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PubMedDate not supplied

[Morphologic characterization and protein analysis of phages isolated from type strains of Bacillus thuringiensis].

The lysogeny of eleven type strains of Bacillus thuringiensis was studied. Eight new phages were isolated from the variants B. thuringiensis var. tochigiensis, yunnanensis, colmeri, shandongiensis, neoleonensis, silo, mexicanensis, and toguchini belonging to the B1 morphotype. The phages that were isolated from B. thuringiensis var. tochigiensis, yunnanensis, shandongiensis, and mexicanensis possessed transverse disks at their tails and were classified into one morphological group. The protein patterns of the isolated phages were determined.

Bacillus thuringiensis↗

PubMed1994

Effect of cholesterol-supplemented diet in heritable hyperlipidemic Yoshida rats: functional and morphological characterization of thoracic aorta.

In this study we have considered the possibility of inducing vascular damage in Yoshida Pittsburg (YOS) rat, an inbred strain which has endogenous hyperlipidemia without vascular atherosclerotic damage. Cholesterol-enriched diet (4% cholesterol plus 1% cholic acid and 0.5% thiouracil) was administered to YOS rats, in order to induce atherogenesis. The results indicate that, despite significant increase in serum (about 2-fold) and aortic tissue cholesterol (about 6-fold), no morphological damage occurred. A reduction in acetylcholine-mediated relaxation (of about 37%) was observed. No inhibition of ATP- or sodium nitrite-induced relaxation, or of contraction induced by norepinephrine was seen. Serum triglyceride concentration did not vary after administration of a cholesterol-enriched diet. Our results suggest that in heritable hyperlipidemic Yoshida rat, after 2 months of cholesterol-enriched diet, despite increased serum cholesterol levels, no atheromatous plaque developed on the aortic wall. Impaired vascular function and reductions in the response to acetylcholine were related to changed endothelial cell function. Administration of a high cholesterol diet to YOS rat may represent a new model of mixed endogenous and exogenous hyperlipidemia that can resemble many human dislipidemic diseases and therefore may become a useful tool for the study of isolated endothelial dysfunction.

Acetylcholine↗

PubMed2003

Electrophysiological and morphological characterization of identified motor neurons in the Drosophila third instar larva central nervous system.

We have used dye fills and electrophysiological recordings to identify and characterize a cluster of motor neurons in the third instar larval ventral ganglion. This cluster of neurons is similar in position to the well-studied embryonic RP neurons. Dye fills of larval dorsomedial neurons demonstrate that individual neurons within the cluster can be reproducibly identified by observing their muscle targets and bouton morphology. The terminal targets of these five neurons are body wall muscles 6/7, 1, 14, and 30 and the intersegmental nerve (ISN) terminal muscles (1, 2, 3, 4, 9, 10, 19, 20). All cells except the ISN neuron, which has a type Is ending, display type Ib boutons. Two of these neurons appear to be identical to the embryonic RP3 and aCC cells, which define the most proximal and distal innervations within a hemisegment. The targets of the other neurons in the larval dorsomedial cluster do not correspond to embryonic targets of the neurons in the RP cluster, suggesting rewiring of this circuit during early larval stages. Electrophysiological studies of the five neurons in current clamp revealed that type Is neurons have a longer delay in the appearance of the first spike compared with type Ib neurons. Genetic, biophysical, and pharmacological studies in current and voltage clamp show this delay is controlled by the kinetics and voltage sensitivity of inactivation of a current whose properties suggest that it may be the Shal I(A) current. The combination of genetic identification and whole cell recording allows us to directly explore the cellular substrates of neural and locomotor behavior in an intact system.

4-Aminopyridine↗

PubMed2000

Morphological characterization of bicontinuous structures in polymer blends and microemulsions by the inverse-clipping method in the context of the clipped-random-wave model.

A method is proposed to determine the spectral function of the clipped-random-wave (CRW) model directly from scattering data. The spectral function f(k) (k is a wave number) gives the distribution of the magnitude of wave vectors of the sinusoidal waves that describes the essential features of the two-phase morphology. The proposed method involves "inverse clipping" of a correlation function to obtain f(k) and does not require any a priori assumptions for f(k). A critical test of the applicability of the inverse-clipping method was carried out by using three-component bicontinuous microemulsions. The method was then used to determine f(k) of the bicontinuous structure of a phase-separating polymer blend. f(k) for the polymer blend turned out to be a multipeaked function, while f(k) for the microemulsions exhibits a single broad maximum representing periodicity of the morphology. These results indicate the presence of the long-range regularity in the morphology of the polymer blend. Three-dimensional (3D) morphology corresponding to the scattering data of the polymer blend was generated using the CRW model together with the multipeaked f(k). Interface curvatures of the 3D morphology calculated from f(k) were measured and compared with those experimentally determined directly from the laser scanning confocal microscopy in the same blend.

Journal Article↗

PubMed2005

Morphological characterization of follicle deviation in Nelore (Bos indicus) heifers and cows.

Follicle diameter deviation is defined as the beginning of the differential change in growth rates between the largest and next largest follicles subsequent to wave emergence and is considered a key component of follicle selection. Follicle selection has been extensively studied in European breeds of cattle (Bos taurus) but has not been critically studied in Zebu breeds (Bos indicus). The objectives of the present study were to determine and compare the morphological characteristics of deviation associated with the first post-ovulatory wave (Wave 1) of the estrous cycle in Nelore heifers (n=8) and nonlactating cows (n=11). Beginning on the day of ovulation (day 0), the three largest follicles (F1-F3, respectively) were individually tracked every 12 h for 6d using transrectal ultrasonography. In individual animals, deviation was determined graphically using visual inspection of the diameter profiles of F1, F2 and sometimes F3 (observed deviation) and mathematically using segmented regression analysis of the diameter differences between F1 and F2 or sometimes F3 (calculated deviation). Mean day of emergence of Wave 1 when F1 reached >3 mm (approximately 1 d after ovulation) and growth rate of F1 during deviation (approximately 1.4 mm/d) were not significantly different between heifers and cows. The results of determining the beginning of deviation within heifers and cows using the observed and calculated methods were not significantly different. Averaged over both methods, diameter deviation occurred 2.8 d after ovulation when F1 reached 5.7 mm in heifers, and 2.4 d after ovulation when F1 reached 6.1 mm in cows. In conclusion, the emergence of Wave 1 and growth rates and diameters of the future dominant follicles at the beginning of deviation were similar in Nelore heifers and nonlactating cows, regardless of the methods used to determine deviation. Relative to Holstein cattle, emergence of Wave 1 appeared to occur about 1 d later and diameter of the future dominant follicle at the beginning of deviation was about 2 mm smaller in Nelore.

Animals↗

PubMed1980

Spontaneous diabetes mellitus in the New Zealand white rabbit: preliminary morphologic characterization.

Electron microscopic studies on a closed colony of rabbits with an 18.5 per cent incidence of spontaneous onset insulin-dependent diabetes mellitus revealed that the beta-cells of the islets of Langerhans of the diabetic animals were hypergranulated. This finding contrasts with most other animal models of spontaneous diabetes mellitus which show degranulation of the beta-cells. There was no evidence of hyperplasia, insulitis, amyloid, or fibrosis of the islets by either light or electron microscopy. Correlation of our morphologic findings with physiologic data suggests a defect in insulin secretion. Rabbits with normal glucose metabolism showed a normal degree of granulation of their beta-cells. The alpha and delta cells were within normal limits in all animals. No other abnormalities associated with diabetes in humans or other animals were noted except for minimal fusion of the glomerular epithelial foot processes and mineralization of the proximal tubules and Bowman's capsules.

Animals↗

PubMed1978

Functional and morphological characterization of isolated bovine adrenal medullary cells.

Single bovine adrenal medullary cells have been obtained by retrograde perfusion of adrenal medullae with a solution of 0.05% collagenase in Ca++-free Krebs Henseleit buffer. Chromaffin cells were obtained in high yield (5 X 10(6) cells/g medulla), and more than 95% of these were viable as shown by exclusion of trypan blue. The isolated cells were capable of respiring at a linear rate for a minimum of 120 min. Ultrastructural examination revealed that the cells were morphologically intact, and two distinct types of adrenal medullary cells were identified, on the basis of the morphology of their electron-dense vesicles, as (a) adrenaline-containing and (b) noradrenaline-containing cells. Biochemical analysis showed that the cells contained catecholamines and dopamine-beta-hydroxylase (DBH). The cells released catecholamines and DBH in response to acetylcholine (ACh), and this release was accompanied by changes in the vesicular and surface membranes observed at the ultrastructural level. The time-course of ACh-stimulated catecholamine and DBH release, and the dependence of this release on the concentration of ACh and extracellular Ca++ have been investigated. The isolated cells were pharmacologically sensitive to the action of the cholinergic blocking agents, atropine and hexamethonium.

Adrenal Medulla↗

PubMedDate not supplied

Extracellular cultivation and morphological characterization of amastigote-like forms of Leishmania panamensis and L. braziliensis.

Two strains of the Leishmania braziliensis complex have been adapted to grow extracellularly at elevated temperature as amastigote-like forms in a cell-free medium. These parasites can be serially cultivated and maintained at 32 degrees C for L. panamensis (WR442; L. braziliensis panamensis) and at 28 degrees C for L. braziliensis (M5052; L. braziliensis braziliensis). Several observations are presented that the forms adapted at elevated temperature are amastigote-like. Morphologically, the amastigote-like organisms appear rounded to ovoid and are immotile and smaller than promastigotes; the flagellum of the amastigote-like forms does not extend beyond the flagellar pocket. In comparison, the promastigotes are very elongated, with a nucleus at mid-cell length and a very long flagellum. By electron microscopy, the short flagellum of the amastigote-like form is within a distended flagellar pocket; the 9 + 2 axonemal configuration is present but the paraxial rod is not observed. By contrast, the flagellum of the promastigote has a paraxial rod which extends from the axosome level. In addition, these amastigote-like forms of Leishmania are able to infect, to survive and to divide within the macrophage cell line J774.

Animals↗

PubMed1989

Functional and morphologic characterization of mast cells recovered by bronchoalveolar lavage from Basenji greyhound and mongrel dogs.

Mast cells are believed to play an important role in the pathogenesis of asthma, and several investigators have suggested that increased numbers of mast cells in the airway lumen or increased releasability of histamine from these mast cells are responsible for chronic airway hyperreactivity. To determine whether mast cells in the lumen of the airways of hyperreactive Basenji greyhound (BG) dogs differ from those of mongrel dogs with normal airway reactivity, we investigated the morphologic and functional characteristics of mast cells recovered by bronchoalveolar lavage (BAL). BAL was performed in five BG and five mongrel dogs with 900 cc of a buffered salt solution. The recovered lavage fluid contained 115 +/- 19 X 10(6) and 116 +/- 14 X 10(6) (mean +/- SEM) cells in BG and mongrel dogs, respectively. The proportion of all mast cells within the recovered cell population as enumerated after fixation with basic lead acetate and staining with alcian blue was not different in BG and mongrel dogs and averaged 0.80 +/- 0.07% and 1.1 +/- 0.3%, respectively. Typical mast cells as identified after fixation with paraformaldehyde were rare; however, significantly more mast cells were found in mongrel (0.03 +/- 0.009%) than in BG dogs (0.004 +/- 0.002%; p less than 0.02). Mast cells recovered from BG and mongrel dogs were not different in their low spontaneous histamine release (2.0 +/- 0.5% and 2.9 +/- 0.8%), their histamine release on stimulation with the calcium ionophore A23187 (maximum release 44.8 +/- 5.7% and 41.5 +/- 3.9%), and their lack of response to compound 48/80 (maximum release 5.8 +/- 1.8% and 6.1 +/- 6.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

Airway Resistance↗

PubMed1985

Selective culture of neurons from rat cerebral cortex: morphological characterization, glutamate uptake and related enzymes during maturation in various culture media.

Dissociated cerebral cortex of fetal rat was grown in a serum-free, chemically defined medium (CDM) (containing insulin, progesterone, estradiol, transferrin, putrescine, selenium and 15 mM KCl) and compared with cultures grown in a medium containing 20% fetal calf serum (SCM). Neurons survived well using either medium, but in the serum-free medium the cellular population was exclusively neuronal (at 96%), while glial cells began to proliferate after one week in the SCM. The various cellular morphologies are described in the present report and the presence of immunological markers characteristic of neurons was investigated. Autoradiographic experiments have been performed after incubation with various putative neurotransmitters and we have shown the presence of a strikingly high proportion of glutamatergic neurons in these cultures. Glutamate high affinity uptake was also greatly increased in neuronal cultures maintained in a CDM compared to a SCM, especially in young cultures. The development of different enzymes involved in the metabolism of glutamate was also studied; the specific activity of glutaminase increased in culture and was found to be higher in a CDM than in a SCM, while the inverse was true for glutamine synthetase. The relative proportion of both enzymes in neurons compared to glial cells was opposite, as neuronal cultures had higher levels of glutaminase and glial cultures were enriched in glutamine synthetase activity. It seems that the proportion of glutamate-neurons increases when cultured in a CDM compared to a SCM and we suggest that this culture procedure may provide a purely neuronal population enriched in mature glutamatergic neurons. It may thus be useful for future in vitro studies on glutamate and GABA metabolism in neurons.

Animals↗

PubMed2005

Morphological characterization of a human glioma cell l ine.

A human malignant continuous cell line, named NG97, was recently established in our laboratory. This cell line has been serially subcultured over 100 times in standard culture media presenting no sign of cell senescence. The NG97 cell line has a doubling time of about 24 h. Immunocytochemical analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP) and S-100 protein, and negative for vimentin. Under phase-contrast microscope, cultures of NG97 showed cells with variable morphological features, such as small rounded cells, fusiform cells (fibroblastic-like cells), and dendritic-like cells. However, at confluence just small rounded and fusiform cells can be observed. At scanning electron microscopy (SEM) small rounded cells showed heterogeneous microextentions, including blebs and filopodia. Dendritic-like cells were flat and presented extensive prolongations, making several contacts with small rounded cells, while fusiform cells presented their surfaces dominated by microvilli.We believe that the knowledge about NG97 cell line may be useful for a deeper understanding of biological and immunological characteristics of gliomas.

Journal Article↗

PubMed1990

Morphologic characterization of osteoblast-like cell cultures isolated from newborn rat calvaria.

Two methods for harvesting osteoblast-like cell populations from newborn (10 days) rat calvaria were compared. The first one consisted in culturing the periosteum-free bones and then trypsinizing the cells on the bone surface. The second one involved the migration of the osteoblasts on glass fragments before trypsinization. Since the plating efficiency, the proportion of alkaline phosphatase-positive cells, the population doubling time, and the calcium deposition were more adequate, the second method was used to further characterize the behavior of the cultures. During the first week of culture, the cells featured shapes similar to those observed in vivo on the surface of periosteum-free calvaria. They formed multilayers and, in the presence of ascorbic acid, synthetized an organic matrix containing exclusively type I collagen. Later, small amounts of type III collagen appeared. The cells were embedded in the matrix and progressively acquired the morphologic phenotype of osteocyte-like cells. The matrix mineralized in the presence of beta-glycerophosphate. The technique of drop-inoculation (high concentration of cells in a small volume of medium) promoted the multilayer formation and the achievement of large mineralized plates (about 1 cm2) in 3 weeks of culture.

Alkaline Phosphatase↗

PubMed1983

Morphological characterization of the cholesteryl ester cycle in cultured mouse macrophage foam cells.

Mouse peritoneal macrophages can be induced to accumulate cholesteryl esters by incubating them in the presence of acetylated low density lipoprotein. The cholesteryl esters are sequestered in neutral lipid droplets that remain in the cell even when the acetylated low density lipoprotein is removed from the culture media. Previous biochemical studies have determined that the cholesterol component of cholesteryl ester droplets constantly turns over with a half time of 24 h by a cyclic process of de-esterification and re-esterification. We have used morphologic techniques to determine the spatial relationship of cholesteryl ester, free cholesterol, and lipase activity during normal turnover and when turnover is disrupted. Lipid droplets were surrounded by numerous 7.5-10.0-nm filaments; moreover, at focal sites on the margin of each droplet there were whorles of concentrically arranged membrane that penetrated the matrix. Histochemically detectable lipase activity was associated with these stacks of membrane. Using filipin as a light and electron microscopic probe for free cholesterol, we determined that a pool of free cholesterol was associated with each lipid droplet. Following incubation in the presence of the exogenous cholesterol acceptor, high density lipoprotein, the cholesteryl ester droplets disappeared and were replaced with lipid droplets of a different lipid composition. Inhibition of cholesterol esterification caused cholesteryl ester droplets to disappear and free cholesterol to accumulate in numerous myelin-like structures in the body of the cell.

Animals↗

PubMed1984

Biochemical and morphological characterization of a plasma membrane-enriched fraction from bovine parathyroid cells.

A fraction of enriched plasma membranes from bovine parathyroid cells has been prepared by differential centrifugation. Biochemical characterization shows that this fraction has a specific activity enrichment of 7.2-fold in ouabain-sensitive Na+-K+ ATPase, and 3.5-fold in 5'-nucleotidase. Less than 4% of the total mitochondria and lysosomes are present within the plasma membranes, while microsomal contamination accounts for 14% of total specific activity. Parathyroid hormone radioimmunoassay also reveals the presence of some secretory granules within the plasma membrane fraction. The characteristic morphological aspect of the unusual surface membrane is shown by freeze-fracture electron microscopy. In the enriched pellets, vesicles identified as having a plasma membrane origin have variable sizes, and 50% show an inside-out conformation. Even though the plasma membrane fraction described herein is not absolutely free from contamination by other subcellular components, this protocol represents the first attempt to purify surface membrane from parathyroid tissue and provide the starting material for understanding, at a molecular level, the properties of extracellular Ca2+ regulation and its coupling with secretion of parathyroid hormone.

5'-Nucleotidase↗

PubMed1998

Structural and morphological characterization of ultralente insulin crystals by atomic force microscopy: evidence of hydrophobically driven assembly.

Although x-ray crystal structures exist for many forms of insulin, the hormone involved in glucose metabolism and used in the treatment of diabetes, x-ray structural characterization of therapeutically important long-acting crystalline ultralente insulin forms has been elusive because of small crystal size and poor diffraction characteristics. We describe tapping-mode atomic force microscopy (TMAFM) studies, performed directly in crystallization liquor, of ultralente crystals prepared from bovine, human, and porcine insulins. Lattice images obtained from direct imaging of crystal planes are consistent with R3 space group symmetry for each insulin type, but the morphology of the human and porcine crystals observed by AFM differs substantially from that of the bovine insulin crystals. Human and porcine ultralente crystals exhibited large, molecularly flat (001) faces consisting of hexagonal arrays of close packed hexamers. In contrast, bovine ultralente crystals predominantly exhibited faces with cylindrical features assignable to close-packed stacks of insulin hexamers laying in-plane, consistent with the packing motif of the (010) and (011) planes. This behavior is attributed to a twofold increase in the hydrophobic character of the upper and lower surfaces of the donut-shaped insulin hexamer in bovine insulin compared to its human and porcine counterparts that results from minor sequence differences between these insulins. The increased hydrophobicity of these surfaces can promote hexamer-hexamer stacking in precrystalline aggregates or enhance attachment of single hexamers along the c axis at the crystal surface during crystal growth. Both events lead to enhanced growth of ¿hk0¿ planes instead of (001). The insulin hexamers on the (010) and (110) faces are exposed "edge-on" to the aqueous medium, such that solvent access to the center of the hexamer and to solvent channels is reduced compared to the (001) surface, consistent with the slower dissolution and reputed unique basal activity of bovine ultralente insulin. These observations demonstrate that subtle variations in amino acid sequence can dramatically affect the interfacial structure of crystalline proteins.

Animals↗

PubMed1998

Morphological characterization of ventricular septal defect with posterior deviation of the outlet septum.

Ventricular septal defect (VSD) with posterior deviation of the outlet septum is frequently associated with left ventricular outflow obstruction and aortic arch anomalies. In this study, 17 patients with large VSD and posterior deviation of the outlet septum were evaluated. The VSD was further classified into 'high' and 'low' types according to the extension of the VSD; the high-type VSD (8 patients) extended mainly to the outlet or infundibular portion and the low-type VSD (9 patients) extended mainly to the trabecular or inlet portion. Related morphological features, such as outlet septum hypoplasia, muscle bundle between aortic-mitral valve junction, pulmonary artery overriding and aortic arch anomalies, were found more frequently in patients with high-type VSD, while tricuspid valve straddling and outlet septum hypertrophy were found more often in patients with low-type VSD. This classification for VSD with posterior deviation of the outlet septum helps to predict the morphological features of left ventricular outflow tract and aortic arch anomalies.

Echocardiography↗

PubMed1986

Morphological characterization of transformed colonies in rat tracheal epithelial cell cultures exposed to carcinogen.

The purpose of the studies described in this and the accompanying paper (D.J. Fitzgerald et al., Cancer Res., 46:4642-4649, 1986) was to define the transformed colonies of the rat tracheal epithelial cell transformation system (see P. Nettesheim and J.C. Barrett, CRC Crit. Rev. Toxicol., 12: 215-239, 1984) in terms of differentiation and growth characteristics. Exposure of low-density primary rat tracheal epithelial cell cultures to N-methyl-N'-nitro-N-nitrosoguanidine results in the development of growth-altered colonies of different sizes and morphologies, which can be readily scored 5 wk after treatment. These colonies were classified into four morphological types (I to IV) based on their light microscopic and ultrastructural features. Colonies designated as types I and II were small, had a low cell density, and were composed principally of large, pale-staining (Giemsa) flattened cells with a low nuclear/cytoplasmic ratio. Examination of the fine structure of these colonies revealed a monolayer of extremely attenuated cells containing well-developed Golgi complexes, endoplasmic reticulum, numerous secondary lysosomes, but few tonofilaments and desmosomes. Colonies of types III and IV were large, basophilic (Giemsa), high-density colonies, consisting primarily of closely packed, small, round cells with high nuclear/cytoplasmic ratio. Analysis of the fine structure of these colonies revealed two to four stratified layers of poorly differentiated cells and cells having features of keratinocyte differentiation, as evidence by abundant tonofilament bundles, well-developed desmosomes, and occasional keratohyaline granules. Differential cell counts were carried out at the ultrastructural level on pooled cells from each colony type (except type I); approximately 90% of the cells from type II colonies were large, electron-lucent cells, while this cell type comprised only 20 to 30% of the cells of colonies of types III and IV. The predominant cell type (approximately 60%) in colonies of types III and IV showed clear signs of keratinocyte differentiation. Type IV colonies contained a significantly larger proportion (greater than 20%) of small, poorly differentiated cells than type III (6%) and type II (1%) colonies. A combined autoradiographic and ultrastructural study revealed that the small, poorly differentiated cells were most active in synthesizing DNA and had a labeling index of 60%. Other cell types were far less frequently labeled, particularly the large, electron-lucent cells which had a labeling index of only 10%. Morphometric measurements on fixed and stained colonies disclosed that each colony type differs markedly in cell population size.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed2003

Morphological characterization of the mycorrhiza formed by Helianthemum almeriense Pau with Terfezia claveryi Chatin and Picoa lefebvrei (Pat.) Maire.

This work presents the first anatomical description of the mycorrhizal systems of Helianthemum almeriense, and of the structure and ultrastructure of the mycorrhizae formed by this plant species with the ascomycetes Terfezia claveryi and Picoa lefebvrei. Four different mycorrhizal systems are described, the club-shaped mycorrhiza being the most abundant. The type of mycorrhiza formed depended on the mycorrhiza culture conditions, but not on the fungal species. For both fungal species, H. almeriense formed an endomycorrhiza in natural field conditions, an ecto- and ectendomycorrhiza without a sheath in pot cultures, and an ectomycorrhiza with a characteristic sheath and Hartig net in in vitro cultures. This is the first report of a typical sheath in Helianthemum-desert truffle mycorrhizal associations. The results support the idea that culture conditions can induce changes in mycorrhiza morphology and that there is no clear barrier between the two main types of mycorrhiza organization in Helianthemum species. The ultrastructural study confirmed the regular presence of T. claveryi intracellular hyphae in direct contact with the host wall, a localization which seems to be a characteristic of the T. claveryi mycorrhiza organization. The P. lefebvrei mycorrhiza organization was characterized by intracellular hyphae with large amounts of electron-dense globules, probably with a lipidic content, and a warty ornamentation on the wall of the root external hyphae.

Ascomycota↗