RESUMO
BACKGROUND: Single parent families are at higher risk of re-report to Child Protective Services (CPS) than two-parent families. Yet, how single-family homes differ in risk from two-parent families remains under researched. OBJECTIVE: To identify heterogenous patterns of child and caregiver factors among CPS-involved families and the subsequent risk for CPS re-report based on child and family characteristics (i.e., sociodemographic information, family structure, and risk indicators). PARTICIPANTS AND SETTING: Data were from the 2017 National Child Abuse and Neglect Data System Child File (N = 249,026). METHODS: We conducted latent class analysis (LCA) to identify discrete patterns (i.e., classes) based on child and caregiver risk indicators (e.g., substance use, behavioral health). We then used logistic regression to examine family structure and other family characteristics and CPS indicators predicted CPS re-report for each class. RESULTS: Results yielded five distinct classes: 1) Financial Stressors (25 % of the sample); 2) Caregiver Substance Use (16 %); 3) Complex Household Stressors (3 %); 4) Child Disabilities (4 %); and 5) Minimal Household Stressors (53 %). Family structure was significantly associated with CPS re-reports for Classes 1, 2, and 5. For Class 1, single father families had increased odds of CPS re-report compared to other family structures. For Classes 2 and 5, single father families' odds of CPS re-reports were greater than those of married families, but lower than single mother families. CONCLUSIONS: Children growing up in single father families have different likelihoods of repeat CPS involvement compared to those in single mother and married families. Financial stressors and parental substance use within single father families should be addressed.
Assuntos
Maus-Tratos Infantis , Serviços de Proteção Infantil , Características da Família , Humanos , Serviços de Proteção Infantil/estatística & dados numéricos , Criança , Feminino , Masculino , Maus-Tratos Infantis/prevenção & controle , Maus-Tratos Infantis/estatística & dados numéricos , Pré-Escolar , Adolescente , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Cuidadores/psicologia , Análise de Classes Latentes , Fatores de Risco , Família Monoparental/estatística & dados numéricos , Lactente , Adulto , Estados Unidos , Estresse Financeiro/psicologia , Estrutura FamiliarRESUMO
Accurate diagnosis of lymphoid malignancies is essential for appropriate therapeutic intervention. In conjunction with other diagnostic determinants, immunophenotypic analysis of differentially expressed cell surface markers, such as CD5, CD20, CD23 and FMC7, is useful in the subclassification of lymphomas and leukemias arising from the B-cell lineage. Recent evidence suggesting that CD20 predicts FMC7 expression has prompted reappraisal of the utility of monitoring both markers. Here, we report that the FMC7 monoclonal antibody (mAb) specifically and strongly recognized CD20 ectopically expressed in hematopoietic and nonhematopoietic cell lines. The reactivity of FMC7 was abolished by mutations in the extracellular domain of CD20. These data confirm the CD20 specificity of FMC7. Like other CD20 mAbs, FMC7 binding was temperature dependent and induced detergent insolubility of CD20. Of significant interest, the CD20 epitope recognized by FMC7 was unusual in that it was exceptionally sensitive to membrane cholesterol. Cholesterol depletion profoundly reduced expression of the FMC7 epitope, whereas cholesterol enrichment enhanced its expression. FMC7 mAb binding thus appears to be a sensitive indicator of the level of plasma membrane cholesterol and reveals a conformational state of CD20 that is regulated by cholesterol.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos CD20/análise , Antígenos CD20/imunologia , Linfócitos B/imunologia , Colesterol/farmacologia , Epitopos , Glicoproteínas/análise , Afinidade de Anticorpos , Antígenos CD20/genética , Linfócitos B/patologia , Epitopos/análise , Glicoproteínas/imunologia , Humanos , Leucemia de Células B/patologia , Linfoma de Células B/patologia , Mutagênese Sítio-Dirigida , Solubilidade , Células Tumorais CultivadasRESUMO
TNF-alpha and IL-1beta promote leukocyte recruitment to arthritic joints and may contribute to cartilage degradation while regulatory cytokines such as IL-4 and IL-1RA may in part determine the course of arthritis. Here we report the pattern of TNF-alpha, IL-1beta, IL-6, IFN-gamma, IL-1RA, and IL-4 mRNA expression, detected by RT/PCR, in the talar joint and draining popliteal lymph node (PLN) of rats with adjuvant arthritis (AA). Levels of TNF-alpha and IFN-gamma mRNA were increased in the PLN before clinical signs of arthritis. This was followed by increases in IL-1beta and IL-1RA mRNA at d9 and IL-6 mRNA at d12. PLN IL-1RA mRNA levels were positively correlated with those of IL-1beta and TNF-alpha throughout d5-d20. IL-4 mRNA levels were highest on days 7 and 20. In the synovium, a small increase in TNF-alpha, IL-1beta, and IL-6 mRNA was detected on d5 then again on d12. Maximal synovial TNF-alpha levels were reached on d20, while IL-1beta peak expression was on d16 and IL-6 on d14. IL-4, IL-1RA, and IFN-gamma mRNA was undetectable in the synovium. Cyclosporin treatment for 4 days, initiated at the height of arthritis, rapidly decreased clinical disease, and decreased migration of neutrophils and T lymphocytes into the joints. Yet no significant effect of CyA was observed on inflammatory cytokine expression, although the correlation between PLN IL-1RA and IL-1beta or TNF-alpha was lost in treated animals. Thus there is a variable pattern of cytokine gene expression in rat AA, the undetectable IL-4 and IFN-gamma mRNA in synovium being analogous to human rheumatoid arthritis.
Assuntos
Artrite Experimental/metabolismo , Ciclosporina/farmacologia , Citocinas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Imunossupressores/farmacologia , Linfonodos/metabolismo , RNA Mensageiro/biossíntese , Membrana Sinovial/metabolismo , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/genética , Artrite Reumatoide/metabolismo , Quimiotaxia de Leucócito/efeitos dos fármacos , Ciclosporina/uso terapêutico , Citocinas/biossíntese , Modelos Animais de Doenças , Humanos , Hipersensibilidade Tardia/imunologia , Imunossupressores/uso terapêutico , Interferon gama/biossíntese , Interferon gama/genética , Proteína Antagonista do Receptor de Interleucina 1 , Interleucina-1/biossíntese , Interleucina-1/genética , Interleucina-4/biossíntese , Interleucina-4/genética , Interleucina-6/biossíntese , Interleucina-6/genética , Masculino , Modelos Animais , Ratos , Ratos Endogâmicos Lew , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sialoglicoproteínas/biossíntese , Sialoglicoproteínas/genética , Tarso Animal/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genéticaRESUMO
The prerequisite for the recruitment of circulating leukocytes to sites of inflammation is adhesion to vascular endothelial cells. Selectins play a significant role in the initiation of this multistep process by mediating "rolling" of the leukocytes on the endothelium. Investigation of selectin-dependent cell interactions using function blocking monoclonal antibodies (MAb) provides insights into the mechanisms involved in leukocyte migration into inflammation. Until now most studies in inflammation models in rats have relied on cross-reactive or polyclonal antibodies against rat E-selectin. In an E-selectin knockout mouse, we aimed to generate an adhesion function blocking MAb to rat E-selectin by immunization with rat E-selectin transfected Chinese hamster ovary cells (RESEC). An IgG1 kappa MAb was identified that reacts with RESEC but not with untransfected Chinese hamster ovary cells, as well as with recombinant mouse E-selectin protein as assessed by ELISA. This MAb is designated RME-1. It does not cross-react with rat L-selectin or rat P-selectin or E-selectin expressed on human umbilical vein endothelium. Adhesion of the HL-60 myeloid cells to immobilized mouse E-selectin was completely inhibited by MAb RME-1 under static conditions and adhesion of rat polymorphonuclear leukocytes to recombinant mouse E-selectin was blocked under rotation condition. This novel antibody thus recognizes a function-related epitope on rodent E-selectin.
Assuntos
Anticorpos Bloqueadores/imunologia , Anticorpos Monoclonais/imunologia , Adesão Celular/imunologia , Selectina E/imunologia , Epitopos/imunologia , Animais , Células CHO , Cricetinae , Ensaio de Imunoadsorção Enzimática , Células HL-60 , Humanos , Hibridomas , Camundongos , Camundongos Knockout , Neutrófilos/imunologia , Ratos , Ratos Endogâmicos LewRESUMO
P-selectin is an important adhesion molecule involved in leukocyte migration. However, to date, no monoclonal antibodies (MAb) generated against rat P-selectin have been identified which block P-selectin mediated leukocyte adhesion. Most studies in the rat have utilized crossreacting antibodies generated against P-selectin in higher species. In a P-selectin deficient mouse we generated an anti-rat/mouse P-selectin MAb, designated RMP-1, by immunization with activated rat platelets. This IgG2a MAb immunoprecipitates a 140 kDa protein under reducing conditions from rat platelet lysate. By ELISA and immunofluorescence flow cytometry, MAb RMP-1 reacts with thrombin-activated but not unactivated rat platelets. In addition, by ELISA MAb RMP-1 binds to activated mouse platelets and recombinant rat and mouse P-selectin. MAb RMP-1 inhibited adhesion of HL-60 myeloid cells to immobilized mouse P-selectin by 97% and to activated rat and mouse platelets by 100% under static conditions, confirming the adhesion function blocking activity of MAb RMP-1. This novel MAb should be useful for studying P-selectin function in vitro and in vivo in both rat and mouse inflammation models.
Assuntos
Anticorpos Bloqueadores/imunologia , Anticorpos Monoclonais/imunologia , Selectina-P/imunologia , Animais , Anticorpos Bloqueadores/química , Anticorpos Monoclonais/química , Plaquetas/imunologia , Western Blotting , Adesão Celular/imunologia , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Células HL-60 , Humanos , Ligantes , Glicoproteínas de Membrana/imunologia , Glicoproteínas de Membrana/metabolismo , Camundongos , Peso Molecular , Ativação Plaquetária/efeitos dos fármacos , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/imunologiaRESUMO
The migration of leucocytes from blood into the joint is a key feature of human and experimental arthritis. Adhesion molecules on leucocytes and vascular endothelium are important in this process and may be therapeutic targets for intervention in arthritis. We investigated whether monoclonal antibody treatment to block the alpha 4 integrin, very late activation antigen-4 (VLA-4), and beta 2 integrins (CD11/CD18) administered to rats during the preclinical (day 5) or clinical phase (day 10+) would modify disease. When treatment was initiated 5 days after induction of disease, development of arthritis was significantly reduced by either anti-alpha 4 (TA-2) or anti-beta 2 (WT.3) monoclonal antibodies (mAb) and the combination of both mAb was even more effective (clinical scores: control 11.4; anti-alpha 4 6.6; anti-beta 2 6.8; anti-alpha 4 + anti-beta 2 3.9). When treatment was delayed until arthritis was apparent (day 10), the anti-alpha 4 + anti-beta 2 mAb combination still significantly diminished the arthritis score on day 14 (control 13; anti-alpha 4 + anti-beta 2 7.9). Treatment with anti-alpha 4 + anti-beta 2 mAb decreased the migration to the joints of blood polymorphonuclear leucocytes (PMNL) by 66-79% and of spleen T lymphocytes by 56-75%, depending on the joint. In contrast, PMNL migration was abolished (> 98%) and T-cell migration markedly (87%) inhibited to dermal inflammatory reactions in the same animals. These findings demonstrate: that blocking mAb to alpha 4 and beta 2 integrins can reduce the severity of adjuvant arthritis, even after joint inflammation has developed; that this treatment can markedly inhibit PMNL and T-lymphocyte migration to the joints; and that yet to be defined mechanisms distinct from alpha 4 (CD49d) and beta 2 (CD11/CD18) integrins, also contribute to leucocyte migration to inflamed joints. Identifying these additional adhesion mechanisms may be required to control joint inflammation further.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Artrite Experimental/terapia , Antígenos CD18/imunologia , Integrinas/imunologia , Receptores de Retorno de Linfócitos/imunologia , Animais , Artrite Experimental/imunologia , Movimento Celular/imunologia , Integrina alfa4beta1 , Articulações/imunologia , Masculino , Neutrófilos/imunologia , Ratos , Ratos Endogâmicos Lew , Pele/imunologia , Linfócitos T/imunologiaRESUMO
OBJECTIVE: To determine the prevalence of autoantibodies to high mobility group (HMG) proteins in systemic sclerosis (SSc). METHODS: One hundred ninety-seven unselected sera from patients diagnosed as SSc (n = 180) or Raynaud's phenomenon (RP) (n = 17) were tested for HMG autoantibodies by ELISA and immunoblotting. RESULTS: Seventy-one of the 180 (39.0%) SSc sera bound to HMG proteins in an ELISA: 56 (31%) to HMG-1 and/or HMG-2; 29 (16%) to HMG-14/17. In the same assay, 7 of 17 RP sera (41%) bound to HMG proteins: 4 (23%) to HMG-1 and/or HMG-2, and 5 (29%) to HMG-14/17. The specificity of HMG binding was confirmed by immunoblotting. CONCLUSION: Antibodies to HMG proteins, particularly to HMG-1 and HMG-2 are found in about 1/3 of SSc Sera. Since HMG-1 and HMG-2 have a role in transcription, these observations further implicate transcriptional complexes as targets of autoantibodies in scleroderma. This is the first published report of HMG autoantibodies in scleroderma.
Assuntos
Autoanticorpos/sangue , Proteínas de Grupo de Alta Mobilidade/imunologia , Doença de Raynaud/imunologia , Escleroderma Sistêmico/imunologia , Autoanticorpos/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , ImmunoblottingRESUMO
OBJECTIVE: To determine the prevalence of autoantibodies to high-mobility group (HMG) proteins in sera from patients with drug-induced lupus (DIL). METHODS: Forty-two patients who developed autoantibodies and/or lupus after treatment with procainamide or other drugs were tested for HMG autoantibodies by immunoblotting and enzyme-linked immunosorbent assay (ELISA). RESULTS: Twenty-eight of the 42 sera (67%) bound HMG-14 and/or HMG-17. In comparison, 9 of 42 (21%) bound HMG-1 and/or HMG-2. There was a good correlation between ELISA results and binding on immunoblots. CONCLUSION: The high prevalence of antibodies to the nucleosomal core HMGs (HMG-14 and HMG-17) in DIL patients adds evidence implicating nucleosomes as immunogens in drug-induced autoimmunity.
Assuntos
Autoanticorpos/sangue , Proteínas de Grupo de Alta Mobilidade/imunologia , Lúpus Vulgar/induzido quimicamente , Acebutolol/uso terapêutico , Doenças Autoimunes/sangue , Doenças Autoimunes/induzido quimicamente , Autoimunidade/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Lúpus Vulgar/sangue , Lúpus Vulgar/tratamento farmacológico , Procainamida/efeitos adversos , Sulfassalazina/uso terapêuticoRESUMO
The prescription drugs procainamide (PA) and hydralazine (HYD) are associated with the induction of autoimmunity and a clinical syndrome called drug-induced lupus. Since PA- and HYD-induced autoantibodies are directed primarily against histones and histones are prime acceptors of poly (ADP-ribose) (PADPR), we have investigated the effects of PA and HYD on the activity of poly (ADP-ribose) polymerase (PADPRP). Control substances, with structures similar to PA and HYD but not known to induce lupus, included N-acetylprocainamide (NAPA) and the amino acids phenylalanine, tryptophan and proline, and their amide derivatives. Wil-2 cells were incubated in 0.5-50 microM PA, NAPA and HYD, which included therapeutic concentrations of these drugs. The mean enhancement of incorporation of [3H]-nicotinamide adenine dinucleotide (NAD) into PADPR was 1.84 (P = 0.005) with PA, with HYD 1.48 (P = 0.029), and with NAPA 1.38 (P = 0.036). This increase was suppressed by 3-aminobenzamide, an inhibitor of PADPRP activity. Little or no increase in [3H]-NAD incorporation was observed with equivalent concentrations of phenylalanine, phenylalaninamide or tryptophan. However, a 1.29-fold increase was noted with 0.5 microM tryptophanamide, a 1.26-fold increase with 0.5 microM prolinamide and a 1.4-fold increase with 50 microM proline. PA increased PADPRP activity in B- and T-cell lines but not in promyelocytic leukemia or epithelial cell lines. Since poly (ADP-ribosylation) is important in the cellular response to various agents, the increased ADP-ribosylation of intracellular molecules may be a key event in the induction of autoantibodies.
Assuntos
Hidralazina/farmacologia , Poli Adenosina Difosfato Ribose/metabolismo , Procainamida/farmacologia , Acecainida/farmacologia , Linhagem Celular Transformada , Humanos , Poli(ADP-Ribose) Polimerases/metabolismo , Células Tumorais CultivadasRESUMO
A residential facility for the aged was evaluated for their illumination needs. Only the shared facility and grounds immediately surrounding it were considered. The facility evaluated is one of the most modern and the lighting design is considered its best feature. The age of residents occupying the facility ranges from 60 to 85 years. The lighting evaluation included administering a lighting adequacy/inadequacy questionnaire and measurement of actual light levels in the various parts of the facility during the day and after dark hours. The results indicated that lighting in approximately half the areas surveyed was substandard. An attempt to develop recommendations for the lighting needs of the elderly was unsuccessful. It was concluded that realistic illumination level recommendations need to be developed for the elderly.
Assuntos
Instituição de Longa Permanência para Idosos , Iluminação/normas , Idoso , Humanos , Matemática , Pessoa de Meia-Idade , Inquéritos e Questionários , Estados UnidosRESUMO
Twelve patients with serologically and histologically defined symptomatic primary biliary cirrhosis were randomized to receive either oral cyclosporin A or vehicle placebo. The cyclosporin A-treated group had improvement in serum alkaline phosphatase and gamma-glutamyltransferase, suggesting a moderating effect on the course of the liver disease. However, other indices of the liver disease, including liver biopsies, did not show significant improvement. The purpose of this study was to examine the effect of the cyclosporin A treatment on serum indicators of bone and mineral metabolism in these patients, as in its later stages, primary biliary cirrhosis is associated with significant osteopenic bone disease. There were no significant changes in serum calcium, phosphate, magnesium, or vitamin D metabolites between the two groups. However, after one year of treatment, mean serum parathyroid hormone level was significantly lower in the cyclosporin A treatment group, and serum osteocalcin rose significantly. There were no significant changes in any parameter of mineral metabolism in the placebo group. The changes in parathyroid hormone and osteocalcin following cyclosporin A therapy suggest a reduction in the secondary hyperparathyroidism commonly seen with primary biliary cirrhosis and also an increase in bone formation, respectively. This study therefore provides preliminary evidence that cyclosporin A therapy seems to have a mild beneficial effect on the abnormalities of mineral metabolism seen with this disorder.
Assuntos
Cálcio/metabolismo , Ciclosporina/uso terapêutico , Cirrose Hepática Biliar/tratamento farmacológico , Doenças Ósseas Metabólicas/etiologia , Doenças Ósseas Metabólicas/prevenção & controle , Calcifediol/sangue , Calcitriol/classificação , Creatinina/sangue , Ciclosporina/farmacologia , Humanos , Cirrose Hepática Biliar/complicações , Cirrose Hepática Biliar/metabolismo , Minerais/sangue , Osteocalcina/sangue , Hormônio Paratireóideo/sangue , Projetos PilotoRESUMO
Parathyroid cysts were found in 2-5% of beef steers and heifers while collecting parathyroids at a local abattoir. The cyst fluid contained parathyroid hormone (PTH), averaging 15 ng/ml of fluid. Reversed-phase high-performance liquid chromatographic (HPLC) analysis of the fluid using an acetonitrile-trifluoroacetic acid solvent system, followed by carboxyl-terminal specific PTH radioimmunoassay of the eluant fractions, revealed two major peaks of PTH immunoreactivity. The first eluted at approximately 25% acetonitrile, a position similar to the commercially available human carboxyl terminal fragment of PTH-(39-84). The second peak, at approximately 37% acetonitrile, corresponded to the elution position for bovine PTH-(1-84). The cyst fluid was tested for enzyme activity by incubation of 125I-labeled bovine PTH with cyst fluid for 30 minutes at 37 degrees C, which resulted in no fragmentation of the labeled peptide. Immunohistochemistry of the cells lining the parathyroid cyst indicated that they contained PTH.
Assuntos
Cistos/metabolismo , Doenças das Paratireoides/metabolismo , Hormônio Paratireóideo/análise , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Cistos/patologia , Técnicas Imunoenzimáticas , Masculino , Microscopia Eletrônica , Doenças das Paratireoides/patologia , RadioimunoensaioRESUMO
We measured serum osteocalcin concentrations in 82 pregnant and 21 nonpregnant women. Osteocalcin values declined in the second trimester, but returned to nonpregnant levels late in the third trimester. The mean serum osteocalcin concentration in 36 women during pregnancy (mean gestation, 26 weeks) of 2.8 ng/mL was significantly lower than that in nonpregnant women (6.4 ng/mL; P less than 0.001) or term pregnant women at delivery (6.1 ng/mL; n = 46). Serum immunoreactive PTH (iPTH) levels were significantly higher during pregnancy than in nonpregnant women [97 +/- 5 vs. 56 +/- 4 ng/L (mean +/- SE); P less than 0.001]. No significant correlations were found between maternal osteocalcin concentrations and serum phosphorus, alkaline phosphatase, or iPTH, but significant negative correlations were found between osteocalcin and total calcium or total protein. Osteocalcin concentrations in midtrimester amniotic fluid were very low (mean, 0.3 +/- 0.1 ng/mL; n = 11). In 29 lactating mothers, the mean serum osteocalcin level was 9.5 +/- 1.5 ng/mL, significantly higher than in any of the other groups (P less than 0.05), but their serum calcium and iPTH levels were normal. There was no correlation between serum osteocalcin and calcium or iPTH concentrations in lactating women. These changes are compatible with a sequence in which bone turnover is reduced during early pregnancy, rebounds in the third trimester, and increases in postpartum lactating women.
Assuntos
Proteínas de Ligação ao Cálcio/sangue , Lactação/sangue , Gravidez/sangue , Adolescente , Adulto , Líquido Amniótico/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Feminino , Humanos , Minerais/metabolismo , Osteocalcina , Segundo Trimestre da Gravidez , Terceiro Trimestre da GravidezRESUMO
Antibodies from 5 patients with systemic sclerosis reacted with an antigen localized to the metaphase chromatin, the cleavage furrow and the midbody of anaphase and telophase HEp-2 cells. The titer of antimidbody antibodies ranged from 1:160 to 1:1280. Four patients had systemic sclerosis and one had idiopathic Raynaud's phenomenon. In situ biochemical characterization of the antigen revealed that it was resistant to DNase I, micrococcal nuclease and RNase A, but was sensitive to trypsin treatment. The antigen remained insoluble in 400 mM acetic acid but was extracted from the cells with 400 mM hydrochloric acid. The antibody was not seen in sera from 2500 normal female blood donors, 120 patients with systemic lupus, 60 patients with rheumatoid arthritis, 15 patients with linear scleroderma or 25 patients with Raynaud's disease.
Assuntos
Antígenos/imunologia , Células/imunologia , Cromatina/imunologia , Escleroderma Sistêmico/sangue , Linhagem Celular , Células/ultraestrutura , Humanos , Metáfase , Escleroderma Sistêmico/imunologiaRESUMO
The role of intracellular calcium in the secretion of fragments of PTH by human parathyroid tissue has not been examined previously. We used an in vitro perifusion system to study this phenomenon. Hyperplastic human parathyroid tissue, obtained at surgery from patients with chronic renal failure, was sliced into small pieces, placed in perifusion chambers, and exposed to low (0.5 mM) or high (2.0 mM) calcium concentrations. PTH secreted into the perifusion medium was analyzed by gel exclusion chromatography and RIA using a carboxyl-terminal antiserum. Under high calcium conditions, proportionally more carboxyl-terminal fragments of PTH (65.3%) were released by the perifused tissue than intact PTH (34.7%). When perifused with low calcium medium, the tissue released more intact PTH (55.0%) than carboxyl-terminal fragments (45.0%). These studies are consistent with the suggestion that calcium-dependent intracellular degradation of PTH plays an important role in the regulation of hormone secretion.
Assuntos
Cálcio/fisiologia , Glândulas Paratireoides/metabolismo , Hormônio Paratireóideo/metabolismo , Fragmentos de Peptídeos/metabolismo , Adulto , Idoso , Cromatografia em Gel , Feminino , Humanos , Hiperplasia/metabolismo , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Perfusão , RadioimunoensaioRESUMO
Antibodies directed against centromeric chromatin characteristically occur in the sera of patients with the CREST variant of scleroderma. We have studied the in situ enzymatic sensitivity and solubility of the centromeric antigen and have isolated an antigenic moiety that reacts with anticentromere antibodies. The centromeric antigen in the human epithelial cell line, HEp-2, was sensitive to DNAase I and micrococcal nuclease but not affected by RNAase A, trypsin or amylase. It was insoluble in 0.15-4 M NaCl but was extracted from the HEp-2 cells by 4 M urea/2 M NaCl. Antigenic activity in a 4 M urea/2 M NaCl extract of rabbit thymus was demonstrated by immunoabsorption. Indirect immunofluorescence of the extract separated by polyacrylamide gel electrophoresis revealed a fluorescent band with a mol. wt of 33,000. Calf thymus and trout testis histone preparations were fractionated by gel electrophoresis and transferred by blotting techniques to diazobenzyloxymethyl cellulose paper for autoradiography. Anticentromere antibodies bound to and were absorbed by trout testis histone 1. We propose that the centromeric antigen may be a variant of histone 1 that is associated with condensed chromatin.