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PubMed1989

Association of Tat protein and viral mRNA with nuclear matrix from HIV-1-infected H9 cells.

The transactivating protein from human immunodeficiency virus type 1 (HIV-1), Tat, was found to bind to the nuclear matrix from uninfected and HIV-1-infected H9 cells. Addition of the Zn2+, Cd2+ and Cu2+ chelator o-phenanthroline destroyed the matrix fibrils and the binding affinity of Tat to the matrix. A sequential treatment of the matrix, first with o-phenanthroline and then with ZnCl2, partially restored the fibrillar-like matrix structure. Infection of H9 cells with HIV-1 resulted in a displacement of cellular mRNA by viral mRNA from the nuclear matrix. Both the matrix-bound host cell and HIV-1 mRNA were found to dissociate from the matrix in the presence of o-phenanthroline. This could be prevented by coincubation with Zn2+ or Cu2+ (but not Mg2+), which stabilize the mRNA containing nuclear matrix structure.

Cell Nucleus↗

PubMed1991

Increased matrix vesicle protein in rachitic rat epiphyseal growth plates.

Extracellular, membrane-bound vesicles are widely regarded to be the initial site of calcification in a variety of tissues under normal and pathological conditions. Alkaline phosphatase is believed to play a vital role in this process by hydrolysing ester phosphates or mineral inhibitors, e.g. inorganic phosphates. In the present study, matrix vesicles from normal and rachitic rat growth plates were compared with regard to specific activity of alkaline phosphatase, total vesicle protein and ultrastructural distribution of alkaline phosphatase activity. Matrix vesicles were released from normal or rachitic growth plates by collagenase digestion and isolated by differential centrifugation. Enzyme cytochemical localization involving a cerium capture method was performed on vesicles collected by vacuum filtration on Millipore filters. SDS gels and Western blots on fractions of both normal and rachitic matrix vesicles showed major proteins to be almost identical and confirmed the presence of alkaline phosphatase in both. Total matrix vesicle protein ((mg total matrix vesicle protein/rat) x 10(2)) per rat was significantly greater for the rachitic animals (9.0 +/- 2.0 vs. 4.0 +/- 1.0), P less than 0.0001. Alkaline phosphatase specific activity (units alkaline phosphatase/mg vesicle protein) in the rachitic and normal matrix vesicles was 25.29 +/- 9.36 and 18.78 +/- 3.37, respectively (0.05 less than P less than 0.1). Electron dense cerium phosphate deposits were localized to the outer membrane surface of matrix vesicles derived from both types of rats. This data, the first to quantify the relationship between rickets, matrix vesicle protein and alkaline phosphatase specific activity, suggests that matrix vesicles from rachitic and normal rats have biochemical and morphological similarity.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

PubMed2005

Epitaxial deposition of calcium oxalate on uric acid rich stone matrix is induced by a 29 kDa protein.

BACKGROUND: Association of macromolecules particularly the role of proteins in urolithiasis has been studied for last few centuries, but still a complete profile of stone matrix proteins that mediate co-precipitation of uric acid and calcium oxalate has not been characterized. We isolated and characterize proteins from uric acid rich stone matrix, which have oxalate binding activity. METHODS: Matrix proteins were isolated from uric acid rich stone matrix using EDTA as a demineralizing agent. The radiolabelled solubilized proteins were fractionated with increasing ionic concentration by DEAE cellulose column chromatography to identify the oxalate binding protein. It was purified using Sephadex G-200 column chromatography. Amino acid composition was determined and monoclonal antibody was produced against the oxalate binding uric acid rich stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers urine, their oxalate binding activity assayed and cross reactivity with the produced monoclonal antibody were checked using ELISA and Western blotting. RESULTS: Matrix on DEAE column chromatography elution yielded 3 protein peaks and they were named as fraction I, II and III among which fraction I had higher oxalate binding activity which was further purified with Sephadex G-200 column which yielded 2 protein peaks designated as Ia and Ib. Fraction Ib with molecular weight 29 kDa exhibited the maximum oxalate binding activity. Forty percent of this 29 kDa protein is comprised of basic amino acids. Monoclonal antibody (IgG1) was produced against the 29 kDa stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers, 4 protein peaks were obtained named as fraction I to IV. Among them, fraction IV having molecular weight of approximately 29 kDa cross reacted up to 85.6% with 29 kDa stone matrix protein. Moreover, urinary 29 kDa protein exhibited oxalate binding activity of 94.16 +/- 6.08 pmol/mg protein at pH 5.5. CONCLUSION: The 29 kDa protein isolated from uric acid rich stone matrix and urine are one and the same, thereby insinuating that 29 kDa protein might play a major role in epitaxial deposition of calcium oxalate over uric acid core, consequently favoring the lithogenic events like uric acid and calcium oxalate nucleation, aggregation and retention.

Animals↗

PubMed2004

Cross-linked alginate-gelatine beads: a new matrix for controlled release of pindolol.

This work is focused on the development of a new particulate drug delivery system using a sodium alginate matrix containing pindolol as a model drug molecule for intestinal drug prolonged release. Calcium alginate beads are known to be unable to control the release of most insoluble drugs. Pindolol-loaded alginate-gelatine beads have been developed using a solvent-free technique that involves a cross-linking reaction. Modifications in matrix structure and physicochemical behaviour caused by the cross-linking reaction were assessed during particle formation and drug release. Several parameters, such as matrix gelling rate, encapsulation efficiency, drug release profile and matrix erosion rate, were investigated. Physicochemical characterisation indicates the formation of a new alginate-gelatine matrix and shows that pindolol does not interfere with the matrix formation process. Matrix swelling of calcium alginate beads induced by phosphate buffer ends up in erosion and destruction. However, for cross-linked beads swelling does not lead to complete erosion, which may be the main cause of pindolol retention within the matrix. The modifications introduced in the initial calcium alginate formulation by means of an appropriate method such as the use of a cross-linking agent successfully changed the matrix performance, allowing the controlled release of pindolol.

Alginates↗

PubMed2006

Comparison of visual field defects using matrix perimetry and standard achromatic perimetry.

PURPOSE: To compare visual field (VF) defects found by Swedish interactive thresholding Algorithm (SITA) perimetry and Matrix perimetry, a new VF device that utilizes frequency doubling technology in a 24-2 test pattern. DESIGN: Prospective cross-sectional study. PARTICIPANTS: Fifty eyes from 50 subjects with SITA field defects were recruited for an observational study. METHODS: Swedish Interactive Threshold Algorithm and Matrix VF testing were performed on patients from a glaucoma practice. To evaluate the learning effect on the performance of the VF, we tested subsets of each group who had previous experience with standard automated perimetry (SAP). MAIN OUTCOME MEASURES: Test duration, mean threshold, mean deviation (MD), pattern standard deviation (PSD), glaucoma hemifield test, and number of abnormal points on the pattern deviation plot were evaluated for each device. RESULTS: Test duration was significantly shorter for Matrix (SITA, 357.0+/-85.6 seconds; Matrix, 319.5+/-16.5 seconds; P = 0.0002, paired t-test). Thirty-six percent of eyes with SITA VF defects showed a normal Matrix field. In 30 of 32 eyes (94%) where both devices showed VF defects, the defects were congruent. Mean threshold value was significantly lower with Matrix compared to SITA (P<0.0001, paired t-test), as was MD (-5.34+/-5.42 dB, -4.14+/-5.29 dB, respectively; P = 0.03, paired t-test). There was no significant difference in PSD between the 2 devices (P = 0.78, paired t-test). Matrix delineated significantly smaller (P = 0.005, Wilcoxon's test) and deeper (P<0.001, Wilcoxon's test) defects than those found with SITA. Similar results were observed in the subgroups with prior SAP experience. CONCLUSIONS: The Matrix examination did not detect 36% of abnormal SITA fields. Matrix field defects were smaller and deeper than those appearing in SITA perimetry.

Adult↗

PubMed2003

Cell orientation determines the alignment of cell-produced collagenous matrix.

In healing ligaments and tendons, the cells are not aligned and collagen matrix is not organized as in normal tissues. In addition, the mechanical properties of the tissues are abnormal. We hypothesized that the lack of alignment of the collagen matrix results from random orientation of the cells seen in the healing area. To test this hypothesis, a novel in vitro model was used in which the orientation of cells could be controlled via microgrooves, and alignment of the collagen matrix formed by these cells could be easily observed. It is known that cells align uniformly along the direction of microgrooves; therefore MC3T3-E1 cells, which produce large amounts of collagen, were grown on silicone membranes with parallel microgrooves (10 microm wide x 3 microm deep) in the surface. As a control, the same cells were also grown on smooth silicone membranes. Cells on both the microgrooved and smooth silicone surfaces produced a layer of readily visible collagen matrix. Immunohistochemical staining showed that the matrix consisted of abundant type I collagen. Polarized light microscopy of the collagen matrix revealed the collagen fibers to be parallel to the direction of the microgrooves, whereas the collagen matrix produced by the randomly oriented cells on the smooth membranes was disorganized. Thus, the results of this study suggest that the orientation of cells affects the organization of the collagenous matrix produced by the cells. The results also suggest that orienting cells along the longitudinal direction of healing ligaments and tendons may lead to the production of aligned collagenous matrix that more closely represents the uninjured state. This may enhance the mechanical properties of healing ligaments and tendons.

3T3 Cells↗

PubMed2006

3. Protein-protein interactions of the developing enamel matrix.

Extracellular matrix proteins control the formation of the inorganic component of hard tissues including bone, dentin, and enamel. The structural proteins expressed primarily in the enamel matrix are amelogenin, ameloblastin, enamelin, and amelotin. Other proteins, like biglycan, are also present in the enamel matrix as well as in other mineralizing and nonmineralizing tissues of mammals. In addition, the presence of sulfated enamel proteins, and "tuft" proteins has been examined and discussed in relation to enamel formation. The structural proteins of the enamel matrix must have specific protein-protein interactions to produce a matrix capable of directing the highly ordered structure of the enamel crystallites. Protein-protein interactions are also likely to occur between the secreted enamel proteins and the plasma membrane of the enamel producing cells, the ameloblasts. Such protein-protein interactions are hypothesized to influence the secretion of enamel proteins, establish short-term order of the forming matrix, and to mediate feedback signals to the transcriptional machinery of these cells. Membrane-bound proteins identified in ameloblasts, and which interact with the structural enamel proteins, include Cd63 (cluster of differentiation 63 antigen), annexin A2 (Anxa2), and lysosomal-associated glycoprotein 1 (Lamp1). These and related data help explain the molecular and cellular mechanisms responsible for the removal of the organic enamel matrix during the events of enamel mineralization, and how the enamel matrix influences its own fate through signaling initiated at the cell surface. The knowledge gained from enamel developmental studies may lead to better dental and nondental materials, or materials inspired by Nature. These data will be critical to scientists, engineers, and dentists in their pursuits to regenerate an entire tooth. For tooth regeneration to become a reality, the protein-protein interactions involving the key dental proteins must be identified and understood. The scope of this review is to discuss the current understanding of protein-protein interactions of the developing enamel matrix, and relate this knowledge to enamel biomineralization.

Amelogenin↗

PubMed2001

Endothelial cell growth factor (ECGF) enmeshed with fibrin matrix enhances proliferation of EC in vitro.

The vascular biomaterials that are currently used for clinical implants have been considered as poor substrates for human endothelial cell adhesion and spreading. Therefore, thrombotic occlusion is the predominant cause for the failure of small diameter vascular grafts made out of Dacron or Teflon. To reduce surface thrombogenicity of material surfaces used for vascular implants, in vitro seeding of endothelial cells using adhesive protein matrix is under evaluation in various laboratories. Evidences suggest that fibrin matrix is a suitable matrix for endothelial cell (EC) adhesion to the currently available vascular graft materials; however, poor proliferation of attached cells seems to be a major limitation. During this study we have also found that fibrin is a better matrix compared to gelatin to support cell attachment and spreading. However, the poor proliferation of initially attached human umbilical cord vein endothelial cell (HUVEC) necessitated modification of the matrix composition to get a monolayer within a limited period. Since fibrin can form a network of protein bundles, an effort is made to incorporate growth factors within the matrix. Endothelial cell growth factor (ECGF) isolated from bovine hypothalamus is immobilized on the surface with fibrin glue (FG) to promote proliferation of HUVEC. The results demonstrate that proteins with similar molecular weights as growth factors (GF) are retained within the matrix and released into the culture medium for 96 h, in quantities that would be sufficient to promote cell proliferation. When cells were seeded on the matrix composed with components of FG and ECGF, the HUVEC proliferated at a significantly higher rate compared to the cells on surfaces coated with gelatin or fibrin. The EC thus grown on the composite (FG + ECGF) resisted the shear stress as compared to the cells grown on gelatin. The HUVEC monolayer grown on the composite seems thromboresistant as adhesion and activation of platelets are negligible after platelet rich plasma is incubated with the monolayer for about 1 h with agitation. Therefore, the composite of fibrin and ECGF can be a suitable matrix for further evaluation of patients' autologous endothelial cell attachment and proliferation for clinical application.

Cell Adhesion↗

PubMed2003

Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS.

In recent years, high-performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection has been demonstrated to be a powerful technique for the quantitative determination of drugs and metabolites in biological fluids. However, the common and early perception that utilization of HPLC-MS/MS practically guarantees selectivity is being challenged by a number of reported examples of lack of selectivity due to ion suppression or enhancement caused by the sample matrix and interferences from metabolites. In light of these serious method liabilities, questions about how to develop and validate reliable HPLC-MS/MS methods, especially for supporting long-term human pharmacokinetic studies, are being raised. The central issue is what experiments, in addition to the validation data usually provided for the conventional bioanalytical methods, need to be conducted to confirm HPLC-MS/MS assay selectivity and reliability. The current regulatory requirements include the need for the assessment and elimination of the matrix effect in the bioanalytical methods, but the experimental procedures necessary to assess the matrix effect are not detailed. Practical, experimental approaches for studying, identifying, and eliminating the effect of matrix on the results of quantitative analyses by HPLC-MS/MS are described in this paper. Using as an example a set of validation experiments performed for one of our investigational new drug candidates, the concepts of the quantitative assessment of the "absolute" versus "relative" matrix effect are introduced. In addition, experiments for the determination of, the "true" recovery of analytes using HPLC-MS/MS are described eliminating the uncertainty about the effect of matrix on the determination of this commonly measured method parameter. Determination of the matrix effect allows the assessment of the reliability and selectivity of an existing HPLC-MS/MS method. If the results of these studies are not satisfactory, the parameters determined may provide a guide to what changes in the method need to be made to improve assay selectivity. In addition, a direct comparison of the extent of the matrix effect using two different interfaces (a heated nebulizer, HN, and ion spray, ISP) under otherwise the same sample preparation and chromatographic conditions was made. It was demonstrated that, for the investigational drug under study, the matrix effect was clearly observed when ISP interface was utilized but it was absent when the HN interface was employed.

Chromatography, High Pressure Liquid↗

PubMed1981

Hybridization of nuclear matrix attached deoxyribonucleic acid fragments.

Annealing studies were performed on DNA fragments associated with rat and mouse liver interphase nuclear matrix and the metaphase scaffold of Chinese hamster DON cells. Matrix and scaffold bound DNA fragments, reassociated with an excess of total genomic DNA, displayed kinetics virtually identical with total nuclear DNA probes. Moreover, both the extent and kinetics of these hybridizations were independent of the matrix DNA fragment size (less than 350--5000 base pairs) and the method of nuclease digestion used in their preparation (DNase I, micrococcal nuclease or endogenous digestion). The repetitive DNA component of the matrix DNA was examined by reacting discrete sizes of matrix DNA fragments (less than 350--5000 base pairs) from mouse liver with a library of cloned repetitive sequence DNA fragments which included mouse major satellite sequences. Our results demonstrate that short DNA fragments anchored to the nuclear matrix contain these cloned sequences is similar proportion of total nuclear DNA and, when viewed in light of the annealing results, indicate that matrix DNA is not enriched in either repetitive or unique sequences. Furthermore, the matrix DNA fragments appear to contain the entire sequence complexity of the genome. Finally, we hybridized both matrix and total nuclear DNA fragments with cDNA to total nuclear polyadenylated RNA. The kinetics and extent of hybridization indicate that most, if not all, of the actively transcribed DNA sequences are present in similar concentrations. We conclude that in the overall organization of eukaryotic DNA within the nucleus, the repeating domains or loops which have been demonstrated by a number of investigators are not anchored at specific attachment sequences in interphase cells or during mitosis. These findings are discussed with regard to current concepts of eukaryotic DNA loop organization.

Animals↗

PubMedDate not supplied

Tissue engineering human placenta trophoblast cells in 3-D fibrous matrix: spatial effects on cell proliferation and function.

Nonwoven polyethylene teraphathalate (PET) fabrics with different porosities and knitted fabric were used as support matrixes to grow human trophoblast cells to study the spatial effects of fibrous matrix on cell adhesion, spatial organization, proliferation, and metabolic functions. In general, cells grown on 2-D surface and knitted fabric had faster metabolic rates and also showed higher proliferation activities as detected by cyclin B assay. For nonwoven PET fibers, matrix porosity had profound effects on cell morphology, spatial organization, and proliferation. Cells grown in a low-porosity fibrous matrix formed small aggregates ( approximately 100 cells per aggregate), whereas cells grown in high-porosity matrix formed big aggregates ( approximately 1000 cells per aggregate). This was attributed to the difference in pore volume or averaged fiber distance, which dictated a cell's ability to cross over and form a bridge between adjacent fibers. The high-porosity matrix had a relatively poor surface accessibility for cells to attach and spread, which are essential for cell proliferation. Dual staining with PI and BrdU showed that 60% of cells in the small aggregates found in the low-porosity matrix were proliferating, while only 18% of cells in the large aggregates found in the high-porosity matrix were proliferating. These results suggest that spatial characteristics of fibrous matrix are important to cell proliferation and function and should be considered in tissue-engineering human cells.

Biocompatible Materials↗

PubMed1977

Identification of four extracellular-matrix enamel proteins during embryonic-rabbit tooth-organ development.

1. Investigations were designed to identify the proteins which characterize the ameloblast phenotype, and to determine to what extent these extracellular-matrix proteins were degraded as a function of enamel matrix mineralization and maturation. 2. The identification of enamel proteins was based on comparisons between the electrophoretic patterns of enamel-containing and non-enamel-containing matrix extracts isolated from specific regions within 26-day embryonic New Zealand White rabbit incisor and molar tooth organs. 3. Since enamel proteins become mineralized on secretion, matrix specimens were demineralized in cold 5% (w/v) trichloroacetic acid, extracted with buffered 6M-urea and reduced with mercaptoethanol, and then the solubilized proteins were fractionated by urea/polyacrylamide-gel electrophoresis. 4. Three enamel-specific electrophoretic components were identified in newly secreted enamel-matrix specimens and this number increased as a function of mineralization and maturation. 5. Antibodies were prepared against embryonic rabbit extracellular matrix containing enamel. Comparison between immunoelectrophoretic patterns demonstrated that two of the three enamel components were antigenic. 6. Polyacrylamide-gel electrophoresis in sodium dodecyl sulphate was used to identify four enamel proteins of mol.wts. (1) 65 000 (2) 58000 (3) 22 000 and (4) 20 000, localized within enamel matrix. Enamel proteins (1) and (3) were phosphorylated, whereas (2) and (4) did not contain detectable phosphate. Labelled proline, leucine, tryptophan and glucosamine were incorporated into each of the four enamel proteins extracted from tooth explants incubated in the presence of radioactive precursors for 6 h. Whereas four proteins were identified in newly secreted enamel matrix, the concentrations of high-molecular-weight proteins (1) and (2) were found to decrease and the number (greater than 10) and concentration of low-molecular-weight polypeptides increased as a function of advanced enamel-matrix mineralization and maturation.

Animals↗

PubMed1993

DNA precursors are channelled to nuclear matrix DNA replication sites.

Studies of replicative DNA synthesis using DNA precursors have shown that the DNA that was replicated most recently is that associated with the nuclear matrix. Consequently, precursors arising via the salvage and the de novo metabolic pathways are first incorporated into a small percentage of the total nuclear DNA that is termed nuclear matrix-associated DNA. These results have been substantiated in cell culture, as well as in intact mammalian systems. Furthermore, when DNA precursors were injected intravenously into regenerating rat liver, a significant lag in the incorporation of orotic acid-derived nucleotides (de novo pathway precursors) into nuclear DNA was observed, when compared with deoxythymidine-derived nucleotides (salvage pathway precursors). This lag in incorporation kinetics was also evident at the nuclear matrix level, although, once incorporated into nuclear matrix-associated DNA, the distribution patterns of both precursors into extra-matrix nuclear DNA fractions were identical. To determine the basis for this kinetic lag, we compared the incorporation kinetics of orotic acid and of deoxythymidine into dTTP and into nuclear matrix-associated DNA, respectively. Orotic acid-derived nucleotides entered the cytosolic dTTP pool before being incorporated into nuclear matrix-associated DNA, that is, traversing the classical metabolic route of DNA precursors. Conversely, deoxythymidine-derived nucleotides by-passed the soluble dTTP cellular pool and engaged directly in DNA synthesis at the nuclear matrix. Not only is this the first evidence for nucleotide channelling in an intact mammalian system, but it also forms direct evidence that salvage pathway DNA precursors are channelled to nuclear matrix-associated sites of DNA replication.

Animals↗

PubMed2002

Evaluation of healing with use of an internal matrix to repair furcation perforations.

AIM: The purpose of this study was to evaluate healing responses following repair of furcation perforations, with and without an internal matrix. Two matrix materials, HAPSET (65% non-resorbable hydroxyapatite and 35% plaster of paris) and hydroxyapatite were compared. METHODOLOGY: Four adult female baboons (Papio anubis) served as experimental models. Furcation perforations were made in the molar and premolar teeth, which were then randomly assigned to one of the five groups, according to the method of perforation repair: 1 Experimental group 1 (16 teeth): The matrix material was HAPSET and the sealing material, amalgam. 2 Experimental group 2 (16 teeth): The matrix material was hydroxyapatite and the sealing material, amalgam. 3 Experimental group 3 (16 teeth): No matrix was placed. The sealing material was amalgam. 4 Positive control group (16 teeth): The perforation was not sealed. 5 Negative control group (16 teeth): No perforation was made. The animals were sacrificed at 1 week and 1, 3 and 7 months. Specimens were prepared for examination with light microscopy. RESULTS: The data revealed that when amalgam was used alone without a matrix, there was marked extrusion of the material into the underlying bone with an associated severe inflammatory response, which continued throughout the observational period. When an internal matrix was used, there was an initial acute inflammatory response that diminished with time such that at 7 months, 75% of these specimens were free of inflammation. There was no difference in the tissue response to the different matrix materials. HAPSET and hydroxyapatite underwent connective tissue encapsulation in the early stages followed by new bone deposition in direct contact with the materials. CONCLUSIONS: Within this animal model healing responses are better when an internal matrix, whether HAPSET or hydroxyapatite, is used in the repair of furcation perforations.

Alveolar Process↗

PubMed2001

Bacitracin inhibits fibronectin matrix assembly by mesangial cells in high glucose.

BACKGROUND: Accumulation of mesangial extracellular matrix is a major characteristic of diabetic nephropathy (DN). Expression of several extracellular matrix proteins is up-regulated in human mesangial cells (HMC) cultured in high glucose. One protein, fibronectin (FN), associates to form an insoluble disulfide-linked matrix and possesses inherent protein-disulfide isomerase (PDI) activity. Bacitracin is a known PDI inhibitor. We tested the hypothesis that inhibiting FN-PDI activity with bacitracin would disrupt excessive FN-matrix assembly by cultured HMCs grown under high glucose conditions. METHODS: The effect of bacitracin on FN-PDI activity was tested using an RNase-refolding assay. High glucose cultures of HMC were labeled with (3)H-leucine, with and without bacitracin, and (3)H-FN immunoprecipitated from the medium and sequential extracts of cell layers to distinguish insoluble FN. FN transcription was assessed by reverse transcription-polymerase chain reaction (RT-PCR). Pericellular FN-matrix was examined by immunohistology. RESULTS: Bacitracin inhibited the PDI activity of FN, with maximal inhibition at 1.0 mmol/L. Treatment of HMC cultures grown in high glucose with bacitracin brought about changes in the distribution of newly synthesized FN. With increasing concentrations of bacitracin there was a significant reduction in the level of FN present as an insoluble matrix of HMC cultures maintained in high glucose, and a corresponding increase in FN in medium. Decreases in FN matrix laid down by HMCs treated with different concentrations of bacitracin were seen by immunohistology. FN mRNA levels were unchanged. CONCLUSION: PDI inhibition of FN reduces its association into an insoluble matrix and potentially provides a new approach to reduce excessive matrix deposition in DN.

Anti-Bacterial Agents↗

PubMed1996

Characterization of the proteins comprising the integral matrix of Strongylocentrotus purpuratus embryonic spicules.

In the present study, we enumerate and characterize the proteins that comprise the integral spicule matrix of the Strongylocentrotus purpuratus embryo. Two-dimensional gel electrophoresis of [35S]methionine radiolabeled spicule matrix proteins reveals that there are 12 strongly radiolabeled spicule matrix proteins and approximately three dozen less strongly radiolabeled spicule matrix proteins. The majority of the proteins have acidic isoelectric points; however, there are several spicule matrix proteins that have more alkaline isoelectric points. Western blotting analysis indicates that SM50 is the spicule matrix protein with the most alkaline isoelectric point. In addition, two distinct SM30 proteins are identified in embryonic spicules, and they have apparent molecular masses of approximately 43 and 46 kDa. Comparisons between embryonic spicule matrix proteins and adult spine integral matrix proteins suggest that the embryonic 43-kDa SM30 protein is an embryonic isoform of SM30. An adult 49-kDa spine matrix protein is also identified as a possible adult isoform of SM30. Analysis of the SM30 amino acid sequences indicates that a portion of SM30 proteins is very similar to the carbohydrate recognition domain of C-type lectin proteins.

Animals↗

PubMed1991

Contact matrices for multipopulation epidemic models: how to build a consistent matrix close to data.

In models of the dynamics of sexually transmitted diseases between and within N interacting populations, it is necessary to specify the matrix of contacts between populations. Mixing matrices have to satisfy a consistency condition that generally will not be satisfied by empirically obtained matrices. The problem of inferring a mixing matrix from data is phrased here as the problem of finding a matrix in a prescribed set that minimizes an opportune distance from the matrix of data. Two different distances that attempt to measure relative errors are suggested. When no constraints are posed on the activity rates of the populations, the author shows that the minimum distance from data is attained at Knox's (1986) matrix. When activity rates are to be preserved, the minimum cannot be found explicitly for N greater than 2. An algorithm proposed by Arcà, Perucci, Spadea, and Rossi (1990) is then investigated, and shown always to converge to a consistent matrix. Through several examples, it is shown that this limiting matrix does not minimize distance from data, but is generally close to the minimum. Finally, the author simulated the collection of data with sampling errors and possible bias and evaluated the performance of this algorithm in approximating the 'true' contact matrix starting from the simulated 'data' matrix.

Algorithms↗

PubMed1986

Characterization of nuclear matrix from cultured normal human fibroblasts.

Nuclear matrix was isolated from cultured human fibroblasts by extraction of nuclei with 2 M NaCl. Electron microscopic observation on the isolated nuclear matrix revealed a fine network structure. The matrix fraction contained approximately 15% of total nuclear DNA and the matrix DNA was about 3- to 4-fold enriched in transcriptionally active collagen I (alpha 2) gene sequences, whereas transcriptionally inactive beta-globin gene sequences were not enriched. The nuclear matrix contained two major proteins of 65,000 and 45,000 daltons (pI 5.9 and 5.6, respectively). The DNA-binding activity of these nuclear matrix proteins was examined by Western blotting or by nitrocellulose filter-binding assay using cloned specific gene probes. The results suggest that there is no base sequence specificity in the binding, and that protein species of 60,000 to 200,000 daltons showed DNA-binding activity. These results indicate that association of transcribing genes with the nuclear matrix may reflect the functional state of the genes and may not be determined solely by the base sequence specificity of DNA binding. The nuclear matrix protein of 65,000 daltons was phosphorylated in vivo, and was the main substrate for protein kinase(s) associated with the nuclear matrix.

Cell Nucleus↗