RESUMO
We have established a new method for allogeneic pancreatic islet (PI) transplantation: relatively low doses of irradiation followed by simultaneous transplantation of PIs and bone marrow cells (BMCs) via the portal vein (PV). In the present study, we have compared this method with intra-bone marrow (IBM)-bone marrow transplantation (BMT), and with a combination of both methods. Streptozotocin (STZ)-induced diabetic-recipient rats, Fischer 344 (F344, RT1A(l), RT1B(l)), were irradiated 1 day before transplantation. PIs of Brown Norway rats (BN, RT1A(n), RT1B(n)) were transplanted into the liver of the diabetic F344 rats via the PV. BMCs from BN rats were injected into the recipients' bone marrow (IBM), PV or intravenously (IV) or by a simultaneous combination of PV plus IBM (PV+IBM). We compared graft survival among the groups of '9 Gy+IBM'(10/10 accepted), '9 Gy+PV'(7/10 accepted), '9 Gy+IV'(0/7 accepted), '9 Gy+PV+IBM'(8/8 accepted), '8.5 Gy+IBM'(4/9 accepted), '8.5 Gy+PV'(0/7 accepted), '8.5 Gy+IV'(0/7 accepted), '8.5 Gy+PV+IBM'(9/12 accepted), '8 Gy+IBM'(2/10 accepted) and '8 Gy+PV+IBM'(2/8 accepted). As we reported previously, PV-BMT is more effective in inducing the acceptance of allogeneic PIs than IV-BMT. However, IBM-BMT requires less pretreatment than PV-BMT. (PV+IBM)-BMT was found to be the most effective in inducing the acceptance of allogeneic PIs. These results suggest that allogeneic PI-transplantation in conjunction with (PV+IBM)-BMT could become a viable strategy.
Assuntos
Transplante de Medula Óssea/métodos , Transplante das Ilhotas Pancreáticas/métodos , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/imunologia , Diabetes Mellitus Experimental/terapia , Feminino , Sobrevivência de Enxerto , Tolerância Imunológica , Transplante das Ilhotas Pancreáticas/patologia , Transplante das Ilhotas Pancreáticas/fisiologia , Masculino , Veia Porta , Ratos , Ratos Endogâmicos ACI , Ratos Endogâmicos BN , Ratos Endogâmicos F344 , Transplante HomólogoRESUMO
AIMS: To find the new microbial parameters explaining the soil fertility from the microbial community viewpoint. METHODS AND RESULTS: Fatty acid methyl ester (FAME) analysis and terminal-restriction fragment length polymorphism (T-RFLP) analysis were carried out using 16 differently treated plots from the same field that had been kept under different fertilizer management systems since 1984. It was found that organic fertilizer application had small impact, whereas chemical fertilizer application, especially ammonium-nitrogen fertilizer, had strong impact on microbial community structures. Principal component analysis was conducted based on soil chemical and physical parameters, crop yields, FAMEs and terminal-restriction fragments (T-RFs) to provide 10 FAMEs and 10 T-RFs showing strong relation with soil fertility. CONCLUSION: We defined these 10 FAMEs and 10 T-RFs as 'keystone' biological parameters explaining soil fertility in the soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the FAMEs and T-RFs related to soil fertility. Both analyses are rapid, inexpensive and reproducible means. As field assessment needs precise and rapid analysis, FAME and T-RFLP analyses and these new parameters are very useful to analyse soil fertility at biological viewpoint.
Assuntos
Agricultura , Ecossistema , Microbiologia do Solo , Amônia , Técnicas de Tipagem Bacteriana , DNA Bacteriano/análise , Fertilizantes , Polimorfismo de Fragmento de RestriçãoRESUMO
BACKGROUND: Sarcoidosis is a systemic granulomatous disease of unknown aetiology. It has been suggested that T helper type 1 (Th1) polarisation is associated with the pathophysiology of sarcoidosis, but the mechanism of skewing towards Th1 has not been elucidated. Dendritic cells (DCs) are known to regulate immune responses. This study was performed to determine whether DCs are involved in the aetiology of sarcoidosis. METHODS: The numbers of peripheral blood DCs in 24 patients with sarcoidosis were analysed and biopsy specimens from four patients were stained immunohistochemically using monoclonal antibodies. RESULTS: The numbers of both myeloid and lymphoid DC subsets were significantly decreased in the blood and mature DCs were found in the granulomas of patients with sarcoidosis. A number of interferon-gamma (IFN-gamma) producing T cells were also detected in the sarcoid granuloma, as well as many interleukin (IL)-4 producing T cells. Double staining of the biopsy specimen using anti-fascin and anti-CD3 antibodies showed an anatomical interaction between DCs and T cells. CONCLUSIONS: These findings suggest that the blood DC subsets may migrate into the affected tissues, contributing to the formation of the granulomas in sarcoidosis. It is hypothesised that the migrating DCs may regulate the T cell response in sarcoidosis, at least in the granulomatous lesions.
Assuntos
Células Dendríticas/patologia , Sarcoidose/patologia , Adulto , Idoso , Feminino , Granuloma/patologia , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estatísticas não ParamétricasRESUMO
The dibenzofuran (DF)-utilizing bacterium strain YY-1 was newly isolated from soil. The isolate was identified as Janibacter sp. with respect to its 16S rDNA sequence and fatty acid profiles, as well as various physiological characteristics. In addition to DF, strain YY-1 could grow on fluorene and dibenzothiophene as sole sources of carbon and energy. It was also able to cometabolize a variety of polycyclic aromatic hydrocarbons including dibenzo- p-dioxin, phenanthrene, and anthracene. The major metabolites formed from DF, biphenyl, dibenzothiophene, and naphthalene were identified by using gas chromatography-mass spectrometry as 2,3,2'-trihydroxybiphenyl, biphenyl-dihydrodiol, dibenzothiophene 5-oxide, and coumarin, respectively. These results indicate that strain YY-1 can catalyze angular dioxygenation, lateral dioxygenation, and sulfoxidation.
Assuntos
Actinomycetales/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Tiofenos/metabolismo , Actinomycetales/classificação , Actinomycetales/enzimologia , Actinomycetales/genética , Benzofuranos/metabolismo , Fluorenos/metabolismo , Filogenia , Hidrocarbonetos Policíclicos Aromáticos/químicaRESUMO
The effects of various fractions of copper (Cu) and zinc (Zn) on soil bacteria were evaluated by the heavy metal tolerance level of the bacterial community (IC50) in soil samples collected near a mine. The IC50 values had no relationship with the total concentrations of Zn and Cu in the soils, but were weakly correlated with the 0.05 M CaCl2-extractable form of each metal in the soils (Cu: R2 = 0.670, p < 0.01; Zn: R2 = 0.453, p < 0.05). It was found that the IC50 correlated strongly with the total concentration of each metal in the extracts from water-saturated soil samples, described below as "soil solution" (Cu: R2 = 0.789, p < 0.01; Zn: R2 = 0.617, p < 0.01). The speciation of these metals in the soil solutions was estimated using an equilibrium thermodynamic computer model, SOILCHEM. Simulated free Cu ion ranged from 18 to 98% of total Cu, and organic complexes of Cu ranged from < 1 to 56%. In all samples, Zn existing as the free ion was estimated to be more than 80% of total Zn in the soil solutions. The IC50 values were also correlated with the estimated free metal ion activities, but with slightly lower correlation coefficients than found for total concentration in the soil solutions (Cu: R2 = 0.735, p < 0.01; Zn: R2 = 0.610, p < 0.01). The results suggest that not only high metal ion activities, but also total dissolved metal concentrations in soil solutions may affect the bacterial community.
Assuntos
Adaptação Fisiológica , Bactérias , Cobre/efeitos adversos , Poluentes do Solo/efeitos adversos , Zinco/efeitos adversos , Cobre/farmacologia , Microbiologia do Solo , Poluentes do Solo/farmacologia , Solubilidade , Zinco/farmacologiaRESUMO
AIMS: To develop oligonucleotide probes for visualizing bacteria belonging to Enterobacteriaceae. METHODS AND RESULTS: 24-mer oligonucleotide probe (probe D) was designed by comparison of 16S rDNA sequences of 35 species of Enterobacteriaceae, eight species of Vibrionaceae and six species of Pasteurellaceae. The sequence of the probe corresponding to the complementary sequence of a position 1251-1274 of Escherichia coli 16S rRNA was found to be a highly conserved region of 16S rDNA sequence in Enterobacteriaceae different from that of Vibrionaceae and Pasteurellaceae. The fluorescent dye-labelled probe was tested for the specificity by in situ hybridization and epifluorescence microscopy. Seventy-six out of 78 strains belonging to Enterobacteriaceae were visualized in an optimal hybridization condition. No bacterial strains belonging to Vibrionaceae (31 strains) and Gram-positive bacteria (three strains) were visualized. CONCLUSIONS: In situ hybridization using probe D allows the detection of bacterial cells belonging to Enterobacteriaceae without false positive reaction. SIGNIFICANCE AND IMPACT OF THE STUDY: In situ hybridization techniques using the probe D are potential tools for detecting Enterobacteriaceae in food and water samples.
Assuntos
Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Hibridização in Situ Fluorescente/métodos , Abastecimento de Água/normas , DNA Bacteriano/análise , DNA Ribossômico/análise , Microbiologia de Alimentos , Sondas de Oligonucleotídeos/genética , RNA Ribossômico 16S/análise , Sensibilidade e Especificidade , Microbiologia da ÁguaRESUMO
It is well known that the P(i) uptake system via the high-affinity P(i) transporter and the organic acid exudation system via PEPC are enhanced in the roots of P(i)-starved plants. In this paper, we compared the expression of these two systems in Sesbania rostrata, a leguminous plant, on whose roots and stems it forms nodules. When S. rostrata plants were transferred to a 0 microM P(i) nutrient solution, the expression of both the high-affinity P(i) transporter and PEPC was enhanced within 2 d. The enhancement of the expression of the high-affinity P(i) transporter genes and the PEPC gene coordinated with the increases in the P(i) uptake rate and the PEPC activity, respectively. This suggests that the expression of the high-affinity P(i) transporters and PEPC is regulated in part at the transcript level. Furthermore, we examined which of the environmental or the endogenous P(i) level regulates the expression of these two systems. The P(i) content in the 6-day-old plants decreased to a lower level than that in the 15-day-old plants when grown in a 30 microM P(i) solution. At that time, the expression of the high-affinity P(i) transporters and PEPC was enhanced only in the 6-day-old plants. Moreover, the P(i) content in plants forming many nodules on their stems decreased. The expression of the high-affinity P(i) transporters and PEPC was then enhanced in the nodulated plants. These facts suggest that the expression of these two systems may be regulated by the P(i) content in the plants, not by the P(i) concentration in the soil.
Assuntos
Fabaceae/metabolismo , Proteínas de Transporte de Fosfato/metabolismo , Fosfatos/metabolismo , Fosfoenolpiruvato Carboxilase/metabolismo , Fabaceae/enzimologia , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Proteínas de Transporte de Fosfato/genética , Fosfoenolpiruvato Carboxilase/genética , Raízes de Plantas/metabolismo , Caules de Planta/metabolismo , Simbiose , Fatores de Tempo , Transcrição GênicaRESUMO
Cepharanthin (CE) is a medicine that contains several biscoclaurine alkaloids. We examined the effects of CE on radiation-induced T cell apoptosis. Radiation induced apoptosis on T cells in a dose-dependent manner, while CE inhibited radiation-induced apoptosis. CE also attenuated the cytotoxic effects of radiation on the proliferative response of T cells. CE inhibited not only the loss of mitochondrial transmembrane potential but also the activation of caspase 3 in irradiated T cells. Radiation plus CE induced the up-regulation of Bax and the down-regulation of Bcl-2 in T cells in comparison with radiation alone. These results suggest that CE inhibits the signal transduction pathway of apoptosis induced by radiation, regardless of the expression of Bcl-2 or Bax.
Assuntos
Alcaloides/farmacologia , Apoptose/efeitos dos fármacos , Benzilisoquinolinas , Protetores contra Radiação/farmacologia , Linfócitos T/efeitos dos fármacos , Adulto , Apoptose/efeitos da radiação , Caspase 3 , Caspases/metabolismo , Células Cultivadas/citologia , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , Células Cultivadas/efeitos da radiação , Relação Dose-Resposta à Radiação , Combinação de Medicamentos , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/efeitos da radiação , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Genes bcl-2/efeitos dos fármacos , Genes bcl-2/efeitos da radiação , Genes p53/efeitos dos fármacos , Genes p53/efeitos da radiação , Humanos , Membranas Intracelulares/efeitos dos fármacos , Membranas Intracelulares/efeitos da radiação , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/efeitos da radiação , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/efeitos da radiação , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/efeitos da radiação , Fito-Hemaglutininas/farmacologia , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Linfócitos T/efeitos da radiação , Proteína Supressora de Tumor p53/biossíntese , Proteína X Associada a bcl-2RESUMO
A strictly anaerobic, irregularly coccoid, methanogenic archaeon, strain MG62T (= JCM 10825T = DSM 13459T), was isolated from paddy field soil in Chikugo, Fukuoka, Japan. The cells stained gram-negative, were 1.0-2.0 microm in diameter, were lysed by SDS and hypotonic solutions and were flagellated. Motility was not observed. The strain was able to use H2/CO2, 2-propanol/CO2, formate, 2-butanol/CO2 and cyclopentanol/CO2 as substrates for methanogenesis, but did not utilize acetate, ethanol, methanol or methylamines. The optimum temperature and pH were 25-30 degrees C and 6.7-7.2. Analysis of lipid component parts (core lipids, phospholipid polar head groups and glycolipid sugar moieties) showed the characteristic pattern of members of the family Methanomicrobiaceae except for the absence of glucose as a glycolipid sugar moiety. The G+C content of the DNA was 62.2 mol %. Sequence analysis of the 16S rDNA revealed that the strain belonged to the genus Methanoculleus. The strain had DNA-DNA hybridization values of less than 50% with type strains of Methanoculleus species. On the basis of phenotypic, genotypic and phylogenetic characteristics, the name Methanoculleus chikugoensis sp. nov. is proposed for strain MG62T (= JCM 10825T = DSM 13459T). The DNA hybridization study also revealed the close relationships of three species, Methanoculleus olentangyi, Methanoculleus bourgensis and Methanoculleus oldenburgensis, among Methanoculleus species.
Assuntos
DNA Arqueal/genética , Metano/metabolismo , Methanomicrobiaceae/classificação , Oryza , Microbiologia do Solo , Composição de Bases , DNA Arqueal/análise , DNA Ribossômico/análise , Japão , Lipídeos/análise , Methanomicrobiaceae/genética , Methanomicrobiaceae/isolamento & purificação , Methanomicrobiaceae/fisiologia , Methanomicrobiaceae/ultraestrutura , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Oryza/crescimento & desenvolvimento , Fenótipo , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The influences of Zn and Cu on soil enzyme activities (acid phosphatase, alkaline phosphatase, arylsulfatase, cellulase, dehydrogenase, protease (z-FLase), urease, beta-D-glucosidase and beta-D-fructofuranosidase (invertase)) and microbial biomass carbon were investigated in agricultural soils amended with municipal sewage sludge or compost since 1978. The trace metals in the soils were fractionated using a sequential extraction method. Long-term application of the sewage sludge and composts caused accumulations of Cu and Zn in the soils, ranging from 140 to 144 and from 216 to 292 mg kg(-1), respectively. The percentage of Cu was highest in the NaOH- and HNO3-extractable fractions (44-51% and 38-46%, respectively), while the percentage of Zn was highest in the HNO3- and EDTA-extractable fractions (65-83% and 11-32%, respectively). Although the percentage of the bioavailable fractions (sum of KNO3 + H2O-, NaOH-, and EDTA-extractable amounts) of Cu (53-64%) was higher than that of Zn (15-37%), the percentage of the most labile fractions (KNO3 + H2O) of Zn (2.1-5.9%) was larger than that of Cu (1.1-2.4%). The size of the microbial biomass carbon increased with the application of sewage sludge or compost. For some enzymes, however, the ratio of the enzyme activity to microbial biomass was lower in the soils amended with sewage sludge or compost than that in the control soil. The soil enzyme activities were more adversely affected by Zn than by Cu. From a multiple regression analysis, it was found that dehydrogenase, urease, and beta-D-glucosidase activities were reduced by the KNO3 + H2O-extractable fraction of Zn in the soils. These microbial activities seem to be sensitive to Zn stress, indicating the possibility that they might be useful bioindicators for evaluation of the toxic effects of Zn on microorganisms in the soils.
Assuntos
Bactérias/efeitos dos fármacos , Cobre/farmacologia , Enzimas/metabolismo , Esgotos/química , Microbiologia do Solo , Zinco/farmacologia , Agricultura , Bactérias/crescimento & desenvolvimento , Biodegradação Ambiental , Biomassa , Cobre/análise , Enzimas/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Eliminação de Resíduos/métodos , Solo/análise , Poluentes do Solo/análise , Estatística como Assunto , Fatores de Tempo , Zinco/análiseRESUMO
Genomic DNA homologies were examined from six Microcystis (cyanobacteria) strains, including five different species, Microcystis aeruginosa, Microcystis ichthyoblabe, Microcystis novacekii, Microcystis viridis and Microcystis wesenbergii. All DNA-DNA reassociation values between two strains of M. aeruginosa and the other four species exceeded 70%, which is considered high enough for them to be classified within the same bacterial species. It is proposed to unify these five species into M. aeruginosa under the Rules of the Bacteriological Code and NIES843T (= IAM M-247T) is proposed as the type strain. Two other species, Microcystis flos-aquae and Microcystis pseudofilamentosa, should be regarded as morphological variations of this unified M. aeruginosa. The current taxonomy of cyanobacteria depends too much upon morphological characteristics and must be reviewed by means of bacteriological methods as well as traditional botanical methods.
Assuntos
Microcystis/classificação , Filogenia , Terminologia como Assunto , DNA Bacteriano/genética , DNA Ribossômico/genética , Evolução Molecular , Geografia , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genéticaRESUMO
Paclitaxel is a chemotherapeutic drug that induces apoptosis in tumor cells by stabilizing microtubules, prevents normal mitosis, and blocks the cell cycle at the G2/M phase. We have previously reported that the activation of caspase-3 and caspase-8 plays a crucial role in paclitaxel-induced apoptosis. Anti-tumor reagents including paclitaxel, irradiation, and other stimuli activate the transcription factor NF-kappaB, which has the ability to suppress the apoptotic potential of those stimuli. Using a human lung adenocarcinoma cell line (LC-2-AD), we therefore examined whether the inhibition of NF-kappaB activity by proteasome inhibitor 1 (PS1) could become a new adjuvant therapy for cancer. A synergistic effect on apoptosis induction was observed with the combination of more than 0.1 microg/ml paclitaxel and 0.5 microM PS1. An increase in the cell number of apoptotic cells is correlated with the loss of Deltaphim and the activation of caspase-3 and caspase-8. Furthermore, augmented apoptosis is related to NF-kappaB activation. Based on these findings, we propose that the combination of paclitaxel with PS1 could be a new strategy for cancer treatment.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Complexos Multienzimáticos/antagonistas & inibidores , Paclitaxel/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Adenocarcinoma/enzimologia , Adenocarcinoma/patologia , Western Blotting , Caspase 3 , Caspases/metabolismo , Ciclo Celular/efeitos dos fármacos , Cisteína Endopeptidases , Sinergismo Farmacológico , Ativação Enzimática , Citometria de Fluxo , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/patologia , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Complexo de Endopeptidases do Proteassoma , Células Tumorais Cultivadas/enzimologia , Células Tumorais Cultivadas/patologiaRESUMO
An autopsy case of giant cell myocarditis (GCM) in a 74-year-old woman is presented. She suffered from hepatic dysfunction, skin eruption and disseminated intravascular coagulation due to the side-effects of a non-steroidal anti-inflammatory drug. After admission, heart failure progressed rapidly, and the patient died suddenly. At autopsy, her heart was slightly enlarged and the heart muscle was thickened with many small whitish nodules. She was diagnosed with GCM because of the infiltration of multinuclear giant cells, histiocytes, eosinophils and lymphocytes into the heart. We did not find any similar lesions in any other organs. Giant cell myocarditis, the etiology of which is not defined, is a rare disease with unfavorable prognosis. This case suggests the possibility of drug-induced GCM.
Assuntos
Anti-Inflamatórios não Esteroides/efeitos adversos , Células Gigantes/patologia , Insuficiência Cardíaca/etiologia , Miocardite/induzido quimicamente , Fenilpropionatos/efeitos adversos , Idoso , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Toxidermias/etiologia , Evolução Fatal , Feminino , Insuficiência Cardíaca/patologia , Humanos , Imuno-Histoquímica , Miocardite/diagnóstico , Miocardite/metabolismo , Miocárdio/metabolismo , Miocárdio/patologia , Radiografia TorácicaRESUMO
This report describes the rapid development of multiple meniscal signs complicating invasive pulmonary aspergillosis in a 53-year-old man receiving chemotherapy for acute leukemia. While undergoing first induction therapy for AML, he developed chest pain, and multiple bilateral infiltrations were seen in chest roentgenograms. Administration of antibiotics, antifungal agents, steroid pulse therapy and G-CSF was begun. Pulmonary cavities with meniscal signs developed. The next day, pneumothorax and hemothorax were noted. Although drainage and mechanical ventilation were performed, the patient died after massive hemoptysis. Invasive pulmonary aspergillosis was diagnosed at autopsy.
Assuntos
Aspergilose/etiologia , Leucemia Mieloide Aguda/tratamento farmacológico , Pneumopatias Fúngicas/etiologia , Aspergilose/diagnóstico , Humanos , Pneumopatias Fúngicas/diagnóstico , Masculino , Pessoa de Meia-IdadeRESUMO
To minimize contamination of bone marrow cells (BMCs) with T cells from the peripheral blood, a new "perfusion method" for collecting BMCs is proposed using cynomolgus monkeys. Two BM puncture needles are inserted into a long bone such as the humerus, femur, or tibia. One needle is connected to an extension tube and the end of the tube is inserted into a culture flask to collect the BM fluid. The other needle is connected to a syringe containing 30 ml of phosphate-buffered saline. The solution is pushed gently from the syringe into the medullary cavity, and the medium containing the BM fluid is collected into the culture flask. There is significantly less contamination with peripheral blood, determined from the frequencies of CD4(+) and CD8(+) T cells, when using this method (<6%) than when using the conventional method (>20%) consisting of multiple BM aspirations from the iliac crest. Furthermore, the number and progenitor activities of the cells harvested using this "perfusion method" are greater than those harvested using the conventional aspiration method. This perfusion method was carried out 42 times using 15 cynomolgus monkeys, and no complications such as pulmonary infarction or paralysis were observed. These findings suggest that the "perfusion method" is safe and simple and would be of great advantage in obtaining pure BMCs, resulting in a less frequent occurrence of acute graft-versus-host-disease in allogeneic BM transplantation.
Assuntos
Células da Medula Óssea/citologia , Separação Celular/métodos , Animais , Antígenos CD/análise , Antígenos de Superfície/análise , Células da Medula Óssea/imunologia , Contagem de Células , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/imunologia , Macaca fascicularisRESUMO
The broad and vague phenotypic definition allowed the genus Pseudomonas to become a dumping ground for incompletely characterized polarly flagellated, gram-negative, rod-shaped, aerobic bacteria, and a large number of species have been accommodated in the genus Pseudomonas. The 16S rRNA sequences of 128 valid and invalid Pseudomonas species, which included almost valid species of the genus Pseudomonas listed in the Approved Lists of Bacterial Names, were obtained: sequences of 59 species were determined and those of 69 species were obtained from the GenBank/EMBL/DDBJ databases. These sequences were compared with the sequences of other species of the Proteobacteria. Fifty-seven valid or invalid species including Pseudomonas aeruginosa (type species of the genus Pseudomonas Migula 1894) belonged to the genus Pseudomonas (sensu stricto). Seven subclusters were formed in the cluster of the genus Pseudomonas (sensu stricto), and the resulting clusters conformed well to the rRNA-DNA hybridization study by Palleroni (1984). The other species did not belong to the genus Pseudomonas (sensu stricto) and were related to other genera, which were placed in four subclasses of the Proteobacteria (alpha, beta, gamma and gamma-beta subclasses). Twenty-six examined species, which were not included in the cluster of the Pseudomonas (sensu stricto) and have not been transferred to other genera as yet, are listed alphabetically: 'Pseudomonas abikonensis', Pseudomonas antimicrobica, Pseudomonas beijerinckii, Pseudomonas beteli, Pseudomonas boreopolis, 'Pseudomonas butanovora', Pseudomonas carboxydohydrogena, Pseudomonas cissicola, Pseudomonas doudoroffii, Pseudomonas echinoides, Pseudomonas elongata, Pseudomonas flectens, Pseudomonas geniculata, Pseudomonas halophila, Pseudomonas hibiscicola, Pseudomonas huttiensis, Pseudomonas iners, Pseudomonas lanceolata, Pseudomonas lemoignei, Pseudomonas mephitica, Pseudomonas pictorum, Pseudomonas saccharophila, Pseudomonas spinosa, Pseudomonas stanier, Pseudomonas syzygii and Pseudomonas woodsii. The phylogenetic affiliations of these 26 pseudomonads species are shown.
Assuntos
Pseudomonas/classificação , Clonagem Molecular , DNA Bacteriano/genética , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , Pseudomonas/genética , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genética , Homologia de Sequência do Ácido NucleicoRESUMO
The autopsy of a 76-year-old Japanese female patient, which revealed thymic carcinoma with various tumor markers such as NSE, CYFRA, and CA-125, is presented. The patient died from hepatic failure because the liver was overtaken by the tumors. At autopsy, the thymic carcinoma was found to have metastased only in the liver. From microscopical analyses and electron microscopical findings, we diagnosed poorly differenciated squamous cell carcinoma of thymic origin. In the histochemical analyses, the tumor cells were positively stained in CA 125, CA 19-9, EMA, NSE, AE 1, AE 3, CEA, S-100, glimerius and Bcl-2. These date suggest that the tumor cells produced various tumor markers. In 222 autopsy cases of thymic malignant tumor observed in Japan over a period of 4 years, the dominant pathohistological image was squamous cell carcinoma. It is interesting that the greatest number of combined malignant tumors with thymic malignancies were thyroid papillary carcinomas.
Assuntos
Biomarcadores Tumorais/análise , Antígeno CA-19-9/análise , Carcinoma de Células Escamosas/secundário , Neoplasias Hepáticas/secundário , Neoplasias do Timo/patologia , Idoso , Antígeno Ca-125/análise , Antígeno Carcinoembrionário/análise , Carcinoma de Células Escamosas/patologia , Feminino , Humanos , Neoplasias Hepáticas/patologia , Fosfopiruvato Hidratase/análiseRESUMO
The (NZW x BXSB)F1 (W/BF1) mouse is known as an autoimmune-prone strain which develops lupus nephritis, thrombocytopenia due to platelet-specific autoantibodies, leukocytosis, and myocardial infarction. In this experiment, we investigated the age-dependent abnormalities of the hematopoietic stem cells (HSCs) and hematopoiesis in this mouse. White blood cell counts (especially Mac-1- or Gr-1-positive cells) in the peripheral blood of 12-week-old W/BF1 mice increased in comparison with those of four-week-old W/BF1 or normal mice. To investigate whether the abnormal hematopoiesis can be attributed to the HSCs of W/BF1 mice, colony-forming unit in spleen (CFU-S) and colony-forming unit in culture (CFU-C) assays were performed. Day 12 CFU-S counts of 12-week-old W/BF1 mice significantly increased in comparison with those of four-week-old W/BF1 mice or normal mice. In the CFU-C assay, CFU-GEMM and CFU-GM counts in 12-week-old W/BF1 mice increased in comparison with those of four-week-old W/BF1 or control mice. The bone marrow cells (BMCs) from 12-week-old W/BF1 mice showed a high level of G-CSF and a low level of GM-CSF in mRNA expression. To examine the effect of HSCs from 12-week-old W/BF1 mice on the onset of autoimmune diseases and the abnormal hematopoiesis, T- and B-cell-depleted BMCs of four-week-old or 12-week-old W/BF1 mice were transplanted to C3H mice. Recipient C3H mice that had received the BMCs from 12-week-old W/BF1 mice showed an earlier onset of autoimmune diseases and a shorter survival rate than those that had received the BMCs from four-week-old W/BF1 mice. These data suggest that the HSCs from 12-week-old W/BF1 mice showing the symptoms of autoimmune diseases have the capacity to induce autoimmune diseases earlier than the HSCs from four-week-old W/BF1 mice.
Assuntos
Envelhecimento/patologia , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/patologia , Nefrite Lúpica/patologia , Trombocitopenia/patologia , Idade de Início , Animais , Modelos Animais de Doenças , Expressão Gênica/imunologia , Fator Estimulador de Colônias de Granulócitos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Hematopoese/imunologia , Células-Tronco Hematopoéticas/química , Contagem de Leucócitos , Leucocitose/mortalidade , Leucocitose/patologia , Leucocitose/terapia , Nefrite Lúpica/mortalidade , Nefrite Lúpica/terapia , Fator Estimulador de Colônias de Macrófagos/genética , Antígeno de Macrófago 1/análise , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos Endogâmicos NZB , Contagem de Plaquetas , RNA Mensageiro/análise , Análise de Sobrevida , Trombocitopenia/mortalidade , Trombocitopenia/terapiaRESUMO
The anticancer drug paclitaxel is well known as an inhibitor of microtubule depolymerization, resulting in mitosis arrest. We investigated the mechanism underlying antitumor effects of paclitaxel on the lung adenocarcinoma cell line LC-2-AD. Less than 10 microg/ml paclitaxel induced mitosis arrest upon LC-2-AD, followed by apoptosis, but more than 30 microg/ml paclitaxel induced apoptosis without mitosis arrest. LC-2-AD with less than 1 microg/ml paclitaxel showed a loss of mitochondrial transmembrane potential (deltapsim), which correlated with antitumor effects. However, LC-2-AD with more than 10 microg/ml paclitaxel showed slight changes in the loss of deltapsim in spite of its ability to induce apoptosis significantly. The cleavage of caspase 3, caspase 8, and DFF45/ICAD was also observed in paclitaxel-induced apoptosis, and the inhibitor of caspase 3 and caspase 8 inhibited both antitumor effects and apoptosis induced by paclitaxel. These results suggest that activation of caspase 3 and caspase 8 plays a crucial role in paclitaxel-induced apoptosis under any concentrations of paclitaxel.