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PubMed2000

The eye and nutrition

Purpose: To examine the effect of vitamins and trace elements on ocular tissue.Materials and Methods: Rats or mice were fed diets deficient in the trace elements Zn, Cu, Mn, Se, Mg, and Cr or in vitamins A, B(12), C, and E. In some rats Al and vitamin A were injected in excessive amounts. We studied the conjunctiva, cornea, retina, and optic nerve with a light microscope, transmission and scanning electron microscopes, an energy dispersive X-ray analyzer, and an ion microscope. Histochemical, cytochemical, and immunohistochemical techniques were applied to the pathological specimens.Results: Deficiencies of Zn, Cu, Mn, and vitamins A, C and E caused a loss of goblet cells in the conjunctiva and a prominent decrease of microvilli and microplicae in the conjunctiva and cornea. The elements in the goblet cells were changed in these conditions. In addition, epithelial cells showed poor fibrous development and abnormal distribution of chromatin in the nucleus.Zn, Cu, Mn, and vitamins A and E deficiencies caused photoreceptor cells to degenerate and disappear. Se deficiency reduced the horizontal and amacrine cells. Vitamin B(12) deficiency reduced nerve fibers in the nerve fiber layer of the retina. Mg deficiency induced multifocal necrosis in the retinal pigment epithelium and apoptotic nuclear changes in the photoreceptor cells. Cr deficiency showed abnormal phagocytosis of the photoreceptor outer segment discs in the retinal pigment epithelium. Vitamin B(12) was found to be related to the circadian rhythm in the retina.Deficiencies of Zn, Cu, Mn, and vitamins A, B(12), and E induced degeneration and disappearance of myelin lamellae in the myelinated optic nerve fibers.In hypervitaminosis A, lipid droplets appeared in the retinal pigment epithelium and alcohol dehydrogenase disappeared in the retinal pigment epithelium and photoreceptor outer segments. Excessive Al was toxic to the retina, which showed disappearance of photoreceptor cells. Al deposits were seen in dendrites and neurons in the outer plexiform layer.Zn seemed to be necessary for corneal epithelial cell wound healing.Discussion: Trace elements usually are contained in enzymes, which have many metabolic functions. They are related to synthesis and breakdown of many substances. Some trace elements such as Zn, Cu, Mn, and Se and vitamins including vitamins A, C, and E prevent peroxidation of lipids. Some vitamins have an affinity for specific tissues such as epithelial cells, nerve fibers, and neuronal cells and are needed for cell differentiation, development, and maintenance.Conclusion: Cu, Zn, Mn, Se, Mg, and Cr and vitamins A, B(12), C, and E are necessary for maintenance of cellular structure and metabolism.

Journal Article↗
PubMedDate not supplied

Development of novel corneal storage medium: first report. Examinations of rabbit cornea.

PURPOSE: To develop a new corneal storage medium with a simple formula and evaluate it by histological methods. METHODS: Two corneal storage media containing minimum essential medium and 2.5% chondroitin sulfate (pH 7.33), with osmolarity of 320 mOsm/kg, were compared to Optisol-GS. The differences in the two media were the molecular weight (MW) and the source of chondroitin sulfate. The MW of Medium I was 27,500 and the MW of Medium II was 33,700. Japanese albino rabbits were used in this study. A cornea with scleral rim obtained from a rabbit was stored in either Medium I or Medium II and the fellow cornea was stored in Optisol-GS for 7 or 14 days at 4 degrees C. Histological examination of corneal endothelial cells was performed both by scanning electron microscopy and by transmission electron microscopy. RESULTS: At day 7, there was no significant difference in histological findings among the rabbit corneas stored in Optisol-GS, Medium I, or Medium II. At day 14, corneas stored in Optisol-GS or Medium I showed similar histological findings. In Medium II, endothelial cells showed marked degeneration. CONCLUSIONS: The results of experiments with rabbit cornea indicated that Optisol-GS and Medium I could preserve endothelial cellular structure better than Medium II. The difference between Medium I and Medium II was only the MW of the chondroitin sulfate used. The MW may be an important factor for determining suitable chondroitin sulfate for use in a corneal storage medium.

Animals↗
PubMedDate not supplied

Lens reconstruction after mature cataract in SCR rat.

PURPOSE: To conduct a long-term observation study of SCR rats that had developed a mature cataract at 11 weeks of age at 3-month intervals until the rats were 12 months old. METHODS: Lenses of 15 rats were examined with both light and electron microscopes. RESULTS: At 12 weeks, opacity was observed in the perinuclear zone and the cortical intermediate layer. Liquefaction of the posterior subcapsular area and regression of cortical superficial fibers were also observed at this stage. Epithelial cells at the anterior polar area were multilayered. At 12 months, the lens recovered as a result of the regenerated lens fibers in the intermediate layer and the cortical superficial layer, although the opacity remained in the perinuclear zone. The multilayered cellular structure in the center of the epithelium returned to its original monolayer form. However, the equatorial epithelial cells became vacuolated and swollen with age, showing regression from the bow region. CONCLUSIONS: These results suggest that the decrease of opacity in SCR rats is merely a temporary phenomenon that reflects the differentiating and metabolizing functions of the epithelial cells. With initiation of epithelial regression, the regeneration of the lens fibers ceased, suggesting that further decrease in opacity was no longer possible.

Animals↗
PubMed2000

The mechanics of cell sorting and envelopment.

Aggregates of embryonic cells undergo a variety of intriguing processes including sorting by histological type and envelopment of cell masses of one type by another. It has long been held that these processes were driven by differential adhesions, as embodied in the famous differential adhesion hypothesis (DAH). Here, we use analytical mechanics to investigate the forces that are generated by various sub-cellular structures including microfilaments, cell membranes and their associated proteins, and by sources of cell-cell adhesions. We consider how these forces cause the triple junctions between cells to move, and how these motions ultimately give rise to phenomena such as cell sorting and tissue envelopment. The analyses show that, contrary to the widely accepted DAH, differential adhesions alone are unable to drive sorting and envelopment. They show, instead, that these phenomena are driven by the combined effect of several force generators, as embodied in an equivalent surface or interfacial tension. These unconventional findings follow directly from the relevant surface physics and mechanics, and are consistent with well-known cell sorting and envelopment experiments, and with recent computer simulations.

Actin Cytoskeleton↗
PubMed1997

Histological changes caused by the rc mutation in chickens.

The rc gene represents a recessive mutation in chickens, known to cause retinal degeneration and blindness, as well as abnormal sarcolemmal membranes of cardiac myocytes associated with reduced choline transport. In this study, the visceral organs from "old" (aged 12 months) and "young" (aged 5 months) homozygous blind (rc/rc), heterozygous (Rc+/rc) and normal (Rc+/Rc+) chickens were examined histologically to investigate whether the primary effect of the mutation was on cellular structure. Homozygous birds showed enlarged thyroids with acidophilic colloids in enlarged and often ruptured follicles, macrovesicular lipid accumulation in the liver, increased numbers of nuclei in the myocardial fibres, hypertrophy of the lobular structure of the medullary portion of the thymus, cloudy swelling of the tubular epithelium of the kidney and slow maturation (in young birds) and degeneration (in old birds) of the gonads. All lesions, except for those of the thymus, were more severe in old than in young birds. Some heterozygous chickens were mildly affected and none of the normal (Rc+/Rc+) birds exhibited these abnormalities.

Age Factors↗
PubMed1989

Modelling the periodicity of cardiac muscle.

The intractable problem of modelling cardiac muscle of arbitrary extent while preserving cellular structure has been solved using an analytical rather than numerical approach with a method called two-scale asymptotic analysis. In this method, the myocardium was modelled as a collection of bundles arranged periodically in space and connected by junctions, and the distribution of the steady-state potential and current density was determined. The potential both along and across fibers was found to contain a distinct periodic component that determines the transmembrane potential. The magnitude of the transmembrane potential depends on the gradient of applied potential, the dimensions of the bundles, and their internal conductivity. Current flows primarily in the extracellular space, and the extracellular pathway also determines the apparent conductivity of cardiac muscle.

Electric Conductivity↗
PubMed1997

In vivo and in vitro studies of glycine- and glutamate-evoked acetylcholinesterase release from spinal motor neurones: implications for amyotrophic lateral sclerosis/motor neurone disease pathogenesis.

To investigate the spinal cellular structures and molecular mechanisms involved in acetylcholinesterase (AChE) release evoked by both glycine (GLY) and glutamate (GLU)--responses that might play a role in chronic neurotoxicity--we analysed AChE histochemistry and histology upon systemic administration of aspartate (ASP), and conducted in vitro experiments in synaptosomes and slices prepared from mouse spinal ventral horns. Upon superfusion and incubation exposure of these preparations to GLY- and GLU-receptor agonists, we assayed both tissue content and release of AChE, butyrylcholinesterase and lactic dehydrogenase. Histochemical reduction of motor neurone (MN) AChE, calcium dependency, decreases in intracellular AChE and the ratio amongst molecular forms released, suggest that both synaptosomal GLY-evoked AChE release (GLY-EAR) and GLU-receptor-elicited AChE release (GEAR) have release sites located at MN presynaptic terminals. These responses exhibited remarkable postnatal regulation. GEAR seems to be mediated through alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate receptors after the fourth postnatal week and through both NMDA and non-NMDA receptors at earlier stages. Sustained rises of extracellular AChE might link acute excitotoxic injury with several long-lasting pathways leading to chronic neurotoxicity, since AChE molecular properties include: (1) the ability to block cholinergic mechanisms that protect MN against overactivity; (2) activation of ATP-dependent potassium channels; (3) promotion of neurite and axon outgrowth; and possibly (4) stimulation of brain macrophage migration and activation.

Acetylcholinesterase↗
PubMed1989

Correlation of altered penile ultrastructure with clinical arterial evaluation.

We investigated the ultrastructural changes in the penile erectile tissue from 32 consecutive patients who underwent penile prosthesis implantation. Because most of the patients had undergone papaverine injection with or without duplex ultrasonography, we compared these results with the electron microscopic findings. In patients with a good arterial response and full erection after papaverine injection the ultrastructural findings were similar to those reported in normal men. In patients with moderate arterial disease a distinct increase in mitochondria with aggregation and cytoplasmic vacuolization in smooth muscle cells was noted. These findings could be interpreted as an active cellular attempt to respond to the altered environmental and nutritive situation. In patients with severe arterial insufficiency the cellular structure was markedly altered, the number of intracavernous smooth muscle cells was reduced and the density of the connective tissue separating individual cells was increased. These changes in the smooth muscle cells consisted of contour irregularity with fragmentation and loss of the basal lamina. The cytoplasm was largely devoid of contractile elements. The nuclei tended to be pleomorphic with unevenly distributed chromatin. The endothelium was also altered significantly in this group. A careful clinical evaluation of penile arterial function should be performed in all patients undergoing penile arterial or venous corrective surgery. If doubt remains, a penile biopsy may be indicated.

Adult↗
PubMed1999

Chemistry, physiology and pathology of free radicals.

The superoxide anion radical and other reactive oxygen species (ROS) are formed in all aerobic organisms by enzymatic and nonenzymatic reactions. ROS arise in both physiological and pathological processes, but efficient mechanisms have evolved for their detoxification. Similarly, reactive nitrogen intermediates (RNI) have physiological activity, but can also react with different types of molecules, including superoxide, to form toxic products. ROS and RNI participate in the destruction of microorganisms by phagocytes, as in the formation of a myeloperoxidase-hydrogen peroxide-chloride/iodide complex which can destroy many cells, including bacteria. It is known that the cellular production of ROS and RNI is controlled by different mechanisms. These free radicals can react with key cellular structures and molecules, thus altering their biological function. An imbalance between the systems producing and removing ROS and RNI may result in pathological consequences.

Animals↗
PubMed1996

Immunohistochemical and histochemical characterisation of epithelial cells of rabbit lacrimal glands in tissue sections and cell cultures.

The purpose of this study was to establish conditions for isolation and long term culture of acinar cells from the Harderian gland, and superior and inferior lacrimal glands of the rabbit and to compare the in vitro growth patterns of cultured cells from these glands. In order to determine the predominant cell type in the cultures, cells and tissue sections were stained using a variety of antibodies to cytokeratins, smooth muscle actin, and neuron specific enolase. Similarly, PAS and alcian blue histochemistry were used to test for the presence of mucins. The glands were excised and cells isolated using enzymatic digestion and then established in long term culture. Different media and substrata were trialed for suitability. When cultured on uncoated Costar plastic in DMEM/10%FBS, the pattern of cell growth was similar for all glands with distinct phases involving aggregation and migration out from the aggregates before cells died between 20 to 30 days. Immunohistochemical staining indicated that the cultures were of acinar cells with a small percentage of ductal cells. The acinar cells of the lacrimal glands in situ and in vitro stained with antibody MNF116 directed against cytokeratins 5, 6, 8 and 17 but did not stain for antibodies to cytokeratin 18. The reverse staining pattern was true for the Harderian gland. Sections from the white lobe of the Harderian gland showed islets of serous secreting cells which showed positive staining when MNF116 was used. In situ, PAS positive cells were found in a small number of demilunes in the superior and inferior lacrimal glands and also in cells of the intercalated ducts. Surprisingly, in culture nearly all cells, including those isolated form the Harderian gland became PAS positive. In this study we have demonstrated that acinar cells from the Harderian and lacrimal glands of rabbit can be isolated and maintained in culture for 20 to 30 days, and that despite dramatic morphological changes, these cells retain their distinctive phenotype as indicated by antibody staining to specific cellular structural proteins such as cytokeratins and actin. However, the cultured cells also begin to produce mucins as indicated by PAS staining.

Animals↗
PubMed1998

Exchangeability of actin in cardiac myocytes and fibroblasts as determined by fluorescence photobleaching recovery.

Rhodamine (Rho)-labeled muscle and non-muscle actins were microinjected into cultured embryonic chicken cardiac myocytes and fibroblasts. After incorporation of the fluorescent actin analog into cellular structures, small areas of labeled structures were photobleached with a laser pulse, and fluorescence recovery (FR) was measured to determine the exchangeability of isoactins in these structures. With both Rho-muscle and Rho-non-muscle actins, the FR rate in any part of stress fibers was consistently faster than that observed in any part of myofibrils. Thus, although non-striated (proximal and terminal) portions of nascent myofibrils are similar in appearance and composition to stress fibers, our data clearly revealed differences in actin stability between these two structures. Further, although cardiomyocytes were incapable of discriminating between the incorporation of muscle and non-muscle actin isoforms into myofibrils, FR after photobleaching of Rho-muscle actin was faster than that of Rho-non-muscle actin in immature non-striated portions. This indicates that actin molecules in cardiac myofibrils cannot be readily exchanged by heterotypic non-muscle actin. Fluorescently labeled actin incorporated into non-striated (proximal and terminal) portions of myofibrils and terminal portions of stress fibers was found to be more stable than alpha-actinin. The relative stability of actin could facilitate the formation of nascent Z-bands of myofibrils and the reorganization of stress fibers at these portions.

Actinin↗
PubMed1997

Cholinergic modulation, visual function and Alzheimer's dementia.

Electrophysiological evidence at a cellular level and in vivo macroelectrode recordings converge in indicating a degree of specificity of acetylcholine action in vision. Acetylcholine (ACh) function is also thought to play a significant role in memory, learning and other cognitive processes. In this respect, ACh action is suggested to serve in both sensory and cognitive processes. The pharmacological blocking of brain muscarinic transmission has been proposed as a model of geriatric memory impairment and Alzheimer's dementia. Visual electrophysiological testing is deemed of diagnostic specificity for this disease. ACh brain neurotransmission, however, mostly contributes to the modulation of nonspecific aspects of cognition, such as arousal or attention. Alzheimer's dementia results from complex neuron alterations [which also affect muscarinic receptors among other (sub)cellular structures] rather than simply reflecting ACh impoverishment. A substantial loss of retinal ganglion cells is documented in patients with Alzheimer's disease and is consistent with electrophysiological observations. However, it is unclear to what extent the dysfunction of the visual system observable in Alzheimer's dementia is qualitatively different from that occurring spontaneously during aging. The dissimilarities between the effect of acute muscarinic blocking (e.g. by scopolamine) and dementia outnumber the similarities. Accordingly, the conventional ACh agonist-antagonist model of dementia now appears questionable, and replacement treatment with compounds enhancing ACh function proved disappointing. It is suggested that (nonspecific) ACh action becomes function-specific, as determined by the architecture of local brain circuits in which it is involved.

Acetylcholine↗
PubMed2003

A review of the biochemistry of heavy metal biosorption by brown algae.

The passive removal of toxic heavy metals such as Cd(2+), Cu(2+), Zn(2+), Pb(2+), Cr(3+), and Hg(2+) by inexpensive biomaterials, termed biosorption, requires that the substrate displays high metal uptake and selectivity, as well as suitable mechanical properties for applied remediation scenarios. In recent years, many low-cost sorbents have been investigated, but the brown algae have since proven to be the most effective and promising substrates. It is their basic biochemical constitution that is responsible for this enhanced performance among biomaterials. More specifically, it is the properties of cell wall constituents, such as alginate and fucoidan, which are chiefly responsible for heavy metal chelation. In this comprehensive review, the emphasis is on outlining the biochemical properties of the brown algae that set them apart from other algal biosorbents. A detailed description of the macromolecular conformation of the alginate biopolymer is offered in order to explain the heavy metal selectivity displayed by the brown algae. The role of cellular structure, storage polysaccharides, cell wall and extracellular polysaccharides is evaluated in terms of their potential for metal sequestration. Binding mechanisms are discussed, including the key functional groups involved and the ion-exchange process. Quantification of metal-biomass interactions is fundamental to the evaluation of potential implementation strategies, hence sorption isotherms, ion-exchange constants, as well as models used to characterize algal biosorption are reviewed. The sorption behavior (i.e., capacity, affinity) of brown algae with various heavy metals is summarized and their relative performance is evaluated.

Adsorption↗
PubMed2003

Coevolution of viruses with hosts and vectors and possible paleontology.

The coevolution of viruses with their hosts and vectors depends on the evolution of the hosts and vectors coupled with factors involved in virus evolution. The long-term perspective involves the origin of life forms, the evolution of host and vector (especially arthropods) kingdoms and families, and changes in biological diversity induced mainly by the last five great extinctions. In the medium term, the diversification of hosts and vectors is important, and in the short term, recent events, especially humans, have had a great impact on virus coevolution. As there are few, if any, examples of conventional fossils of viruses, evidence for their evolution related to host and vector evolution is being found from other sources, especially virus-induced cellular structures and recent developments in molecular biology. Recognizing these other sources is becoming important for paleontologists gaining an understanding of the influence that viruses have had on the development of higher organisms.

Animals↗
PubMed2004

The effects of tyrphostine Ag 556 on experimental spinal cord ischemia reperfusion injury.

BACKGROUND: To investigate the effects of Tyrphostin Ag 556 on spinal cord ischemia reperfusion injury. METHODS: The inhibition of tyrosine kinase may represent a novel approach in the treatment of spinal cord ischemia reperfusion injury. Recently, a family of tyrosine kinase inhibitors, the tyrphostins, has been successfully used in models of endotoxemia, peritonitis, and hypovolemic shock. MATERIALS AND METHODS: Twenty-four Wistar rats were used in the study. Rats were divided into 4 groups of 6 animals. The groups were named as sham operated group, injury group, vehicle group, and treatment group. Clamping of the abdominal aorta was performed for 45 minutes with all of the groups except sham-operated group. All of the rats were sacrificed 24 hours after the operation for biochemical and ultrastructural studies. RESULTS: Tyrphostin Ag 556 treatment was found effective on experimental spinal cord ischemia reperfusion injury. The Malondialdehyde (MDA) values of the treatment group were statistically significant lower then the other reperfusion injury groups. The histologic examination showed better cellular structure in the treatment group than the other reperfusion injury groups. The neurologic scores of the treatment group also improved after treatment. CONCLUSIONS: Tyrphostin Ag 556 alters spinal cord ischemia reperfusion injury by inhibiting protein kinases. Further investigations will be required to determine the long-term effects of this drug.

Animals↗
PubMed1999

GFP variants for multispectral imaging of living cells.

Unlike enzyme markers, green fluorescent protein can be visualized at high resolution in living cells using confocal microscopy. The images are not prone to fixation or staining artifacts, and can be of exceptional clarity. Moreover, the activities of living cells, such as cytoplasmic streaming, are clearly evident during microscopy. Ordinarily, movement within a sample is a nuisance, placing constraints on the use of sometimes lengthy techniques for noise reduction during confocal microscopy, such as frame averaging. However, it is possible to monitor dynamic events by time-lapse confocal microscopy, and this combination of a vital fluorescent reporter with high-resolution optical techniques shows much promise for use in cell biological and physiological experiments. Genetic systems such as that of Arabidopsis provide a large resource of potentially informative mutants, and there has been much recent improvement in techniques for determining the molecular basis of a particular phenotype. The use of fluorescent proteins will provide further tools for examining the biology of mutant cells. The precision with which particular cellular structures can be decorated with GFP and the ease with which subcellular traffic can be monitored indicate that this approach will be very useful for cell biological and physiological observations, particularly for detailed examination of plant mutant phenotypes.

Animals↗
PubMed2001

Human preadipocytes seeded on freeze-dried collagen scaffolds investigated in vitro and in vivo.

Currently, there is no adequate implant material for the correction of soft tissue defects such as after extensive deep burns, after tumor resection and in hereditary and congenital defects (e.g. Romberg's disease, Poland syndrome). The autologous transplantation of mature adipose tissue has poor results. In this study human preadipocytes of young adults were isolated and cultured. 10(6) preadipocytes were seeded onto collagen sponges with uniform 40 microm pore size and regular lamellar structure and implanted into immunodeficient mice. Collagen sponges without preadipocytes were used in the controls. Macroscopical impression, weight, thickness, histology, immunohistochemistry (scaffold structure, cellularity, penetration depth of the seeded cells) and ultrastructure were assessed after 24 h in vitro and after explantation at 3 and 8 weeks. Preadipocytes penetrated the scaffolds 24 h after seeding at a depth of 299+/-55 microm before implantation. Macroscopically after 3 and 8 weeks in vivo layers of adipose tissue accompanied by new vessels were found on all preadipocyte/collagen grafts. The control grafts appeared unchanged without vessel ingrowth. There was a significant weight loss of all grafts between 24 h in vitro and 3 weeks in vivo (p < 0.05), whereas there was only a slight weight reduction from week 3 to 8. The thickness decreased in the first 3 weeks (p < 0.05) in all grafts. The preadipocyte/collagen grafts were thinner but had a higher weight than the controls at this point in time. The histology showed adipose tissue and a rich vascularisation adherent to the scaffolds under a capsule. The control sponges contained only few cells and a capsule but no adipose tissue. Human-vimentin positive cells were found in all preadipocyte/collagen grafts but not in the controls, penetrating 1188+/-498 microm (3 weeks) and 1433+/-685 microm (8 weeks). Ultrastructural analysis showed complete in vivo differentiation of viable adipocytes in the sponge seeded with preadipocytes. Formation of extracellular matrix was more pronounced in the preadipocyte/collagen grafts. The transplantation of isolated and cultured preadipocytes within a standardised collagen matrix resulted in well-vascularised adipose-like tissue. It is assumed that a pore size greater than 40 microm is required, as preadipocytes enlarge during differentiation due to incorporation of lipids.

Adipocytes↗