological demonstration of H-2 K, D antigen showed that tumors induced by F-2 cells mainly consisted of the inoculated F-2 cells. These results indicate that F-2 presents a good experimental system for investigation of vascular endothelial cell tumorigenesis and differentiation.

Animals↗

Morphological characterization of skin ganglion-like cells in Djungarian hamsters (Phodopus sungorus).

Characteristic ganglion-like cell proliferation observed in the skin of Djungarian hamsters was investigated using 24 male and 24 female hamsters, 1-6 months of age, to examine the anatomic location of these ganglion-like cells and their morphologic features. One abdominal skin tumor composed of these cells and resembling proliferative fasciitis in humans was also examined. Skin ganglion-like cells were rarely observed in young animals but increased in number and extent with age, especially in males. These cells were frequently seen in the ventral and medial regions of the trunk and legs rather than in the dorsal and lateral regions. Light microscopic examination of these ganglion-like cells revealed abundant vesicular basophilic cytoplasm with delicate intracytoplasmic silver stain-positive fibrils. Ultrastructurally, these cells contained abundant rough endoplasmic reticulum and Golgi complexes with dilated cisternae; intracellular collagen fibrils were present within these cisternae. Heat shock protein 47, beta-tubulin, and androgen receptor were expressed in these cells. The morphologic features of cells of one tumor resembling human proliferative fasciitis were identical to those observed in ganglion-like cells. The results of the present study suggest that these ganglion-like cells are derived from intrinsic undifferentiated mesenchymal cells in the dermis or subcutaneous adipose tissue and that any tumor-like lesion they form should be regarded as an abnormal proliferative lesion of skin ganglion-like cells rather than as proliferative fasciitis or fibroma.

Abdominal Neoplasms↗

Avian model for 13-cis-retinoic acid embryopathy: morphological characterization of ventricular septal defects.

In developing an avian model for 13-cis-retinoic acid (13cisRA) embryopathy, we found 13cisRA induced cardiovascular defects, especially Type I ventricular septal defects (VSDs) (Hart et al.: Teratology 41:463-472, '90). As the first step of investigating possible mechanisms, we have examined the light microscopic morphology of RA-induced cardiovascular defects in chick embryos. Fertilized eggs were injected via yolk sac with 150 micrograms 13cisRA in dimethylsulfoxide (DMSO), DMSO or mock injection on embryonic day 5 (E5). On E6, E7, or E8, surviving embryos were sacrificed and fixed in 10% formalin. Thoracic blocks were exised, embedded in paraffin and serially sectioned through the heart, base to apex. Slides were stained, screened for tissue orientation, then coded and evaluated without knowledge of treatment group. Examination of serial sections permitted qualitative evaluation of conotruncal ridge volume, mesenchymal organization, necrosis and extent of fusion. Extent of fusion was the only parameter influenced by 13cisRA treatment. On E6, ridge fusion was incomplete in all groups at comparable levels. On E7, ridge fusion in 13cisRA-treated embryos had not progressed as far proximally as in controls. By E8, there was a significant difference in the extent of fusion between 13cisRA-treated and non-RA-treated groups. We conclude 13cisRA-induced VSDs resulted from defective conotruncal ridge fusion and that the fusion defects did not result from decreased tissue volume, altered mesenchymal organization or increased necrosis.

Animals↗

Epidemic neuropathy in Cuba: morphological characterization of peripheral nerve lesions in sural nerve biopsies.

More than 50,000 patients were affected in Cuba during an epidemic outbreak of peripheral neuropathy from January 1992 until September 1993. The disease presented as either a retrobulbar optic neuropathy, a predominantly sensory peripheral neuropathy, a dorsolateral myeloneuropathy, or as mixed forms. The morphological findings in sural nerve biopsies from 34 patients with various forms of the disease are presented here. Frozen, paraffin and semi-thin sections were prepared for light and electron microscopy, immunohistochemistry and morphometric analysis. Every case presented morphological alterations ranging from mild axonal dystrophy (9 cases, or 27%) to moderate and severe axonal damage (25 cases, or 73%). In 6 cases (18%), axonal damage was accompanied by perineural fibrosis and vascular abnormalities. Axonal regeneration was noted in 8 cases (23%) and remyelination in 9 (26%). Morphometric analysis showed a predominant loss of myelinated fibers in 92% of the patients. Quantification of myelinated fiber loss in 11 patients revealed a remarkable decrease in large caliber fibers. Scarce mononuclear cells were observed in 17 cases. No virus-like elements were seen. The morphological features found in this study indicate that, regardless of the clinical presentation, peripheral nerve lesions of the epidemic neuropathy in Cuba correspond to an axonal neuropathy. These lesions are compatible with nutritional, toxic, or metabolic etiologies. An inflammatory etiology would be unusual with these lesions.

Adult↗

Genetic and morphological characterization of the Aedes (Ochlerotatus) dorsalis (Diptera: Culicidae) group in North America.

An examination of the electrophoretically detectable variation among the North American members of the Aedes (Ochlerotatus) dorsalis group revealed large genetic differences among all 4 species. At least 9 of 18 loci examined (50%) were diagnostic for each species pair. However, morphological variation observed among species was low. Only Aedes canadensis (Theobald) was separated readily from the other members of this group [Aedes dorsalis (Meigen), Aedes melanimon Dyar and Aedes campestris Dyar & Knab] in all life stages. Characters traditionally used to separate the remaining 3 species were less reliable. In the adult female, Ae. melanimon may be distinguished from Ae. campestris by the scaling patterns of the wings and abdomen, but Ae. dorsalis could not be distinguished reliably by these characters. Adults of Ae. dorsalis may be separated reliably from those of Ae. campestris and Ae. melanimon only by the length of the subapical tooth relative to the length of the tarsal claw. Ae. melanimon was identified in the larval stage by the short mesothoracic hair 1. Eight larval characters differed between Ae. dorsalis and Ae. campestris. However, the ranges of these characters overlapped and no character was truly diagnostic. Genetic variation within species was low as measured by average heterozygosity and Nei's genetic distance coefficients. No allozymes were diagnostic for coastal and inland populations of Ae. dorsalis, and the pattern of genetic differentiation within this species did not correspond to the geographic location of the populations examined. Therefore, the genetic data did not support the hypothesis that Ae. dorsalis represents a complex of 2 or more cryptic species.

Abdomen↗