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At least 739 records · Page 41Linked to original sources
PubMed2007

Relation between corneal innervation with confocal microscopy and corneal sensitivity with noncontact esthesiometry in patients with dry eye.

PURPOSE: An alteration in corneal innervation has been described in dry eye associated with diabetes mellitus, contact lens use, and LASIK. The purpose of this study was to evaluate whether dry eye not related to Sjögren's syndrome (NSDE) and dry eye related to primary Sjögren's syndrome (PSDE) are associated with an alteration of the corneal nerves and sensation. METHODS: Twenty-one patients with dry eye (10 NSDE and 11 PSDE) and 20 healthy volunteers were studied. Healthy volunteers were divided into two groups: one younger than 60 years (N<60) and the other 60 years of age or older (N> or =60). The study of the epithelium, stroma, and subbasal corneal nerves was performed with a confocal microscope. Mechanical, chemical, and thermal sensation was evaluated using the Belmonte noncontact esthesiometer. RESULTS: A statistically significant decrease in the number and density of subbasal nerves (P < 0.0001) and the density of superficial epithelial cells (P < 0.0001) was observed in dry eyes. The number and density of subbasal nerves was higher in the N<60 group. A significant decrease was found with respect to mechanical, chemical, and thermal sensitivity (P < 0.0001). Sensibility was better in the healthy eyes. A strong correlation was found between the density of superficial epithelial cells and the nerves and between the number and density of subbasal nerves and sensation (P < 0.001). CONCLUSIONS: The use of confocal microscopy and noncontact esthesiometry allow the detection of the presence of corneal neuropathy in patients with dry eye.

Adult↗
PubMed1995

[Flow cytometry and laser scanning confocal microscopy analysis of the receptor distribution].

To study the physiological regulation of the receptor protein, a fluorescent probe and detection system for alpha 1B-adrenergic receptors have been developed. By using the anti-peptide antibody developed against the alpha 1B-adrenergic receptor NH2-terminus, we have examined the agonist-regulated alpha 1B-adrenergic receptor redistribution in desensitized cells. Flow cytometry analysis showed that anti-peptide antibody against alpha 1B-adrenergic receptor specifically identifies the receptor in CHO cells, COS-7 cells that were transfected with alpha 1B-adrenergic receptor cDNA and rat hepatocytes. Using a fluoro-labeled receptor ligand, BODIPY FL-prazosin, as a probe, cell surface alpha 1-adrenergic receptor subtypes can be detected by flow cytometry. Laser scanning confocal microscopy visualized the agonist-regulated redistribution process of alpha 1B-adrenergic receptor in living cells; thus, following phenylephrine (10(-6) M) stimulation, receptor antigen at the cell surface rapidly internalized and clustered together in a cell within 30 min. The results showed that the antibody and fluoro-labeled ligand are valuable tools for studying the localization and functional role of the alpha 1-adrenergic receptor subtype.

Animals↗
PubMed1998

Detection of low copy numbers of HPV DNA by fluorescent in situ hybridization combined with confocal microscopy as an alternative to in situ polymerase chain reaction.

In genital lesions infected by human papillomavirus (HPV), histological criteria and HPV DNA typing are of prognostic value. Therefore, non-radioactive methods such as in situ hybridization are used extensively since they preserve the histological organization of the tissue, and allow the detection and characterization of HPV DNA. However, the sensitivity of these methods is often limited to detection of low copy numbers of HPV DNA in isolated cells or in tissue sections, and therefore alternative techniques have been explored. In the present study, 1-2 copies of HPV DNA were visualized in SiHa cells either by in situ amplification of nucleic acid sequences with the polymerase chain reaction (PCR) or by fluorescent in situ hybridization (FISH) associated with observation by laser scanning confocal microscopy (LSCM). The latter procedure was evaluated for use on histological tissue sections to identify low copy numbers of HPV DNA. Genital lesions which were negative by enzymatic in situ hybridization and FISH but histologically suspected of HPV infection were investigated, and intense signals were obtained both with in situ PCR and with the combined use of FISH and LSCM. Therefore, the combination of FISH with LSCM examination may be as valuable as in situ PCR to detect viral genes present in small amounts in isolated cells and in tissue sections.

Condylomata Acuminata↗
PubMed2006

GRIF-1-kinesin-1 interactions: a confocal microscopy study.

GRIF-1 [GABA(A) (gamma-aminobutyric acid(A)) receptor interacting factor-1] is a member of a coiled-coil family of proteins thought to function as adaptors in the anterograde trafficking of organelles utilizing the kinesin-1 motor proteins to synapses. To study in more detail the molecular interaction between GRIF-1 and the kinesin-1 family member KIF5C, fluorescent yellow- and fluorescent cyan-tagged GRIF-1, KIF5C, the KIF5C MD (motor domain) and the KIF5C NMD (non-motor domain) fusion proteins were generated. Each was characterized with respect to size and ability to co-associate by immunoprecipitation following expression in HEK-293 (human embryonic kidney 293) cells. Further, their distribution in transfected HEK-293 and transformed African green monkey kidney (COS-7) cells was analysed by confocal microscopy. The fluorescent GRIF-1 and KIF5C fusion proteins were all found to behave as wild-type. Double GRIF-1/KIF5C transfectants revealed co-localization. The GRIF-1/KIF5C and GRIF-1/KIF5C NMD double transfectants showed different subcellular distributions compared with single GRIF-1, KIF5C or KIF5C NMD transfections. These studies confirm the association between GRIF-1 and kinesin-1 NMDs. Fluorescence resonance energy transfer studies are ongoing to characterize this interaction in more detail.

Animals↗
PubMed1999

Time-lapsed confocal microscopy reveals temporal and spatial expression of the lysine epsilon-aminotransferase gene in Streptomyces clavuligerus.

To investigate the temporal and spatial expression patterns of the gene (lat ) encoding lysine epsilon-aminotransferase (LAT) for cephamycin C biosynthesis, a mutant form of green fluorescent protein (mut1GFP) was integrated into the Streptomyces clavuligerus chromosome (strain LH369), resulting in a translational fusion with lat. LAT activity and fluorescence profiles of the recombinant protein paralleled the native LAT enzyme activity profile in wild-type S. clavuligerus, which peaked during exponential growth phase and decreased slowly towards stationary phase. These results indicate that the LAT-Mut1GFP fusion protein retains both LAT and GFP functionality in S. clavuligerus LH369. LH369 produced wild-type levels of cephamycin C in minimal medium culture conditions supplemented with lysine. Time-lapsed confocal microscopy of the S. clavuligerus LH369 strain revealed the temporal and spatial characteristics of lat gene expression and demonstrated that physiological development of S. clavuligerus colonies leading to cephamycin C biosynthesis is limited to the substrate mycelia.

Blotting, Southern↗
PubMed2005

Reconstruction and exploration of three-dimensional confocal microscopy data in an immersive virtual environment.

An immersive virtual environment for interactive three-dimensional reconstruction and exploration of confocal microscopy data is presented. For some structures automatic alignment of serial sections can lead to geometric distortions. The superior visual feedback of a Virtual Reality system is used to aid in registering and aligning serial sections interactively. An ImmersaDesk Virtual Reality display system is used for display and interaction with the volumetric confocal data. Detailed methods for handling both single-section and multi-section confocal data are described.

Humans↗
PubMed1994

Elastic extracellular matrix of the embryonic chick heart: an immunohistological study using laser confocal microscopy.

The "elastic matrix" constitutes a specialized component of the extracellular matrix which confers resiliency to tissues and organs subjected to repeated deformations. The role of the elastic matrix in living organisms appears to be of key importance since diseases characterized by expression of defective inherited genes which encode components of the elastic matrix lead to premature death. While the elastic matrix of adult organs has received a great deal of attention, little is known about when it first appears in embryonic tissues or its possible role in developing organs. In the present study we have performed an immunohistochemical study of the distribution of elastin and three additional components often associated with elastic matrices in adult tissues (i.e., fibrillin, emilin, and type VI collagen) during the development of the chicken embryonic heart. The three-dimensional arrangement of these components was established through the observation of whole-amount specimens with scanning laser confocal microscopy. Our results revealed three different periods of heart development regarding the composition of the elastic matrix. Prior to stage 21 the embryonic heart lacks elastin but exhibits a matrix scaffold of fibrillin and emilin associated with the endocardium and the developing cardiac jelly. Between stages 22 and 29 the heart shows a transient elastic scaffold in the outflow tract which contains elastin, fibrillin, and emilin. Elastin-positive fibrillar material is also observed during these stages in the base of the atrioventricular cushion adjacent to the myocardial wall. In addition, emilin-positive material appears to be associated with the zones of formation of ventricular trabeculae. Collagen type VI was not detected during these early stages. From stage 30 to stage 40 a progressive modification of the pattern of distribution of elastin, fibrillin, emilin, and collagen type VI is observed in association with the formation of the definitive four-chambered heart. The distribution of the elastic scaffold in the outflow tract appears to be rearranged and becomes restricted to the roots of the main arteries. Each of the components studied here is also deposited at increasing levels in the developing valvular apparatus including the valve leaflets and the chordae tendinea. The components are also present in the subendocardial space where they form aligned fibrillar tracts, an arrangement suggestive of a role in ventricular contractile function. The epicardium constitutes an additional region of elastic matrix deposition during these later stages and contains elastic, fibrillin, and collagen type VI.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2003

Efficient center-line extraction for quantification of vessels in confocal microscopy images.

In this paper we present a novel method for the three-dimensional (3-D) centerline extraction of elongated objects such as vessels. This method combines the basic ideas in distance transform-based, thinning, and path planning methods to extract thin and connected centerlines. This efficient approach needs no user interaction or any prior knowledge of the object shape. We used the path planning approach, which has exclusively been used in the virtual endoscopy or robotics, to obtain the medial curve of the objects. To make our approach fully automated, a distance transform mapping is used to identify the end points of the object branches. The initial paths are also constructed on the surface of the object, traversing the same distance map. Then a thinning algorithm centralizes the paths. The proposed approach is especially efficient for centerline extraction of the complex branching structures. The method has been applied on the confocal microscopy images of rat brains and the results confirm its efficiency in extracting the medial curve of vessels, essential for the computation of quantitative parameters.

Algorithms↗
PubMed2005

Investigation of the stability of polyelectrolyte multilayer coatings in open-tubular capillary electrochromatography using laser scanning confocal microscopy.

A simple polyelectrolyte multilayer (PEM) coating procedure was used for the development of stable modified capillaries. PEM coatings were constructed in fused-silica capillaries using alternating rinses of cationic and anionic polyelectrolytes. The multilayer coatings investigated in this study consisted of two and twenty layer pairs, or bilayers. A bilayer is one layer of a cationic polymer and one layer of an anionic polymer. Poly(diallyldimethylammonium chloride) was used as the cationic polymer, and the polymeric surfactant poly(sodium N-undecanoyl-L-leucylvalinate) was used as the anionic polymer. Previous studies for both chiral and achiral separations have shown that PEM-coated capillaries have excellent reproducibilities, remarkable endurance, and strong stabilities against extreme pH values when used in open-tubular capillary electrochromatography (OT-CEC). In this study, the stability of the coatings was further investigated after exposure to 0.1 M and 1.0 M NaOH. Structural changes of these coatings were monitored using laser scanning confocal microscopy (LSCM) after flushing the capillaries with NaOH. This technique allowed observation of the degradation of the coatings. Observations are discussed in terms of separations using OT-CEC. Electropherograms obtained from the chiral separation of 1,1'-binaphthyl-2,2'-dihydrogenphosphate in OT-CEC showed a decrease in selectivity and an increase in electroosmotic mobility after long exposure to NaOH. The ability to recover the capillaries by exposure to NaOH was also demonstrated. Measurements of electroosmotic mobility and selectivity showed that 2-bilayer and 20-bilayer PEM coatings could be completely removed from the capillary surface after approximately 3.5 and 9.5 h, respectively, of continuous exposure to 1 M NaOH.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed1998

Assessment of stress fiber orientation during healing of radial keratotomy wounds using confocal microscopy.

Radial keratotomy (RK) is a refractive surgical procedure in which partial thickness incisions are made in the cornea in order to alter its shape. Previous studies suggest that RK wounds undergo changes in wound gape in response to the ingrowth of myofibroblasts which mediate subsequent wound contraction and may modulate changes in corneal curvature seen after RK. A recent quantitative analysis of f-actin organization in full-thickness incisional wounds in the rabbit demonstrated that microfilament bundles (stress fibers) present in myofibroblasts align parallel to the long axis of the wound during wound contraction. To investigate whether the same pattern of alignment occurs after RK, a similar analysis of f-actin organization was undertaken using the cat RK model. Radial keratotomy wounds were studied from 10 to 28 days after surgery using en block staining with fluorescein isothiocyanate (FITC) phalloidin, and three-dimensional (3-D) datasets (z-series of en face optical sections) were collected using laser confocal microscopy at various regions within the wound. In addition, conventional en face sections were double-labeled using combinations of phalloidin and antibodies to fibronectin and alpha 5 beta 1 integrin. Myofibroblast ingrowth started in the bottom of the wound and progressed anteriorly. At 10 to 14 days, f-actin was predominantly distributed in long, thick bundles (stress fibers) within the wound. These fibers appeared to be randomly oriented anteriorly, but became progressively more aligned with the long axis of the wound posteriorly. At 21 days, the stress fibers were predominantly oriented parallel to the long axis of the wound at all levels. F-actin, fibronectin and integrin were coaligned at both the 14 and 21 day time points. Since the majority of wound closure occurs between 14 and 28 days after surgery, we conclude that parallel alignment of the actin filament-fibronectin-integrin assembly in the cat RK model is associated with wound contraction.

Actin Cytoskeleton↗
PubMed2006

Diffusion delays and unstirred layer effects at monolayer cultures of Chinese hamster ovary cells: radioligand binding, confocal microscopy, and mathematical simulations.

Cells grown in monolayer culture offer a convenient system for binding and other experiments under conditions that preserve the complexity of the living state. Kinetics experiments, however, may be distorted by the time course of drug penetration into even so simple a "tissue" as the monolayer. The impediments include unstirred layers both above and between the cells, the congregation of receptors within the confined space between cells, and nonspecific binding to membrane components. The contributions of these factors were investigated in cultures of Chinese hamster ovary (CHO) cells either nontransfected or stably transfected with mu opioid receptors. The dissociation of [3H]naloxone was four times faster under displacement than under infinite dilution conditions, clearly demonstrating the "retention effect" of receptors confined in space. Even the penetration of this ligand between nontransfected cells showed salient delays with respect to diffusion into a slab, indicating that nonspecific, low-affinity binding to membrane components was arresting its progress. The optical sectioning capabilities of confocal microscopy demonstrated that the kinetics of two fluorescent antagonists depended on the vertical plane, providing direct evidence for slowed diffusion down a single cell depth. Modeling shows that kinetic errors increase with receptor density, forward rate constant, and the thickness of the unstirred layer.

Animals↗
PubMed1997

Appositions made by axons of descending interneurons in the guinea-pig small intestine, investigated by confocal microscopy.

There are four major classes of descending interneurons in the myenteric plexus of the guinea-pig small intestine. In this study, the connections made by two of these classes of descending interneurons with other interneurons and with inhibitory motor neurons have been investigated using confocal, conventional fluorescence and electron microscopy. The terminals of descending interneurons known to contain both bombesin (BN) and nitric oxide synthase (NOS) were identified by BN immunoreactivity (IR). Cholinergic interneurons known to contain somatostatin (SOM) were identified by SOM-IR. The connections of these two groups of interneurons with the following three classes of nerve cell bodies were examined: those with NOS-IR that also contain gamma-aminobutyric acid (GABA) (inhibitory motor neurons), those with only NOS-IR (descending interneurons and inhibitory motor neurons) and those with only GABA-IR (motor neurons). The BN-IR and SOM-IR interneurons were found to form connections with each other, and both types of interneurons provided inputs to motor neurons. Most previous analyses of interconnections in the enteric plexuses have been by conventional fluorescence microscopy and electron microscopy. In the present work these are compared with confocal microscopy. BN-IR varicosities formed pericellular baskets around each class of nerve cell that were easily identifiable with all techniques. Using confocal microscopy, BN-IR varicosities that were in contact with NOS-IR and GABA-IR nerve cells were quantified. Confocal microscopy demonstrated over twice as many contacts as were shown by a previous electron microscopic study. In contrast, conventional fluorescence microscopy showed little indication that SOM-IR varicosities formed inputs to NOS-IR or GABA-IR nerve cells, despite the fact that confocal microscopy revealed direct appositions and electron microscopy revealed synapses. This study has shown that confocal analysis is a valuable adjunct to conventional fluorescence microscopy for determining neuronal circuitry. Moreover, it allows a more rapid collection of data than does electron microscopy. It is concluded that chains of BN-IR and SOM-IR interneurons from descending pathways in the small intestine and that both types of interneuron connect with muscle motor neurons.

Animals↗
PubMed2001

The developing neurovascular anatomy of the embryo: a technique of simultaneous evaluation using fluorescent labeling, confocal microscopy, and three-dimensional reconstruction.

The close spatial relationship between peripheral nerves and blood vessels in the adult is well known. However, evidence supporting the congruent development of these structures in embryos remains anecdotal. Neurovascular relationships also have been shown to be conserved in other vertebrates. This homology suggests that either peripheral nerves or blood vessels, or both, might have fundamental morphogenetic roles during embryologic development. Both peripheral nerves and blood vessels have been independently implicated as etiologic agents in the pathogenesis of congenital disabilities, and several congenital anomalies fit their distribution patterns. This article presents a technique for the simultaneous visualization of peripheral nerves and blood vessels at different stages in the developing embryo. The forelimbs of 310 quail embryos were dissected over a 1-year period. Peripheral nerves were labeled with the neural crest and axon antibody, HNK-1, followed by fluorescein-conjugated secondary antibodies. Blood vessels were labeled by a perfusion technique using the fluorescent dye, dioctadecyl-tetramethylindocarbocyanine. Specimens were processed and imaged in whole-mount with confocal microscopy, and images were reconstructed using three-dimensional modeling software. Both nerves and blood vessels seem to undergo a highly stereotypic sequence of development in the embryonic quail forelimb. Furthermore, the existence of a close spatial relationship between nerves and blood vessels suggests either a high degree of developmental interdependence or shared patterning mechanisms. This technique permits further evaluation of the possible role peripheral nerves and blood vessels might play in the pathogenesis of congenital disabilities and provides a starting point for further studies aimed at elucidating the means by which peripheral nerves and blood vessels are patterned in the forelimb of the avian embryo.

Animals↗
PubMed1996

Efficient Monte Carlo simulation of confocal microscopy in biological tissue.

A variance-reduction technique is described that greatly improves the efficacy of Monte Carlo simulations of reflection-mode confocal microscopy in anisotropically scattering media. The efficiency gain is large enough that the performance of confocal microscopes probing as deep as 5 scattering lengths can be simulated with a desktop computer. We use the technique to simulate the response of a true confocal microscope probing biological tissue, a problem that has been impractical to undertake by using conventional Monte Carlo methods. Our most important finding is that operation of a confocal microscope in the true confocal mode enables much more effective rejection of undesired scattered light than operation in the partially coherent mode, but the maximum probing depths of microscopes operated in either mode are similar (2-3) scattering lengths) in practice because of sensitivity limitations.

Animals↗
PubMed2006

Impact of del32-71-GH (exon 3 skipped GH) on intracellular GH distribution, secretion and cell viability: a quantitative confocal microscopy analysis.

BACKGROUND: Familial isolated growth hormone deficiency (IGHD) is a disorder with about 5-30% of patients having affected relatives. Among those familial types, IGHD type II is an autosomal dominant form of short stature, associated in some families with mutations that result in missplicing to produce del32-71-GH, a GH peptide which cannot fold properly. The mechanism by which this mutant GH may alter the controlled secretory pathway and therefore suppress the secretion of the normal 22-kDa GH product of the normal allele is not known in detail. Previous studies have shown variance depending on cell type, transfection technique used, as well as on the method of analysis performed. AIM: The aim of our study was to analyse and compare the subcellular distribution/localization of del32-71-GH or wild-type (wt)-GH (22-kDa GH), each stably transfected into AtT-20, a mouse pituitary cell line endogenously producing ACTH, employed as the internal control for secretion assessment. METHODS: Colocalization of wt- and del32-71 mutant GH form was studied by quantitative confocal microscopy analysis. Using the immunofluorescent technique, cells were double stained for GH plus one of the following organelles: endoplasmic reticulum (ER anti-Grp94), Golgi (anti-betaCOP) or secretory granules (anti-Rab3a). In addition, GH secretion and cell viability were analysed in detail. RESULTS/CONCLUSIONS: Our results show that in AtT-20 neuroendocrine cells, in comparison to the wt-GH, the del32-71-GH has a major impact on the secretory pathway not only affecting GH but also other peptides such as ACTH. The del32-71-GH is still present at the secretory vesicles' level, albeit in reduced quantity when compared to wt-GH but, importantly, was secretion-deficient. Furthermore, while focusing on cell viability an additional finding presented that the various splice site mutations, even though leading eventually to the same end product, namely del32-71-GH, have different and specific consequences on cell viability and proliferation rate.

Adrenocorticotropic Hormone↗
PubMed2005

Quantitative assessment of ophthalmic viscosurgical device retention using in vivo confocal microscopy.

PURPOSE: To develop and apply a new laboratory method for in vivo quantitative assessment of the retention of ophthalmic viscosurgical devices (OVDs) following phacoemulsification. SETTING: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. METHODS: Studies of both eyes of New Zealand White rabbits were performed. Six OVDs were evaluated: Provisc and Healon (both sodium hyaluronate 1%), Healon5 (sodium hyaluronate 2.3%), Amvisc Plus (sodium hyaluronate 1.6%), Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%), and a new viscous-dispersive OVD, DisCoVisc (sodium hyaluronate 3%-chondroitin sulfate 4%). The OVD was injected to fill the anterior chamber and a phacoemulsification needle inserted with the tip positioned just anterior to the lens capsule. Simulated phacoemulsification was performed for 1 minute using flow rates of 20, 40, and 60 mL/min; a vacuum level of 300 mm Hg; and ultrasound power of 60% using a Legacy phacoemulsification unit. The needle was removed, and silicone oil (1000 centistokes) was injected into the anterior chamber. The distance between the corneal endothelium and the OVD-silicone oil interface was measured using in vivo confocal microscopy through-focusing (CMTF). RESULTS: Significant differences in residual thickness were found between the OVDs tested. Specifically, the residual thickness of both DisCoVisc (mean 324.5 microm +/- 163.7 [SD]) and Viscoat (251.4 +/- 100.9 microm) was significantly greater than that of Provisc (9.5 +/- 16.7 microm), Healon (3.8 +/- 11.3 microm), Healon5 (0.6 +/- 2.4 microm), and Amvisc Plus (65.6 +/- 134.0 microm) (P < .05, Dunn test). Ophthalmic viscosurgical device retention was greatest with DisCoVisc; however, there was no statistically significant difference between DisCoVisc and Viscoat in residual thickness. The flow rate did not have a significant effect on the residual thickness (Friedman 2-way analysis of variance by ranks). CONCLUSIONS: Residual OVD thickness following simulated phacoemulsification could be quantitatively measured using in vivo CMTF. The results are consistent with human surgical experience in that the dispersive OVD (Viscoat) was better retained than the cohesive compounds. DisCoVisc, a new viscous-dispersive OVD, also showed retention compared with Viscoat under the experimental conditions.

Animals↗
PubMed1999

The TATA-box binding protein of Entamoeba histolytica: cloning of the gene and location of the protein by immunofluorescence and confocal microscopy.

A 309 bp DNA fragment from Entamoeba histolytica was amplified by PCR using primers derived from the Acanthamoeba castellanii consensus TATA-box binding protein amino acid sequence. The amplified fragment was used to isolate cDNA and genomic DNA clones containing an ORF encoding the complete E. histolytica TATA-box binding protein (Ehtbp, 702 bp, 234 aa, molecular mass 26 kDa). The EhTBP functional domain showed 55% sequence identity to that of Homo sapiens, 54% to A. castellanii and 37% to Plasmodium falciparum TBPs. In Southern blot experiments we detected a single Ehtbp band, which was transcribed as a 1.3 kb mRNA containing a 420 nt 5' untranslated region. However, the probe hybridized with the 0.8 and 1.5 Mb chromosomes, suggesting that this sequence is diploid. In situ PCR assays showed two signals in 95% of trophozoites, one located in the nucleus and another in EhkO, the novel DNA-containing organelle recently reported. The recombinant E. histolytica TATA-box binding protein was expressed in Escherichia coli. Antibodies against it recognized two proteins of 26 and 29 kDa in E. histolytica nuclear extracts. Confocal microscopy immunofluorescence analysis located the protein in both the nucleus and EhkO.

Amino Acid Sequence↗
PubMed2003

Comparative study of human keratocyte density after corneal grafting by using confocal microscopy in vivo.

PURPOSE: In a case control study we determined keratocyte density and size of nuclear area in the central segment of the corneal stroma. METHODS: We compared 20 corneas after keratoplasty with 24 eyes of normal healthy individuals. Both groups were matched according to age. Keratocyte density and nuclear area were analyzed using Nidek Confoscan 2 separately for each group. In corneal graft recipients we studied how the mentioned variables were influenced by age of corneal graft donor (ranged from 4- to 71-years old) and by the time from surgery (8 to 77 months). RESULTS: The comparison of healthy controls and patients with keratoplasty revealed no changes in keratocyte density and the size of nuclear area in central stromal layers. In patients after keratoplasty donor age influenced an increase in keratocyte nuclei area only in the posterior stroma layer (p = 0.042). No such changes were observed in anterior and midstroma layers. Donor age was not found to be significant for keratocyte density in any of the layers. Time from surgery neither influenced changes in keratocyte density nor in keratocyte nuclei area. CONCLUSIONS: In our study using confocal microscopy in vivo we found that corneal grafting does not influence keratocyte density and nuclear area in individual layers of the central corneal stroma segment (anterior, midstroma and posterior layers). Donor age influenced an increase in keratocyte nuclei area only in the posterior stroma layer.

Adolescent↗