Biochemical and immunocytochemical characterization of mineral binding proteoglycans in rat bone.

We examined biochemically and immunocytochemically the type and distribution of mineral binding proteoglycans (PGs) in rat mid-shaft subperiosteal bone using three monoclonal antibodies (MAb 1-B-5, 9-A-2, and 3-B-3) which specifically recognize unsulfated chondroitin, chondroitin 4-sulfate (C4-S) and dermatan sulfate (DS), and chondroitin 6-sulfate. Bone proteins were extracted from fresh specimens with a three-step technique: 4 M guanidine HCl (GdnCl), aqueous EDTA without GdnCl (E-extract), followed by GdnCl. Western blot analysis of SDS-polyacrylamide gel electrophoresis revealed that E-extract after chondroitinase ABC digestion reacted strongly with MAb 9-A-2 but not with MAb 1-B-5 or 3-B-3. After adehyde fixation, ethanolic trimethylammonium EDTA was used as a demineralizing agent for light and electron immunocytochemistry. This provided good retention of water-soluble PGs in the specimens. After chondroitinase ABC pre-treatment of tissue sections, MAb 9-A-2 specifically stained C4-S and/or DS in the walls of osteocyte lacunae and bone canaliculi in the mineralized matrix as well as in the unmineralized matrix such as pre-bone, vascular canals, and pericellular matrix surrounding osteocytes; the remainder of the mineralized matrix lacked staining. These results indicate that mineral binding PGs contain C4-S and/or DS and are exclusively localized in the walls of the bone lacuna and canaliculus.

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Monoclonal antibodies to type V collagen for immunohistological examination of new tissue deposition associated with biomaterial implants.

Journal of Histochemistry & Cytochemistry ◽

10.1177/39.9.1918939 ◽

1991 ◽

Vol 39 (9) ◽

pp. 1215-1220 ◽

Cited By ~ 18

Author(s):

J A Werkmeister ◽

J A Ramshaw

Keyword(s):

Monoclonal Antibodies ◽

Polyacrylamide Gel Electrophoresis ◽

Experimental Models ◽

Type V Collagen ◽

Tissue Sections ◽

Novel Approach ◽

Wide Range ◽

Conventional Elisa ◽

Pre Treatment ◽

Type V

We developed a panel of highly specific monoclonal antibodies (MAb) against dog Type V collagen. Each antibody showed differential reactivities towards Type V collagen from other species. All the antibodies were highly reactive in conventional ELISA, as well as with electroblots of collagen after polyacrylamide gel electrophoresis using non-denaturing conditions. The MAb were shown to be suitable for the immunohistological detection of Type V collagen in tissue sections, although this normally required pre-treatment of sections with 50 mM acetic acid. In particular, the antibodies were shown to be useful for examining samples of a collagen-based biomaterial, a vascular prosthesis, after explant from evaluation in an animal model. This showed that Type V collagen was most prominent in regions of new tissue formation within the neointima, close to the inner surface of the prosthesis. The broad spectrum of differential reactivities allows the antibodies to be used for a wide range of experimental models. These MAb therefore provide a novel approach for the evaluation of biomaterial performance, particularly for collagen-based implants.

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Immunocytochemical localization of protein antigens in large sections of tissues embedded in water-soluble embedding media.

Journal of Histochemistry & Cytochemistry ◽

10.1177/34.4.3512701 ◽

1986 ◽

Vol 34 (4) ◽

pp. 535-538 ◽

Cited By ~ 7

Author(s):

T M Duello ◽

F C Gumkowski

Keyword(s):

Water Soluble ◽

Placental Lactogen ◽

Protein Antigens ◽

Ph Change ◽

Binucleate Cells ◽

Tissue Sections ◽

Pre Treatment ◽

Ethanol Pretreatment ◽

Large Sections ◽

Absolute Etoh

JB4 and Immunobed are water-soluble embedding media used for embedding large blocks of tissue. Immunobed was specifically designed for immunocytochemistry because ethanol extraction of an additive in the monomer of the resin is reported to render tissue sections permeable to immunoglobulins. We have modified the manufacturer's protocol to accomplish localization of two protein antigens in tissues embedded in either JB4 or Immunobed. Luteinizing hormone-beta (LH beta) was localized in sections of rat and bovine pituitary tissues and bovine placental lactogen (bPL) was localized in sections of placentomes from bovine placentas. Sections received one of the following pre-treatments: absolute EtOH; NaHCO3 buffer, pH 6-10; EtOH followed by NaHCO3 buffer; one of several enzymes; EtOH followed by enzyme; NaHCO3 buffer followed by enzyme. Anti-LH beta stained only pituitary gonadotrophs and anti-bPL stained only placental binucleate cells, as assessed by absorption controls. Pre-treatment with enzyme was required for staining of sections, but an alkaline pH change (NaHCO3) had little or no effect. Ethanol pretreatment had little or no effect alone or in conjunction with NaHCO3 or enzyme. Sections were sufficiently thin (1.5 micron) to afford resolution of structure, but suitably large (approximately 2 cm2) to minimize problems of sampling.

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Nature and distribution of mineral-binding, keratan sulfate-containing glycoconjugates in rat and rabbit bone.

Journal of Histochemistry & Cytochemistry ◽

10.1177/40.11.1431063 ◽

1992 ◽

Vol 40 (11) ◽

pp. 1779-1788 ◽

Cited By ~ 9

Author(s):

M Maeno ◽

M Taguchi ◽

K Kosuge ◽

K Otsuka ◽

M Takagi

Keyword(s):

Animal Species ◽

Polyacrylamide Gel Electrophoresis ◽

Molecular Size ◽

Keratan Sulfate ◽

Preferential Localization ◽

Rabbit Bone ◽

Mineral Binding ◽

Mineralized Matrix ◽

Beta Galactosidase ◽

Rat Bone

The presence of keratan sulfate (KS) and KS proteoglycans in bone has been demonstrated in birds and rabbits but comparison with other animal species has not been investigated. The nature and distribution of mineral-binding, KS-containing glycoconjugates in rat and rabbit bone were investigated with a monoclonal antibody (MAb 5D4) specific for KS. Mineral-binding proteins were extracted from the mineralized bone with 0.4 M EDTA without guanidine-HCl (E-extract). On Western blot analysis of SDS-polyacrylamide gel electrophoresis, rat E-extract gave a weak 5D4-reactive band, M(r) 66,000-68,000, whereas rabbit E-extract produced two major reactive populations of small and large molecular size; one population consisted of two closely spaced bands at M(r) 61,000-63,000 and 66,000-68,000, and the other population consisted of one band at approximately M(r) 200,000. The identity of KS chains was further established by the sensitivity of these bands to keratanase II (Bacillus sp. Ks 36) and endo-beta-galactosidase. Immunocytochemistry with MAb 5D4 showed that, in rat bone, staining associated with the mineral phase was limited to the walls of osteocytic lacunae and bone canaliculi, whereas the remainder of the mineralized matrix lacked staining. In contrast, in rabbit bone the staining was distributed over the entire portion of the mineralized matrix with focal accumulation of staining in the wall of the lacunocanalicular system. These results indicate that rat bone contains a mineral-binding, KS-containing glycoconjugate with preferential localization in the wall of the lacunocanalicular system, whereas rabbit bone contains at least two or possibly three types of KS-containing glycoconjugates distributed over the entire portion of the mineralized matrix.

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Metal-catalyzed oxidation renders silver intensification selective. Applications for the histochemistry of diaminobenzidine and neurofibrillary changes.

Journal of Histochemistry & Cytochemistry ◽

10.1177/34.12.3537114 ◽

1986 ◽

Vol 34 (12) ◽

pp. 1667-1672 ◽

Cited By ~ 54

Author(s):

F Gallyas ◽

J R Wolff

Keyword(s):

Hydrogen Peroxide ◽

Oxidative Polymerization ◽

Silver Coating ◽

Neurofibrillary Changes ◽

Iodide Ions ◽

Tissue Sections ◽

Metal Catalyzed Oxidation ◽

Pre Treatment ◽

Metal Catalyzed ◽

Catalyzed Oxidation

Physical developers can increase the visibility of end products of certain histochemical reactions, such as oxidative polymerization of diaminobenzidine and selective binding of complex silver iodide ions to Alzheimer's neurofibrillary changes. Unfortunately, this intensification by silver coating is generally superimposed on a nonspecific staining originating from the argyrophil III reaction, which also takes place when tissue sections are treated with physical developers. The present study reveals that the argyrophil III reaction can be suppressed when tissue sections are treated with certain metal ions and hydrogen peroxide before they are transferred to the physical developer. The selective intensification of Alzheimer's neurofibrillary changes requires a pre-treatment with lanthanum nitrate (10 mM/liter) and 3% hydrogen peroxide for 1 hr. The diaminobenzidine reaction can be selectively intensified when physical development is preceded by consecutive treatments with copper sulfate (10 mM/liter, pH 5, 10 min) and hydrogen peroxide (3%, pH 7, 10 min). In peroxidase histochemistry, this high-grade intensification may help to increase specificity and reduce the threshold of detectability in tracing neurons with horseradish peroxidase or in immunohistochemistry when the peroxidase-antiperoxidase method is used.

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Interaction of autoantibodies to thyrotropin receptor with a hydrophilic subunit of the thyrotropin receptor

Biochemical Journal ◽

10.1042/bj2280111 ◽

1985 ◽

Vol 228 (1) ◽

pp. 111-117 ◽

Cited By ~ 10

Author(s):

E Davies Jones ◽

F A Hashim ◽

Y Kajita ◽

F M Creagh ◽

P R Buckland ◽

...

Keyword(s):

Gradient Centrifugation ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Water Soluble ◽

Tsh Receptor ◽

Human Thyroid ◽

Thyrotropin Receptor ◽

Freezing And Thawing ◽

Sucrose Density Gradient Centrifugation ◽

A Subunit

Reduction of human thyroid membranes with dithiothreitol caused the release of a water-soluble glycoprotein which neutralized the thyrotropin (TSH) receptor-binding and thyroid-stimulating activities of Graves‘ serum. Analysis of the protein by gel filtration and sucrose density gradient centrifugation allowed estimates of 3.45 nm for the Stokes’ radius, 3.6 S for the s20,w and 47 000 +/- 5000 (mean +/- S.D.; n = 4) for the Mr. The material released by dithiothreitol treatment could be crosslinked to 125I-labelled TSH coupled to N-hydroxysuccinimidyl 4-azidobenzoate (125I-HSAB-TSH), suggesting that it contained a component of the TSH receptor. Furthermore, analysis of the crosslinked material by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated that it contained the TSH receptor A subunit (Mr 50 000). Several factors suggested therefore that the glycoprotein released by dithiothreitol treatment of human thyroid membranes was the TSH receptor A subunit. In particular, (a) both preparations were hydrophilic and were released from membranes by reduction, (b) they had similar Mr values and (c) both preparations crosslinked to 125I-HSAB-TSH. Material similar to the TSH receptor A subunit was released from thyroid membranes by treatment with papain, probably as a result of cleavage of the receptor A subunit at a site close to the interchain disulphide bridge. A similar mechanism, involving thyroid proteinases, was probably involved in release of material with similar properties to the TSH receptor A subunit during freezing and thawing of human thyroid homogenates.

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Preparation and application of different size materials on the cotton yarn and investigating the effect of sizing on the tensile properties of cotton yarn

Bangladesh Journal of Scientific and Industrial Research ◽

10.3329/bjsir.v49i1.18850 ◽

2014 ◽

Vol 49 (1) ◽

pp. 25-30

Author(s):

S Sultana ◽

MZ Haque ◽

HP Nur

Keyword(s):

Tensile Properties ◽

Water Soluble ◽

Woven Fabric ◽

Seed Kernel ◽

Cotton Yarn ◽

Textile Production ◽

Tamarind Seed ◽

Different Types ◽

Cotton Yarns ◽

Pre Treatment

Sizing of the cotton yarn is essential to reduce breakage of the yarn due to abrasion during weaving process. The sizing agent on the woven fabric after weaving needs to remove completely before the next textile production process of dyeing and finishing. So, water soluble sizing agent is easy to handle and easy to desizing for pre-treatment of woven fabric. In this work, different types of water soluble tamarind seed kernel based sizing formulations (assigned by A, B and C) were made and applied on cotton yarn to investigate the effect on the tensile properties of sized and unsized cotton yarns. Cotton yarn treated with size B formulation shows the better tensile properties than the application of size A and size C formulation. The effect of lubricant has also been investigated and shows that the addition of lubricant decreases the tensile properties of the cotton yarn. DOI: http://dx.doi.org/10.3329/bjsir.v49i1.18850 Bangladesh J. Sci. Ind. Res. 49(1), 25-30, 2014

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STUDY OF PROTEIN FRACTIONS OF HIGH-TEMPERATURE HYDROLYZATES OBTAINED FROM SMOKED SPRAT HEADS

Fisheries ◽

10.37663/0131-6184-2020-2-113-117 ◽

2020 ◽

Vol 2020 (2) ◽

pp. 113-117

Author(s):

Olga Mezenova ◽

Vladimir Wolkov ◽

Larisa Baydalinova ◽

Natalia Mezenova ◽

Svetlana Agafonova ◽

...

Keyword(s):

Molecular Weight ◽

Chemical Composition ◽

Raw Materials ◽

Weight Fraction ◽

Water Soluble ◽

Protein Fractions ◽

Raw Material ◽

Organoleptic Characteristics ◽

Pre Treatment ◽

Peptide Fractions

The authors study three fractions obtained as a result of hydrolysis of smoked sprat heads (under temperature of 130oC and presser of 0.25 MPa) – fat, protein water-soluble, and protein-and-mineral ones. Waste from sprat production of two fish canning complexes of the Kaliningrad Region - “RosCon” and “Kolkhoz for the Motherland” - was used as raw material. Hydrolysis was carried out in an aqueous medium in two ways - with preliminary separation of fat and without this operation. The protein fraction was sublimated and its quantitative and qualitative indices were examined - mass yield, solubility, chemical composition and molecular fractional composition of the obtained peptide fractions were determined. The output of sublimated protein fractions is practically independent of the type of raw material and the method of pre-treatment and is 6.47.9% of the mass of raw materials. The chemical composition of protein fractions varies widely in terms of fat (1.4–8.3%), minerals (9.8–13.4%) and proteins (72.1–80.2%). The solubility of the peptide fractions ranged from 91-98%. The molecular weight assessment results showed a high content of a low molecular weight fraction of peptides with an MM of less than 10 kDa in all experimental samples (about 38%). This indicates a high digestibility and biological value of the obtained peptide compositions. Sublimated peptide compositions had typical organoleptic characteristics, pleasant aroma and taste of smoked fish. Ключевые

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Application of a Sequential Extraction Method for Analyzing Cu Distribution in Pre-Treated Mine Tailings after Electrodialytic Remediation

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph16040584 ◽

2019 ◽

Vol 16 (4) ◽

pp. 584 ◽

Cited By ~ 2

Author(s):

Andrea Lazo ◽

Henrik Hansen ◽

Pamela Lazo ◽

Claudia Gutiérrez

Keyword(s):

Sequential Extraction ◽

Mine Tailings ◽

Extraction Method ◽

Constant Electric Field ◽

Extraction Procedure ◽

Water Soluble ◽

Copper Compounds ◽

Leaching Solution ◽

Sequential Extraction Method ◽

Pre Treatment

Mine tailings have been analyzed by a sequential extraction procedure after their pre-treatment with a leaching solution for 24 h and electrodialytic remediation during 15 days with a constant electric field of 2.7 V cm−1. Four leaching solutions were tested: H2SO4 + HNO3 (2:1 vol.) pH = 1.9; H2SO4 + HNO3 (2:1 vol) pH = 4.2; NH4Cl 0.8M, pH = 5.5 and 30% H2O2 adjusted to pH 2 with HNO3 1M + HCl 1M. After the treatment, the tailings were divided in six slices from anode to cathode. The highest removal efficiency of copper was obtained with H2SO4 + HNO3 pH = 1.9, which allows one to remove 67% of the copper in the total cell and 85% of the copper in the slice closest to anode. The same solution with pH = 4.2 allows one to remove 62% of the total copper. The analysis realized by the sequential extraction method indicates the easy removal of water-soluble and exchangeable fractions in all experiments, moreover, residual and sulfide are the less mobile fractions. The general trend was the movement of copper associated to different fractions from anode to cathode and its accumulation closest to the cathode in the case of exchangeable, Fe-Mn oxides and acid soluble fractions, possibly due to some precipitation of copper compounds associated with less acidic conditions.

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Immunohistochemical localization of glycosaminoglycans and proteoglycans in predentin and dentin of rat incisors.

Journal of Histochemistry & Cytochemistry ◽

10.1177/38.3.1689334 ◽

1990 ◽

Vol 38 (3) ◽

pp. 319-324 ◽

Cited By ~ 55

Author(s):

M Takagi ◽

H Hishikawa ◽

Y Hosokawa ◽

A Kagami ◽

F Rahemtulla

Keyword(s):

Dermatan Sulfate ◽

Core Protein ◽

Polyclonal Antibodies ◽

Keratan Sulfate ◽

Immunohistochemical Localization ◽

Immunoperoxidase Staining ◽

Weak Staining ◽

Dentin Matrix ◽

Pre Treatment ◽

Definite Difference

We examined immunocytochemically the type and distribution of glycosaminoglycans and proteoglycans (PG) in predentin and dentin demineralized with EDTA after aldehyde fixation of rat incisors using (a) four monoclonal antibodies (1-B-5,9-A-2,3-B-3, and 5-D-4) which recognize epitopes in unsulfated chondroitin (C0-S), chondroitin 4-sulfate (C4-S), chondroitin 6-sulfate (C6-S), and keratan sulfate (KS) associated with the PG, and (b) monoclonal (5-D-5) and polyclonal antibodies specific for the core protein of large and small dermatan sulfate (DS) PG. Light microscope immunoperoxidase staining after pre-treatment of tissue sections with chondroitinase ABC localized the majority of stainable PG (C4-S, KS, DSPG, C0-S, and C6-S) in predentin and, to a lesser extent (C4-S and small DSPG), in the dentin matrix. The former site demonstrated relatively homogeneous PG distribution, whereas the latter site revealed that strong staining of C4-S and small DSPG was confined mostly to dentinal tubules surrounding odontoblastic processes, with only weak staining in the rest of the dentin matrix. These results indicate that there is not only a definite difference between PG of predentin and dentin but also a selective decrease in the concentration or alteration of these macromolecules during dentinogenesis and mineralization.

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