RESUMO
Lymphomas are common spontaneous tumors in nonhuman primates but remain poorly characterized in Japanese macaques (Macaca fuscata). This study examined 5 cases of spontaneous malignant lymphoma in Japanese macaques, focusing on the immunophenotypes and presence of simian lymphocryptoviruses, which are Epstein-Barr virus-related herpesviruses in nonhuman primates. The macaques with lymphoma were 5 to 28 years old, indicating that lymphomas develop over a wide age range. The common macroscopic findings were splenomegaly and enlargement of lymph nodes. Histologic and immunohistochemical analyses revealed that all cases were non-Hodgkin type and exhibited a T-cell phenotype, positive for CD3 but negative for CD20 and CD79α. The lymphomas exhibited diverse cellular morphologies and were subdivided into 3 types according to the World Health Organization classification. These included 3 cases of peripheral T-cell lymphoma, not otherwise specified; 1 case of T-cell prolymphocytic leukemia; and 1 case of an unclassifiable T-cell lymphoma. Positive signals were detected by in situ hybridization in 2 of the 4 examined cases using probes for the Epstein-Barr virus-encoded small RNA (EBER). Furthermore, the presence of M. fuscata lymphocryptovirus 2, a macaque homolog of Epstein-Barr virus, was demonstrated in EBER-positive cases by polymerase chain reaction amplification followed by direct sequencing. Immunohistochemistry using antibody to the Epstein-Barr virus-encoded nuclear antigen 2 was negative, even in the EBER-positive cases. The present study suggests that T-cell lymphoma is more common than B-cell lymphoma in Japanese macaques and that M. fuscata lymphocryptovirus 2 is present in some cases.
Assuntos
Linfoma/veterinária , Doenças dos Macacos/patologia , Animais , Feminino , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/veterinária , Hibridização In Situ/veterinária , Leucemia Prolinfocítica de Células T/diagnóstico , Leucemia Prolinfocítica de Células T/patologia , Leucemia Prolinfocítica de Células T/veterinária , Leucemia Prolinfocítica de Células T/virologia , Linfonodos/patologia , Lymphocryptovirus , Linfoma/complicações , Linfoma/patologia , Linfoma/virologia , Linfoma de Células T/diagnóstico , Linfoma de Células T/patologia , Linfoma de Células T/veterinária , Linfoma de Células T/virologia , Linfoma de Células T Periférico/diagnóstico , Linfoma de Células T Periférico/patologia , Linfoma de Células T Periférico/veterinária , Linfoma de Células T Periférico/virologia , Macaca , Masculino , Doenças dos Macacos/diagnóstico , Doenças dos Macacos/virologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Esplenomegalia/etiologia , Esplenomegalia/patologia , Esplenomegalia/veterinária , Infecções Tumorais por Vírus/diagnóstico , Infecções Tumorais por Vírus/patologia , Infecções Tumorais por Vírus/veterinária , Infecções Tumorais por Vírus/virologiaRESUMO
STUDY OBJECTIVES: Cheyne-Stokes respirations occur in 40% of patients with heart failure. Orthopnea is a cardinal symptom of heart failure and may affect the patient's sleeping angle. The objective of this study was to assess the respiratory and hemodynamic response to sleeping angle in a group of subjects with stable heart failure. DESIGN: Twenty-five patients underwent overnight polysomnography with simultaneous and continuous impedance cardiographic monitoring. Sleeping polysomnographic and impedance cardiographic data were recorded. SETTING: The study was conducted in a sleep center. PATIENTS: All 25 patients had clinically stable heart failure and left ventricular ejection fractions < 40%. INTERVENTIONS: The patients slept at 0 degrees, 15 degrees, 30 degrees, and 45 degrees in random order. MEASUREMENTS AND RESULTS: Seventeen patients had Cheyne-Stokes apneas (index > 5/h) and 23 patients had hypopneas (index > 5/h). The hypopnea index showed no response to sleeping angle. The Cheyne-Stokes apnea index decreased with increasing sleeping angle (P < 0.001). This effect was seen only during supine sleep and non-rapid eye movement sleep and was absent in non-supine sleep, rapid eye movement sleep, and during periods of wakefulness. Thoracic fluid content index and left ventricular hemodynamics measured by impedance cardiography showed no response to sleeping angle. CONCLUSIONS: Changing the heart failure patient's sleeping angle from 0 degrees to 45 degrees results in a significant decrease in Cheyne-Stokes apneas. This decrease occurs on a constant base of hypopneas. The changes in Cheyne-Stokes apneas are not related to changes in lung congestion and left ventricular hemodynamics.
Assuntos
Respiração de Cheyne-Stokes/etiologia , Respiração de Cheyne-Stokes/terapia , Insuficiência Cardíaca/complicações , Insuficiência Cardíaca/fisiopatologia , Postura/fisiologia , Sono/fisiologia , Idoso , Cardiografia de Impedância , Respiração de Cheyne-Stokes/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Polissonografia , Sistema Respiratório/fisiopatologia , Método Simples-Cego , Volume Sistólico , Resultado do TratamentoRESUMO
Osteoblasts and their precursors respond to specific cytokines, growth factors, and hormones. One facet of this response includes the secretion of additional cytokines, some of which are part of the circuitry involved in the regulation of osteoblast and osteoclast function. Therefore, understanding which cytokines are able to activate osteoblastic cells and the consequences of that activation are central to understanding normal and pathologic bone remodeling. Oncostatin M (OSM) is a glycoprotein belonging to a new subfamily of cytokines related by sequence and structural homology and the use of the signal transducing receptor component gp130. Osteoblastic cells secrete and respond to leukemia-inhibiting factor (LIF) both in vitro and in vivo, suggesting that LIF is an autocrine regulatory factor. OSM is closely related to LIF, and therefore we hypothesized that OSM should regulate the function of cells in the osteoblastic lineage. Primary neonatal murine or fetal rat calvarial osteoblastic cultures were treated with OSM or LIF and a series of biochemical and biological parameters were determined. In these cultures, OSM induced proliferation, collagen synthesis, and interleukin-6 secretion, whereas it inhibited alkaline phosphatase activity. Bone resorption was also inhibited by OSM. These data represent the first report of OSM's effects on bone cell function and indicate that, like some other members of the LIF/interleukin-6 subfamily, OSM has potent bone regulatory activity.
Assuntos
Reabsorção Óssea/prevenção & controle , Citocinas/farmacologia , Osteoblastos/efeitos dos fármacos , Peptídeos/farmacologia , Fosfatase Alcalina/antagonistas & inibidores , Animais , Divisão Celular/efeitos dos fármacos , Citocinas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Oncostatina M , Osteoblastos/fisiologia , Biossíntese de Proteínas , Ratos , Ratos Sprague-DawleyRESUMO
A hybrid gene encoding for a polypeptide consisting of the first 33 N-terminal amino acid (aa) residues of transforming growth factor-alpha (TGF-alpha) and a C terminus consisting of 20 aa residues of vaccinia growth factor (VGF) was chemically synthesized and expressed as a fusion protein in Escherichia coli. The primary structure of the hybrid gene product maintained the same positioning of the three disulfide bonds found in each parent molecule thus conserving the first two loop regions of TGF-alpha and the third loop region of VGF. After cleavage with CNBr its renatured biological activity was found to be comparable to TGF-alpha and VGF with respect to binding to the epidermal growth factor receptor, stimulation of DNA synthesis and induction of anchorage-independent growth of NRK cells in the presence of TGF-beta. Thus, we suggest that similar domains can be interchanged within the same family of molecules and equivalent functionality maintained.
Assuntos
Biossíntese Peptídica , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Divisão Celular/efeitos dos fármacos , DNA Recombinante , Escherichia coli/genética , Escherichia coli/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Dados de Sequência Molecular , Peptídeos/genética , Peptídeos/farmacologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/farmacologia , Fatores de Crescimento TransformadoresRESUMO
HYPOTHESIS: Transforming growth factor-beta1 (TGF-beta1) plus demineralized bone matrix (DBM) will reconstruct a critical mandibular defect devoid of periosteum in a canine model. STUDY DESIGN: Randomized, blinded, placebo-controlled, prospective animal pilot study. METHODS: Canine critical mandibular defects devoid of periosteum were reconstructed with DBM (group 1, n = 3) and DBM plus TGF-beta1 (250 microg TGF-beta1/g DBM) (group 2, n = 3). Radiologic, histologic, and biomechanical testing was performed on the test group and control group specimens at 12 weeks after implantation. RESULTS: A palpable bone bridge was present in the group 2 subjects 5 to 6 weeks after implantation and was never present in the group 1 subjects. Radiologic and histologic examination at the time of harvest (12 weeks after implantation) demonstrated a solid bone bridge in the group 2 subjects and a fibrous union in the group 1 subjects. Group 2 specimens demonstrated failure in four-point bending testing at an average maximum moment of 9.9 +/- 2.2 N-m. This value was 9.4% of the maximum moment of the contralateral nonoperated side. Group 1 specimens were palpably flexible on plate removal and had a biomechanical strength of 0. The difference in strength between group 1 and group 2 was statistically significant (P < 0.02), supporting the hypothesis that the addition of TGF-beta1 to the DBM carrier resulted in the formation of significantly stronger bone in the critical gap. CONCLUSION: The addition of TGF-beta1 to DBM results in healing of a critical bone defect devoid of periosteum in a higher mammalian model.
Assuntos
Mandíbula/cirurgia , Osteogênese/efeitos dos fármacos , Procedimentos de Cirurgia Plástica/métodos , Fator de Crescimento Transformador beta/uso terapêutico , Animais , Matriz Óssea , Cães , Periósteo , Distribuição AleatóriaAssuntos
Transtorno Autístico , Educação Inclusiva , Criança , Pré-Escolar , Humanos , Nova Zelândia , Relações Pais-FilhoAssuntos
Crime , Gravidez , Adolescente , Fatores Etários , Feminino , Humanos , Nova Zelândia , Controle Social FormalRESUMO
Multifocal submucosal stromal tumors were diagnosed in a 5.5-year-old rhesus macaque (Macaca mulatta) experimentally infected with simian immunodeficiency virus, strain SIVsmE660, and CD4+ T cell depleted. The animal was negative for simian retroviruses, SRV-1, -2, and -5. Polymerase chain reaction analysis of DNA from tumor and spleen tissue revealed abundant, preferential presence of retroperitoneal fibromatosis herpesvirus, the macaque homologue of the Kaposi sarcoma-associated herpesvirus (human herpesvirus-8), in the tumors. This was corroborated by demonstration of viral latent nuclear antigen-1 in the nuclei of a majority of the spindeloid tumor cells. Low levels of an additional macaque herpesvirus, rhesus rhadinovirus, were also detected in the spleen and tumor tissues. The spindeloid cells labeled positively for vimentin and CD117 but were negative for CD31, CD68, desmin, and smooth muscle cell actin. Collectively, these findings suggest a relation to but not absolute identity with simian mesenchymoproliferative disorders (MPD) or typical gastrointestinal stromal tumors (GISTs).
Assuntos
Tumores do Estroma Gastrointestinal/veterinária , Macaca mulatta , Doenças dos Macacos/patologia , Doenças dos Macacos/virologia , Síndrome de Imunodeficiência Adquirida dos Símios/complicações , Vírus da Imunodeficiência Símia , Animais , Antígenos Nucleares/metabolismo , Primers do DNA , Tumores do Estroma Gastrointestinal/complicações , Tumores do Estroma Gastrointestinal/patologia , Herpesvirus Humano 8/metabolismo , Imuno-Histoquímica/veterinária , Reação em Cadeia da Polimerase/veterinária , Retrovirus dos Símios/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Vimentina/metabolismoRESUMO
Oncostatin M (OSM), a glycoprotein of Mr approximately 28,000 produced by activated monocyte and T-lymphocyte cell lines, was previously identified by its ability to inhibit the growth of cells from melanoma and other solid tumors. We have detected significant similarities in the primary amino acid sequences and predicted secondary structures of OSM, leukemia-inhibitory factor (LIF), granulocyte colony-stimulating factor (G-CSF), and interleukin 6 (IL-6). Analysis of the genes encoding these proteins revealed a shared exon organization, suggesting evolutionary descent from a common ancestral gene. Using a panel of DNAs from somatic cell hybrids, we have shown that OSM, like LIF, is located on human chromosome 22. We have also demonstrated that OSM has the ability to inhibit the proliferation of murine M1 myeloid leukemic cells and can induce their differentiation into macrophage-like cells, a function shared by LIF, G-CSF, and IL-6. We propose that OSM, LIF, G-CSF, and IL-6 are structurally related members of a cytokine family that have in common the ability to modulate differentiation of a variety of cell types.
Assuntos
Citocinas/química , Peptídeos/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Divisão Celular/efeitos dos fármacos , Cromossomos Humanos Par 22 , Citocinas/genética , Éxons , Genes , Glicoproteínas/química , Glicoproteínas/genética , Fator Estimulador de Colônias de Granulócitos/química , Fator Estimulador de Colônias de Granulócitos/genética , Inibidores do Crescimento/química , Inibidores do Crescimento/genética , Haplorrinos , Humanos , Interleucina-6/química , Interleucina-6/genética , Fator Inibidor de Leucemia , Linfocinas/química , Linfocinas/genética , Camundongos , Dados de Sequência Molecular , Oligonucleotídeos/química , Oncostatina M , Peptídeos/genética , Conformação Proteica , Alinhamento de SequênciaRESUMO
T4 RNA ligase will catalyze the addition of nucleoside 3', 5'-bisphosphates onto the 3' terminus of tRNA resulting in tRNA molecule one nucleotide longer with a 3' terminal phosphate. Under appropriate conditions the reaction is quantitative and, if high specific radioactivity bisphosphates are used, it provides an efficient means for in vitro labeling of tRNA. Although the 3' terminal hydroxyl is a good acceptor, the 5' terminal phosphate in most tRNA's is not an effective donor in the RNA ligase reaction. This poor reactivity is due to the secondary structure of the 5' terminal nucleotide. If E. Coli tRNAf Met is used, the 5' phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.
Assuntos
Polinucleotídeo Ligases/metabolismo , RNA Ligase (ATP)/metabolismo , RNA de Transferência/metabolismo , Colífagos/enzimologia , Dimetil Sulfóxido , Cinética , Metionina , Fenilalanina , Especificidade por Substrato , TemperaturaRESUMO
An efficient procedure for the replacement of the anticodon and the adjacent hypermodified nucleotide (residues 34-37) of yeast tRNAPhe with any desired oligoribonucleotide sequence has been developed. The four residues are removed by chemical cleavage at Y-37 and partial ribonuclease A digestion at U-33. An oligonucleotide is inserted in three steps by using T4 RNA ligase and T4 polynucleotide kinase. When different oligonucleotides are inserted, both the size of the loop and the sequence of nucleotides in the anticodon region of this tRNA can be varied. The ability of the different anticodon loop substituted tRNAs to be aminoacylated by yeast phenylalanyl-tRNA synthetase is dependent upon the sequence of the oligonucleotide inserted. This suggests that there is an important interaction between the anticodon region of yeast tRNAPhe and its synthetase.
Assuntos
Aminoacil-tRNA Sintetases/metabolismo , Anticódon , Fenilalanina-tRNA Ligase/metabolismo , Aminoacil-RNA de Transferência/biossíntese , RNA de Transferência/metabolismo , Sequência de Bases , Fenilalanina , Polinucleotídeo 5'-Hidroxiquinase/metabolismo , Ligação Proteica , RNA Ligase (ATP)/metabolismo , Saccharomyces cerevisiae , Relação Estrutura-AtividadeRESUMO
Thirteen different yeast tRNAPhe variants with single nucleotide changes in positions 34-37 in the anticodon region were prepared by an enzymatic procedure described previously. Aminoacylation kinetics using purified yeast phenylalanyl-tRNA synthetase revealed that the level of aminoacylation was very different for different sequences inserted. The low level of aminoacylation was the result of a steady state between a slow forward reaction rate and spontaneous deacylation of the product. Aminoacylation kinetics performed at higher synthetase concentrations revealed that substitution at position 34 in tRNAPhe decreased the Km nearly 10-fold but only had a small effect on Vmax. Similar substitutions at positions 35, 36, and 37 had a lesser effect. These data suggest a sequence-specific contact between the anticodon of yeast tRNAPhe and the cognate synthetase.
Assuntos
Aminoacil-tRNA Sintetases/metabolismo , Anticódon/metabolismo , Fenilalanina-tRNA Ligase/metabolismo , RNA Fúngico/metabolismo , RNA de Transferência/metabolismo , Acilação , Sequência de Bases , Cinética , Aminoacil-RNA de Transferência , Saccharomyces cerevisiae/enzimologia , Especificidade por SubstratoRESUMO
Oncostatin M (OSM) is a glycoprotein cytokine that was recently demonstrated to be structurally and functionally related to the leukemia inhibitory factor (LIF). We have investigated the binding of each cytokine to a variety of cellular receptors including those on solid tumor lines, leukemic cells, endothelial cells, macrophages, and cells transfected with the recently cloned low-affinity LIF receptor, and to a soluble form of the LIF receptor. LIF is incapable of binding either high- or low-affinity OSM receptors, yet OSM is capable of binding the high-affinity but not the low-affinity LIF receptor. Since the presence of high-affinity LIF receptors correlates with the biological activity of LIF on a wide range of target cells, we predict that OSM should have similar effects on LIF-responsive cells.
Assuntos
Citocinas/metabolismo , Interleucina-6 , Peptídeos/metabolismo , Receptores de Citocinas , Receptores Imunológicos/metabolismo , Animais , Clonagem Molecular , Inibidores do Crescimento , Humanos , Fator Inibidor de Leucemia , Subunidade alfa de Receptor de Fator Inibidor de Leucemia , Linfocinas , Oncostatina M , Receptores Imunológicos/genética , Receptores de OSM-LIF , Células Tumorais CultivadasRESUMO
The expression of certain normal genes requires a specific ribosomal frameshift event because the mRNA has the coding information for one protein in two different reading frames. One of several possible mechanisms for this involves recognition of a nontriplet codon by a noncognate tRNA. The AGUC-decoding Escherichia coli tRNASer3 reads a GCA alanine codon to cause a -1 frameshift. Replacement of the anticodon of tRNAPhe with the anticodon of tRNASer3 allows the constructed tRNA to cause this frameshifting. By altering the anticodon loop nucleotides at positions 33-36 in the constructed tRNAPhe molecules, the tRNA was found to recognize a 2-base codon. Instead of the usual anticodon, positions 34-36, the nucleotides in positions 34 and 35 form essential base pairs with the first two positions of the alanine codon. The uridine in position 36 is also required but not for base pairing.
Assuntos
Anticódon/genética , RNA Bacteriano/genética , Aminoacil-RNA de Transferência/genética , RNA de Transferência/genética , Sequência de Bases , Escherichia coli/genética , Regulação da Expressão Gênica , Mutação , Conformação de Ácido Nucleico , Ribossomos/metabolismoRESUMO
Oncostatin M (OSM) was initially identified as a polypeptide cytokine which inhibited the in vitro growth of cells from melanoma and other solid tumors. OSM shows significant similarities in primary amino acid sequence and predicted secondary structure to leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), granulocyte colony-stimulating factor (G-CSF), interleukin 6 (IL-6), and interleukin 11 (IL-11). Analysis of the genes encoding these proteins reveals a shared exon organization suggesting evolutionary descent from a common ancestral gene. Recent data indicates that OSM also shares a number of in vitro activities with other members of this cytokine family. The overlapping biological effects appear to be explained by the sharing of receptor subunits.
Assuntos
Citocinas/fisiologia , Peptídeos/fisiologia , Receptores de Citocinas , Sequência de Aminoácidos , Animais , Sequência de Bases , Citocinas/genética , DNA/genética , Humanos , Dados de Sequência Molecular , Estrutura Molecular , Oncostatina M , Peptídeos/genética , Peptídeos/farmacologia , Receptores de Superfície Celular/fisiologia , Receptores de Oncostatina M , Homologia de Sequência de Aminoácidos , Transdução de Sinais , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Oncostatin M (OSM) is a 28-kDa glycoprotein produced by stimulated macrophages and T lymphocytes that inhibits the proliferation of a number of different cell lines derived from solid tumors. Analysis of both amino acid sequence and gene structure has demonstrated that OSM is a member of a cytokine family that includes leukemia inhibitory factor (LIF), IL-6, and granulocyte colony-stimulating factor (G-CSF). We demonstrate that, like LIF, IL-6 and G-CSF, OSM can induce the differentiation of the myeloblastic M1 murine leukemia cells into macrophage-like cells. The morphologic and functional changes induced by OSM are more similar to those observed with LIF and IL-6 than those induced with G-CSF. OSM can also induce the differentiation of the histiocytic U937 human leukemia cells in the presence of granulocyte-macrophage CSF, a property shared with LIF and IL-6. In murine M1 cells, binding of labeled OSM is completely inhibited by excess LIF or OSM, reflecting the binding of OSM to the high affinity form of the murine LIF receptor. In contrast, the binding of labeled OSM to human U937 leukemia cells is inhibited by OSM, but the inhibition by LIF is significantly less. These results suggest that, in human leukemia cells, OSM may act through the LIF receptor and an OSM-specific receptor. The existence of an OSM-specific receptor was confirmed by both growth inhibition and competition binding assays on A375 human melanoma cells. The growth of human A375 cells was inhibited by OSM and IL-6 but not LIF or G-CSF. Neither LIF, G-CSF, nor IL-6 could compete with the binding of labeled OSM to A375 cells.