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[Origin of stromal bone marrow mechanocytes according to the results of typing them by isoantigens and chromosomal markers].

The bone marrow of radiochimaeras and heterotopic bone marrow transplants were used to study the origin of precursors of the fibroblasts growing in the monolayer cultures of hemopoietic tissue. In the bone marrow explants of the (C57BL/6 X CBA) F1 mice, in which the CBA bone marrow was transplanted following the lethal irradiation, the fibroblasts grown in the colonies were of recipient origin judging by isoantigens in the reaction of indirect immunofluorescence with the anti-C57BL/6-serum. At the same time in the bone marrow explants from heterotopic transplants (CBA leads to CBA X C57BL/6) the fibroblasts grown in colonies were of donor origin. The cultures of hemopoietic cells of the bone marrow of females heterotopically transplanted in the singenic male (guinea pigs Huston) contained only fibroblasts which were of donor origin judging by sex chromosomes in the metaphase plates of dividing cells. Hence, the bone marrow precursors of fibroblasts do not depend histogenetically on hemopoietic cells and are not replaced at the expense of repopulating cells of the second partner.

Animals

Gene markers in human bone marrow transplantation.

One or more differences between donors and recipients were found in polymorphic red blood cell antigens and enzymes in each of 56 bone marrow transplant sibling pairs. These results identify those polymorphic traits which are potentially most informative for detection of donor cells in a recipient following transplantation.

Blood Group Antigens

Lymphocyte chimerism after bone marrow transplantation. Surface markers and in vitro function of donor and recipient lymphocyte subpopulations.

Specific HLA antibodies were used to eliminate donor and recipient cells, respectively, from lymphocyte suspensions prepared from the blood of a child who had been transplanted with bone marrow from an HLA-A- and HLA-B-incompatible, HLA-D-compatible donor. About 70% of the lymphocytes were of donor HLA type, the remaining of recipient type. The phytohemagglutinin-responsive lymphocytes were exclusively limited to the lymphocyte population carrying donor-type HLA antigens. Membrane immunofluorescence investigations of the donor and recipient populations showed a low percentage of IgM-positive lymphocytes in the donor population and an extremely high proportion of IgM-positive lymphocytes in the recipient population. About 90% of the donor lymphocytes were T cells, as judged by their capacity to form rosettes between sheep erythrocytes and T lymphocytes; no cells in the recipient cell population expressed this ability.

B-Lymphocytes

The Anti-Osteoporosis Effects of Panax japonicus via Downregulation of Inflammatory Factors: A Network Pharmacology and Ovariectomized Rat Model Study.

OBJECTIVE: Osteoporosis is a major and growing public health problem characterized by decreased bone mineral density and destroyed bone microarchitecture. Panax japonicus has been clinically used in the treatment of bone diseases, especially osteoporosis. However, there is a lack of study on the mechanism of osteoporosis treatment with Panax japonicus. MATERIALS AND METHODS: A network pharmacology approach was employed to identify the targets of osteoporosis and Panax japonicus. Cytoscape 3.7.2 and DAVID were used to visualize the pharmacological mechanism of Panax japonicus in treating osteoporosis by building up compound-target and protein-protein interaction (PPI) networks and conducting Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. An ovariectomized SD rat osteoporosis model was used to assess the potential therapeutic effect of Panax japonicus in vivo. The biomechanical properties, pathological changes, inflammatory cytokines, bone density, and bone microstructural parameters in rat bone tissue were carefully measured. The biochemical markers of bone metabolism in serum were detected by Enzyme-Linked Immunosorbent Assay (ELISA). RESULTS AND DISCUSSION: Fifty-two active components and sixty-five target genes of Panax japonicus involved in the treatment of osteoporosis were identified. The PPI network revealed IL-6, TNF, NR3C1, IL-1β, CASP3, ESR1, PGR, and AR to be involved in the treatment of osteoporosis with Panax japonicus. Chikusetsusaponin IVa and Radix ginsenoside-Ro were the main saponins found in Panax japonicus. Panax japonicus was found to exert potent preventive effects on osteoporosis by maintaining biomechanical properties, increasing bone mineral density, and protecting the trabecular microstructure in an ovariectomized rat osteoporosis model. Panax japonicus hindered the initiation of osteoporosis induced by ovariectomy by regulating bone metabolism and downregulating the expression of IL-6 and TNF-α. CONCLUSION: Panax japonicus was found to contain 52 compounds and 65 targets in the treatment of osteoporosis. The administration of Panax japonicus could mitigate osteoporosis in rats induced by ovariectomy, and one of the mechanisms was associated with downregulating the expression of inflammatory factors.

Animals

Influence of calcitonin treatment on the osteocalcin concentration in the algodystrophy of bone.

Algodystrophy (AD) attacks all tissues in the affected region and results in the rapid demineralization of bones. Osteocalcin (OC) and alkaline phosphatase (AP) are markers of bone turnover. Calcitonin is the treatment of choice of AD. Two groups of patients were studied: Group I (n = 8)--acute stage of AD (before and during the calcitonin treatment), Group II (n = 5)--late chronic stage of AD. In the acute stage of AD both OC level and AP activity were increased. They were normal in the chronic stage of AD. During the calcitonin treatment OC level normalized after 14 days and then increased again. During the treatment, AP activity temporarily increased and then returned to the initial level. We confirm that an increased bone turnover is observed in the acute stage of AD. Discrepancy between OC level and AP activity reflects the local metabolic disturbances. Salmon calcitonin inhibits the algodystrophic process and probably contributes to the activation of the skeletal restoration.

Acute Disease

Potential induction of osteogenesis by systemic administration of bovine bone proteins.

This study shows, apparently for the first time, that the administration of bone derived proteins (putative bone growth factors) obtained from bovine demineralized maxillaries has a direct effect on osteogenesis, affecting significantly some markers of bone formation such as lactate dehydrogenase activity and serum osteocalcin. Also, collagen deposition and bone protein turnover were markedly increased by the treatment, which may have important biological and clinical applications.

Animals

[Osteocalcin and hyperthyroidism].

Osteoporosis may be induced by hyperthyroidism through an increase of bone turnover, because bone resorption exceeds formation in this condition. Also therapy with 1-thyroxine, especially by TSH-suppressive doses, may induce a reduction in bone mineral content. Circulating osteocalcin (sBGP) significantly increases both in endogenous and exogenous hyperthyroxinemia and is considered a reliable non invasive marker of bone turnover. In this study an extra-increase of sBGP in hyperthyroid post-menopausal women towards pre-menopausal is reported, the persistence of high sBGP levels in patients affected by any type of hyperthyroidism after four months of therapy and a positive relationship with thyroid hormones (fT4). Therefore monitoring of this serum marker may be suggested also in patient chronically treated with 1-thyroxine to avoid, if possible, overzealous therapy.

Adult

Plasma Proteomics Identifies Thousand-and-One-Amino Acid Kinase 3 as a Potential Biomarker of Rheumatoid Arthritis Activity and a Novel Therapeutic Target.

OBJECTIVE: Bone destruction associated with active rheumatoid arthritis (RA) remains a major therapeutic challenge, with a lack of reliable molecular markers reflecting bone injury. This study aims to identify novel biomarkers linked to bone destruction in active RA through proteomic analysis, providing new strategies for precise monitoring and targeted therapy. METHODS: Data-independent acquisition mass spectrometry was used for proteomic quantification and bioinformatic analysis on plasma samples from 160 patients with RA and 40 healthy controls. Key proteins associated with bone destruction were screened by integrating Sharp scores with synovial single-cell RNA sequencing data and subsequently validated in two independent cohorts (N1 = 50 and N2 = 10) using enzyme-linked immunosorbent assay and multiplex immunohistochemistry. Functional studies were conducted using fibroblast-like synoviocytes (FLSs) in vitro and a collagen-induced arthritis (CIA) mouse model in vivo. RESULTS: A total of 4,998 plasma proteins were identified, with 506 showing significant differential expression between active and remitted RA. Thousand-and-one-amino acid kinase 3 (TAOK3) levels were positively associated with Sharp scores and markedly elevated in patients with active RA. Combining TAOK3 with C-reactive protein improved diagnostic accuracy for active RA (area under the curve = 0.915). High TAOK3 expression was also associated with increased relapse frequency. Functional studies showed that TAOK3 knockdown suppressed the tumor-like phenotype of FLSs and down-regulated matrix metalloproteinase 1/2/3 and cathepsin K, whereas TAOK3 overexpression promoted pannus cell-mediated bone erosion, mitigated by TAOK3-targeted inhibitor. In vivo, its inhibition showed therapeutic effects in CIA mice. CONCLUSION: TAOK3 serves as a potential biomarker for bone destruction in active RA and as a therapeutic target for precision monitoring and intervention.

Arthritis, Rheumatoid

Changes in shape, ossification and quality of bones in children with spina bifida.

Changes in the cross-sectional shape, size, bone mass and amount of unmineralised osteoid tissue were studied in 17 dissected tibiae from spina-bifida babies who died with paralysis and foot deformities and in 14 tibiae from non-spina bifida controls of matching age. In addition, 12 tibiae from young experimental rats with myotomy of foot dorsiflexors and foot plantiflexors were double-labelled with bone-seeking markers and studied in order to find the role of experimental muscle imbalance in the dynamic remodelling of the developing long bones. It was found that in tibiae from spina-bifida children with paralysis the total area of cortical bone, its thickness, number of Haversian systems and number of large remodelling cavities are diminished. Significant changes in the cross-sectional shape of the midshaft of the tibia were found, ranging from the triangular shape seen in normal babies and in those with spina bifida and calcaneus-type foot deformity, to the circular shape of tibiae from babies with spina-bifida paralysis and no foot deformity or with spina bifida and equinovarus-type of deformity. Results of experimental myotomy on growing rats showed the direct influence of working muscles on the remodelling process of growing tibiae. On the side of myotomy the flat cortex resumed a bulging convex shape and the centre of gravity shifted towards the myotomised side. These principles cannot on their own explain the specific changes in the shape of human tibiae found during anatomical studies. There is, however, a common denominator in these apparently contradictory findings. This is the combined action of two factors previously reported: the combination of paralysis of the growing limb and mechanical intra-uterine pressure acting on it. The findings in the present study also indicate that they played a major role in the production of deformities. The total amount of osteoid tissue in spina-bifida paralysed bone is increased. This delay of mineralisation of newly laid-down bone matrix would lead to softening of the new bone matrix and osteoid-rich subepiphyseal and metaphyseal regions. This 'paralytic rickets', together with the diminished total bone mass found, could probably be the cause of the common spina-bifida fractures in these regions.

Animals

B- and T-cell markers in lymphoproliferative disorders with blood and bone marrow involvement.

B- and T-cell surface markers were determined in 26 adult patients with lymphoproliferative malignancies who had bone marrow and blood involvement. The patients in whom more than 60% of the abnormal cells were immunoglobulin-bearing cells were considered to have B-cell lymphoproliferative malignancy. The diagnosis of a T-cell disorder was made in those patients in whom more than 70% of the cells formed E rosettes (T cells). Those patients in whom 30% and more of the abnormal cells did not show B- and T-cell surface markers were regarded as suffering from "null" cell disorder. The B-cell type of lymphoproliferative malignancy was the most commonly encountered (63%) with an equal male to female ratio. In addition, 4 male patients with "hairy-cell" leukaemia were shown to have a B-cell disorder. T-cell disorders were found in 3 male patients; 1 patient had acute lymphoblastic leukaemia and in the other 2 the histopathological diagnosis was well-differentiated lymphoma and Sternberg sarcoma. Four patients with "null" cell lymphoma were found, the histopathological diagnoses in all were poorly-differentiated lymphocytic lymphoma, and 3 of the 4 patients were males. Further haematological investigations of the B-cell and the non-B-cell disorders showed that although the bone marrow was equally extensively infiltrated in both groups, the non-B-cell disroders were more commonly associated with complications of anemia and thrombocytopenia. It is therefore postulated that the abnormal lymphoid cell involved in B-cell disorders is an end-stage cell and not the haemopoietic stem cell.

Adult

Distinctive distribution of HLA class II presenting and bone marrow derived cells in the anterior segment of human eyes.

Cells of bone marrow origin that normally occupy the stroma of the murine iris and ciliary body have been implicated in the immune phenomenon, anterior chamber associated immune deviation (ACAID). Following injection of antigen into the anterior chamber, cells of this type deliver an ACAID inducing signal into the systemic circulation, presumably through the outflow tract. In an effort to identify such cells in man, anterior chambers of 34 human donor eyes of different age groups were stained immunocytochemically with monoclonal antibodies directed at HLA class II molecules, CD 45 (a molecular marker of bone marrow-derived cells) and macrophage-associated membrane molecules (CD 68, CD 14). Within the outflow tissue, the cells of the filtering trabecular meshwork stained with none of those reagents. However, infrequent single, dispersed, dendritic cells were positively stained in the intertrabecular spaces. More numerous labelled cells were found in the anterior- and posterior-most portions of the non-filtering part of the trabecular meshwork. These cells were continuous with stained cells adjacent to the outer wall of Schlemm's canal and to the collector channels. Numerous labelled cells were seen in the vicinity of the intra- and episcleral vessels, the ciliary meshwork, the stroma of the ciliary muscle and epithelial processes, and the iris stroma. With advancing age, increasing numbers of CD 45+, HLA class II expressing cells appeared to accumulate in the so-called uveoscleral pathway. These results indicate that bone marrow-derived cells with the potential to function of ACAID induction reside within human eyes, and that cells of this type are located not only in the stroma of iris and ciliary body, but within the non-filtering portions of the trabecular meshwork and the uveoscleral pathway. The appearance of rare CD 45+ cells "in transit" in the filtering trabecular meshwork is compatible with the view that cells carrying ACAID-inducing signals to the systemic immune apparatus escape from the eye by this route.

Adult

Nafcillin-associated granulocytopenia.

Three patients treated with nafcillin developed granulocytopenia. Only three other such cases have been reported. This complication is most likely due to a direct toxic effect on the bone marrow; markers for an immunologically mediated mechanism have not been found. Nafcillin-associated granulocytopenia usually occurs during the third week of therapy and remits spontaneously with prompt cessation of nafcillin administration.

Adolescent

Conditions influencing the dose-response effect of growth hormone on longitudinal bone growth in the hypophysectomized rat.

Various factors that might influence the dose-response effect of growth hormone on the longitudinal bone growth were investigated with tetracycline as intravital marker of the bone growth in hypophysectomized rats. These are of practical importance for the bioassay of growth hormone. The growth response was found to be almost the same for subcutaneous and intraperitoneal injection, whereas the intravenous route resulted in significantly lower growth response. The administration of growth hormone in various volumes subcutaneously did not significantly influence the dose-dependent growth response. Freezing of the dissolved growth hormone or addition of NaOH to the solution had no significant effect on growth response. Cortisone acetate 0.5 mg/kg given at hypophysectomy increased the post-operative survival somewhat, without having any post-operative depressing influence on the growth hormone-induced longitudinal bone growth. The present investigation showed that in standardized conditions the dose-dependent growth response is constant.

Animals

Effect of administration frequency of growth hormone on longitudinal bone growth in the hypophysectomized rat.

The effect of the administration frequency of growth hormone on longitudinal bone growth was investigated with tetracycline as intravital marker of the bone growth of the proximal tibia in hypophysectomized rats. The total dose of growth hormone (NIH-GH-B16) and the administration period were the same in all compared experiments. It was possible to achieve an optimum growth response for a certain total dose of growth hormone by increasing the injection frequency. The period of hormone administration was 10 or 5 days followed by a 10 days withdrawal period. When the growth hormone was administered alone or in association with L-thyroxine for 10 days, the optimum injection frequency for growth hormone was found to be 1 inj./day in hypophysectomized rats and 2 inj./day in thyroxine-treated hypophysectomized rats. When the administration period was 5 days for growth hormone given in association with L-thyroxine, the growth stimulation induced by one daily growth hormone injection was the same as that induced by two or four daily injections of the same total dose. An increase in the administration frequency for a total daily dose of thyroxine from 1 to 2 inj./day did not increase the longitudinal bone growth either when thyroxine was given alone or in association with growth hormone.

Animals