RESUMO
Small B-cell lymphocytic lymphoma/chronic lymphocytic leukemia, which typically affects elderly people, is a group of conditions that are not clinically uniform. It has been suggested that using the combined activity of the monoclonal antibody anti-CD20 (rituximab) and Listeria monocytogenes toxin listeriolysin O (LLO) for this condition could produce an enhanced treatment effect. Here, we tested the effect of the joint activity of rituximab and LLO, which is a cell membrane toxin, in human leukemia cell lines. The human B-leukemia Raji cell line, which expresses CD20, and the T-cell Jurkat cell line, which does not express CD20, for comparison were used in model tests. Cell cytotoxicity of rituximab or LLO and both applied jointly to the cell lines was compared in the presence of human plasma complement. Optimal cytotoxic effects dependent on rituximab or LLO concentration were tested separately. LD50 values were determined and used for optimal application of a mixture of the two factors. The cytotoxic effect on Raji cells of both rituximab and LLO was more than 2.5 times that of LLO alone and 1.5 times that of rituximab alone. At the highest tested concentrations, a mixture of the tested factors had a non-specific cytotoxic effect on the Jurkat cell line, as well. The rituximab and LLO binding sites appear to be in a similar region of the Raji leukemia cell membrane, suggesting an effective interaction of both factors. The joint interaction of these compounds in cell membrane pore formation suggests an explanation for the more effective cytotoxic activity that their combination was observed in this experiment.
Assuntos
Anticorpos Monoclonais/farmacologia , Linfócitos B/efeitos dos fármacos , Toxinas Bacterianas/farmacologia , Proteínas de Choque Térmico/farmacologia , Proteínas Hemolisinas/farmacologia , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Rituximab/farmacologia , Antígenos CD20/metabolismo , Linhagem Celular Tumoral , Proteínas do Sistema Complemento/metabolismo , Humanos , Imunossupressores/farmacologiaRESUMO
Bacterial toxins can exhibit anticancer activities. Here we investigated the anticancer effects of the listeriolysin O toxin produced by Listeria monocytogenes. We found that supernatants of Listeria monocytogenes strains (wild type, 1189, and 1190) were cytotoxic to the Jurkat cell line and human peripheral blood mononuclear cells (PBMC) in a concentration-dependent manner. The supernatant of strain 1044, not producing listeriolysin O, was inactive. The supernatants of Listeria strains were also cytotoxic toward B cells of chronic leukemia patients, with no significant differences in activities between strains. We also tested supernatants of Bacillus subtilis strains BR1-90, BR1-S, and BR1-89 producing listeriolysin O. BR1-S and BR1-89 were cytotoxic to PBMC and to Jurkat cells, the latter being more sensitive to the supernatants. BR1-90 was not hemolytic or cytotoxic to PBMC, but was cytotoxic to Jurkat cells in the concentration range of 10-30%, suggesting that listeriolysin O is selectively effective against T cells. Overall, the response of human peripheral blood mononuclear and human leukemia cell lines to bacteria supernatants containing listeriolysin O depended on the bacteria strain, target cell type, and supernatant concentration.
Assuntos
Bacillus subtilis/metabolismo , Toxinas Bacterianas/administração & dosagem , Citotoxinas/administração & dosagem , Leucócitos/fisiologia , Listeria monocytogenes/metabolismo , Neoplasias Experimentais/tratamento farmacológico , Toxinas Bacterianas/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citotoxinas/metabolismo , Proteínas de Choque Térmico , Proteínas Hemolisinas , Humanos , Células Jurkat , Leucócitos/efeitos dos fármacosRESUMO
Implantable bioartificial liver has been investigated for patients suffering from liver insufficiency after mass liver resection or acute liver failure. Liver cells are implanted as free cell suspension, in microencapsulation systems or using microcarriers. To exhibit their typical functions, hepatic cells need a three-dimensional environment that is much more physiological than a flat one. The aim of our study was in vivo evaluation of spongy polyethersulfone membranes as a synthetic support for hepatic cells grown three dimensionally and transplanted to SCID/NOD mice. Spongy membranes were prepared using phase inversion from membrane-forming mixtures containing the following: polyethersulfone (based polymer), dimethylformamide (solvent), polyvinylpyrrolidone MW 10000 (small pore precursor), and cellulose (large pore precursor). We observed that polyethersulfone membranes were well tolerated by C3A cells and we did not observe any toxic effect, resulting in viability of cells >95%. Use of collagen gel as a support for cells on the scaffold gives the opportunity to increase 10 times the number of cells seeded on the membrane. Heparin addition to collagen gel did not influence albumin production in SCID/NOD mice. We observed an increase of albumin production after 7 and 14 days after implantation. Use of collagen gels in combination with polymer scaffolds allows preparation of bioartificial organs possessing high cell concentration for transplantation purposes.
Assuntos
Transplante de Células/métodos , Hepatócitos/citologia , Animais , Técnicas de Cultura de Células/métodos , Corantes , Hepatopatias/terapia , Membranas Artificiais , Camundongos , Camundongos Endogâmicos NOD , Polímeros , SulfonasRESUMO
INTRODUCTION: Autologous osteochondral transplantation is one method that can be used to create hyaline or hyaline-like repair in a defect area. The purpose of the present study was to repair full-thickness articular cartilage defects in 9 rabbit knee joints with autologous cultured chondrocytes. METHODS: An articular cartilage defect was created on the patellar groove of the femur. The defect was filled with chondrocytes cultured in vitro and placed into the knee on a polysulphonic membrane. At 8 weeks after the operation, the reparative tissue was analyzed macroscopically and histologically. RESULTS: At 8 weeks after the operation, the surfaces of the reparative tissue were smooth, and the defects were filled with mature hyaline cartilage in 5 cases. In 2 cases, the reparative hyaline cartilage was immature and there was worse integration of grafted tissue into the adjacent normal cartilage. In 2 cases, the surface of the grafted area was irregular, and the reparative tissue was disintegrated and incompletely differentiated. CONCLUSION: The results suggest that transplantation of autologous chondrocytes cultured in vitro and placed into the knee on polysulphonic membrane is effective in repairing an articular cartilage defect.
Assuntos
Cartilagem Articular/cirurgia , Condrócitos/transplante , Polímeros , Sulfonas , Animais , Materiais Biocompatíveis , Células Cultivadas , Condrócitos/citologia , Articulação do Joelho , Membranas Artificiais , Modelos Animais , CoelhosRESUMO
BACKGROUND: Philadelphia cells are human chronic myelogenous leukemia (CML) cells that contain the BCR/ABL oncogene (a fusion of the BCR and ABL genes). Selective eradication of these cells in vitro can be achieved by combined treatment with antisense phosphorothioate oligodeoxynucleotides ([S]ODNs) specifically targeted to this oncogene (bcr/abl [S]ODNs) and a suboptimal (for use as a single agent) dose of mafosfamide (the in vitro active form of cyclophosphamide). PURPOSE: We evaluated the ability of bcr/abl antisense [S]ODNs, alone or subsequent to treatment with a single injection of cyclophosphamide, to suppress the leukemic process induced in severe combined immunodeficient (SCID) mice by Philadelphia cells (i.e., primary CML-blast crisis [CML-BC] cells). In addition, we studied potential mechanisms that might explain the efficacy of the bcr/abl antisense [S]ODN-mafosfamide combination against Philadelphia cells in vitro. METHODS: The effects of treating leukemic mice with cyclophosphamide (25 mg/kg body weight; 25% of the dose required to eradicate evidence of leukemia in SCID mice) and/or bcr/abl antisense [S]ODNs were assessed by analysis of survival, by examination of bone marrow for the presence of leukemia cells (using a colony formation assay or using coupled reverse transcription and the polymerase chain reaction to screen for bcr/abl messenger RNA), and by examination of a variety of tissues for the presence of infiltrating leukemia cells. The induction of apoptosis (a cell death program) in vitro in primary CML-BC cells following treatment with bcr/abl antisense [S]ODNs plus or minus prior treatment with mafosfamide was monitored by use of a commercial assay. Relative cellular uptake of [S]ODNs by CML-BC cells treated in vitro with or without prior treatment with mafosfamide was determined by use of confocal microscopy and flow cytometry (for fluorescent [S]ODNs) or by use of blotting techniques that employed radioactively labeled probes (for extracted, unlabeled [S]ODNs). Levels of specific proteins in treated and untreated cells were determined by use of western blotting methods. Reported P values are two-sided. RESULTS: The disease process in leukemic mice was retarded substantially by combination treatment with cyclophosphamide and specific bcr/abl antisense [S]ODNs (P < .001, relative to treatment with specific antisense [S]ODNs alone, cyclophosphamide alone, or cyclophosphamide plus nonspecific [i.e., control] antisense [S]ODNs); 50% of the mice treated with cyclophosphamide and specific antisense [S]ODNs appeared to be cured of leukemia. The combination treatment was associated with increased induction of apoptosis. In addition, cellular uptake of bcr/abl antisense [S]ODNs appeared to be increased twofold to sixfold by prior treatment with mafosfamide. This increased uptake of [S]ODNs was associated with enhanced suppression of p210bcr/abl protein levels. CONCLUSIONS AND IMPLICATIONS: Combination therapy with antisense [S]ODNs targeted to specific oncogenes and less toxic doses of anticancer drugs may represent a rational strategy to purpose for the treatment of human leukemias.
Assuntos
Antineoplásicos/uso terapêutico , Ciclofosfamida/análogos & derivados , Proteínas de Fusão bcr-abl/efeitos dos fármacos , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Oligonucleotídeos Antissenso/uso terapêutico , Cromossomo Filadélfia , Tionucleotídeos/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Medula Óssea/efeitos dos fármacos , Células da Medula Óssea , Ciclofosfamida/uso terapêutico , Sondas de DNA , Citometria de Fluxo , Proteínas de Fusão bcr-abl/biossíntese , Proteínas de Fusão bcr-abl/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Camundongos , Camundongos SCID , Reação em Cadeia da Polimerase , Análise de Sobrevida , Resultado do Tratamento , Células Tumorais CultivadasRESUMO
The purpose of the observations was the viability and quality evaluation of E. coli bacteria encapsulated in hollow fiber membranes (HF) in short in vivo and in vitro experiments. A polypropylene, surface-modified hollow fiber was applied for immunoisolation of E. coli bacteria transfected with a green fluorescent protein (E. coli GFPI). The presence of GFP fluorescence of organisms was assessed with the use of flow cytometry. The E. coli GFPIs were then observed for the period of 5 days in in vitro experiments in the culture medium. A single IPTG (isopropyl beta-D-1-thiogalactopyranoside) induction of GFP gene appeared to be adequate for an expression of GFP protein for 5 days. The GFP expression values observed for E. coli GFPs encapsulated in HF during culture in different culture media were comparable. The survival of E. coli GFPIs encapsulated in HF after 1, 2, 4, or 5 days of subcutaneous implantation into mice was evaluated. The explanted E. coli GFPIs exhibited mean expression 603 +/- 17 (n = 32) units of fluorescence during the implantation period. The values obtained were comparable for selected days of observation. It was observed that the membranes applied ensured the bacteria growth within the HF's space only.
Assuntos
Escherichia coli/fisiologia , Polipropilenos , Animais , Biotecnologia/métodos , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Citometria de Fluxo , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Camundongos , Camundongos SCIDRESUMO
The efficacy of five disinfecting agents was tested by a new microbiological method, i.e. the flow cytometry. The method is based on kinetic measurements of the effects of a disinfecting agent on the percent of live/dead cells detected with propidium iodide. E. coli ATCC52922 strain and S. aureus ATCC 29213 strain were used in the experiment. From the measurements the killing time 50 (KT50) was estimated as the period of time needed to kill 50% of microorganisms in the disinfected volume. KT50 for ethanol 70% was 11s and it was the most efficient disinfecting agent among all examined. Other commercial preparations were compared with ethanol 70% and were traced throughout the period of 5 min. The results were obtained rapidly, frequently in less than 10 min. In conclusion, the effectiveness of a particular antibacterial disinfectant preparation may be estimated quantitatively within a few minutes by the flow cytometry. The method proved to be very useful for a fast comparison of the effectiveness of various disinfectant preparation against pathological microorganisms.
Assuntos
Desinfetantes/farmacologia , Escherichia coli/efeitos dos fármacos , Citometria de Fluxo/métodos , Staphylococcus aureus/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Propídio/metabolismo , Staphylococcus aureus/crescimento & desenvolvimento , Fatores de TempoRESUMO
Albumin is rarely used for electrospinning because it does not form fibres in its native globular form. This paper presents a novel method for electrospinning human albumin from a solution containing pharmaceutical grade protein and 25% polyethylene oxide (PEO) used as the fibre-forming agent. After spontaneous cross-linking at body temperature, with no further chemicals added, the fibres become insoluble and the excess PEO can be washed out. Albumin deposited along the fibres retains its native characteristics, such as its non-adhesiveness to cells and its susceptibility for degradation by macrophages. To demonstrate this we evaluated the mechanical properties, biocompatibility and biodegradability of this novel product. After subcutaneous implantation in mice, albumin mats were completely resorbable within six days and elicited only a limited local inflammatory response. In vitro, the mats suppressed cell attachment and migration. As this product is inexpensive, produced from human pharmaceutical grade albumin without chemical modifications, retains its native protein properties and fulfils the specific requirements for anti-adhesive dressings, its clinical use can be expedited. We believe that it could specifically be used when treating paediatric patients with epidermolysis bullosa, in whom non-healing wounds occur after minor hand injuries which lead to rapid adhesions and devastating contractures.
Assuntos
Materiais Biocompatíveis/farmacologia , Teste de Materiais/métodos , Engenharia Tecidual , Albuminas/ultraestrutura , Animais , Dicroísmo Circular , Feminino , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Leucócitos/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Microscopia de Força Atômica , Projetos Piloto , Polietilenoglicóis/química , Implantação de Prótese , Solubilidade , SoluçõesRESUMO
BALB/cxDBA/2Wf F1 (CD2F1) mice were lethally irradiated and reconstituted with syngeneic bone marrow cells untreated or treated with 75 micrograms/ml of mafosfamide. One day after bone marrow transplantation some groups of mice were injected with syngeneic splenocytes, peripheral blood leukocytes, or thymocytes. Seven days after marrow grafting the anti-L1210 leukemia immunization of mice, consisting of four i.p. injections of 10(6) L1210-Maf cells (L1210 cells treated in vitro with mafosfamide for inhibition of their growth in vivo), was started. Strong resistance against leukemia could be obtained only in mice receiving splenocytes or peripheral blood leukocytes, not in mice injected with thymocytes or in those not receiving any cells. In vitro elimination of various subpopulations from among the splenocytes before their injection into the mice made it possible to deduce which are necessary for early induction of antitumor resistance after bone marrow transplantation in mice. These cells are: Thy 1.2-, Ig-, AsGM 1-, Mac 1+, 1-Ad+/-, are adherent and nonsusceptible to carrageenan toxicity.
Assuntos
Leucemia Experimental/imunologia , Animais , Antineoplásicos/farmacologia , Células da Medula Óssea , Transplante de Medula Óssea , Adesão Celular , Ciclofosfamida/análogos & derivados , Ciclofosfamida/farmacologia , Feminino , Imunidade Inata , Imunoterapia Adotiva , Leucemia L1210/imunologia , Transfusão de Leucócitos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos , Fatores de Tempo , Transplante Isogênico , Irradiação Corporal TotalRESUMO
BALB/c x DBA/2 F1 (CD2F1) mice were lethally irradiated and reconstituted with syngeneic bone marrow cells (SBMC) obtained from normal or previously immunized (against L1210 lymphatic leukemia) donors. These recipient mice are called TBI + SBMT or TBI + Imm-SBMT mice, respectively. TBI + Imm-SBMT, but not TBI + SBMT mice, were able to develop strong immune resistance against L1210 leukemia, but not against MOPC 104E plasmacytoma, if the immunization procedure (four i.p. injections at weekly intervals of immunogenic L1210 cells) was started as early as 7 days posttransplantation. Incubation of Imm-SBMC with mafosfamide (ASTA Z7654) before grafting abrogated the ability of the recipient mice to develop early resistance against the leukemia. Treatment of Imm-SBMC with monoclonal or polyclonal antibodies plus complement showed that two or three subpopulations of Imm-SBMC were necessary for the transfer of immune information against leukemia: T lymphocytes with phenotype Thy 1.2+, Lyt 1+2-, I-Ad-, macrophages with phenotype Mac-1+, I-Ad-, and probably asialo-GM 1+ cells. Recipient mice immunized against L1210 leukemia before TBI + SBMT do not develop early resistance to the leukemia.
Assuntos
Transplante de Medula Óssea/imunologia , Leucemia L1210/cirurgia , Animais , Anticorpos/administração & dosagem , Antígenos de Neoplasias/administração & dosagem , Antineoplásicos/administração & dosagem , Transplante de Medula Óssea/métodos , Proteínas do Sistema Complemento/administração & dosagem , Ciclofosfamida/administração & dosagem , Ciclofosfamida/análogos & derivados , Imunização , Imunização Passiva , Técnicas In Vitro , Leucemia L1210/imunologia , Masculino , Camundongos , Camundongos Endogâmicos , Plasmocitoma/imunologia , Plasmocitoma/cirurgiaRESUMO
We have tested the immunologic status and hematologic parameters of mice 2 months (short-term survivors) or 18 months (long-term survivors) after lethal total body irradiation and syngeneic bone marrow transplantation (SBMT), and of normal mice of corresponding age. Long-term SBMT survivors showed significantly lowered bone marrow and spleen cellularities, decreased numbers of CFU-S in hemopoietic organs and severe impairment in the formation of CFU-F colonies compared with short-term SBMT survivors and normal mice. The peripheral blood parameters (hematocrit, erythrocytes, reticulocytes, platelets, white blood cells and granulocyte counts), however, remained unaltered. In long-term SBMT survivors we also observed a relative increase of Lyt-2+ lymphocytes (CD8+, cytotoxic/suppressor) and Mac-1+ cells among splenocytes. At the same time the L3T4+/Lyt-2+ ratio (CD4+/CD8+) was decreased. Relative contents of Ig+, Thy-1+ and L3T4+ cells were unchanged. The ability of splenocytes to generate IL-2R+ cells after in vitro stimulation with concanavalin A was greatly diminished. In summary, the immunohematopoietic status after initial normalization is again impaired in long-term SBMT survivors.
Assuntos
Transplante de Medula Óssea/imunologia , Células-Tronco Hematopoéticas/imunologia , Animais , Células Sanguíneas/citologia , Células Sanguíneas/imunologia , Células Sanguíneas/efeitos da radiação , Medula Óssea/efeitos da radiação , Medula Óssea/cirurgia , Células da Medula Óssea , Transplante de Medula Óssea/mortalidade , Transplante de Medula Óssea/patologia , Concanavalina A/farmacologia , Hematócrito , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/fisiologia , Antígeno de Macrófago 1/imunologia , Masculino , Camundongos , Receptores de Interleucina-2/efeitos dos fármacos , Baço/citologia , Baço/efeitos da radiação , Baço/ultraestrutura , Linfócitos T Auxiliares-Indutores/citologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Reguladores/citologia , Linfócitos T Reguladores/imunologia , Fatores de Tempo , Transplante Isogênico , Irradiação Corporal TotalRESUMO
The immunohematopoietic reconstitution of mice lethally irradiated (TBI) and reconstituted with syngeneic bone marrow cells untreated or treated with mafosfamide (ASTA Z 7654) [TBI + SBMT or TBI + SBMT-Maf mice, respectively] was examined. The number of CFU-S was greatly reduced in TBI + SBMT-Maf mice compared with those in TBI + SBMT mice. The recovery of blood parameters (hematocrit, reticulocytes, erythrocytes, white blood cells, granulocytes, platelets) and of bone marrow and spleen cells, but not of peritoneal exudate cells, was slightly delayed in TBI + SBMT-Maf mice compared with those in TBI + SBMT mice. The time for immune system regeneration was, however, considerably longer in TBI + SBMT-Maf than in TBI + SBMT mice, as measured by the incidence of Ig+, Thy-1.2+, L3T4+, Lyt-2+, and IL-2R+ cells in the spleens. The appearance of Mac-1+ and asialo-GM 1+ cells was only slightly prolonged or unchanged, respectively.
Assuntos
Transplante de Medula Óssea , Ciclofosfamida/análogos & derivados , Células-Tronco Hematopoéticas/imunologia , Animais , Anticorpos Monoclonais/uso terapêutico , Contagem de Células Sanguíneas , Ciclofosfamida/farmacologia , Ciclofosfamida/uso terapêutico , Transplante de Células-Tronco Hematopoéticas , Masculino , Camundongos , Transplante Isogênico , Irradiação Corporal TotalRESUMO
Cladribine is a lymphocytotoxic purine nucleoside with potential for treatment of autoimmune diseases. However, optimal administration regimens remain to be established. Twenty multiple sclerosis patients enrolled into this study were given 30 intermittent 2-hour cladribine infusions (0.07 mg/kg per infusion) each. Ten patients received cycles of 5 consecutive daily infusions at 5-week intervals (clustered dosage) on an inpatient basis; the other 10 patients received 1 infusion weekly (nonclustered dosage) on an outpatient basis. Red blood cell (RBC), platelet, and total white blood cell (WBC) counts were assessed at 5-week intervals during the treatment and at 13-week intervals during a 26-week follow-up period. Major WBC and lymphocyte subsets were assessed cytometrically at 15-week intervals during the treatment and at 13-week intervals thereafter. The clustered dosage produced a lasting decline in granulocyte count, a delayed decrease in monocyte count, and a transient decrease in RBC count. The nonclustered dosage caused a larger and persistent decline in RBC count, a smaller (P = .051. compared over the study period) decrease in monocyte count, and no change in granulocyte count. Both regimens transiently reduced natural killer and B-cell subsets (by 40%-60% and >80%, respectively) and caused lasting declines in CD4+ T-cell subsets (by >50%). No significant change was found in CD8+ T-cell subsets. These results show similar potency of these regimens with respect to major lymphocyte subsets, while suggesting that the nonclustered dosage is less toxic to myeloid precursors and more toxic to erythroid lineage precursors.
Assuntos
Cladribina/administração & dosagem , Esclerose Múltipla/tratamento farmacológico , Adulto , Contagem de Células Sanguíneas , Cladribina/farmacologia , Esquema de Medicação , Feminino , Humanos , Subpopulações de Linfócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/sangueRESUMO
A 3000--6000-fold purified kallikrein was obtained from human serum in 10--25% yield by chromatography on QAE-Sephadex A-50, Molselect CM-50 and on soybean trypsin inhibitor (SBTI)-AH-Sepharose 4-B. The enzyme had a specific activity of 14--23 U, as measured by BAEE hydrolysis. Some properties of highly purified kallikrein are described.
Assuntos
Calicreínas/isolamento & purificação , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Enzimas Imobilizadas , Humanos , Concentração de Íons de Hidrogênio , Calicreínas/sangue , Cinética , Temperatura , Inibidor da Tripsina de Soja de KunitzRESUMO
A highly purified human serum kallikrein immobilized on CH-Sepharose 4-B was obtained. KM values for N-benzoyl-L-arginine ethyl ester and N-tosyl-L-arginine methyl ester hydrolysis of this preparation were 1.10 x 10(-3) M and 3.6 x 10(-4) M, respectively; pH optimum of hydrolysis of these esters were found to be 8.2 and 8.5, respectively. The immobilized kallikrein possessed kininogenase activity and was capable of activating prekallikrein.
Assuntos
Calicreínas/metabolismo , Arginina/análogos & derivados , Arginina/metabolismo , Enzimas Imobilizadas , Humanos , Concentração de Íons de Hidrogênio , Calicreínas/isolamento & purificação , Tosilarginina Metil Éster/metabolismoRESUMO
Glycosaminoglycans were determined in the blood plasma of rabbits as a function of time after intravenous, intramuscular and intragastric administration of chondroitin sulfate preparation. After intravenous administration two phases of chondoitin sulfate elimination from the blood with T1/2 131-135 min and 8.5-9.5 min, respectively were observed. The level of the blood plasma glycosaminoglycans did not increase after intragastric introduction of the chondroitin sulfate preparation. Excretion of glycoaminoglycans in the urine was also determined.
Assuntos
Sulfatos de Condroitina/sangue , Condroitina/análogos & derivados , Administração Oral , Animais , Sulfatos de Condroitina/administração & dosagem , Feminino , Injeções Intramusculares , Injeções Intravenosas , Masculino , Coelhos , Ácidos Urônicos/sangueRESUMO
Patients with scleroderma (systemic sclerosis-SSc) frequently develop an interstitial lung disease. The role of lymphocytes in fibrosing alveolitis preceding lung fibrosis has been established. The purpose of this work was to evaluate cell profiles and lymphocyte phenotypes in the bronchoalveolar lavage (BAL) fluid and to correlate them with depression in lung function tests detected by depletion of diffusing capacity (DLCO). BAL was carried out in 25 untreated, non-smoking patients suffering from diffuse scleroderma and in 12 healthy non-smoking volunteers. For the analysis of lymphocyte sub-sets flow cytometry and monoclonal antibodies were used. The following cell sub-types were counted: T lymphocytes, B lymphocytes, helper lymphocytes, suppressor/cytotoxic lymphocytes, natural killer cells, cytotoxic T lymphocytes and activated T lymphocytes. The total cell count was higher in the group of patients with mild and moderate impairment in DLCO. The percentage of lymphocytes was greater in patients with DLCO lower than 65% of the predicted value since neutrophilia was found in patients with severe DLCO depletion, i.e. significant when compared with healthy subjects. The proportions of suppressor/cytotoxic lymphocytes and of activated T lymphocytes were higher in patients than in controls. The statistical analysis revealed significant differences between patients with moderate and mild changes in DLCO and the healthy volunteers. A decreased helper/suppressor ratio was noticed in these patients. We concluded that the BALF lymphocyte phenotype analysis may reflect the features of alveolitis in patients with SSc.
Assuntos
Líquido da Lavagem Broncoalveolar/imunologia , Capacidade de Difusão Pulmonar , Fibrose Pulmonar/etiologia , Escleroderma Sistêmico/complicações , Subpopulações de Linfócitos T , Adulto , Idoso , Relação CD4-CD8 , Estudos de Casos e Controles , Feminino , Humanos , Imunofenotipagem , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Neutrófilos , Fibrose Pulmonar/imunologia , Escleroderma Sistêmico/imunologia , Estatísticas não Paramétricas , Linfócitos T Auxiliares-Indutores , Linfócitos T ReguladoresRESUMO
The concentration of plasma prekallikrein (PK) in five patients with hepatocellular carcinoma (HCC) has been measured and related to levels in 18 patients with liver cirrhosis (LC) and 30 healthy subjects. It was found that the mean PK level was significantly increased in patients with HCC, while patients with LC demonstrated lower concentrations, as compared with healthy subjects. The results indicate that PK might be useful in screening cirrhotic patients for HCC. Longitudinal studies of PK in a larger group of patients at risk of developing HCC are therefore recommended.
Assuntos
Carcinoma Hepatocelular/sangue , Cirrose Hepática/sangue , Neoplasias Hepáticas/sangue , Pré-Calicreína/análise , Adulto , Idoso , Testes de Coagulação Sanguínea , Carcinoma Hepatocelular/diagnóstico , Feminino , Humanos , Neoplasias Hepáticas/diagnóstico , Masculino , Pessoa de Meia-Idade , Projetos PilotoRESUMO
A trypsin inhibitor was isolated from mouse lymphocytic leukemia L 1210 cells by ammonium sulphate precipitation and preparative isoelectric focusing. A 39-fold purification was attained. The inhibitor is a protein since its activity is destroyed by pronase and it binds to insolubilized trypsin. Two main forms of the inhibitor were found of pH 4.8 and 5.3. The inhibitor is copurified with DNA, although neither DNase II nor RNase A change its activity.
Assuntos
DNA/isolamento & purificação , Leucemia L1210/metabolismo , Proteínas/isolamento & purificação , Sulfato de Amônio , Animais , Cromatografia de Afinidade , Eletroforese em Gel de Poliacrilamida , Precipitação Fracionada , Concentração de Íons de Hidrogênio , Focalização Isoelétrica , Camundongos , Peso Molecular , Proteínas/metabolismo , Inibidores de Serina Proteinase , Espectrofotometria Ultravioleta , Tripsina/metabolismo , Inibidores da TripsinaRESUMO
The thymocytes were analyzed on the 7th day after i.p. inoculation of 10(6) leukemia L 1210 cells to syngeneic DBA/2 Wf mice. A three-fold decrease of the total number of thymocytes was found as well as 1.7-fold decrease of the per cent of thymocytes with Lyt 2+ phenotype, while the per cent of cells with phenotypes Thy 1.2+ and Lyt 1+ was unchanged.