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At least 325 records · Page 18Linked to original sources

PubMed2001

High density culturing of porcine hepatocytes immobilized on nonwoven polyurethane-based biomatrices.

OBJECTIVE: Hepatocytes are increasingly used as functional units in bioartificial liver devices. The objective of the present study was to investigate the feasibility of culturing porcine hepatocytes in high density on a novel polyurethane-based nonwoven three-dimensional matrix. We investigated (1) the optimal cell density within this culture configuration, (2) the maintenance of liver-specific morphology and cell functions over long-term periods and (3) the necessity to apply an additional extracellular matrix component (collagen gel). METHODS: Nonwoven polyurethane matrices were manufactured by a specially developed fiber extrusion technology. Pig hepatocytes were cultured at various cell densities of 0.1, 0.25, 0.5, 0.75, 1 and 2 x 10(6) cells/cm(2) on three-dimensional networks of nonwoven polyurethane matrices and cell adhesion as well as functional parameters (DNA of nonattached/attached cells, lactate dehydrogenase release and cytochrome P450 activity) were determined. To assess the performance of cells within this configuration albumin and urea excretion was measured over 8 days. The potentially beneficial effect of an additional extracellular matrix configuration was evaluated by comparing the average albumin synthesis in groups of identical cell numbers. RESULTS: The optimal cell density in this three-dimensional culture configuration was 1 x 10(6) cells/cm(2). The functional capacity of hepatocytes was stable for 8 days at an average level of 53.7 +/- 5.6 ng/h/microg DNA and of 1.8 +/- 0.14 microg/h/microg DNA for albumin and urea excretion, respectively. The supplementation of an extracellular matrix configuration did not improve functional activity of cells. Average albumin synthesis was 35.6 ng/h/microg DNA (28.7, 42.8) and 32.7 ng/h/microg DNA (23.4, 49.2) for collagen-immobilized and control cultures, respectively. CONCLUSION: The results of the study indicate that nonwoven polyurethane sheets supply a biocompatible support structure for functionally active high density cultures. Thus, nonwoven polyurethane matrices should be further investigated on with respect to their role in the development, optimization and design of bioartificial liver systems.

Animals↗

PubMed2000

Messenger ribonucleic acid expression of the MyoD gene family in muscle tissue at slaughter in relation to selection for porcine growth rate.

Livestock meat production capacity is related to muscle fiber numbers and growth. Muscle fibers develop during early embryonic development from proliferating and differentiating myoblasts. Post-natal muscle growth requires satellite cell proliferation and differentiation. Myoblast and satellite cell proliferation and differentiation is regulated by the genes of the MyoD gene family (myogenin, myf-5, myf-6, and MyoD1). Our aim was to study the mRNA expression of these genes in postnatal muscle tissue in relation to porcine selection for growth rate or leanness. Five boars from a line selected for fast growth (F-line) and five boars from a line selected against backfat thickness (L-line) were slaughtered, and biopsies were taken from 12 muscles. Between-line effects, within-line effects in relation to the performance of the pigs, and muscle-specific effects were studied. Comparing the F-line with the L-line revealed significantly greater myogenin, myf-5, and MyoD1 mRNA expression in some muscles of the F-line. The expression of myf-6 showed a tendency for the opposite effect in some muscles. Muscles were ordered by their muscle-specific growth rate (b-value). Within-line evaluation of the data revealed a systematic muscle effect for the myf-6 expression level in the F-line because higher b-values correlated with increased myf-6 expression level. Backfat thickness was negatively related to myogenin expression in the F-line. A relationship was found between myogenin:MyoD1 mRNA expression ratio and meat color/muscle fiber type composition in the L-line. Furthermore, the myogenin:MyoD1 ratio was greater in muscles from F-line boars than in muscles from L-line boars, which relates to the difference between the lines in muscle fiber type. We conclude that the mRNA levels of the MyoD genes in porcine muscle tissue at slaughter showed different relationships to selection for growth rate when evaluated between selection lines and within selection lines.

Animals↗

PubMed2005

Cellular and molecular tunnels surrounding the forebrain commissures of human fetuses.

Glial cells and extracellular matrix (ECM) molecules surround developing fiber tracts and are implicated in axonal pathfinding. These and other molecules are produced by these strategically located glial cells and have been shown to influence axonal growth across the midline in rodents. We searched for similar cellular and molecular structures surrounding the telencephalic commissures of fetal human brains. Paraffin-embedded brain sections were immunostained for glial fibrillary acidic protein (GFAP) and vimentin (VN) to identify glial cells; for microtubule-associated protein-2 (MAP-2) and neuronal nuclear protein (NeuN) to document neurons; for neurofilament (NF) to identify axons; and for chondroitin sulfate (CS), tenascin (TN), and fibronectin (FN) to show the ECM. As in rodents, three cellular clusters surrounding the corpus callosum were identified by their expression of GFAP and VN (but not MAP-2 or NeuN) from 13 to at least 18 weeks postovulation (wpo): the glial wedge, the glia of the indusium griseum, and the midline sling. CS and TN (but not FN) were expressed pericellularly in these cell groups. The anterior commissure was surrounded by a GFAP+/VN+ glial tunnel from 12 wpo, with TN expression seen between the GFAP+ cell bodies. The fimbria showed GFAP+/VN+ cells at its lateral and medial borders from 12 wpo, with pericellular expression of CS. The fornix showed GFAP+ cells somewhat later (16 wpo). Because these structures are similar to those described for rodents, we concluded that the axon guiding mechanisms postulated for commissural formation in nonhuman mammals may also be operant in the developing human brain.

Axons↗

PubMed1989

Intracellular pH and cell-to-cell transmission in sheep Purkinje fibers.

Intracellular pH (pHi) is a significant modifier of cell-to-cell communication in some tissues but its role is uncertain in heart tissue. The present studies examined the effect of cytosolic protons on electrotonic spread and conduction velocity in cardiac Purkinje fibers. Cable analysis provided values for internal longitudinal resistance (ri) and pH-selective microelectrodes monitored pHi during CO2 and HCO3- alterations. Resting fibers developed changes in ri that were proportional to intracellular free proton concentration ([H+]i) during CO2 changes at constant [HCO3-]. However, the effects on ri were small between pHi 6.9-7.8 and predicted only a 2.2% increase in ri per 10 nM increase in [H+]i. Other findings suggested that titration of cytosolic protons may not directly produce the changes in ri: (a) For an equal change in [H+]i, the effects on ri were roughly three times greater (6.8% increase per 10 nM rise in [H+]i) if bicarbonate was lost during CO2 changes. (b) pH-associated changes in ri were preceded by a time delay (1-5 min) producing hysteresis in the [H+]i-ri relation during successive perturbations. (c) The same CO2 variations modified the direction and magnitude of ri differently during pacing than at rest. The cumulative results suggest that the action of protons on ri in the heart may be subordinate to another regulator or mediated by another pH-dependent substance or reaction.

Animals↗

PubMed2004

Characterization of the brassinosteroid insensitive 1 genes of cotton.

Suppression of brassinosteroid (BR) biosynthesis in cotton ovules by treatment with brassinazole inhibits fiber formation, indicating that BR plays an important role in cotton fiber development. Plant responses to brassinosteroids (BR) are mediated through a plasma membrane-bound leucine-rich repeat (LRR) receptor-like protein kinase known as BRI1. Mutations in the BRI1 genes of several species result in dwarfed plants with reduced sensitivity to BR. A single expressed sequence tag (EST) from cotton with strong sequence similarity to Arabidopsis BRI1 ( AtBRI1 ) was identified in a search of publicly available databases. With this EST as a starting point, full-length cDNAs and genomic coding sequences from upland cotton ( Gossypium hirsutum ) BRI1 ( GhBRI1 ) were obtained and characterized. Ectopic expression of this coding sequence in BR-insensitive Arabidopsis plants resulted in recovery of normal growth indicating that GhBRI1 is a functional homologue of AtBRI1. G. hirsutum is an allotetraploid (AADD) derived from diploid ancestors. Analysis of several GhBRI1 cDNAs showed two distinct sequences indicating the presence of two GhBRI1 genes, denoted GhBRI1-1 and GhBRI1-2. Sequence comparisons between these GhBRI1 coding sequences and those from related A and D genome diploid Gossypium species ( G. arboreum and G. thurberi ) indicated that GhBRI1-1 is likely to the A sub-genome orthologue while GhBRI1-2 is from the D sub-genome.

Amino Acid Sequence↗

PubMed1988

QTU prolongation and polymorphic ventricular tachyarrhythmias due to bradycardia-dependent early afterdepolarizations. Afterdepolarizations and ventricular arrhythmias.

Polymorphic ventricular tachyarrhythmias occurred spontaneously during bradycardia in dogs given the inotropic polypeptide anthopleurin-A (AP-A). The arrhythmia was investigated in in vitro and in vivo experiments. In in vitro experiments, AP-A (50 micrograms/l) produced bradycardia-dependent prolongation of action potential duration that was more pronounced in Purkinje than in muscle fibers. Only Purkinje fibers developed early afterdepolarizations (EAD) and triggered activity. These effects could be abolished by rapid pacing, lidocaine (4 mg/l), or tetrodotoxin (1 mg/l). In vivo experiments were conducted in anesthetized healthy dogs with simultaneous recording of surface ECG, monophasic action potentials from the endocardial and epicardial surface of the left ventricle by contact electrode catheter technique, and transmembrane action potentials from the epicardial surface of the left ventricle with a floating microelectrode technique. AP-A in a dose comparable to that used in vitro (4 micrograms/kg, i.v. bolus) resulted in bradycardia-dependent marked prolongation of both monophasic and transmembrane action potentials. An EAD gradually appeared on both recordings but was more marked in endocardial monophasic action potentials. Eventually, a premature ventricular depolarization arose from or very close to the peak of the EAD. The prolongation of action potentials was associated with similar prolongation of the QTU interval in surface ECG, and in some experiments, the EAD corresponded to a distinct prominent U wave. A ventricular premature depolarization arose from the U or TU complex and initiated polymorphic ventricular tachyarrhythmias that terminated spontaneously or degenerated into ventricular fibrillation. These effects were reversed by rapid pacing or lidocaine (1 mg/kg). The present study provides evidence in support of the hypothesis that AP-A-induced ventricular tachyarrhythmias are due to bradycardia-dependent EAD and triggered activity.

Action Potentials↗

PubMed2000

DeltaFosB-induced cataract.

PURPOSE: The objective of this study was to investigate a possible relationship between posterior subcapsular cataract (PSC) formation and expression of the transcription factor DeltaFosB. METHODS: Western blot analysis was performed on bitransgenic NSE-tTA, TetOp-DeltaFosB, and single-transgenic NSE-tTA control mice to determine the pattern of DeltaFosB expression within the eye. Light and scanning electron microscopy and biochemical analyses were also performed. RESULTS: In mice expressing DeltaFosB, cataract developed that initially appeared to be posterior subcapsular and gradually matured to involve the entire lens. The enlarged posterior ends of developing secondary fibers curved away from the visual axis to form an elevated opaque posterior plaque. As a result, posterior suture formation did not occur. At a later time, the attenuated posterior capsule overlying the plaque ruptured and the lens nucleus subluxated into the vitreous. Retinal damage was also observed but only from postnatal day 65, a time when extensive lens degeneration had already occurred. DeltaFosB expression was observed well before the detection of morphologic change in both the lens and the retina. Within the lens, DeltaFosB expression was found in both the epithelium and fibers. The development of cataracts was a direct consequence of DeltaFosB expression and was not due to the disruption of an endogenous gene by transgene integration since cataracts could be prevented by silencing expression of DeltaFosB by feeding bitransgenic animals doxycycline (Dox). Moreover, cataracts were observed in bitransgenic mice derived from two independent TetOp-DeltaFosB founder lines but not in single NSE-tTA transgenic controls. Cataractogenesis was not a consequence of abnormal development, because mice conceived and raised on Dox to prevent expression of DeltaFosB also were subject to formation of PSC when expression of DeltaFosB was turned on in adult animals by removing Dox. Examination of biochemical parameters indicated that the earliest change observed was the disruption of calcium homeostasis with a significant increase in Ca(2+) influx, followed by a gradual but marked decrease in protein content. Significant changes in certain metabolic parameters and protein composition were also observed. CONCLUSIONS: The DeltaFosB-induced cataract in which the major morphologic early event was the disruption of normal posterior fiber formation, may be a good model for PSC. By identifying DeltaFosB-regulated target genes, it should be possible to achieve a better understanding of the molecular mechanisms through which PSC is formed.

Animals↗

PubMed1992

Development of collagen fibers and vasculature of the fetal TMJ.

Using 12 human fetuses, histological development and changes in connective fiber structure and fine vascular patterns have been investigated in various fetal gestational stages by light and scanning electron microscopy. The main arterial supply of the articular disc was from the bilaminar region and pterygoideus lateralis muscle. The vascular network on the disc surface was related with fluid secretion. When the bilaminar region was compressed, it caused ischemia and fibrosis as the main pathological changes in TMJ derangement. A decrease in fluid from blood vessels might occur in TMJ degeneration. Collagen fibers in the disc passed mainly anteroposteriorly. In the anterior and posterior bands, muscular tendon fibers came from the pterygoideus lateralis muscle and superior stratum of the bilaminar region. In the posterior band three-dimensional structures of collagen fibers suitable for load bearing were observed. The compass network and process on the disc showed the normal structure that is formed gradually and has functions including dispersion, pressure bearing, friction-proofing and storage of the synovial fluid. Attachments of the disc were suitable for disc function. Large elastic fibers in the posterolateral part of the superior stratum of the bilaminar region may be antagonistic to the upper head of the pterygoideus lateralis muscle fibers passing medioanteriorly, indicating that this antagonism is available for disc function.

Collagen↗

PubMed1988

Periapical innervation of the ferret canine and the local retrograde neural changes after pulpectomy.

The amputation of the dental pulp severs a population of axons that are predominantly in the A delta and C fiber size range and are principally involved in nociception. Local periapical neuromas, if they are formed after pulpectomy, may be the sites of spontaneous nervous activity that may, in some circumstances, be involved in the genesis of chronic pain. The periapical tissues from the mandibular canines of four ferrets were examined 3 months after pulpectomies. Silver-stained paraffin sections were examined in three dimensions at the light microscope level. Ultrathin sections were examined at the electron microscope level. Compared with contralateral and independent controls, the principal changes were the loss of the periodontal plexus around the root apex, the extension of damage well below the apical foramen, and the persistence of inflammation 12 weeks postoperatively. While a somewhat disorderly mass of nerve fibers develops subapically, the arrangement has only some of the features usually associated with neuromas.

Animals↗

PubMed2005

pH-Responsive shrinkage/swelling of a supramolecular hydrogel composed of two small amphiphilic molecules.

A pH-responsive volume-change function was successfully introduced into a supramolecular hydrogel that contained GalNAc-appended (GalNAc=N-acetylgalactosamine) glutamate ester 1 by the simple mixing of it with an appropriate amount of 2 a or 2 b amphiphilic carboxylic acid. In the 1:1 mixture (1:2), the hydrogel swelled under neutral pH conditions, but shrank to almost half of its original volume under acidic pH conditions. The structure and pH response of the mixed hydrogel were characterized by using X-ray diffraction (XRD), confocal laser scanning microscopy (CLSM), transmission or scanning electron microscopy (TEM, SEM), and Fourier transform IR (FTIR) spectroscopy. Well-developed fibers formed a stable hydrogel by self-assembly, and under acidic conditions the charge of the carboxylic acid terminal (from the carboxylate anion) was neutralized and then these fibers became densely packed. This macroscopic pH response was also applied to the pH-triggered release of bioactive substances. In this mixed supramolecular hydrogel, the hydrogelator 1 provides a stable hydrogel structure and the additive 2 acts as a commander that is sensitive to an environmental pH signal. The present supramolecular copolymerization strategy should be useful for the construction of novel, stimuli-responsive, soft materials.

Journal Article↗

PubMed1993

Immunocytochemical demonstration of early appearing astroglial structures that form boundaries and pathways along axon tracts in the fetal brain.

During normal development of the mammalian forebrain, the paired cerebral hemispheres are initially separated midsagittally by the connective tissue-filled longitudinal fissure. During subsequent stages, the hemispheres fuse as basal lamina is remodeled and fibroblasts are eliminated from the fissure to create new central nervous system (CNS) territory in the midline. Two axon pathways, the corpus callosum and dorsal callosal stria, eventually use this region as part of their pathway. In order to assess the possible role of glial cells in the fusion process and in the guidance of axons in this and several other areas of the forebrain, we have analyzed the developing brain in timed cat and mouse embryos with immunohistochemical and morphological techniques. With the use of astroglial-specific antibodies and electron microscopy, we have visualized two distinct, primitive astroglial structures associated with the cerebral midline, and seven more associated with other specific brain regions. The way in which one of these structures moves as a column along the hemispheric midline in synchrony with seam formation suggests the possibility that during morphogenesis of the telencephalon, astrocytes may aid in the fusion process. In addition, the compact assemblage, early appearance and location of this and the other glial structures in relation to well defined neuroanatomical landmarks or axon pathways suggest that they may transiently compartmentalize relatively large regions of the CNS and organize certain developing fiber systems by acting as guides or barriers at critical stages of ontogeny.

Animals↗

PubMed1991

Why are slow and fast muscles differentially affected during prenatal undernutrition?

The biceps brachii and soleus muscles (fast and slow twitch muscles, respectively) were taken from guinea-pig fetuses, undernourished via maternal dietary intake to 60% of ad lib, and examined for differences from their controls. At birth, muscle fiber number was found to be reduced by 26% in the biceps brachii but was unaltered in the soleus muscle. An examination of the muscles at ages through gestation revealed that any reduction in fiber number was associated with a reduction in the secondary/primary fiber ratio. There was a stage in soleus myogenesis at which fiber number was significantly lower than the control. The deficit was corrected by the continuation of secondary fiber hyperplasia beyond the time of usual cessation. A similar delay was found in the biceps brachii but it did not enable recovery to a full complement of fibers. The difference between the 2 muscles was thought to be related to the relative proportions of primary fibers in the muscles, primary fiber development being apparently unaffected by undernutrition in utero.

Animals↗

PubMed1999

Neutral and acidic glycosphingolipids in glucocorticoid-induced cataract in chick lens.

Administration of glucocorticoids induces transient cataract in 15-day-old chick embryos within 48 hr, and the opaque lens again becomes clear within the subsequent 48 hr. Oxidative stress is likely to be involved in the process of cataract formation, resulting in the appearance of numerous vacuoles around the perinuclear region. Chick lens contained low amounts of glycosphingolipids, which mainly consists of GM3, GD3, sialyl-LewisX gangliosides and glucosylceramide. Most lens gangliosides were immunohistochemically detected in lens epithelia, annular pads and developing fibers, but not in perinuclear and nuclear regions. Since cell surface gangliosides, for example GM3 and sialyl-LewisX gangliosides, are involved in cell adhesion, weak cell-to-cell interactions in the perinuclear and nuclear regions may allow vacuole formation in steroid-induced cataractogenesis.

Animals↗

PubMed1982

Tissue culture of chick embryonic choroidal cells: cell aggregation and pigment accumulation.

Cells originating from the chick embryo choroid were grown in culture for up to 1 month. The cells did not form coherent monolayer colonies but rather formed large transparent cell aggregates surrounded by a random, loose network of cells. The cell aggregates avidly took up and retained pigment granules while the individual cells did not. Choroidal cell aggregates were not similar to the "lentoid" bodies described by Okada and coworkers in cultures from other ocular tissues; they did not develop fiber cells and did not synthesize delta-crystallin. Thus aggregation of ocular cells appears to be a necessary but insufficient step for the formation of a "lentoid" body.

Animals↗

PubMed1988

Formation of a new fibrous attachment to human dental roots. A new vitro model for studying periodontal regeneration.

This study was performed to improve currently employed in vitro models for the study of periodontal regeneration by using a porous filter upon which periodontal ligament cells were grown. Periodontal ligament cells were harvested and 0.3 mm root discs cut from three partially erupted and extracted third molar teeth of one patient. Experimental culturing was performed by seeding periodontal ligament cell suspensions on Puropor-200 filters supported by wire-mesh grids in Grobstein Petri dishes. The following day, an interdental space of 0.1 to 0.3 mm was created by gently placing two dental root discs upon the filter. Cultures were terminated after 42, 56, 112 and 124 days, and processed for light- and electron microscopy. Collagen fibril diameters were measured. Adjacent and often attached to large areas of cementum-lined root discs, a dense fiber fringe developed. This fiber fringe was not found on dentin-lined root discs. Although less organized, older cultures demonstrated a similar disc-culture interface, which depended upon the presence or absence of original root cementum. Collagen fibrils of early cultures had a mean diameter of about 42 nm, while in older cultures the diameters ranged from 47 to 68 nm. It is concluded that the fibrous matrix attached to cementum-lined root discs somewhat resembles the initial stages of the formation of dental root cementum in vivo.

Adult↗

PubMed1976

Brain tissue transplanted to the anterior chamber of the eye. 4. Drug-modulated transmitter release in central monoamine nerve terminals lacking normal postsynaptic receptors.

Small pieces of fetal rat brain selected to contain a high number of noradrenaline (NA), dopamine (DA), or 5-hydroxytryptamine (5-HT) neuroblasts were transplanted to the anterior chamber of the eye of adult rats. The sympathetic ground plexus of the host iris was removed by superior cervical ganglionectomy so that transmitter mechanisms of the different central monoamine fibers innervating the iris could be selectively studied after intraocular maturation. Such irides, containing NA, DA, or 5-HT nerve terminals were incubated with radiolabelled transmitters and then stimulated by an electrical field while superfused, to investigate the spontaneous and stimulation-induced release of amine, both in drug-free buffer and buffer containing drugs acting on monoamine receptors. The central monoamine neurons of all three types were able to take up exogenous amines and release them upon stimulation by an electrical field, in much the same way as corresponding nerves in situ in slices of cerebral cortex (NA, 5-HT) or olfactory tubercle (DA). The alpha-adrenergic receptor blocking agent phentolamine increased the stimulation-induced release of 3H-NA from central NA fibers on the iris significantly. The dopamine receptor stimulating agent apomorphine decreased the stimulation-induced release of 3H-DA from central DA fibers on the iris. Pimozide, a DA receptor blocking drug tended to increase the 3H-DA release. The 5-HT receptor stimulating agent ergocornine tended to reduce the stimulation-induced release of 3H-5-HT from central 5-HT fibers on the iris. It was concluded that all three types of central monoamine nerve fibers develop essentially normal transmitter storage and release mechanisms also in an environment completely devoid of normal postsynaptic receptors. The drug experiments add strong support to the view that there are presynaptic monoamine receptors ("autoreceptors") able to modulate transmitter release present on the monoamine nerve terminals.

Animals↗

PubMed1975

[Scanning electron microscopy of the zonula apparatus of higher primates. A further approach to a new theory of accommodation (author's transl)].

The architecture of the zonula apparatus of two species of monkeys is studied by scanning electron microscopy. Some specimens were treated with pilocarpine respectively atropine immediately before enucleation. The zonula consists of two functionally different groups of fiber strands, i.e. span and holding bundles (after Rohen and Rentsch), the course and structure of which is described in detail. After application of atropine the "holding bundles" appear to be stretched in the area between the ciliary processes and the lens, whereas after pilocarpine application they appear loose and irregularly arranged. Screw-like, spiral fiber strands were often seen in the scanning electron microscope. It is assumed that the spiral fibers develop in the stage of relaxation of the zonula apparatus.

Accommodation, Ocular↗

PubMed1977

The obsolescent renal glomerulus--collapse, sclerosis, hyalinosis, fibrosis. A light- and electron microscopical study on human biopsies.

By light and electron microscopical examination it is shown that four structural components can contribute to obsolescent glomeruli: capillary basement membranes, enriched mesangium matrix, "vascular" hyalin and collagen fibers. Each of these components can bring about glomerular damage alone. One non-reactive form--a glomerular collapse with only basement membrane remnants--can be separated from three reactive forms: the accumulation of mesangium matrix (sclerosis or matrix-sclerosis), deposition of vascular hyalin (hyalinosis in the narrow sense), and fiber development within the former urinary space (fibrosis or fibro-sclerosis). The use of the term "fibrinoid" in place of the descriptive term "hyalin" is not supported by objective results. Knowledge of the various constituents which accumulate in the reactive types of glomerular obsolescence might be important in the diagnosis of the underlying disease, though mixed pictures were often observed. To avoid terminological overlap we suggest that the term "hyalinization" is replaced by "obsolescence" or "scarring" with specification of the structural components involved.

Adult↗