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A novel aldehyde dextran sulfonate matrix for affinity biosensors.

Aldehyde dextran sulfonate (ADS), a modified oligosaccharide polymer, was used to prepare a new matrix structure for affinity biosensors. The principal difference between the ADS matrix and similar structures developed previously results from presence of two active functional groups in the matrix, namely, aldehyde and sulfonate. These groups perform two different functions in the matrix. The aldehyde group is responsible for covalent bonding in the biomaterials, and the negatively charged sulfonate group provides electrostatic attraction of the positively charged biomolecules. By varying the ratio between the aldehyde and sulfonate groups in the matrix, one can control contributions from the two binding modes (covalent and electrostatic). A number of oligosaccharides, such as simple dextran, aldehyde dextran (AD), aldehyde dextran sulfonate (ADS) and aldehyde ethylcellulose (AEC), were used for preparation of matrix structures. The properties of the obtained matrices were analysed and compared. Surface plasmon resonance (SPR) was used as the main technique to characterize the matrix structures.

Aldehydes↗

Transscleral permeability and intraocular concentrations of cisplatin from a collagen matrix.

This study determined the in vitro permeability of cisplatin through isolated human sclera as delivered by a collagen matrix vehicle. Short-term and long-term intraocular levels of cisplatin were also measured in the rabbit eye after a subconjunctival injection. Cisplatin in either a collagen matrix vehicle or a control balanced salt solution (BSS) vehicle was applied to human sclera mounted in a specially designed in vitro perfusion chamber. The amount of cisplatin that diffused across the sclera was measured in hourly samples for 24 h using atomic absorption spectrometry. In vivo studies were also performed in Dutch Belted rabbits given subconjunctival injections of cisplatin in collagen matrix or in BSS. Eyes were enucleated at 1.5 h and 2 weeks after injection, frozen, and dissected to determine the intraocular cisplatin concentrations. Cisplatin had a peak in vitro scleral permeability constant of 8.3+/-1.2 x 10(-6) and 20.1+/-1.8 x 10(-6) cm/s, delivered in collagen matrix and in BSS, respectively (mean+/-S.D.). At the end of the in vitro experiments, 35.9+/-4.6% of the cisplatin remained in the collagen matrix, while 0.8+/-0.2% remained in the BSS vehicle. Subconjunctival injection of cisplatin in the collagen matrix vehicle achieved 3.3+/-0.1 microg/ml in the vitreous humor at 1.5 h and 0.1+/-0.1 microg/ml at 2 weeks. This vehicle also achieved a cisplatin concentration of 73.5+/-23.9 microg/mg in the choroid and retina at 1.5 h and 3.2+/-1.3 microg/mg at 2 weeks. Compared to BSS, the collagen matrix vehicle provided a more controlled release of cisplatin, and after subconjunctival injection into rabbits, attained higher drug levels in several ocular tissues.

Animals↗

Expression patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-alpha and TGF-beta1.

BACKGROUND/AIMS: Although matrix metalloproteinases (MMPs) and their specific inhibitors (TIMPs) play an essential role in liver injury associated with tissue remodeling, the cellular origin of MMPs/TMPs within the liver remains to be clarified. METHODS: Different liver cell populations were analysed with respect to their expression by reverse transcription-polymerase chain reaction, Northern blot analysis and zymography. RESULTS: MMP and TIMP coding transcripts were detectable in all liver cell types by reverse transcription-polymerase chain reaction; however, the cellular expression levels were markedly different as assessed by Northern blot analysis. Gelatinase-B was predominantly expressed in Kupffer cells, gelatinase-A in hepatic stellate cells and rat liver myofibroblasts and stromelysins-1, -2 as well as collagenase in hepatic stellate cells. Membrane type-1 MMP (MMP-14) was found in significant amounts in all liver cells. TIMP-1 coding m-RNAs were present mainly in hepatic stellate cells and rat liver myofibroblasts, TIMP-2 additionally in Kupffer cells, while TIMP-3 expression was detectable only in hepatocytes. During in vitro activation of hepatic stellate cells, MMP expression was mostly downregulated, while TIMP expression was enhanced, thereby providing an explanation for matrix accumulation co-localised with these cells during chronic liver injury. In general, TNF-alpha stimulated both MMP and TIMP expression of hepatic stellate cells, while TGF-beta1 induced TIMP expression only. CONCLUSIONS: Collectively these data demonstrate that all resident liver cells are involved in matrix degradation to some extent and that hepatic stellate cells play an important role in matrix breakdown in addition to matrix synthesis. The cytokine-specific regulation of MMP/TIMP expression in hepatic stellate cells suggests that the initial matrix breakdown following liver injury might be enhanced by TNF-alpha, while diminished matrix degradation during chronic tissue injury might be due to the action of TGF-beta1 through TIMP induction.

Animals↗

Characterization of cultured nail matrix cells.

BACKGROUND: Cultures of epidermal cells are commonly used to study skin biology and differentiation. Recently a method to culture nail matrix cells has been established. OBJECTIVE: We report the biologic characteristics of nail matrix cells in vitro compared with those of epidermal keratinocytes. METHODS: Human nail matrix cells were isolated and cultured in defined medium. Electron-microscopic examination, growth rate, integrin expression and keratin synthesis pattern were evaluated. In addition, the cells were cultured in serum-containing medium. RESULTS: Nail matrix cells appear to be larger than human epidermal keratinocytes and, at the ultrastructural level, they contain a higher euchromatin/heterochromatin ratio and a lower nucleus/cytoplasm ratio and have a higher growth rate. The synthesis of "hard" keratins was detected at all calcium concentrations. Immunofluorescence analyses showed the expression of alpha 2, alpha 3, and alpha 6 integrin subunits. When cultured in serum-containing medium, nail matrix cells produced an outgrowth of epithelium and a spontaneous migration phenomenon associated with a tendency to stratify in a semilunar area that resembles the architecture of the nail matrix. The pluristratified epithelium showed characteristic markers of nail differentiation. CONCLUSION: Culture of nail matrix cells may represent a useful model to study the biologic properties of nail structure, alterations in some nail diseases and the effects of drugs.

Cell Division↗

Nail matrix nevi: a clinical and histopathologic study of twenty-two patients.

BACKGROUND: Because most dermatologists do not regularly perform biopsies of longitudinal melanonychia, even when the pigmentation presents as a single band, the true prevalence of nail matrix nevi is unknown. OBJECTIVE: Our purpose was to determine the prevalence of nail matrix nevi in white patients with longitudinal melanonychia involving a single digit and to determine whether longitudinal melanonychia caused by a nail matrix nevus can be clinically distinguished from longitudinal melanonychia from other causes. METHODS: From January 1989 to December 1994 we performed a nail biopsy on 100 of 128 consecutive white patients who had a single band of "idiopathic" longitudinal melanonychia. RESULTS: A nail matrix nevus was detected in 22 patients. A junctional nevus was found in 19 specimens and a compound nevus in three specimens. CONCLUSION: Nail matrix nevi in Caucasian patients are uncommon but not exceptional. The number of nevi presenting with longitudinal melanonychia exceeded that of melanoma. The diagnosis of nail matrix nevi is impossible clinically and always requires histopathologic study. The pathologic features of nail matrix nevi are similar to those of skin nevi except for their architectural pattern, which reflects the peculiar anatomy of the nail unit.

Adolescent↗

Efficacy and tolerability of a new estradiol delivering matrix patch (Estraderm MX) in postmenopausal women.

OBJECTIVE: To examine the efficacy and tolerability of a new matrix patch delivering estradiol (E2 Matrix) at doses of 0.05 and 0.10 mg per day (Estraderm MX 50, 100) in the treatment of moderate to severe postmenopausal symptoms. METHODS: A total of 254 postmenopausal women were randomized to receive treatment with E2 Matrix 0.10 mg (N = 86), E2 Matrix 0.05 mg (N = 82), or placebo (N = 86) in a double-blind, double-dummy fashion for a period of 12 weeks continuously. Patches were applied twice weekly to the buttocks with each patient wearing two patches at all times. The primary efficacy criterion was the difference from baseline of the mean number of moderate to severe hot flushes per 24 h during the last 2 weeks of treatment. Other efficacy variables included reduction in hot flushes at 4 and 8 weeks, reduction in daytime flushing and night sweats, and Kupperman Index at 4, 8, and 12 weeks. RESULTS: E2 Matrix 0.10 and 0.05 mg were both significantly superior to placebo in reducing hot flushes per 24 h after 4, 8, and 12 weeks of treatment (P < 0.001). Also, for all other efficacy parameters studied, both dosage strengths of E2 Matrix were statistically significantly superior to placebo at all time points (P < 0.001). Local tolerability was good in both groups. A slight increase in estrogen related adverse effects (breast tenderness, leukorrhoea) was seen with the 0.10 mg patch. Adhesion of patches and compliance were good. Overall systemic tolerability was good in both treated groups. However, a 4.8% overall incidence of endometrial hyperplasia was observed in patients with an intact uterus. CONCLUSIONS: This new matrix patch offers an effective and well tolerated dosage form for delivery of 0.05 and 0.1 mg estradiol per day. It may be particularly suitable for those women who experience local sensitivity to alcohol-containing systems. In light of the observed hyperplasia after treatment in five patients, estrogen therapy should as yet be supplemented monthly with a progestogen in women with an intact uterus.

Administration, Cutaneous↗

Regulation of glutathione S-transferase enzymes in primary cultures of rat hepatocytes maintained under various matrix configurations.

Primary rat hepatocytes were cultured under various matrix and media conditions and examined after 1 week for the expression and regulation of cytosolic glutathione S-transferase (GST) enzymes. Striking effects on cell morphology were observed in relation to the different matrix conditions, whereas media effects were less prominent. Hepatocytes cultured in serum-free Dulbecco's modified Eagle's medium (DMEM) or modified Chee's medium (MCM) maintained similar levels of total GST protein regardless of the matrix configuration or corresponding cell integrity. However, HPLC analysis showed a differential expression pattern of individual GST subunits in both a time- and medium-dependent fashion. A variable, but pronounced, matrix and medium effect was observed on the induction of total GST expression by various prototypical inducers. Dexamethasone (10 microM) induced subunits A2, M1 and M2 in a medium- and matrix-dependent fashion, whereas phenobarbital (100 microM) induced significantly only subunit A2. beta-Naphthoflavone (50 microM) suppressed all GST subunit expression except subunit P1, which was induced in a matrix- and medium-dependent fashion. These studies show that total basal level expression of GSTs in vitro is reflective of a concomitant increase in mu and pi class subunits and a decrease in alpha class subunits. Moreover, the matrix and medium conditions influence both the basal and inducible expression of GST subunits in cultured rat hepatocytes.

Animals↗

Characterization and quantitation of wound matrix in the fetal rabbit.

Fetal response to injury has been characterized by the deposition of a matrix that is not primarily collagen. This study was designed to identify this matrix, in order to better understand the fetal mechanism of tissue repair. Silastic/polyvinyl alcohol sponge (PVA) wound implants were placed paravertebrally in 24-day gestation (31 days = term) fetal (n = 65) and adult (n = 43) rabbits and then harvested from one hour to 6 days post-wounding. Histologic analysis of the fetal wound matrix deposited in the PVA implants suggested the presence of glycosaminoglycans (GAG) rather than the collagen found in adult wound implants. To further analyze the fetal wound matrix, the GAG content was quantitated using an Alcian Blue dye-binding assay. Results showed significantly increased (p less than 0.05) GAG deposition on days 2-6 in the fetal wound compared to the adult wound. Fetal GAG levels were approximately three times those of the adult during this period. Separation of individual GAG species by cellulose acetate electrophoresis demonstrated that the GAG matrix of the fetal wound was composed predominantly of hyaluronic acid. This finding was confirmed by selective enzymatic digestion of separated GAG species using highly specific polysaccharidases. These observations of hyaluronic acid deposition in the fetal wound may be ascribed an important physiologic role by providing a more fluid and malleable matrix rather than a restrictive matrix composed of collagen. This new evidence coupled with earlier findings of the lack of an acute inflammatory response in the fetus further supports the hypothesis that the fetal response to injury is significantly different from the adult response.

Alcian Blue↗

Peritrophic matrix proteins.

The peritrophic matrix (or peritrophic membrane) lines the gut of most insects at one or more stages of the life cycle. It has important roles in the facilitation of the digestive processes in the gut and the protection of the insect from invasion by microorganisms and parasites. The traditional view of the peritrophic matrix as a relatively insert sieve, composed largely of proteins and glycosaminoglycans embedded in a chitinous matrix, is under revision as more is learned about the molecular characteristics of the peritrophic matrix proteins. This review summarizes emerging knowledge of the main protein constituents of the peritrophic matrix. The availability of the first sequences of integral peritrophic matrix proteins has coincided with the explosion of information in sequence databases. It is therefore possible to examine common structural themes in this family of proteins as well as in proteins of unknown location and function from a variety of other insects, nematodes and viruses. The review concludes with speculation about the biological functions of the proteins in this matrix.

Animals↗

The intrinsic peritrophic matrix protein peritrophin-95 from larvae of Lucilia cuprina is synthesised in the cardia and regurgitated or excreted as a highly immunogenic protein.

The intrinsic peritrophic matrix glycoprotein, peritrophin-95, from the midgut of larvae of Lucilia cuprina can only be solubilized from the matrix using strong denaturants. This suggests that the protein has a structural role in the matrix. Consistent with this is the finding that immuno-gold and immuno-fluorescence localizations of the protein showed a uniform distribution within the peritrophic matrix. RT-PCR demonstrated that expression of peritrophin-95 mRNA was restricted to the larval cardia, a small organ located in the anterior midgut from which the type 2 peritrophic matrix originates. Immuno-blots and ELISAs demonstrated that the sera from sheep infested naturally or artificially with these larvae recognised peritrophin-95. This indicates that peritrophin-95 stimulates the ovine immune system during larval infestation even though the protein is firmly attached to the peritrophic matrix in the larval midgut and seemingly "concealed" from the ovine immune surveillance system. Analyses of larval regurgitated or excreted material by immuno-blots, immuno-affinity purification and amino-terminal sequencing demonstrated the presence of soluble monomeric peritrophin-95. These results indicate that peritrophin-95, a candidate vaccine antigen for use in sheep is not a "concealed" antigen as previously thought. The presence of soluble peritrophin-95 in the regurgitated/excreted material from larvae suggests that this protein may be involved in a maturation phase of peritrophic matrix production, a by-product of which is the excretion or regurgitation of soluble peritrophin-95.

Amino Acid Sequence↗

Reduction in matrix-related signal suppression effects in electrospray ionization mass spectrometry using on-line two-dimensional liquid chromatography.

The effect of liquid chromatographic separation on matrix-related signal suppression in electrospray ionization mass spectrometry (LC-ESI-MS) was investigated. A method incorporating on-line two-dimensional liquid chromatography mass spectrometry (LC/LC-MS) was developed to compensate for matrix effects and signal suppression in qualitative and quantitative analysis. The LC/LC-MS(MS) approach was successfully applied for single-component and multicomponent analysis in a variety of complex matrixes. It was demonstrated that matrix-related signal suppression could be induced solely by (i) column overload, (ii) matrix component-analyte coelution, or a combination of each. Application of on-line orthogonal LC/LC separations can be effective in reducing both causes of matrix-related signal suppression effects i.e., column overload and matrix-analyte coelution for a variety of LCn/MSn applications.

Animals↗

Internal energy of ions generated by matrix-assisted laser desorption/ionization.

To provide an objective measure of the correlation between the internal energy content of ions generated by matrix-assisted laser desorption/ionization (MALDI) and the matrix properties, a series of well-characterized benzyl-substituted benzylpyridinium salts were used as thermometer molecules (TMs). To determine the internal energy variations of analyte ions, the survival yields of TM molecular ions were measured in three different matrixes, alpha-cyano-4-hydroxycinnamic acid (CHCA), 3,5-dimethoxy-4-hydroxycinnamic acid (sinapinic acid, SA), and 2,5-dihydroxybenzoic acid (DHB). Statistical analysis of extensive survival yield data indicated that there were discernible differences among the studied matrixes. The experimental survival yields of the TM ions were used to calculate the unimolecular decomposition rate coefficient. Corresponding theoretical reaction rate coefficients were calculated based on the Rice-Ramsperger-Kassel-Marcus (RRKM) theory for different internal energies of the TMs. The internal energies of the ions were obtained by projecting the experimental rate coefficient values onto the theoretical curves obtained by the RRKM calculations. Molecular ions of the analytes showed decreasing survival yields and consequently increasing internal energies in the three matrixes in the following order: CHCA, SA, and DHB with "cold", "intermediate", and "hot" characteristics, respectively. Qualitatively, this could be interpreted as a significant departure from earlier observations suggesting an opposite trend. The classification as hot and cold matrixes should be further qualified by accounting for the influence of laser pulse energy and the nature of the analyte. Higher laser pulse energy led to an elevated level of energy transferred to the analyte, which in turn resulted in a diminished survival yield of the analyte molecular ion. It is quite possible that the assignment of hot and cold reverses as the analyte or the laser energy changes. These findings can help predict the outcome of postsource decay experiments and clarify the concept of hot and cold matrixes in MALDI mass spectrometry.

Journal Article↗

Nuclear matrix-bound deoxyribonucleic acid synthesis: an in vitro system.

A nuclear matrix-bound in vitro replicational system that utilizes alpha-polymerase and matrix-attached DNA has been isolated from regenerating rat liver and characterized for in vitro requirements. beta-Polymerase is a minor component of the matrix in vitro system. Evidence is presented for the native association of alpha-polymerase with the nuclear matrix and for the parallel induction of matrix-bound alpha-polymerase and in vivo DNA synthesis during the proliferative response following partial hepatectomy. Our data suggest that the attachment of alpha-polymerase to the matrix may be an important step in the assembly of functional replisomes. We also demonstrate prereplicative and late-replicative rises in nuclear and matrix-bound beta-polymerase activity and discuss these results in terms of possible genome screening.

Animals↗

Properties of adenoviral DNA bound to the nuclear matrix.

The association of adenoviral DNA with the high salt (2 M NaCl) resistant nuclear fraction, termed the nuclear matrix, has been investigated in HeLa cells at different times after infection with adenovirus type 5. When nuclear matrices were prepared in the absence of exogenously added nucleases, Ad5 DNA was quantitively associated with the matrix throughout the infection period (0-24 h). Moreover, early in infection (0-10 h) Ad5 DNA was severalfold enriched in DNase I digested nuclear matrices (8-15% of total nuclear DNA) compared to the high salt soluble chromatin fraction (85-90% of total nuclear DNA). At later times after infection, progressively more Ad5 DNA appeared in the chromatin fraction until, at 24 h, the nuclear matrix was strikingly depleted in Ad5 DNA. A large proportion of the Ad5 DNA in nuclear matrices prepared early in infection, e.g., 4 h, was full length in size. At later times (12-24 h) most of the viral DNA was fragmented to a size equivalent to total matrix DNA (100-1000 base pairs). The apparent switch of the matrix-associated viral DNA from a relatively DNase I resistant to sensitive state was initiated approximately at the time when viral DNA replication began (12 h). Since no discrete portion of the Ad5 genome was significantly enriched at the sites of attachment to the nuclear matrix throughout the infection period, the switch in DNase I sensitivity is not mediated by a change in DNA sequence attachment to the matrix.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenoviruses, Human↗

Association of globin ribonucleic acid and its precursors with the chicken erythroblast nuclear matrix.

Nuclear matrix was prepared from both erythroblasts and erythrocytes of chicken red blood cells. Greater than 90% of the globin nuclear RNA remains bound to the erythroblast nuclear matrix. There are approximately 1000 copies of globin RNA in the nucleus per cell, and most of these contain a poly(A) tail. Precursor beta globin RNA exists in four high molecular weight forms, some of which are larger than the natural beta globin gene. Most of the ribosomal RNA is lost during the preparation of an erythroblast nuclear matrix. In contrast, some of the snRNAs are specifically enriched in the erythroblast nuclear matrix. There is little or no globin nuclear RNA in the erythrocyte nuclear matrix. There appears to be no selective attachment of the globin genes to the erythroblast nuclear matrix. The nuclear matrix is postulated to be a platform for the differential processing of nuclear RNA.

Animals↗

Soy matrix drug delivery systems obtained by melt-processing techniques.

The aim of this study was to develop new soy protein drug delivery matrix systems by melt-processing techniques, namely, extrusion and injection moulding. The soy matrix systems with an encapsulated drug (theophylline, TH) were previously compounded by extrusion performed at two different pH values, (i) pH 4 (SIpDtp) and (ii) pH 7 (SIDtp), and further injection-moulded into a desired shape. During the extrusion process the matrixes SIDtp were also cross-linked with glyoxal (0.6X-SIDtp) and reinforced with a bioactive filler, hydroxylapatite (SI-HADtp). The obtained mouldings were used to study the drug-release mechanisms from the plastic soy-TH matrixes. In an isotonic saline solution (ISS) buffered at pH 5.0 (200 mM acetate buffer), the resulting release kinetics could be described using the Fick's second law of diffusion. Because the diffusion coefficients were found to be constant and the boundary conditions to be stationary, these systems are drug-diffusion controlled. Conversely, the dominant phenomena in an isotonic saline solution buffered at pH 7.4 (200 mM Tris/HCl buffer) are more complex. In fact, because of the higher polymer solubility, the resulting matrix is time-variant. So, the drug release is affected by swelling, drug diffusion, and polymer dissolution, being faster when compared to ISS-200 mM acetate buffer, pH 5.0. The changes in the formulation composition affecting the correspondent release rates were also investigated. At pH 7.4, increasing the cross-linking degree of the polymer matrix (via reaction with glyoxal or heat treatment) or decreasing the net charge (extruding at pH near its isoelectric point) led to lower release rates. The incorporation of ceramic filler caused the opposite effect. Because of the low solubility of the matrix at pH 5.0, no significant variations were detected with variations in the selected formulations. These systems, based on a nonstandard protein-based material, seem to be very promising to be used as carriers for drug delivery.

Biocompatible Materials↗

Controlled release of a herbicide from matrix granules based on solvent-fractionated organosolv lignins.

An organosolv (Alcell) lignin was fractionated with both sequential solvent-extraction and solvent-mixture precipitation using various organic solvent systems. The resulting fractionated lignins were used to prepare matrix granules with bromacil as a model compound using a melt process. The release rates of bromacil in vitro from the granular matrix system were influenced by both the lignin used as carrier and the solution fractionation. The variations in release rates were related to the high proportion of low-molecular-weight fraction and its associated water-soluble lignin in the fractionated lignins. Fickian diffusion was a predominant factor in controlling the release of bromacil from the matrix granules based on the low-molecular-weight fraction (L(fr1)). For the matrix granules based on the other fractionated lignins, the release of bromacil was of super-case II transport. The fine structure of the matrix granules before and after release was also studied. An unique release pattern of bromacil was observed for the matrix granules based on the low-molecular-weight fraction (L(fr1)), showing a fast release followed by zero-order release, which was a result of remarkable changes in the structure of the matrix granules during the course of release.

Herbicides↗

The complexes between CH3OH and CF4. Infrared matrix isolation and theoretical studies.

The complex formed between methanol and tetrafluoromethane has been identified in argon and neon matrixes by help of FTIR spectroscopy. Three fundamentals (nu(OH), nu(FCF), and nu(CO)) were observed for the complex isolated in the two matrixes, and the OH stretch was red shifted in a neon matrix and blue shifted in an argon matrix with respect to the corresponding vibration of the methanol monomer. The theoretical studies of the structure and spectral characteristics of the complexes formed between CH(3)OH and CF(4) were carried out at the MP2 level of theory with the 6-311+G(2df,2pd) basis set. The calculations resulted in three stationary points from which two (I-1, I-2) corresponded to structures involving the O-H...F hydrogen bond and the third one (I-3) to the non-hydrogen-bonded structure. The topological analysis of the distribution of the charge density (AIM theory) confirmed the existence of the hydrogen bond in I-1, I-2 complexes and indicated weak interaction between the oxygen atom of CH(3)OH and three fluorine atoms of CF(4) in the I-3 complex. The comparison of the experimental and theoretical data suggests that in the matrixes only the non-hydrogen-bonded complex I-3 is trapped. The blue/red shift of the complex OH stretching vibration with respect to the corresponding vibration of CH(3)OH in argon/neon matrixes is explained by the different sensitivity of the complex and monomer vibrations to matrix material. The ab initio calculations performed for the ternary CH(3)OH-CF(4)-Ar systems indicated a negligible effect of an argon atom on the binary complex frequencies.

Journal Article↗