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Biomedical subjects

A Malmström

Publications and source records attributed to A Malmström.

At least 55 records · Page 3Linked to original sources
PubMed1991

Proteoglycan metabolism in the connective tissue of pregnant and non-pregnant human cervix. An in vitro study.

Profound changes occur in the cervix during pregnancy. In particular, the connective tissue is remodelled. To elucidate the mechanisms behind this process, the metabolism of cervical connective tissue was studied using tissue cultures. Cervical biopsies from non-pregnant and pregnant women were incubated with [35S]sulphate. The proteoglycans of the tissue specimens were purified by ion-exchange and gel chromatography and characterized by SDS/PAGE and by enzymic degradation. In the non-pregnant cervix, the incorporation of [35S]sulphate into the proteoglycans was linear for 48 h. During the first 6 h of incubation the accumulation of chiefly one small labelled proteoglycan (apparent Mr 110,000) substituted with dermatan sulphate was recorded. This is in accordance with the known proteoglycan composition of non-pregnant cervical tissue. In addition, small amounts of two larger radioactive dermatan/chondroitin sulphate proteoglycans (apparent Mr values 220,000 and greater than 500,000) were recorded. After longer periods of incubation the proportion of heparan sulphate proteoglycans increased considerably. The pregnant tissue showed a clearly different composition of labelled proteoglycans. An increased accumulation of the two larger dermatan/chondroitin sulphate proteoglycans was seen in addition to the dominant small dermatan sulphate proteoglycan of the non-pregnant cervix. The rate of accumulation of these two proteoglycans was about 3 times higher in the pregnant tissue, whereas that of the small dermatan sulphate proteoglycan was only increased 2-fold. The fact that the concentration of proteoglycans in the pregnant cervix is approximately one-half of that in the non-pregnant cervix indicates that the turnover of proteoglycans in pregnant cervical tissue is significantly increased. The major effect of this profound change of metabolism was a 50% decrease in proteoglycan content and a 2-fold increased proportion of a dermatan sulphate proteoglycan with an apparent Mr of 220,000.

Adult↗
PubMed1991

Proliferation of cultured fibroblasts is inhibited by L-iduronate-containing glycosaminoglycans.

We have modified a method (Gilles et al: Anal. Biochem., 159:109-113, 1986) for measuring cell number, that is based on the binding of crystal violet to cell nuclei and used it to assay effects of various glycosaminoglycans on growth of human lung fibroblasts. The procedure was modified by growing cells in microcultures (96 well microplates) and by measuring the amount of adsorbed dye using a microplate photometer after solubilisation of the cells with detergent. There was a linear relationship between absorbance and cell number measured by a Coulter Counter. The method is rapid and sensitive and can be used for screening many samples as well as measuring growth rates at low initial cell densities. Even the low growth rates obtained in the absence of serum can be detected. Human lung fibroblasts were growth arrested by serum deprivation and then grown for periods of up to 4 days in the presence of serum and exogenously added glycosaminoglycans (range, 0.1-100 micrograms/ml). Hyaluronan, chondroitin sulfate, and dextran sulfate were without effects, whereas dermatan sulfate, certain heparan sulfates, and heparin suppressed growth (20%-50% inhibition). The antiproliferative activity of dermatan sulfate increased with increasing iduronate content. Certain heparan sulfates, with moderately high sulfate and L-iduronate contents were better inhibitors than heparin despite the fact that the latter glycan has even higher sulfate and L-iduronate contents. The antiproliferative effect of exogenous glycans appeared after a lag period of 3-4 days, suggesting that they interfered with factors produced by the cells. Furthermore, the degree of inhibition was greater when the initial cell density was low. Above a certain level of seeded cells (approx. 10,000 cells/well), there was no inhibition by any of the glycosaminoglycans. It is possible that exogenous glycans cannot overcome endogenous growth-promoting effects in densely seeded cultures.

Cell Count↗
PubMed1991

Oligosaccharide mapping of proteoglycan-bound and xyloside-initiated dermatan sulfate from fibroblasts.

The copolymeric structure of dermatan sulfate chains synthesized by skin fibroblasts has been examined. Chains initiated onto exogeneous p-nitrophenyl-beta-D-xylopyranoside or attached to protein in a large proteoglycan, PG-L, and two small proteoglycans, PG-S1 and PG-S2, have been compared by using high resolution electrophoresis and gel chromatography of oligosaccharides generated by specific enzymatic or chemical degradations. The results confirm that chains attached to PG-L are glucuronate-rich, whereas novel findings indicate that chains attached to either of the two PG-S variants yield closely similar oligosaccharide maps, have approximately equal glucuronate and iduronate content and contain over 90% 4-sulfated disaccharide repeating units. Dermatan sulfate chains built onto xyloside at concentrations of 50 microM and below have a copolymeric structure similar to that of chains from the two PG-S variants. These findings indicate that the polymer-modifying machinery can generate chains with extended iduronate-containing repeats also when the xylose primer is not linked to core protein.

Carbohydrate Sequence↗
PubMed1991

The synthesis of a family of structurally related proteoglycans in fibroblasts is differently regulated by TFG-beta.

Fibroblasts synthesize a variety of proteoglycans among which is a family of structurally related small proteoglycans, i.e. PG-S1 (biglycan) and PG-S2 (decorin). Fibromodulin, which is present in some tissues as a keratan sulfate proteoglycan, also belongs to this family. We have used primary fibroblasts from fetal skin and bovine sclera in culture to study the metabolism of proteoglycans. In particular the regulatory effect of transforming growth factor-beta (TGF-beta), interleukin-1 (IL-1) platelet-derived growth factor (PDGF) and dexamethasone was determined by studies of mRNA levels for these structurally related proteoglycans. Furthermore the synthesis and secretion of these macromolecules was studied using radioactive precursors. TGF-beta induced a 3-fold increase of mRNA for PG-S1, collagen I and III in both types of fibroblasts. mRNA for PG-S2 increased only slightly (1.7-fold) in human skin fibroblasts; while no effect was noticed in sclera fibroblasts. The expression of fibromodulin mRNA was not effected in any of the cells investigated. IL-1, PDGF and dexamethasone had no significant effects on the levels of proteoglycan and collagen mRNA, respectively. Synthesis and secretion of PG-S1, -S2 and fibromodulin wa studied by labeling with [3H]-leucine and [35S]-sulfate. Final separation of PG-S1 and -S2 was achieved by hydrophobic interaction chromatography. TGF-beta induced a 3- to 6-fold increase of [3H]- and [35S]-labeled PG-S1; while PG-S2 only increased 1.3- to 1.4-fold in both types of fibroblasts. No effect on synthesis and secretion of immunoprecipitated fibromodulin was noted.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Alteration of proteoglycan metabolism during the differentiation of 3T3-L1 fibroblasts into adipocytes.

3T3-L1 fibroblasts were induced to differentiate to 3T3-L1 adipocytes by dexamethasone, isobutyl-methylxanthine, and insulin. To study how differentiation affects extracellular matrix production, the accumulation of proteoglycans was studied by labeling the 3T3-L1 cells with [35S]sulphate for 24 h. The labeled proteoglycans were isolated from the medium and cell layer extracts by anion-exchange chromatography. They were then taken to gel filtration chromatography on Superose 6 before or after chondroitin ABC lyase digestion. Hyaluronan was determined by radioimmunoassay. The rate of accumulation of proteoglycans and hyaluronan in the control 3T3-L1 fibroblasts increased with time whereas it decreased slightly in the age matched adipocytes where the differentiation had proceeded, as judged by the change of morphology and increase of the activity of the adipose conversion markers glycerol-3-phosphate dehydrogenase and hormone sensitive lipase. The main change noted was that the adipocytes accumulated 50-70% less amount of small proteoglycans (decorin) in the medium than the fibroblasts did. The amount of large chondroitin/dermatan sulphate proteoglycans was also decreased but to a considerably smaller extent (30%). In the cell layer, heparan sulphate proteoglycan decreased by 60% as compared with the control cells. Thus, the differentiation of 3T3-L1 fibroblasts into adipocytes, which changes the morphology and the function of the cells, is also accompanied by a decreased net production especially of proteoglycans typical of fibrous connective tissue.

1-Methyl-3-isobutylxanthine↗
PubMed1991

Insufficient remodelling of the uterine connective tissue in women with protracted labour.

OBJECTIVE: To investigate the association between a slow progress of labour and insufficient remodelling of the uterine connective tissue. DESIGN: An open comparative study. SETTING: Danderyd Hospital, Department of Obstetrics and Gynaecology, referral centre. SUBJECTS: Eleven women (study group) in oxytocin augmented labour but with an unripe cervix in whom vaginal delivery could not be accomplished and 12 women (normal labour group) in normally progressing spontaneous labour and a favourable cervix but who needed to be delivered by caesarean section due to signs of fetal distress. INTERVENTIONS: At caesarean section tissue specimens were obtained from the fundus, the isthmus and the cervix uteri. MAIN OUTCOME MEASURES: Collagen concentration and extractability, collagenolytic activity expressed as DNP-peptide hydrolytic activity and the concentrations of sulphated glycosaminoglycans (S-GAG) and hyaluronic acid (HA) in the tissue specimens. RESULTS: Statistically significantly higher concentrations and lower extractability of collagen in the isthmus and the cervix uteri was found in women with slow progress of labour compared with those with normal labour. CONCLUSIONS: An insufficient remodelling of the connective tissue in the cervix and isthmus uteri may contribute to slow progress of labour.

Adult↗
PubMed1990

Inventory of human skin fibroblast proteoglycans. Identification of multiple heparan and chondroitin/dermatan sulphate proteoglycans.

Heparan sulphate and chondroitin/dermatan sulphate proteoglycans of human skin fibroblasts were isolated and separated after metabolic labelling for 48 h with 35SO4(2-) and/or [3H]leucine. The proteoglycans were obtained from the culture medium, from a detergent extract of the cells and from the remaining 'matrix', and purified by using density-gradient centrifugation, gel and ion-exchange chromatography. The core proteins of the various proteoglycans were identified by electrophoresis in SDS after enzymic removal of the glycosaminoglycan side chains. Skin fibroblasts produce a number of heparan sulphate proteoglycans, with core proteins of apparent molecular masses 350, 250, 130, 90, 70, 45 and possibly 35 kDa. The major proteoglycan is that with the largest core, and it is principally located in the matrix. A novel proteoglycan with a 250 kDa core is almost entirely secreted or shed into the culture medium. Two exclusively cell-associated proteoglycans with 90 kDa core proteins, one with heparan sulphate and another novel one with chondroitin/dermatan sulphate, were also identified. The heparan sulphate proteoglycan with the 70 kDa core was found both in the cell layer and in the medium. In a previous study [Fransson, Carlstedt, Cöster & Malmström (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 5657-5661] it was suggested that skin fibroblasts produce a proteoglycan form of the transferrin receptor. However, the core protein of the major heparan sulphate proteoglycan now purified does not resemble this receptor, nor does it bind transferrin. The principal secreted proteoglycans are the previously described large chondroitin sulphate proteoglycan (PG-L) and the small dermatan sulphate proteoglycans (PG-S1 and PG-S2).

Chondroitin Sulfate Proteoglycans↗
PubMed1990

TGF-beta enhances the production of hyaluronan in human lung but not in skin fibroblasts.

Transforming growth factor-beta (TGF-beta) enhances the production of extracellular matrix components, such as type I and type III collagen, fibronectin, proteoglycans, in various cell types. The effect on hyaluronan synthesis in relation to proteoglycan synthesis has not been investigated. Human lung or skin fibroblast cultures were treated with TGF-beta in serum-free medium for various periods of time. 35SO4 or [3H]glucosamine was then added to the cultures in the absence of TGF-beta for up to 48 h. Hyaluronan and proteoglycans were isolated by ion-exchange chromatography and quantitated. TGF-beta induced a three- to fourfold increase in hyaluronan production by lung cells but had no effect on skin fibroblasts. In contrast, proteoglycan synthesis was enhanced in both cell types, although skin fibroblasts responded at lower concentrations of TGF-beta. Increased accumulation of hyaluronan was noted only in the cell medium, whereas proteoglycan accumulation was observed both in the medium and in the cell layer. The ED50 for TGF-beta on hyaluronan accumulation in lung cells was the same as that for proteoglycan accumulation, i.e., 40 pM. In skin fibroblasts the ED50 was considerably lower (4 pM). The induction time needed to attain full effect of TGF-beta was 6 h for both hyaluronan and proteoglycan synthesis. These results indicate that TGF-beta has tissue-specific effects on matrix production which may be of importance for control of cell proliferation in various disease states.

Cells, Cultured↗
PubMed1989

Shared and distinct structural features of interstitial proteoglycans from different bovine tissues revealed by electron microscopy.

Large and small interstitial proteoglycans were purified from different bovine tissues, i.e. cartilage, sclera, tendon, aorta, cornea, and bone. The structure of the molecules was compared using the glycerol spraying/rotary shadowing technique for electron microscopy. Large proteoglycans from sclera and tendon have a core protein with a domain structure similar to that previously reported for cartilage proteoglycans (Paulsson, M., Mörgelin, M., Wiedemann, H., Beardmore-Gray, M., Dunham, D., Hardingham, T., Heinegård, D., Timpl, R., and Engel, J. (1987) Biochem. J. 245, 763-772). It is comprised of a pair of globules at one end of the molecule, connected by a short extended segment, followed by a long extended domain which is terminated by a third globular domain. Large aorta proteoglycans show a somewhat different structure, with only one globular domain at each end of a long extended segment. Large sclera and aorta proteoglycans form aggregates with hyaluronate and cartilage link protein in a manner similar to that of large cartilage proteoglycans. The large proteoglycans show considerable tissue variability with regard to number, length, and spacing of glycosaminoglycan side chains. The small proteoglycans reveal a small globular core protein to which one or two glycosaminoglycans are attached. Although the main structural features do not differ, proteoglycans of the S1 class have an average glycosylation close to two glycosaminoglycans/molecule, while that of the S2 class is close to one. Differences in glycosaminoglycan length were observed between tissues and between the S1 and S2 class of proteoglycan derived from a single tissue.

Animals↗
PubMed1989

Changes in the connective tissue of corpus and cervix uteri during ripening and labour in term pregnancy.

The composition of the connective tissue of human cervix and corpus uteri was studied in tissue specimens from seven nonpregnant women and 14 pregnant women, delivered at term by section, to examine spontaneous cervical ripening and labour-induced changes in both the uterine and the cervical connective tissue. The main finding in both the cervix and the corpus was a large (40-60%) decrease of the collagen concentration. The collagen extractability, obtained by pepsin digestion, was increased twofold, suggesting a change of the organization of the collagen fibrils. This reorganization process could also be demonstrated by a large increase of the collagenolytic activity demonstrated with an artificial DNP-peptide substrate. The concentrations of sulphated glycosaminoglycans was lower in pregnant women than in non-pregnant women. The results show that both the cervix and the corpus uteri contain substantial amounts of connective tissue components (collagen, sulphated glycosaminoglycans and hyaluronic acid) and that during ripening, reconstruction of the connective tissue components occurs in both sites. This indicates that the cervical state reflects that of the myometrium.

Adolescent↗
PubMed1987

Glucocorticoids change the nucleotide and sugar nucleotide pool sizes in cultured human skin fibroblasts.

Fibroblasts in culture were treated with the glucocorticoid budesonide. The nucleotide and sugar nucleotide pools were quantitated after separation by isotachophoresis. Steroid treatment induced a 40% increase of the UDP-N-acetylhexosamine pool and a 30% increase of the UDP-glucuronic acid pool whereas the UTP pool was diminished. These effects became apparent after 24 h of incubation and a new steady state was attained after 48 h of incubation. The 3'-phosphoadenosine-5'-phosphosulfate pool was probably not influenced by the glucocorticoid treatment. The half-life of the UDP-N-acetylhexosamine pool was considerably longer in treated than in control cells. The efflux from the UDP-N-acetylhexosamine pool was also lowered in the treated cells. The changed efflux may be due to a decreased glycoconjugate synthesis induced by glucocorticoid treatment. The rate of equilibration of [14C]glucose and [3H]glucosamine with the UDP-N-acetylhexosamine pool was changed by glucocorticoid treatment; especially that of [3H]glucosamine was decreased.

Budesonide↗
PubMed1987

Different biochemical composition of connective tissue in continent and stress incontinent women.

The collagen content in biopsies from skin and ligamentum rotundum of 7 women with a long history of stress incontinence was compared with that of continent controls. The collagen was extracted with 0.5 M acetic acid, followed by digestion with pepsin and quantitated as hydroxyproline. The skin of stress incontinent women contained 40% less collagen than that of continent women. The findings for ligamentum rotundum were similar. These results suggest a deteriorated connective tissue in stress-incontinent women and cast new light on the etiology of the disease.

Adult↗
PubMed1987

Biochemical characterization and tissue distribution of the scleredema in a case of Buschke's disease.

Biopsies from a patient with a longstanding form of scleredema adultorum Buschke were analysed for morphological and biochemical changes in the dermal connective tissue. By light microscopy the tissue changes were located to the deep part of the reticular dermis. Therefore dermal tissue was separated into a superficial and a deep part and analysed biochemically. By this procedure it was possible to show that the concentration of hyaluronan in the deep part of the dermis was increased. The urinary excretion of methylimidazole acetic acid, an indicator of the mast cell mass in the body, was also elevated.

Diabetes Mellitus, Type 2↗
PubMed1986

Structure of proteoheparan sulfates from fibroblasts. Confluent and proliferating fibroblasts produce at least three types of proteoheparan sulfates with functionally different core proteins.

[3H]Leucine- and [35S]sulfate-labeled proteoheparan sulfates were isolated from postconfluent or proliferating cultures of human skin fibroblasts. Cell layers were solubilized by Triton X-100, and transferrin-binding macromolecules were isolated by affinity chromatography. Proteoglycans with no affinity for transferrin were purified by using ion-exchange and gel permeation chromatography. Postconfluent cells synthesize a proteoheparan sulfate of Mr 350,000 (as determined by gel permeation chromatography) which has affinity for transferrin as well as for octyl-Sepharose. Its core protein (Mr 180,000) consists of two disulfide-bonded polypeptides of Mr 90,000. This species was not detected in cultures of proliferating cells. Proliferating and confluent cells also synthesize other forms of proteoheparan sulfates (Mr 200,000-400,000) which have no affinity for transferrin. However, most of them have affinity for octyl-Sepharose. The core protein of proteoheparan sulfates made by proliferating cells has Mr 50,000. A smaller form (Mr 250,000) of this proteoglycan was solubilized by Triton X-100, whereas a larger form (Mr 400,000) remained associated with the pericellular matrix. A third type of proteoheparan sulfate (Mr 200,000) without affinity for transferrin nor octyl-Sepharose was associated with postconfluent cell layers but not with proliferating ones. Its core protein has Mr 35,000. Heparan sulfate oligosaccharides (Mr 6,000 or higher) were found in proliferating cells but not in postconfluent ones.

Cell Division↗
PubMed1986

The functions of the heparan sulphate proteoglycans.

Heparan sulphate (HS)-containing proteoglycans (HS-PGs) are present at the surface of nearly all adherent mammalian cells. The principal mode of attachment is by way of the protein core which is inserted into the plasma membrane. Other forms of HS-PG may be components of pericellular matrices, notably basement membranes. The core proteins of HS-PGs can be small (35K) as in hepatocytes, intermediate (50K) as in many mesenchymal cells, or very large (400K) as in basement membranes. A special case is the HS-PG synthesized by postconfluent fibroblasts. This proteoglycan has a core protein that closely resembles the transferrin receptor glycoprotein. It is possible that this HS-PG is a pro-form of the receptor. Low molecular weight, carbohydrate-rich HS-PG forms are probably derived from larger forms by partial degradation. The HS side-chains can contain 24 different disaccharides in an unknown number of arrangements. The biosynthetic machinery can impose considerable restrictions; for example, the extent of N-sulphation rarely exceeds 40-50%, whereas O-sulphation may range from 20% to 75% of potential sites. Nevertheless, the informational capacity of HS is formidable. By way of the HS chains, HS-PG at the surface of endothelial cells can interact specifically or selectively with a number of plasma proteins. HS-PG at the surface of matrix-producing cells is similarly in a position to interact with matrix proteins, notably collagen, fibronectin and laminin. As the cytoplasmic portion of the HS-PG core protein can bind actin, this proteoglycan can provide a connection between extracellular matrices and the cytoskeleton. A number of studies support a role for HS-PGs in the control of cell growth, and this could be one of their major functions. Whether the HS side-chains or the core protein or both are carrying out such a function remains to be determined.

Animals↗
PubMed1986

Cervical collagen: an important regulator of cervical function in term labor.

This study provides the first clear evidence of a close correlation between the biochemical composition of the cervix and the clinical course of delivery in terms of cervical dilatation. Cervical biopsy specimens were obtained from three groups of patients: Group A, ten women with favorable cervix and spontaneous labor; group B, 12 women with unfavorable cervix given 0.5 mg prostaglandin E2 in gel intracervically for cervical priming and induction of labor; and group C, five women with unfavorable cervix and spontaneous labor. Cervical dilatation time was significantly longer (18 hours) for women in group C compared with women in group A (6.7 hours) and in group B (5.0 hours; P less than .001). The total amount of cervical collagen was significantly higher in women in group C at 8.58 micrograms/mg compared with 6.7 micrograms/mg in women in group A and 5.47 micrograms/mg in prostaglandin E2-treated women in group B. The amount of nonextractable collagen also was significantly higher in women in group C, 23.6% compared with 11.3% in group A, and 12.4% in group B (P less than .01). The collagenolytic activity was significantly increased in cervical biopsy specimens from prostaglandin E2 gel-treated patients--520 U/100 mg wet weight compared with 380 U/100 mg wet weight in untreated patients in group A (P less than .05). From these results it is concluded that cervical collagen is an important regulator of cervical function in late pregnancy and term labor; that prostaglandin E2 is involved in cervical priming, initiation, and progress of term labor.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1985

Domain structure of proteoheparan sulphate from confluent cultures of human embryonic skin fibroblasts.

Radiolabelled proteoheparan sulphates were isolated from confluent monolayers of fibroblasts and from their spent media. The cell-surface-associated proteoglycan (Mr 350 000) has a core protein of Mr 180 000 that is cleaved by reduction of disulphide bonds into polypeptides of Mr 90 000, both of which can bind transferrin [Fransson, Carlstedt, Cöster & Malmström (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 5657-5661]. Thrombin digestion of the proteoglycan yielded two major fragments. The larger one contained the heparan sulphate chains and glycoprotein-type oligosaccharides, whereas the smaller one contained interchain disulphide bond(s) and had affinity for transferrin as well as for octyl-Sepharose. The larger thrombic fragment was cleaved by trypsin into fragments containing the heparan sulphate chains and the oligosaccharides respectively. The smaller proteoheparan sulphate derived from the culture medium (Mr 150 000) had a core protein of Mr 30 000, which contained heparan sulphate-attachment and oligosaccharide-attachment regions, but no domains for binding of transferrin or for hydrophobic interactions.

Alkylation↗