RESUMO
Thalidomide is an effective drug for chronic inflammatory diseases, but the mechanism underlying its immunomodulatory action remains uncertain. Thalidomide has been reported to clinically improve chronic inflammatory granulomatous disorders. In such disorders, the granulomas consist of epithelioid cells, scattered lymphocytes and multinucleated giant cells (MNGC; Langhans-type cells). The present experimental approach permitted the reproduction of MNGC formation from peripheral blood monocytes and examination of thalidomides effect on it. MNGC can be effectively generated from monocytes cultured in the presence of interleukin-4 (IL-4) and macrophage colony-stimulating factor(M-CSF) for 14 days. Thalidomide can inhibit the formation of MNGC in a dose-dependent manner. MNGC formation was partly inhibited by the presence of neutralizing TNF-alpha antibody in the responses induced by IL-4 and M-CSF. Autocrinal TNF-alpha production and modulation of cadhelin expression to regulate cell adhesion might be involved in this inhibitory action of thalidomide. Our results support thalidomides clinical efficacy in the treatment of chronic granulomatous disorders (granulomatosis).
Assuntos
Anti-Inflamatórios/farmacologia , Transdiferenciação Celular/efeitos dos fármacos , Células Gigantes de Langhans/efeitos dos fármacos , Granuloma/tratamento farmacológico , Monócitos/efeitos dos fármacos , Talidomida/farmacologia , Anticorpos , Comunicação Autócrina/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Células Gigantes de Langhans/imunologia , Células Gigantes de Langhans/patologia , Granuloma/imunologia , Granuloma/patologia , Humanos , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-4/metabolismo , Fator Estimulador de Colônias de Macrófagos/metabolismo , Monócitos/imunologia , Monócitos/patologia , Interferência de RNA , Receptores do Fator de Necrose Tumoral/genética , Receptores do Fator de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
AIMS: To measure antibacterial activity of the semi-synthetic flavonoid 3-O-octanoyl-(-)-epicatechin and investigate the mechanism of action. METHODS AND RESULTS: MICs determined by the broth microdilution method were 50 microg ml(-1) for beta-lactam sensitive and resistant Staphylococcus aureus, and 100 microg ml(-1) for vancomycin sensitive and resistant enterococci. In time-kill studies, 100 microg ml(-1) 3-O-octanoyl-(-)-epicatechin reduced colony forming unit numbers of antibiotic sensitive and methicillin-resistant Staph. aureus below detectable levels within 120 min. Bacterial aggregation was not observed when cells exposed to 3-O-octanoyl-(-)-epicatechin were examined by light microscopy. It was also shown that 50 microg ml(-1) 3-O-octanoyl-(-)-epicatechin is capable of reducing colony forming unit numbers of high cell density Staph. aureus populations by 80-fold within 60 min incubation, and inducing leakage of 50% of their internal potassium within just 10 min. CONCLUSIONS: 3-O-Octanoyl-(-)-epicatechin is active against Gram-positive bacteria, has bactericidal activity against both antibiotic sensitive and resistant strains, and is likely to exert its primary antibacterial effect by damaging the cytoplasmic membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: 3-O-Octanoyl-(-)-epicatechin has significant antibacterial activity and additional structural modification and/or formulation studies may allow this to be potentiated.
Assuntos
Antibacterianos/farmacologia , Catequina/análogos & derivados , Enterococcus/efeitos dos fármacos , Flavonoides/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Catequina/farmacologia , Agregação Celular/efeitos dos fármacos , Farmacorresistência Bacteriana/fisiologia , Enterococcus/citologia , Testes de Sensibilidade Microbiana , Staphylococcus aureus/citologiaRESUMO
New series histone deacetylase inhibitors comprising a hydroxamic acid or 2-aminobenzamide group as a zinc-chelating function were synthesized and evaluated for antiproliferative activities against a panel of human cancer cells. The 2-aminobenzamide series inhibitors generally had the potency in cell growth inhibitions comparable to that of MS-275. Among them, the compound having a (3,4-difluorobenzyl)(2-hydroxyethyl)amino group at one end and a 2-aminobenzamide group at the other of molecule showed the most promising profile as an anticancer drug candidate, since it had a comparatively low toxicity as did MS-275 against a normal fibroblast cell CCD-1059SK. Additionally, the derivative exhibited a high recovery in human plasma stability test.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Inibidores de Histona Desacetilases , Ácidos Hidroxâmicos/síntese química , Ácidos Hidroxâmicos/farmacologia , ortoaminobenzoatos/síntese química , ortoaminobenzoatos/farmacologia , Antineoplásicos/sangue , Inibidores Enzimáticos/sangue , Humanos , Ácidos Hidroxâmicos/sangue , Espectroscopia de Ressonância Magnética , Espectrometria de Massas/métodos , Espectrofotometria Infravermelho , ortoaminobenzoatos/sangueRESUMO
For the further characterization of bovine leukemia virus (BLV)-induced leukemogenesis, we investigated the association between polymorphism of ovine leukocyte antigen (OLA)-DRB1 gene and tumor development after infection of sheep with BLV. We infected 28 sheep with BLV and cloned exon 2 of the OLA-DRB1 gene from asymptomatic animals and from animals with lymphoma Sequence analysis revealed that, among 12 healthy sheep without any evidence of tumor, ten (83.3%) carried DRB1 alleles encoding Arg-Lys (RK) at positions beta70/71 as compared with only 6 (37.5%) of the 16 sheep with lymphoma, which suggested that alleles encoding the RK motif might protect against development of tumors after infection by BLV. By contrast, alleles encoding Ser-Arg (SR) at positions beta70/71 were present at a significantly elevated frequency in sheep with lymphoma as compared with the healthy carriers, which indicated that OLA-DRB1 alleles encoding the SR motif might be positively related to susceptibility to tumor development. The two amino acids in these motifs line a pocket that accommodates the side chain of a bound peptide according to a model of the crystal structure of human leukocyte antigen (HLA)-DR1. To analyze immunoreactions of sheep with alleles that encoded RK or SR at beta70/71, we selected sheep with either the RK/SR genotypes or the SR/SR genotypes and immunized them with a mixture of multiple synthetic antigenic peptides that corresponded to T-helper, T-cytotoxic, and B-cell epitopes of the BLV envelope glycoprotein gp51. Two weeks after the last immunization, all of the sheep were challenged with BLV. Sheep with the RK/SR genotype produced neutralizing antibodies against BLV; they eliminated BLV completely within 28 weeks of the BLV challenge, and they gave strong lymphocyte-proliferative responses to the peptides used for immunization. Moreover, such animals did not develop lymphoma. By contrast, sheep with the SR/SR genotype continued to produce BLV throughout the experimental period and developed terminal disease. Our results indicate that the differences in immunoresponse were due to differences in major histocompatibility complex class II alleles and reflected the risk of BLV-induced leukemogenesis. In addition, it appears that susceptibility to tumor development may be determined to some extent by polymorphic residues binding to antigenic peptides directly within the binding cleft of the OLA-DR molecule.
Assuntos
Alelos , Leucose Enzoótica Bovina/imunologia , Antígenos HLA-DR/genética , Linfoma/imunologia , Sequência de Aminoácidos , Animais , Bovinos , Suscetibilidade a Doenças , Genótipo , Cadeias HLA-DRB1 , Imunização , Vírus da Leucemia Bovina/imunologia , Dados de Sequência Molecular , OvinosRESUMO
We examined membrane fluidity of bovine adrenal chromaffin cells and chromaffin granules using cationic trimethylammonium derivative of diphenylhexatriene (TMA-DPH) as a fluorescence probe. After adding TMA-DPH to the suspension of chromaffin cells and that of granules, it first bound to the outer layer of the plasma membrane of the cells and that of the granule membrane, then gradually penetrated the inner layer of each membrane and distributed to both leaflets of the respective membranes. Accompanying increases in the ratio of incorporated probe on the cytoplasmic side of the chromaffin cell membrane, its fluorescence anisotropy gradually decreased. However, in chromaffin granules, the fluorescence anisotropy gradually increased with increases in the ratio of incorporated probe. These findings suggest that the inner layer of the plasma membrane and outer layer of the granular membrane are more fluid than the corresponding side of each membrane, which is suitable for the fusion between both membranes. We also examined the effect of trichosporin-B-VIa, a fungal ion channel forming alpha-aminoisobutyric acid-containing peptide, on the fluidity of chromaffin cells using TMA-DPH. The peptide decreased the fluorescence anisotropy and increased the fluorescence intensity in the concentration range that induced Ca2+ dependent catecholamine secretion, suggesting that a change in lipid dynamics of the lipid bilayer of the plasma membrane was induced by this peptide.
Assuntos
Antibacterianos/química , Membrana Celular/química , Células Cromafins/química , Células Cromafins/ultraestrutura , Grânulos Citoplasmáticos/química , Fluidez de Membrana , Peptídeos , Animais , Peptídeos Catiônicos Antimicrobianos , Bovinos , Fluorescência , Canais Iônicos/químicaRESUMO
Two hemorrhagic principles (Bitis arietans hemorrhagin a and b: abbreviated as BHRa and BHRb) were purified from the venom of the viperous snake Bitis arietans (puff adder) by gel filtration, ion-exchange and absorption chromatography. A 10-fold purification was achieved for BHRa and 7-fold for BHRb with an overall yield of 6.4% of hemorrhagic activity. The hemorrhagins were homogeneous according to disc- and SDS-polyacrylamide gel electrophoresis and immunodiffusion. BHRa and BHRb consist of 623 and 685 amino-acid residues and their apparent molecular weights were 68,000 and 75,000, respectively. They were also immunologically distinct. The purified hemorrhagins express proteolytic activity with heat-denatured casein and hide powder azure. The proteolytic activity with heat-denatured casein was almost the same as that of the crude venom, but that with hide powder azure was less than one-tenth of that of the crude venom. The purified hemorrhagins were free of arginine esterase and phospholipase A2 activities and they are acid labile hemorrhagic toxins. Their hemorrhagic activity was inhibited by EDTA, cysteine and by polyvalent anti-snake serum, but not by phenylmethanesulfonyl fluoride or soybean trypsin inhibitor.
Assuntos
Proteínas de Répteis , Venenos de Víboras/química , Venenos de Víboras/isolamento & purificação , Aminoácidos/análise , Animais , Caseínas/metabolismo , Cromatografia , Cromatografia em Gel , Cromatografia por Troca Iônica , Compostos Cromogênicos/metabolismo , Cisteína/farmacologia , Ácido Edético/farmacologia , Eletroforese em Gel de Poliacrilamida , Hemorragia/induzido quimicamente , Concentração de Íons de Hidrogênio , Hidrólise , Soros Imunes/farmacologia , Imunodifusão , Peso Molecular , Compostos Orgânicos , Coelhos , Venenos de Víboras/metabolismoRESUMO
Trichosporin-B-VIa (TS-B-VIa) has a Pro14-kinked helical structure which is considered to be important for the formation of peptaibol-type ion-channels in lipid bilayer membranes. TS-B-VIa and its analog [Aib14]TS-B-VIa with Pro-->Aib substitution at position 14, resulting in a straight helical structure, were tested for ion-channel-forming activity in planar lipid bilayer membranes and for ability to induce catecholamine secretion from cultured bovine adrenal chromaffin cells. Voltage-dependent multi-channel conductance, which is characteristic of TS-B-VIa, was also observed for [Aib14]TS-B-VIa. In single-channel measurements, current fluctuations induced by [Aib14]TS-B-VIa had a shorter life-time and showed fewer substates than those induced by TS-B-VIa. Catecholamine secretion induced by these peptides at low concentrations is completely Ca(2+)-dependent. At high concentrations, TS-B-VIa-induced secretion was partly independent of external Ca2+, but this was not the case for the analog. The differences of behavior can be explained in terms of the differences of hydrophobicity, and magnitude of dipole moment due to the conformational changes around position 14 and the C-terminal domain caused by the Pro-->Aib substitution.
Assuntos
Antibacterianos/química , Catecolaminas/metabolismo , Canais Iônicos , Peptídeos , Prolina , Glândulas Suprarrenais/efeitos dos fármacos , Glândulas Suprarrenais/metabolismo , Sequência de Aminoácidos , Animais , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos , Cálcio/farmacologia , Bovinos , Células Cultivadas , Sistema Cromafim/efeitos dos fármacos , Sistema Cromafim/metabolismo , Condutividade Elétrica , Dados de Sequência Molecular , Estrutura Secundária de Proteína , Relação Estrutura-AtividadeRESUMO
Our previous study shows that 6-O-acyl derivatives of L-ascorbic acid inhibits more markedly cell growth of mouse Ehrlich carcinoma than ascorbic acid. The present study shows that 6-O-palmitoyl ascorbic acid but not ascorbic acid prolongs the lifespan of mice into which tumors such as Meth A fibrosarcoma, MM46 mammary carcinoma, Ehrlich carcinoma and sarcoma 180 are implanted. The potentiated cytotoxicity of 6-O-palmitoyl ascorbic acid is not due to an increase in duration time of the cytotoxic action, because 6-O-palmitoyl ascorbic acid is gradually inactivated during contact with tumor cells and exhibits a similar action time curve to that of ascorbic acid as shown by clonal growth assay. Cytotoxicity of 6-O-palmitoyl ascorbic acid is markedly diminished by combined addition of catalase and superoxide dismutase (SOD), as shown by dye exclusion assay, whereas the cytotoxicity was slightly reduced by either enzyme alone but not by the specifically inactivated or heat-denatured enzymes. In contrast, cytotoxicity of ascorbic acid is abolished by catalyse but not SOD. Autooxidation of 6-O-palmitoyl ascorbic acid was not inhibited by catalase plus SOD. The results indicate that cytotoxicity of 6-O-palmitoyl ascorbic acid is attributed at least partly to both hydrogen peroxide (H2O2) and superoxide (O2-.) generated at the early stage. Cytotoxicity of 6-O-palmitoyl ascorbic acid is also appreciably attenuated by singlet oxygen (1O2) scavengers such as hydroquinone, 1,4-diazobicyclo-2,2,2-octane or sodium azide, but not by hydroxyl radical scavengers including butylated hydroxytoluene, D-mannitol, benzoic acid and ethanol. Thus, in contrast to cytotoxicity of ascorbic acid mediated entirely by H2O2 initially generated, acylated ascorbic acid produces a diversity of active oxygen species including H2O2, O2-. and other species secondarily generated via disproportion, which may be additively involved in the enhanced cytotoxic action.
Assuntos
Antineoplásicos/uso terapêutico , Ácido Ascórbico/análogos & derivados , Neoplasias Experimentais/tratamento farmacológico , Animais , Ácido Ascórbico/farmacologia , Ácido Ascórbico/uso terapêutico , Ácido Ascórbico/toxicidade , Catalase/antagonistas & inibidores , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Radicais Livres , Humanos , Leucemia Experimental/tratamento farmacológico , Neoplasias Pulmonares , Camundongos , Oxirredução , Superóxido Dismutase/antagonistas & inibidoresRESUMO
Trichosporin (TS) -B-VIa, a fungal alpha-aminoisobutyric acid (Aib) -containing peptide consisting of 19 amino acid residues and a phenylalaninol, produced both 45Ca2+ influx into bovine adrenal chromaffin cells and catecholamine secretion from the cells. The secretion induced by TS-B-VIa at lower concentrations (2-5 microM) was completely dependent on the external Ca2+, while that induced by TS-B-VIa at higher concentrations (10-30 microM) was partly independent of the Ca2+. The concentration-response curves (2-5 microM) for the TS-B-VIa-induced Ca2+ influx and secretion correlated well. The TS-B-VIa (at 5 microM) -induced secretion was not antagonized by diltiazem, a blocker of L-type voltage-sensitive Ca2+ channels. The treatment of fura-2-loaded C6 glioma cells with TS-B-VIa (2-5 microM) led to an increase in the intracellular free Ca2+ concentration ([Ca2+]i) in a concentration-dependent manner but the stimulatory effects of TS-B-VIa on [Ca2+]i were only slightly observed in Ca(2+)-free medium, indicating that TS-B-VIa causes Ca2+ influx from the external medium into the C6 cells. The TS-B-VIa-induced increase in [Ca2+]i in the C6 cells was not antagonized by diltiazem and by SK&F 96365, a novel blocker of receptor-mediated Ca2+ entry. High K+ increased neither [Ca2+]1 in the C6 cells nor Mn2+ influx into the cells, while TS-B-VIa increased Mn2+ influx. Also in other non-excitable cells, bovine platelets, similar results were obtained. These results strongly suggest that the mechanism of Ca2+ influx by TS-B-VIa at the lower concentrations is distinct from the event of Ca2+ influx through receptor-operated or L-type voltage-sensitive Ca2+ channels in both excitable cells (the chrornaffin cells) and non-excitable cells (the C6 cells and the platelets) and that TS-B-VIa per se may form Ca(2+)-permeable ion channels in biological membranes. On the other hand, the peptide at the higher concentrations seems to damage cell membranes.
Assuntos
Ácidos Aminoisobutíricos/análise , Antibacterianos/farmacologia , Cálcio/metabolismo , Proteínas Fúngicas/farmacologia , Peptídeos , Trichoderma/química , Glândulas Suprarrenais/efeitos dos fármacos , Glândulas Suprarrenais/fisiologia , Animais , Antibacterianos/análise , Peptídeos Catiônicos Antimicrobianos , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Catecolaminas/metabolismo , Bovinos , Células Cultivadas , Sistema Cromafim/efeitos dos fármacos , Sistema Cromafim/fisiologia , Diltiazem/farmacologia , Endotelinas/farmacologia , Proteínas Fúngicas/análise , Glioma/metabolismo , Manganês/metabolismo , Potássio/farmacologia , Células Tumorais CultivadasRESUMO
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is likely to be caused by continuous imperfection of bone healing after surgical treatments in patients with long-term administration of nitrogen-containing bisphosphonates (NBPs). NBPs inhibit osteoclastic bone resorption by impairing the mevalonic acid sterol pathway in osteoclasts. Thus, we hypothesized that exogenous mevalonic acid metabolites restore the inhibitory effects of NBPs on osteoclastogenesis and bone remodeling. To clarify the effects of mevalonic acid metabolites, especially geranylgeranyl pyrophosphate (GGPP) and geranylgeranyl transferase substrate geranylgeranyl acid (GGOH), we examined the effects of zoledronic acid with or without GGOH or GGPP on osteoclast differentiation, multinucleation, and bone mineral deposition in tooth-extracted sockets. Zoledronic acid decreased the number of tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cells derived from mouse osteoclast precursors treated with receptor activator of nuclear factor-κB ligand and macrophage colony-stimulating factor. Zoledronic acid simultaneously suppressed not only the expressions of osteoclastic differentiation-related molecules such as TRAP, cathepsin K, calcitonin receptor, and vacuolar H-ATPase but also those of multinucleation-related molecules such as dendrocyte-expressed 7 transmembrane proteins and osteoclast stimulatory transmembrane protein. Treatment with GGOH or GGPP, but not farnesyl acid, restored the zoledronic acid-inhibited number of TRAP-positive multinuclear cells together with the expressions of these molecules. Although intraperitoneal administration of zoledronic acid and lipopolysaccharide into mice appeared to induce BRONJ-like lesions with empty bone lacunae and decreased mineral deposition in tooth-extracted socket, both GGOH and GGPP partially restored the inhibitory effects on zoledronic acid-related mineral deposition. These results suggest the potential of mevalonic acid metabolites as therapeutic agents for BRONJ.
Assuntos
Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Imidazóis/farmacologia , Ácido Mevalônico/farmacologia , Osteoclastos/efeitos dos fármacos , Fosfatase Ácida/análise , Proteínas Adaptadoras de Transdução de Sinal/efeitos dos fármacos , Animais , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/etiologia , Remodelação Óssea/efeitos dos fármacos , Calcificação Fisiológica/efeitos dos fármacos , Catepsina K/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Diterpenos/farmacologia , Farneseno Álcool/farmacologia , Isoenzimas/análise , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Masculino , Maxila/efeitos dos fármacos , Proteínas de Membrana/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Fosfatos de Poli-Isoprenil/farmacologia , Receptores da Calcitonina/efeitos dos fármacos , Salmonella , Fosfatase Ácida Resistente a Tartarato , Alvéolo Dental/efeitos dos fármacos , ATPases Vacuolares Próton-Translocadoras/efeitos dos fármacos , Ácido ZoledrônicoRESUMO
Recent investigations have revealed the crucial role of von Willebrand factor (vWF) in platelet thrombus formation under flow conditions. The plasma concentrations of vWF were measured together with various hemodynamic and hemostatic parameters in 51 cases of acute myocardial infarction. In 10 randomly selected cases, the plasma concentrations and distribution of multimers vWF were serially determined after reperfusion therapy by percutaneous transluminal coronary angioplasty (PTCA). The vWF concentration at the onset of the acute myocardial infarction was significantly higher than in an age-matched control group (vWF AG: 18.7 +/- 1.2 microg/ml vs. 10.3 +/- 0.5 microg/ml, p = 8.43 x 10-(12), mean +/- SE). Simultaneous determination of hemodynamic and hemostatic parameters revealed that the only two parameters that were significantly correlated with the patients' plasma vWF concentrations were their pulmonary capillary wedge pressure (PCWP) and heart rate, suggesting a relationship between hemodynamic changes induced by the onset of myocardial infarction and the vWF plasma concentrations. Serial determinations revealed that the vWF concentrations had not changed 1 h after reperfusion therapy, but that they significantly increased by 24 to 72 h. The distribution of the larger multimers of vWF also increased in the acute and subacute phase. The vWF concentration and multimer distribution normalized 14 days after the onset of the myocardial infarction. Our findings suggest that the vWF concentration increased in acute myocardial infarction patients, possibly in association with the hemodynamic deterioration that occurs in acute myocardial infarction.
Assuntos
Infarto do Miocárdio/sangue , Fator de von Willebrand/metabolismo , Adulto , Idoso , Angioplastia Coronária com Balão , Trombose Coronária/sangue , Dimerização , Fibrinolíticos/metabolismo , Hemodinâmica , Hemostáticos/metabolismo , Humanos , Modelos Lineares , Análise por Pareamento , Pessoa de Meia-Idade , Reperfusão Miocárdica , Stents , Fatores de TempoRESUMO
The distribution of thymic tissue at the anterior mediastinum was examined histologically in 18 cases following the removal of the adipose tissue which was located outside the thymic capsule at the time of thymectomy for myasthenia gravis. The gross adipose tissue revealed the presence of histological thymic tissue, including Hassall's bodies, in 13 of 18 cases. In order to be sure of extirpating all of the thymic tissue, the adipose tissue at the anterior mediastinum as well as the gross thymus should be removed.
Assuntos
Mediastino/anatomia & histologia , Timectomia/métodos , Timo/anatomia & histologia , Tecido Adiposo/anatomia & histologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miastenia Gravis/cirurgia , Timo/embriologiaRESUMO
BACKGROUND: Hyperlipidemia has recently received attention as being involved in the progression of diabetic nephropathy (DN). Low-density lipoprotein apheresis (LDL-A) can remove a large amount of plasma lipid directly from the patients in a short time. METHODS: Fifteen type 2 diabetic patients with overt nephropathy received LDL-A in two different manners: short-term intensive therapy (SIT) for nine patients and long-term intermittent therapy (LIT) for six patients. RESULTS: The changes in the monthly decline rates of reciprocal serum creatinine (1/Cr) were -0.035 +/- 0.020 in the three-month period before SIT, 0.047 +/- 0.041 during and until two weeks after SIT, and -0.035 +/- 0.015 after a period of two weeks from the therapy. The mean duration of LIT in six patients was 8.2 +/- 7.4 months, and the mean monthly decline rates of 1/Cr significantly decreased during the period of LIT as compared with the six-month period before the treatment. CONCLUSION: LDL-A can retard the progression of overt DN, especially when it is performed repeatedly for a long period at two-week intervals.
Assuntos
Remoção de Componentes Sanguíneos , Nefropatias Diabéticas/terapia , Hiperlipidemias/sangue , Lipoproteínas LDL/sangue , Idoso , Colesterol/sangue , Creatinina/sangue , Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/patologia , Progressão da Doença , Feminino , Humanos , Rim/patologia , Rim/fisiopatologia , Lipídeos/sangue , Lipoproteínas VLDL/sangue , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Triglicerídeos/sangueRESUMO
The mass spectral characteristics of selected peptaibols were investigated using electrospray ionization in conjunction with low-energy collision-induced dissociation (CID). Protonated and sodiated molecular species were selected as precursor ions and the resulting CID spectra are discussed with respect to fragmentation characteristics related to the structures of peptaibols. The CID spectra of peptaibols with acetylated N-terminus and with the sub-sequences of Aib-Pro were similar. The CID spectra of [M + 2Na]2+ provided structural information more than those of [M + 2H]2+ and [M + H + Na]2+. The CID of [M + 2Na]2+ can be used to deduce complete sequence information for peptaibols.
Assuntos
Antibacterianos/química , Fungos/metabolismo , Espectrometria de Massas , Fragmentos de Peptídeos/química , Sequência de Aminoácidos , Antibacterianos/análise , Dados de Sequência MolecularRESUMO
The antihemorrhagic activity of muscle extract of hedgehog, Erinaceus europaeus, was tested on various snake venoms with hemorrhagic activity. The extract inhibited strongly hemorrhagic activity of venoms from Bitis arietans, Bothrops jararaca and Vipera latastei gaditana, and remarkably that of venoms from Agkistrodon halys blomhoffi, Bitis gabonica rhinoceros, Bitis nasicornis, Bothrops atrox asper, Crotalus horridus horridus and Vipera berus. The antihemorrhagic activity against eight other snake venoms was below the detection level.
Assuntos
Antivenenos/farmacologia , Ouriços/metabolismo , Hemorragia/tratamento farmacológico , Músculos/metabolismo , Venenos de Serpentes/toxicidade , Animais , Antivenenos/isolamento & purificação , Antivenenos/uso terapêutico , Proteínas Sanguíneas/isolamento & purificação , Proteínas Sanguíneas/farmacologia , Proteínas Sanguíneas/uso terapêutico , Bothrops , Venenos de Crotalídeos/toxicidade , Hemorragia/induzido quimicamente , Venenos de Víboras/toxicidadeRESUMO
The ability of purified antihemorrhagic factor isolated from the serum of Japanese Habu (Trimeresurus flavoviridis) was tested on 17 snake venoms to inhibit their hemorrhagic activity. The factor strongly inhibited that of the venoms of Crotalus horridus horridus and Vipera latastei gaditana in addition to that of the homologous (T. flavoviridis) venom. Hemorrhagic activity of Agkistrodon halys blomhoffi, Agkistrodon contortrix contortrix, Bothrops atrox asper and Crotalus atrox venoms was also remarkably inhibited, but that of Vipera mauretanica to less extent. The hemorrhagic activities of nine other snake venoms were not inhibited.
Assuntos
Antivenenos/farmacologia , Proteínas Sanguíneas/farmacologia , Hemorragia/prevenção & controle , Venenos de Serpentes/antagonistas & inibidores , Trimeresurus , Animais , Antivenenos/isolamento & purificação , Antivenenos/uso terapêutico , Proteínas Sanguíneas/isolamento & purificação , Proteínas Sanguíneas/uso terapêutico , Venenos de Crotalídeos/antagonistas & inibidores , Venenos de Crotalídeos/toxicidade , Relação Dose-Resposta a Droga , Hemorragia/induzido quimicamente , Coelhos , Pele/efeitos dos fármacos , Venenos de Serpentes/toxicidadeRESUMO
An antihaemorrhagic factor named erinacin was purified from the skeletal muscle extract of the European hedgehog, Erinaceus europaeus, by ammonium sulfate precipitation followed by various steps of ion-exchange (DEAE-cellulose), absorption chromatography (hydroxylapatite), and gel filtration (cellofine gel). A 625-fold purification was achieved with an overall yield of 19% antihaemorrhagic activity. The protein effectively inhibited the activity of Bothrops jararaca venom haemorrhagin and did not inhibit the enzymatic activity of trypsin and chymotrypsin. Erinacin is a large molecule (about 1,000,000 mol. wt). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of two subunits: one with an apparent mol. wt of 35,000 forming a larger subunit (350,000) by cross-linking with disulfide bridges, and a second with a mol. wt of 39,000 without disulfides. Dissociation of erinacin into its subunits resulted in complete loss of its antihaemorrhagic activity.
Assuntos
Antivenenos/isolamento & purificação , Proteínas Sanguíneas/isolamento & purificação , Proteínas Musculares/isolamento & purificação , Animais , Antivenenos/química , Antivenenos/farmacologia , Proteínas Sanguíneas/química , Proteínas Sanguíneas/farmacologia , Ouriços , Peso Molecular , Proteínas Musculares/química , Proteínas Musculares/farmacocinética , Proteínas Musculares/farmacologia , Inibidores de Proteases/isolamento & purificação , Inibidores de Proteases/farmacologiaRESUMO
Antihemorrhagic activities of skeletal muscle extracts from various animals were compared in inhibiting the hemorrhagic activity of Bothrops jararaca venom. The muscle extracts of the European hedgehog (Erinaceus europaeus) exhibited the strongest activity, followed by those of other insectivores such as the shrew (Crocidura russula) and mole (Talpa europaea). The antihemorrhagic activities of muscle extracts from experimental animals such as mice, rats, guinea-pigs, hamsters and rabbits were negligible.
Assuntos
Hemorragia/prevenção & controle , Músculo Esquelético , Venenos de Serpentes/antagonistas & inibidores , Extratos de Tecidos/uso terapêutico , Animais , Bothrops , Cricetinae , Cobaias , Ouriços , Camundongos , Toupeiras , Coelhos , Ratos , Musaranhos , Venenos de Serpentes/toxicidade , Especificidade da EspécieRESUMO
The rabbit pyrogen test (specified in the pharmacopeia) and the Limulus amebocyte lysate (LAL) test are influenced by high concentrations of certain antibiotics. Therefore, it has not been possible to detect trace amounts of endotoxin which may contaminate these antibiotics. To detect trace amounts of endotoxin in injectable sodium ampicillin, an ultrafiltration technique was utilized which removed the antibiotic and left a solution which contained predominantly the endotoxin. After ultrafiltration, a trace amount of pyrogen (which otherwise could not be detected) was found using both the rabbit pyrogen and the LAL tests. The endotoxin was also determined quantitatively using a chromogenic endotoxin reagent which is made by combining the Limulus amebocyte lysate and a synthetic substrate with a suitable chromophore.