The effects of intrathecally administered opiates (morphine sulfate and meperidine), alpha-adrenergic agonists (clonidine ST-91) baclofen were examined on the shock titration threshold macaque monkeys chronically prepared with intrathecal (I) or epidural (E) catheters. Spinal produced a long-lasting analgesia which was antagonized by naloxone. order potency I morphine greater than meperidine E morphine. Clonidine ST-91, also dose-dependent, elevation in threshold, phentolamine, but not...

Intrathecal administration of norepinephrine (NE) and alpha adrenergic agonists in rats with chronic spinal catheters produced a significant elevation the nociceptive threshold as measured by hot plate tail flick. The intrathecal NE effect was dose-dependent antagonized competitive fashion pretreatment phentolamine (alpha antagonist) but not propranolol (beta antagonist). isoproterenol agonist) did alter threshold. Effective doses produce demonstrable motor effects. Doses 20 times greater...

Osteoclasts (OCLs) in Paget's disease are markedly increased number and size, have numbers of nuclei per multinucleated cell, demonstrate resorption capacity sensitivity to 1,25-(OH)2D3, the active form vitamin D. These cells also contain nuclear inclusions, reminiscent those seen paramyxovirus-infected cells, which cross-react with antibodies measles virus nucleocapsid (MVNP) antigen. To elucidate role MV abnormal OCL phenotype disease, we transduced normal precursors retroviral vectors...

Paget disease is the most exaggerated example of abnormal bone remodeling, with primary cellular abnormality in osteoclast. Mutations p62 (sequestosome 1) gene occur one-third patients familial and a minority sporadic disease, P392L amino acid substitution being commonly observed mutation. However, it unknown how p62(P392L) mutation contributes to development this disease. To determine effects expression on osteoclasts vitro vivo, we introduced either or WT into normal osteoclast precursors...

Abstract A potent interleukin-6 (IL-6) antagonist (Sant 5), which binds tightly to the IL-6α receptor but has impaired gp130 heterodimerization, been developed recently by site-directed mutagenesis of human IL-6. We report here that Sant 5 inhibits IL-6-stimulated osteoclast-like multinucleated cell (MNC) formation in marrow cultures also stimulatory effects IL-1 or tumor necrosis factor alpha (TNF-α these cultures. further show a neutralizing antibody IL-6 TNF-α In contrast, had no effect...

Ultrastructural, immunocytochemical, and in situ hybridization studies have suggested that paramyxoviruses, such as measles virus (MV), are present Pagetic osteoclasts may contribute to the abnormality osteoclast function. However, little additional information is known about potential viruses osteoclasts. As there increased numbers of precursors among marrow mononuclear cells Paget's patients, we used reverse transcriptase-polymerase chain reaction amplify nucleocapsid sequence MV from...

Abstract It has been debated for almost 30 years whether Paget's disease of bone results from paramyxoviral infection osteoclasts (OCs). Paramyxoviral-like nuclear inclusions are found in OCs patients with disease, and measles virus (MV) or canine distemper (CDV) messenger RNA (mRNA) transcripts have detected by situ hybridization cells pagetic lesions. Furthermore, immunocytochemical studies shown the presence several species disease. However, others unable to detect samples marrow cultures...

Autocrine products of osteoclasts such as interleukin-6 may play an important role in normal osteoclast formation and activity.To identify novel stimulatory factors for osteoclasts, we have prepared a mammalian cDNA expression library generated from highly purified human osteoclast-like multinucleated cells (MNC) formed long term bone marrow cultures screened this autocrine that enhance MNC formation.A candidate clone which stimulated was isolated.Sequence analysis showed encoded annexin I1...

Annexin II (AXII), a calcium-dependent phospholipid-binding protein, has been recently found to be an osteoclast (OCL) stimulatory factor that is also secreted by OCLs. In vitro studies showed AXII induced OCL formation and bone resorption. However, the mechanism of action which acts as soluble extracellular protein induce unknown. this paper, we demonstrate gene expression upregulated 1,25-dihydroxyvitamin D3 [1, 25-(OH)2D3] addition significantly increased OCL-like multinucleated cell...

Abstract We targeted the MVNP gene to OCL lineage in transgenic mice. These mice developed abnormal OCLs and bone lesions similar those found Paget's patients. results show that persistent expression of can induce pagetic-like vivo. Introduction: disease (PD) is second most common disease. Both genetic viral factors have been implicated its pathogenesis, but their exact roles vivo are unclear. previously reported transfection normal human osteoclast (OCL) precursors with measles virus...

Abstract Our previous studies suggested that increased osteoclast formation and activity in Paget's disease may be related part to responsiveness of highly purified precursors 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. However, the basis for this enhanced sensitivity 1,25-(OH)2D3 is unclear. To address question, we examined 24-hydroxylase receptor (VDR) messenger RNA (mRNA) expression during human differentiation from normal subjects patients with response as well VDR content affinity....

Abstract Paget disease of bone is characterized by abnormalities in all phases remodeling, but the fundamental cellular abnormality resides osteoclast (OCL). Osteoclasts involved contain viral-like nuclear and cytoplasmic inclusions that react with antibodies directed against paramyxovirus nucleocapsid proteins, such as measles virus, respiratory syncytial or canine distemper virus. However, identity virus mechanisms responsible for its persistence pathologic role unclear. Furthermore,...

Abstract Tartrate-resistant acid phosphatase (TRAP) is an iron-binding protein that highly expressed in osteoclasts. To characterize the regulation of TRAP gene expression, progressive 5′ and 3′ deletions a 1.8 kb fragment containing 5′-flanking sequence were fused to luciferase reporter gene. Two nonoverlapping regions this had promoter activity. The upstream (P1) was located within region from –881 bp –463 relative ATG, while downstream (P2) between –363 –1 we have previously shown be...

Abstract Spinal muscular atrophy (SMA) is characterized by degenerating lower motor neurons and an increased incidence of congenital bone fractures. Survival neuron (SMN) levels are significantly reduced due to deletions/mutations in the telomeric SMN1 gene these patients. We utilized Smn −/− SMN2 mouse model SMA determine functional role for SMN remodelling. µCT analysis lumber vertebrae, tibia femur bones from mice revealed osteoporotic phenotype. Histological demonstrated a thin porous...

Pagetic osteoclasts (OCLs) are abnormal in size and contain paramyxoviral-like nuclear inclusions that cross-react with antibodies to measles virus (MV). However, the role MV infection plays Paget's disease is unknown, because no animal model of available. Therefore, we targeted a cellular receptor, human CD46 (hCD46), cells OCL lineage transgenic mice using mouse tartrate-resistant acid phosphatase (TRAP) gene promoter. In vitro precursors from hCD46 significantly increased formation bone...

Evidence indicates that astronauts experience significant bone loss in space. We previously showed simulated microgravity (μXg) using the NASA developed rotary cell culture system (RCCS) enhanced resorbing osteoclast (OCL) differentiation. However, mechanism by which μXg increases OCL formation is unclear. RANK/RANKL signaling pathway critical for Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) has been shown to increase osteoclastogenesis. hypothesize TRAIL may play an...

Osteoclast (OCL) precursors from patients with Paget's disease (PD) and normal OCL transduced the measles virus nucleocapsid protein gene (MVNP) are hyperresponsive to 1alpha,25-dihydroxyvitamin D(3) [1alpha,25-(OH)(2)D(3)] can form OCLs at physiologic concentrations of 1alpha,25-(OH)(2)D(3). This hyperresponsivity 1alpha,25-(OH)(2)D(3) is due increased expression TATA box-associated factor II-17, a potential coactivator vitamin D receptor. Hyperresponsivity may permit formation in PD low...

Tartrate-resistant acid phosphatase (TRAP) was first identified in cells from patients with hairy cell leukaemia. Subsequently, it has been found other leukaemias, B-lymphoblastoid lines, osteoclasts and subsets of normal lymphocytes, macrophages, granulocytes. Recent data indicate that TRAP porcine uteroferrin, a placental iron-transport protein, represent single gene product. However, the intracellular role is unknown. We used full-length human cDNA probe to examine expression peripheral...

Abstract In contrast to normal OCL precursors, pagetic precursors express MVNP and form at physiologic concentrations of 1,25(OH)2D3, as do transfected with the gene. Using a GST-VDR chimeric protein, we identified TAFII-17 VDR binding protein expressed by transduced precursors. was in part responsible for increased 1,25(OH)2D3 responsivity Introduction: Pagetic osteoclasts (OCLs) their measles virus nucleocapsid (MVNP) large numbers OCLs low 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]....

Factor X (FX) Vorarlberg is a congenital FX deficiency characterized clinically by mild bleeding tendency. Homozygous individuals have activity of less than 10% in the extrinsic system and 25% intrinsic system. antigen 20%. Using molecular techniques, two point mutations were detected coding sequence gene: G----A at base pair 160 exon II resulting change Gla14 (GAA) to Lys (AAA); 424 V from Glu102 (GAG) (AAG). The abolished TaqI restriction site an MnlI V. To determine whether these are...