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1.
J Low Temp Phys ; 193(3): 518-524, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30839748

RESUMO

We present the development of a frequency-domain multiplexing readout of kinetic inductance detectors (KIDs) for pulse signals with a self-trigger system. The KIDs consist of an array of superconducting resonators that have different resonant frequencies individually, allowing us to read out multiple channels in the frequency domain with a single wire using a microwave-frequency comb. The energy deposited to the resonators break Cooper pairs, changing the kinetic inductance and, hence, the amplitude and the phase of the probing microwaves. For some applications such as X-ray detections, the deposited energy is detected as a pulse signal shaped by the time constants of the quasiparticle lifetime, the resonator quality factor, and the ballistic phonon lifetime in the substrate, ranging from microseconds to milliseconds. A readout system commonly used converts the frequency-domain data to the time-domain data. For the short pulse signals, the data rate may exceed the data transfer bandwidth, as the short time constant pulses require us to have a high sampling rate. In order to overcome this circumstance, we have developed a KID readout system that contains a self-trigger system to extract relevant signal data and reduces the total data rate with a commercial off-the-shelf FPGA board. We have demonstrated that the system can read out pulse signals of 15 resonators simultaneously with about 10 Hz event rate by irradiating α particles from 241 Am to the silicon substrate on whose surface aluminum KID resonators are formed.

3.
Vox Sang ; 112(2): 114-121, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28070919

RESUMO

BACKGROUND AND OBJECTIVES: In previous studies, we demonstrated that the basophil-activating effects of supernatants found in residual-transfused platelet concentrates (PC-SNs) on whole blood basophils in cases of allergic transfusion reactions (ATRs) could be assessed by the basophil activation test (BAT) in terms of allergen/IgE dependency. However, in these studies, the basophils were derived from third-party healthy volunteers. In this study, we performed BAT using patients' own blood basophils to analyse ATRs. MATERIALS AND METHODS: The BAT was performed in two cases of severe ATRs using residual PC-SNs and the patients' own basophils in the presence and absence of dasatinib, an inhibitor of IgE-mediated basophil activation. RESULTS: In both cases, PC-SNs exhibited basophil-activating activity against the patients' basophils, but not against basophils from third-party healthy volunteers. In addition, basophil activation was inhibited in the presence of dasatinib, indicating that the basophils were activated in an allergen/IgE-dependent manner. Of note, the basophils in Case 2, but not in Case 1, were activated by PC-SNs from some unrelated non-haemolytic transfusion reaction cases. CONCLUSION: This pilot study indicates that the BAT may be useful in clarifying the causal relationship between ATRs and transfused blood as well as in elucidating the mechanisms behind ATRs considering the allergen/IgE-dependent pathway.


Assuntos
Basófilos/imunologia , Transfusão de Plaquetas/efeitos adversos , Reação Transfusional/etiologia , Basófilos/citologia , Basófilos/efeitos dos fármacos , Dasatinibe/farmacologia , Feminino , Humanos , Imunoglobulina E/metabolismo , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Índice de Gravidade de Doença , Reação Transfusional/patologia , Triptases/sangue
4.
Vox Sang ; 109(3): 239-47, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25953437

RESUMO

BACKGROUND AND OBJECTIVES: We developed a hollow-fibre column system specifically adapted to prepare washed platelet concentrates (WPCs). This study was performed to evaluate the efficacy of the hollow-fibre column system for preparing WPCs. MATERIALS AND METHODS: First, the percentages of platelet (PLT) recovery and remaining plasma proteins were calculated by determining the PLT count, volume and plasma protein levels in both the prewash and postwash. Secondly, washed PLTs and unwashed control PLTs were stored for 5 days, and the changes during this 5-day storage of in vitro PLT characteristics were determined. RESULTS: The hollow-fibre column system effectively removed >98% of plasma in platelet concentrates (PCs), and the PLT recovery was 97% on an average. The CD62P-expression level on washed PLTs immediately after washing was approximately twofold higher than that on prewashed PLTs as well as on PLTs washed via manual methods or cell washing devices. Until day 5 during storage, PLT aggregability, hypotonic shock response and swirling scores of washed PLTs were not significantly different from those of the control PCs. CONCLUSION: Our novel hollow-fibre column system proved valuable in preparing washed PLTs with <2% of residual plasma proteins and high recovery of PLTs.


Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Plaquetas/metabolismo , Preservação de Sangue/instrumentação , Glucose/metabolismo , Humanos , Ácido Láctico/metabolismo , Fatores de Tempo
5.
Vox Sang ; 108(4): 425-7, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25557060

RESUMO

The Am and Bm phenotypes are characterized by weak expression of the A or B antigens, respectively, by red blood cells with a normal expression by the saliva of secretors. Deletion of the regulatory element in the first intron of the ABO gene and disruption of the GATA motif in the element were found to be responsible. In this study, we identified a novel mutation within the GATA motif (G>C substitution at position c.28 + 5830) in the regulatory element of the A allele that might diminish transcription activity causing the generation of the Am B phenotype.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Células Eritroides/metabolismo , Fenótipo , Mutação Puntual , Sequências Reguladoras de Ácido Nucleico , Alelos , Sequência de Bases , Sítios de Ligação , Doadores de Sangue , Fatores de Transcrição GATA/metabolismo , Humanos , Íntrons , Dados de Sequência Molecular , Deleção de Sequência
6.
Vox Sang ; 106(4): 368-71, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24387678

RESUMO

BACKGROUND AND OBJECTIVES: CD36 antibody (Ab) causes several disorders: neonatal alloimmune thrombocytopenia, platelet transfusion refractoriness and non-haemolytic transfusion reactions. However, there is no gold-standard test for CD36 Ab. MATERIALS AND METHODS: We developed a transfectant panel cell line-based MoAb-independent antigen capture assay system for detection of CD36 Ab and compared it with the monoclonal antibody-specific immobilization of platelet antigens (MAIPA) system in terms of sensitivity and specificity. RESULTS: Our new system was characterized by (1) gene-transfected cell lines, but not panel platelets; (2) not being hampered by HLA Abs; and (3) no need to use CD36 MoAbs to ensure the antigen specificity of this detection system. In addition, it showed a much better receiver operating characteristic curve than the MAIPA system. CONCLUSIONS: The present results indicate that our new system permits highly sensitive and specific detection of CD36 Ab.


Assuntos
Anticorpos Monoclonais/química , Autoanticorpos/sangue , Antígenos CD36/imunologia , Antígenos de Plaquetas Humanas/imunologia , Autoanticorpos/isolamento & purificação , Linhagem Celular , Humanos , Curva ROC , Transfecção
7.
Transfus Med ; 24(1): 39-44, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24152224

RESUMO

BACKGROUND: Blood-group genotyping arrays have been widely used in Caucasian and African American populations, but have not been thoroughly tested in Japanese subjects. AIM: To evaluate, using the BLOODchip(®) Reference genotyping system, the concordance of previously typed samples with expected phenotypes and the coverage of the Japanese variants. METHODS: Blood samples from 100 Japanese donors were obtained. DNA was extracted with QIAsymphony (Qiagen, Hilden, Germany). Samples were typed by serological methods and processed with the BLOODchip(®) . When a non-concordant result was identified, further sequencing by polymerase chain reaction-single specific primer (PCR-SSP) was performed. RESULTS: Concordance between systems was 98% (736/751), and 98.8% (742/751) if only non-software-related non-concordances were considered. In the ABO group, 6 'No Call' (NC, inability of the BLOODchip(®) to assign a result) were ascribed to a variant of blood subtype A1 (A102; 467C>T), a common subtype in Asian populations, whereas three NC presented additional polymorphisms not contained in the BLOODchip(®) (A102/A205, A102/O06 and A204/O02). In the RhD group, one discrepancy was correctly genotyped as RHD*1227A (Del phenotype) by the BLOODchip(®) (phenotyped as partial D, RHD*DIVb). Another was phenotyped as D+ by the BLOODchip(®) (phenotyped weak D by serology) and confirmed as RHD*D-CE(2)-D heterozygous by sequencing. The 3 RhD NC can be solved by further software update. For RhCE, one discrepancy was correctly genotyped for both systems; however, only the BLOODchip(®) was able to detect RHCE*CX allele. CONCLUSIONS: By programming the A102 ABO variant into the system software with the new allele combinations, the BLOODchip(®) Reference is a suitable genotyping tool to be applied to Asian samples.


Assuntos
Antígenos de Grupos Sanguíneos/genética , Tipagem e Reações Cruzadas Sanguíneas , Técnicas de Genotipagem , Adulto , Povo Asiático , Antígenos de Grupos Sanguíneos/sangue , Tipagem e Reações Cruzadas Sanguíneas/instrumentação , Tipagem e Reações Cruzadas Sanguíneas/métodos , Feminino , Técnicas de Genotipagem/instrumentação , Técnicas de Genotipagem/métodos , Humanos , Recém-Nascido , Japão , Masculino , Sensibilidade e Especificidade
8.
Vox Sang ; 105(3): 183-95, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23763621

RESUMO

BACKGROUND AND OBJECTIVES: Noninfectious and nonhaemolytic transfusion reactions are the most common type of transfusion reactions. Several new tests have been made, helping diagnosis and understanding of their pathogenesis. This manuscript provides a review of the literature on currently available tests in association with the approach in Japan. MATERIALS & METHODS: Primarily by using key words, more than 100 pertinent articles in the Medline database were identified and reviewed. RESULTS: Numbers of laboratory tests are available including those for plasma protein levels, plasma protein antibodies, leucocyte and platelet antibodies, serum N-terminal-pro-brain natriuretic peptide levels, serum tryptase levels and genetic microchimerism. Cross-match tests, such as basophil activation test and neutrophil activation test, are also available to determine a causal relationship between the reaction and transfusion. CONCLUSIONS: Several tests should help to confirm diagnosis and determine causal relationship between adverse reactions and transfusion and to gain an insight into the mechanism of the reaction in some cases, although some of the recently developed tests have not been completely validated.


Assuntos
Lesão Pulmonar Aguda/diagnóstico , Doença Enxerto-Hospedeiro/diagnóstico , Testes Hematológicos/métodos , Hipersensibilidade/diagnóstico , Púrpura Trombocitopênica/diagnóstico , Reação Transfusional , Lesão Pulmonar Aguda/etiologia , Anticorpos/sangue , Circulação Sanguínea , Feminino , Doença Enxerto-Hospedeiro/etiologia , Humanos , Hipersensibilidade/etiologia , Japão , Leucócitos/imunologia , Masculino , Ativação de Neutrófilo , Púrpura Trombocitopênica/etiologia
9.
Cancer Gene Ther ; 19(5): 358-66, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22402627

RESUMO

We previously reported that polyamidoamine STARBURST dendrimer (generation 3, G3) (dendrimer) conjugate with α-cyclodextrin (α-CyD) having an average degree of substitution of 2.4 of α-CyD (α-CDE) provided remarkable aspects as novel carriers for DNA and small-interfering RNA. To develop novel α-CDE derivatives with tumor cell specificity, we prepared folate-appended α-CDEs (Fol-α-CDEs) and folate-polyethylene glycol (PEG)-appended α-CDEs (Fol-PαCs) with the various degrees of substitution of folate (DSF), and evaluated in vitro and in vivo gene transfer activity, cytotoxicity, cellular association and physicochemical properties. In vitro gene transfer activity of Fol-α-CDEs (G3, DSF 2, 5 or 7) was lower than that of α-CDE (G3) in KB cells, folate receptor (FR)-overexpressing cancer cells. Of the three Fol-PαCs (G3, DSF 2, 5 or 7), Fol-PαC (G3, DSF 5) had the highest gene transfer activity in KB cells. The activity of Fol-PαC (G3, DSF 5) was significantly higher than that of α-CDE (G3) in KB cells, but not in A549 cells, FR-negative cells. Negligible cytotoxicity of the plasmid DNA (pDNA) complex with Fol-PαC (G3, DSF 5) was observed in KB cells or A549 cells up to a charge ratio of 100/1 (carrier/pDNA). The cellular association of the pDNA complex with Fol-PαC (G3, DSF 5) could be mediated by FR on KB cells, resulting in its efficient cellular uptake. Fol-PαC (G3, DSF 5) had a higher binding affinity with folate-binding protein than α-CDE (G3), although the physicochemical properties of pDNA complex with Fol-PαC (G3, DSF 5) were almost comparable to that with α-CDE (G3), although the onset charge ratio and the compaction ability of Fol-PαC (G3, DSF 5) were slightly different. Fol-PαC (G3, DSF 5) tended to show a higher gene transfer activity than α-CDE (G3) 12 h after intratumoral administration in mice. These results suggest that Fol-PαC (G3, DSF 5), not Fol-α-CDEs, could be potentially used as a FR-overexpressing cancer cell-selective DNA carrier.


Assuntos
DNA/genética , Dendrímeros/química , Ácido Fólico/genética , Técnicas de Transferência de Genes , Poliaminas/química , alfa-Ciclodextrinas/química , Animais , DNA/administração & dosagem , Dendrímeros/administração & dosagem , Ácido Fólico/administração & dosagem , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Poliaminas/administração & dosagem , Transfecção/métodos
10.
BJOG ; 119(3): 340-7, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22239415

RESUMO

OBJECTIVE: To investigate the prevalence and risk factors of third- and fourth-degree perineal lacerations in 24, mainly developing, countries. DESIGN: Analysis using cross-sectional data from the WHO Global Survey on Maternal and Perinatal Health. SETTING: Seven African, nine Asian and eight Latin American countries. POPULATION: Women at admission to hospital for delivery in 373 facilities between 2004 and 2008. METHODS: We estimated the country-wise prevalence of third- and fourth-degree perineal lacerations, and conducted region-wise multivariate logistic regression analyses to identify its risk factors. MAIN OUTCOME MEASURES: Prevalence and risk factors of third- and fourth-degree perineal lacerations. RESULTS: A total of 214,599 women who underwent vaginal delivery were analysed. The prevalence of third- and fourth-degree perineal lacerations ranged widely across countries [from 0.1% (China, Cambodia, India) to 15.0% (Philippines)] and facilities (from null to 76.3%). After the deletion of facilities reporting no third- or fourth-degree perineal lacerations, and also highly outlying facilities, the range in prevalence was 0.1% (Uganda) to 1.4% (Japan). Forceps-assisted delivery, nulliparity and high birthweight were significant risk factors in all three regions. Vacuum-assisted delivery was also a significant risk factor in Africa and Asia. CONCLUSIONS: Misdiagnosis of third- and fourth-degree perineal lacerations in developing countries may be common. Correct recognition and diagnosis may lead to timely treatment and fewer sequelae. Risk factors of third- and fourth-degree perineal lacerations in developing countries were similar to those previously reported from developed countries.


Assuntos
Lacerações/epidemiologia , Complicações do Trabalho de Parto/epidemiologia , Períneo/lesões , Adulto , África/epidemiologia , Ásia/epidemiologia , Peso ao Nascer , Estudos Transversais , Países em Desenvolvimento , Extração Obstétrica/efeitos adversos , Feminino , Humanos , Lacerações/diagnóstico , Lacerações/etiologia , América Latina/epidemiologia , Modelos Logísticos , Análise Multivariada , Complicações do Trabalho de Parto/diagnóstico , Complicações do Trabalho de Parto/etiologia , Paridade , Parto , Gravidez , Prevalência , Fatores de Risco
11.
Vox Sang ; 102(2): 171-4, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21883271

RESUMO

There is an international need for a large-scale human neutrophils antigen (HNA) antibody screening platform to minimize the risk of antibody-mediated transfusion-related acute lung injury. However, sourcing a substantial, reliable source of HNA, as well as the scarcity of well-characterized HNA antisera for validating new screening platforms, remain as major obstacles. This short communication presents an improved protocol for the effective use of HNA-expressing KY cells as a screening platform using eight well-characterized HNA antisera of a single defined specificity.


Assuntos
Anticorpos/sangue , Imunofluorescência/métodos , Isoanticorpos/sangue , Isoantígenos/biossíntese , Neutrófilos/imunologia , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Bovinos , Linhagem Celular , Humanos , Isoanticorpos/imunologia , Isoantígenos/sangue , Isoantígenos/imunologia , Transfecção
12.
Transfus Med ; 21(3): 199-204, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21208305

RESUMO

BACKGROUND: A panel of platelets expressing various human platelet antigens (HPAs) for a platelet antibody screening assay is difficult to prepare because some antigens are rarely expressed. Therefore, an alternative method without using platelets would be helpful in detecting HPA antibodies. This study describes the establishment of cell lines that stably express specific HPAs and their application for detecting specific antibodies. METHODS: Wild-type ß3, HPA-1b, -6b, -7b and -7 variant cDNA as well as wild-type αIIb and HPA-3b cDNA were individually co-transduced with wild-type αIIb and ß3 cDNA in the K562 cell line. We performed an immunobead monoclonal antibody immobilisation of platelet antigens (MAIPA) assay to evaluate this cell line panel for antibody detection using identified sera containing HPA antibodies, whose specificities had been determined by the mixed passive haemagglutination test. RESULTS AND CONCLUSION: Of the 12 sera containing HPA-1a (n = 2), HPA-3a (n = 6), HPA-6b (n = 3) or HPA-7 variant (n = 1) antibodies, all antibodies were detected and determined by our new method, except for two HPA-3a antibodies. One of the two antibodies was also negative for conventional platelet MAIPA, suggesting that the cell line panel might be used as an alternative source of platelet antigens in the MAIPA assay.


Assuntos
Antígenos de Plaquetas Humanas , Imunoensaio/métodos , Isoanticorpos/análise , Anticorpos Monoclonais , Linhagem Celular , Humanos , Isoanticorpos/sangue
13.
Transfus Med ; 21(3): 166-74, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21208306

RESUMO

AIMS/OBJECTIVES: Platelets undergo structural and biochemical alterations during in vitro storage and these are collectively called platelet storage lesions (PSL). The mitochondrion is an important cell organelle involved not only in energy production but also in the regulation of cellular functions and viability. This implies that some platelet functions may be regulated by mitochondria; hence, preservation of mitochondrial functions may be important for the maintenance of platelet quality in stored platelet concentrates (PCs). This work describes the effects of various compounds on mitochondrial functions important for the maintenance of platelet quality in in vitro stored PCs. METHODS: PCs were stored at 22 °C with gentle agitation in the presence or absence of 2,4-dinitrophenol, antimycin A, acetyl-l-carnitine and ascorbic acid. The effects of these products on platelet quality were assessed by analysing glucose and lactate concentrations, pH of the storage medium, shape of the platelets, mitochondrial membrane potential and depolarisation, surface expression of CD62P and collagen-induced platelet aggregation. RESULTS: 2,4-Dinitrophenol and antimycin A increased PSL levels, whereas acetyl-l-carnitine reduced the level of changes in pH and mitochondrial depolarisation. Ascorbic acid in the storage medium resulted in improved levels of collagen-induced platelet aggregation. However, none of the examined reagents suppressed CD62P expression in platelets. CONCLUSIONS: These results suggest that preservation of mitochondrial function is fundamental, but not fully sufficient, for the maintenance of platelet in vitro quality during storage. Further research is necessary to develop methods for preserving both mitochondrial and platelet functions in in vitro stored PCs.


Assuntos
Preservação de Sangue/métodos , Mitocôndrias/fisiologia , Ativação Plaquetária , 2,4-Dinitrofenol/farmacologia , Acetilcarnitina/farmacologia , Antimicina A/farmacologia , Ácido Ascórbico/farmacologia , Humanos , Mitocôndrias/efeitos dos fármacos , Ativação Plaquetária/efeitos dos fármacos
15.
Transfus Med ; 16(2): 111-8, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16623917

RESUMO

Although flow cytometric (FCM) analysis is one of the most widely used approaches to screen the presence of leucocyte antibodies, it has several drawbacks. First, neutrophils and, especially, monocytes exhibit high background reactivity. Second, to determine antibody specificity, it is often necessary to examine not only neutrophils and monocytes but also other lineage cells including T cells, B cells and platelets. Therefore, we attempted to establish an FCM analysis system in which four lineages of leucocytes and platelets are simultaneously tested with low background. FCM analysis was performed using ethylene diamine tetraacetic acid-anticoagulated whole blood as cell sample without any cell preparation. Discrimination of five cell lineages was carried out based on the differences in forward vs. side scatter distribution and in the expression of CD4, CD20 and CD14. When anti-HNA (human neutrophil antigen) 1b antiserum was applied to HNA 1b-positive blood samples, only neutrophils were unambiguously positive. When anti-Naka (anti-CD36) antiserum was applied, only platelets and monocytes were positive. The background reactivity of neutrophils and monocytes was low enough. When anti-human leucocyte antigen (HLA) class II antiserum was tested, only B-lymphocytes and monocytes were positive. When anti-HLA class I antiserum was tested, all the five-lineage cells were positive.


Assuntos
Especificidade de Anticorpos , Antígenos CD/análise , Citometria de Fluxo , Leucócitos/citologia , Anticorpos/química , Anticorpos/imunologia , Especificidade de Anticorpos/imunologia , Antígenos CD/imunologia , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Humanos , Leucócitos/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
16.
Immunohematology ; 22(1): 23-9, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16563048

RESUMO

cDNAs for alpha 1,4 galactosyltransferase (A4GALT) have been isolated. To explore the molecular basis of the p phenotype in Japanese donors, we analyzed the A4GALT gene sequences of normal and p phenotype samples. The coding region in the A4GALT gene for DNA sequencing was amplified by PCR amplification. A4GALT expression vectors for individual were constructed by PCR amplification of the coding region using primers and subsequent subcloning into an expression vector. The expression of Gb3/CD77 antigen on the cell surface was evaluated by flow cytometry and by immunochemical techniques. All individuals with the p phenotype were found to have a single base insertion (A4GALT/insC) at the same nucleotide position. Neither the transfectant cells with a mutant gene (A4GALT/insC) of donor origin or those with a synthesized mutant gene (A4GALT/insC-Mu) expressed Gb3 antigen indicating that the presence of A4GALT/insC diminished the A4GALT enzyme activity. In addition, an allele-specific PCR (ASP) system was developed in which of the p phenotype with A4GALT/insC can be unambiguously discriminated from normal donors. Based on the finding that a single base insertion (A4GALT/insC) diminishes A4GALT activity, an ASP assay was developed to detect individuals with this particular p phenotype.


Assuntos
Antígenos Glicosídicos Associados a Tumores/biossíntese , Galactosiltransferases/genética , Regulação Enzimológica da Expressão Gênica/genética , Mutagênese Insercional , Sistema do Grupo Sanguíneo P/biossíntese , Antígenos Glicosídicos Associados a Tumores/genética , Linhagem Celular , Mutação da Fase de Leitura , Galactosiltransferases/análise , Galactosiltransferases/biossíntese , Humanos , Mutagênese Insercional/métodos , Sistema do Grupo Sanguíneo P/genética , Reação em Cadeia da Polimerase/métodos , Transfecção/métodos
17.
Man Ther ; 8(1): 42-5, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12635636

RESUMO

Muscle therapy, a form of manual therapy, was applied to control pain persisting for more than 1 week following posterolateral thoracotomy, and its efficacy for the alleviation of pain was investigated. Eight patients who underwent posterolateral thoracotomy and lung resection for cancer (n=7) or emphysema (n=1) received manual therapy to incised muscles and the muscles inserting into the ribs in the affected area for an average of 17 days postoperatively. Pressure-friction and stretching techniques were used. Treatment was continued until the intensity of the pressure-friction technique reached a level at which the patient complained of pain and a decrease in muscle tone was detected. Treatment was performed once a week for 3 weeks. Pain severity was measured using a visual analog scale (VAS) (0-10). Before the first treatment, the VAS was set at 10, and changes of the score were observed before and after the treatment as well as over time. After three sessions, all patients showed a decrease in pain from 10 to an average of 1.9 (range 1.3-2.6).


Assuntos
Osteopatia/métodos , Dor Pós-Operatória/etiologia , Dor Pós-Operatória/terapia , Toracotomia/efeitos adversos , Adulto , Idoso , Feminino , Humanos , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Medição da Dor , Enfisema Pulmonar/cirurgia , Índice de Gravidade de Doença , Fatores de Tempo , Resultado do Tratamento
19.
J Pharm Sci ; 91(11): 2382-9, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12379923

RESUMO

The hemolytic behavior of a novel cytoprotective agent, DY-9760e (3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate) was investigated using rabbit erythrocytes. Further, the effects of water-soluble cyclodextrin derivatives, such as 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and sulfobutyl ether of beta-cyclodextrin (SBE-beta-CyD), on the hemolytic activity of DY-9760e were studied. DY-9760e induced hemolysis at concentrations >0.2-0.3 mM in phosphate buffered saline (PBS) of pH 4.0 and 6.0, where DY-9760e is predominantly in dicationic and monocationic forms, respectively. The hemolytic activity of the monocationic DY-9760e was higher than that of the dicationic species, and the hemolysis at pH 4.0 involved the formation of methemoglobin. DY9760e induced the morphological change of erythrocytes towards membrane invagination at both pH 4.0 and 6.0. SBE7-beta-CyD significantly suppressed the DY-9760e-induced hemolysis and morphological change at both pH 4.0 and 6.0, as well as the formation of methemoglobin at pH 4.0. On the other hand, HP-beta-CyD suppressed only the hemolysis, but neither the morphological change nor the formation of methemoglobin. In addition, the inhibitory effect of SBE7-beta-CyD on the hemolysis was greater than that of HP-beta-CyD. The superior inhibitory effect of SBE7-beta-CyD on the DY-9760-induced hemolysis, the morphological change, and the formation of methemoglobin may be attributable to the formation of a stable inclusion complex with DY-9760e and to the weaker hemolytic activity of SBE7beta-CyD than HP-beta-CyD. These results suggest potential use of SBE7-beta-CyD as a parenteral carrier for DY-9760e.


Assuntos
Ciclodextrinas/farmacologia , Hemólise/efeitos dos fármacos , Indazóis/farmacologia , beta-Ciclodextrinas , Animais , Citoproteção/efeitos dos fármacos , Citoproteção/fisiologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/fisiologia , Hemólise/fisiologia , Indazóis/antagonistas & inibidores , Indazóis/química , Coelhos
20.
Cytotherapy ; 4(2): 109-18, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12006206

RESUMO

BACKGROUND: In vitro maintenance and expansion of human hematopoietic stem cells is crucial for many clinical applications, and investigators have been using xenogeneic, especially murine, stromal cells for stem-cell expansion. In addition, many such culture systems utilize FCS-containing medium or serum-free medium that contains human- or animal-derived proteins. However, the possible transmission of infectious diseases has led to a debate about the safety of the delivery of grafts expanded in culture using cells and proteins of allogeneic or xenogeneic origin. Using primary human BM stromal cells, we have established an AB serum-based co-culture system to expand human primitive progenitors and transplantable stem cells. METHODS: Cord blood CD34+ cells were cultured on a monolayer of human BM-derived primary stromal cells with thrombopoietin (TPO), stem-cell factor (SCF) and flt3/flk2 ligand (FL) in the presence of either FCS or AB serum. One to three weeks later, cells were examined for total cells, CD34+ cells, CD34+ CD38- cells, and clonogenic progenitors. SCID mouse reconstituting cell (SRC) activity was also studied. RESULTS: Three weeks of culture with TPO, SCF, and FL supported more than a 250-fold expansion of CD34+ cells, CD34+ CD38- cells and CFU-C, regardless of the kind of serum used. SRC assay revealed that transplantable stem cells were moderately expanded as well. DISCUSSION: This ex vivo expansion system should prove valuable in clinical settings in which stromal cells and serum are available from recipients or stem-cell donors.


Assuntos
Técnicas de Cultura de Células/métodos , Sangue Fetal/citologia , Células-Tronco Hematopoéticas , ADP-Ribosil Ciclase/metabolismo , ADP-Ribosil Ciclase 1 , Animais , Antígenos CD/metabolismo , Antígenos CD34/metabolismo , Células Cultivadas , Células Clonais , Técnicas de Cocultura , Meios de Cultura Livres de Soro , Glicoproteínas de Membrana , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Fator de Células-Tronco/metabolismo , Transplante de Células-Tronco , Células Estromais , Trombopoetina/metabolismo
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