RESUMEN
A study was conducted to compare the efficiency and toxicity of two peripheral blood stem cell (PBSC) mobilization procedures for newly diagnosed patients with multiple myeloma. Patients from group 1 (n=51) were treated by high-dose cyclophosphamide (HD-CY) plus G-CSF (5 microg/kg/day), and the second group (n=31) by VAD regimen plus G-CSF administration (10 microg/kg/day). Successful mobilization, defined by a minimal count of 2.5 x 10(6) CD34(+) cells/kg collected, was achieved in 96 and 90% of patients in groups 1 and 2, respectively (P=0.15). The mean peripheral blood CD34(+) cells concentration and the mean CD34(+) cells/kg collected were higher in group 2 than in the group 1 (P=0.05). The mean number of leukaphereses necessary to collect a count of 2.5 x 10(6) CD34(+) cells/kg was reduced in group 2 compared to group 1. Adverse events, blood products consumption and time spent in the hospital were significantly greater after HD-CY. In conclusion, VAD plus a G-CSF dose of 10 microg/kg administration seems preferential to HD-CY plus a G-CSF dose of 5 microg/kg for PBSC collection because of equivalent or better efficiency in stem cell mobilization, strong favorable toxicity profile and reduced cost.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Ciclofosfamida/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Movilización de Célula Madre Hematopoyética/métodos , Mieloma Múltiple/terapia , Antígenos CD34/biosíntesis , Separación Celular , Ciclofosfamida/metabolismo , Dexametasona/uso terapéutico , Doxorrubicina/uso terapéutico , Femenino , Citometría de Flujo , Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Trasplante de Células Madre Hematopoyéticas/métodos , Humanos , Masculino , Persona de Mediana Edad , Mieloma Múltiple/metabolismo , Células Madre/citología , Factores de Tiempo , Resultado del Tratamiento , Vincristina/uso terapéuticoRESUMEN
The effects of adriamycin (ADM), arabinosyl-cytosine (ARA-C), and etoposide (VP16) were studied on human bone marrow mononucleated cells using colony formation in agar, a modified liquid culture system, and flow cytometry analysis of the cell cycle. Drug concentrations tested during a 1-h incubation ranged from 0.1 to 4 micrograms/ml for ADM, from 0.3 to 30 micrograms/ml for VP16, and from 10(-7) to 10(-3) M for ARA-C. Regression analysis of the dose-response curves was used to assess the drug concentration that inhibited 90% +/- 5% (LD90) of colony growth. LD90s were 0.4 microgram/ml for ADM, 20 micrograms/ml for VP16, and 10(-4) M for ARA-C. LD90-surviving cells were cultured in liquid medium for 3 weeks. Surviving cells over this time were 13% of the control for ADM, 22% for VP16, and 95.7% for ARA-C. Although cells decreased drastically in ADM- and VP16-treated samples, granulocyte-macrophage colony-forming units (CFU-GM) per 10(5) surviving cells rose to twice the control for ADM, to 60% for VP16, and to 150% for ARA-C. Flow cytometry analysis of the cell cycle was performed at day 0 and at day 4 after treatment with the LD90 dose. It showed a rapid and reversible effect of ARA-C on cells in the S-phase, whereas the action of VP16 concerned all cells, regardless of their cycle phase. We conclude that the direct effects of the three drugs on CFU-GM in agar are poorly predictive of hematopoietic reconstitution capacity, except for VP16. Liquid culture gives a much more accurate appraisal of the long-term damage and recovery due to anticancer drugs.
Asunto(s)
Células de la Médula Ósea , Citarabina/farmacología , Doxorrubicina/farmacología , Etopósido/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Ciclo Celular , Supervivencia Celular , Medios de Cultivo , Granulocitos/citología , Granulocitos/efectos de los fármacos , Humanos , Monocitos/citología , Monocitos/efectos de los fármacosRESUMEN
OBJECTIVE: The aim of this study was to analyze bone marrow lymphocyte subsets and CD34 cell dose and their influence on the outcomes of bone marrow transplantation. MATERIALS AND METHODS: Forty-eight patients (median age 30 years, range 5-54) receiving HLA-identical sibling bone marrow transplantation for hematologic malignancies were analyzed. RESULTS: Median number (range) of nucleated cells and CD34+ cells infused were 2.4 (0.4-6.0) x 10(8)/kg and 3.5 (0.5-13.0) x 10(6)/kg, respectively. Probability of neutrophil recovery was 97%. In a multivariate analysis, time to neutrophil recovery was shortened when a higher number of CD3/CD8 cells was infused (> or =1.0 x 10(7)/kg) (hazard ratio [HR] = 2.13, p = 0.018); when the patient was female or had negative cytomegalovirus serology (HR = 2.03, p = 0.03; HR = 0.41, p = 0.009; respectively). The incidence of grade II to IV acute graft-vs-host disease (GVHD) was 47%. Infusion of >1 x 10(7) CD4 infused/kg increased the risk of acute GVHD (HR = 2.86, p = 0.03). Nineteen of 40 patients at risk experienced chronic GVHD, the risk of which was increased by diagnosis of chronic leukemia (p = 0.03), <2.0 x 10(8) nucleated cells infused/kg (p = 0.05), and a low number of all lymphocyte subsets, except CD19. Estimated 3-year survival rate was 54%. Risk of death was increased in patients receiving <3.5 x 10(6)CD34 infused/kg (HR = 0.37, p = 0.02). Only six patients relapsed. CONCLUSIONS: A high cell dose of CD3/CD8 is associated with faster neutrophil recovery, whereas a high cell dose of CD4+ cells increases the incidence of acute GVHD. A high number of nucleated cells and CD34+ cells infused was associated with decreased risk of chronic GVHD and improved survival, respectively.
Asunto(s)
Trasplante de Médula Ósea , Subgrupos Linfocitarios/trasplante , Enfermedad Aguda , Adolescente , Adulto , Antígenos CD34/análisis , Trasplante de Médula Ósea/mortalidad , Complejo CD3/análisis , Linfocitos T CD4-Positivos/trasplante , Linfocitos T CD8-positivos/trasplante , Niño , Preescolar , Enfermedad Crónica , Comorbilidad , Infecciones por Citomegalovirus/epidemiología , Femenino , Francia/epidemiología , Supervivencia de Injerto , Enfermedad Injerto contra Huésped/epidemiología , Enfermedad Injerto contra Huésped/inmunología , Enfermedad Injerto contra Huésped/prevención & control , Neoplasias Hematológicas/mortalidad , Neoplasias Hematológicas/terapia , Histocompatibilidad , Humanos , Incidencia , Recuento de Leucocitos , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Neutrófilos , Núcleo Familiar , Riesgo , Análisis de Supervivencia , Donantes de Tejidos , Trasplante HomólogoRESUMEN
A novel retroviral vector has been designed based on a Friend-murine leukemia virus (Fr-MuLV) FB29 strain. The latter has been selected according to characteristics of pathogenicity in mice where it induces a disease of the haemopoietic system affecting all lineages. Higher infectivity has also been demonstrated as compared to other strains. In accordance with these findings, the amphotropic producer clone used in this study carrying along the neomycine resistance gene (FOCH-Neo), harbors viral titers over 10(7) cfu/ml. To investigate the potential of genetically engineering hematopoietic precursors, CD34+ progenitors were selected from cord blood, bone marrow, and peripheral blood mobilized stem cells (patients + solid tumors) and transduced with FOCH-Neo. High transduction rates were achieved using virus supernatant and minimal doses of hematopoietic growth factors during pretransduction and transduction steps. A polymerase chain reaction (PCR) assay investigating the presence of both neomycin-encoding and viral vector sequences tested positive in 45-90% of granulocyte-macrophage colony-forming units (CFU-GM) generating cells (bone marrow and peripheral blood derived cells) following transduction. An average of 35% colonies showed resistance to G418. Such levels of transduction proved reproducible using only supernatants harboring over 10(7) cfu/ml. In those experiments where long-term in vitro cultures could be maintained over 5 weeks (all cord blood and 5 among 23 PBSC), efficient transduction of long-term culture initiating cell (LTC-IC) hematopoietic progenitors was demonstrated on the basis of both resistance to G418 and virus integration. In the latter case, the PCR assay tested positive in as much as 35-60% of late unselected CFU-colonies. This novel retroviral vector harbors interesting features toward genetic modification of hematopoietic progenitors.
Asunto(s)
Antígenos CD34 , Virus de la Leucemia Murina de Friend/genética , Vectores Genéticos/genética , Células Madre Hematopoyéticas/metabolismo , Transducción Genética , Células 3T3 , Animales , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/virología , Sistema Libre de Células/virología , Técnicas de Cocultivo , Sangre Fetal/citología , Virus de la Leucemia Murina de Friend/crecimiento & desarrollo , Vectores Genéticos/efectos de los fármacos , Vectores Genéticos/metabolismo , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/virología , Humanos , Ratones , Factor de Células Madre/farmacología , Células Madre/metabolismo , Células Madre/virología , Transducción Genética/efectos de los fármacos , Trasplante HomólogoRESUMEN
Umbilical cord blood T cells are less functional. Different explanations have been proposed. In this study we analyze the Vbeta T cell cord blood repertoire. All the Vbeta families are expressed. We found only the overexpression of three Vbeta: Vbeta 5-1, Vbeta 5-2, Vbeta 21-2.
Asunto(s)
Sangre Fetal/citología , Linfocitos T/citología , Adulto , Sangre Fetal/química , Humanos , Recién Nacido , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Linfocitos T/químicaRESUMEN
Positive selection of CD34+ cells is being increasingly performed to support hematological reconstitution following high-dose and dose-intensive chemotherapy and to reduce the non-target cell content of transplants. The present study was designed to evaluate the performance of an immunomagnetic cell selection system, including comparison of enzyme and peptide releasing agents and of semi-automated and fully automated selection systems. A total of 74 immunomagnetic CD34+ cell selection procedures were performed involving 55 subjects, the majority of whom had hematologic malignancies. Median CD34+ cell purity with a newly developed specific octapeptide releasing agent (98.5%; 81.0-99.0%) was significantly higher (P = 0.002) than that with chymopapain (85.8%; 28.1-99.7%). No significant differences were observed between semi-automated and fully automated systems in CD34+ cell purity or yield or time to WBC or platelet recovery. Immunomagnetic selection was found to provide highly purified populations of CD34+ cells in sufficient numbers for use in transplantation procedures. CD34+ cell transplants supported rapid and reliable hematologic reconstitution. Use of a fully automated system markedly reduced the time and labor required for immunomagnetic selection, potentially affording more standardized and reproducible positive selection of CD34+ cells.
Asunto(s)
Antígenos CD34/análisis , Trasplante de Células Madre Hematopoyéticas , Separación Inmunomagnética , Oligopéptidos/farmacología , Adulto , Eliminación de Componentes Sanguíneos , Quimopapaína/farmacología , Femenino , Citometría de Flujo , Movilización de Célula Madre Hematopoyética , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
Donor potential to exert NK cell alloreactivity has been shown to confer survival advantage in haploidentical hematopoietic cell transplantation for hematological malignancies. We investigated killer immunoglobulin receptor (KIR) ligand incompatibility in 40 children receiving haploidentical transplantation for primary immunodeficiencies. The conditioning regimen consisted of busulfan and cyclophosphamide. T-cell depletion of the graft used complement-dependent lysis or CD34+ selection. Two patients died in the first month. The remaining 38 patients were divided into those with (n=13) and those without (n=25) donor potential to exert NK cell alloreactivity. Engraftment was similar in the two groups (61.5 and 64%, respectively). The incidence of grade II-IV acute graft-versus-host disease (GVHD) tended to be lower in the group with donor potential to exert NK cell alloreactivity, but the difference was not significant. In conclusion, in this series of patients with primary immunodeficiencies, donor potential to exert NK cell alloreactivity was not associated with significant advantages in engraftment and prevention of acute GVHD.
Asunto(s)
Enfermedad Injerto contra Huésped/inmunología , Trasplante de Células Madre Hematopoyéticas , Células Asesinas Naturales/inmunología , Inmunodeficiencia Combinada Grave/terapia , Acondicionamiento Pretrasplante , Preescolar , Femenino , Enfermedad Injerto contra Huésped/prevención & control , Humanos , Depleción Linfocítica , Masculino , Receptores Inmunológicos/inmunología , Receptores KIR , Inmunodeficiencia Combinada Grave/inmunología , Trasplante HomólogoRESUMEN
Harvesting of peripheral blood stem cells (PBSCs) following chemotherapy and G-CSF administration is currently performed for hematological therapies. However, a procedure based on the use of a large quantity of G-CSF is relatively costly. Therefore, we retrospectively compared the effects of two PBSC mobilization procedures in a population with recently diagnosed multiple myeloma. The first procedure consisted of chemotherapy and systematic G-CSF administration (group 1: 24 patients). The second consisted of chemotherapy alone, G-CSF having been administered only in the case of failure of PBSC mobilization or delayed white blood cell (WBC) recovery (group 2: 28 patients). Leukapheresis was performed when WBC recovery reached 1 x 10(9)/l if the peripheral blood CD34+ cell count was over 10/microl. Leukapheresis was maintained until a total of 2.5 x 10(6) CD34+ cells/kg was harvested. A significant difference was observed between the two groups only in regard to the median period of WBC recovery (delayed for group 2) and the number of CD34+ cells/kg collected on the first leukapheresis (higher for group 1) but not to the proportion of patients with failure of PBSC collection. Ten group 2 patients, who had insufficient CD34+ cells after WBC recovery or delayed WBC recovery, received G-CSF which resulted in sufficient PBSC harvesting in nine. To obtain a sufficient CD34+ cell level, the patients without systematic G-CSF administration had more leukaphereses (2.1 vs 1.5) but the mean consumption of G-CSF per patient was eight times less than in the other group. Nonsystematic use of G-CSF before WBC recovery or preferentially its introduction just after, could be an interesting economical alternative in PBSC mobilization but should be assessed by a prospective controlled study of cost/efficacy.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Células Sanguíneas , Factor Estimulante de Colonias de Granulocitos/farmacología , Movilización de Célula Madre Hematopoyética/métodos , Células Madre Hematopoyéticas , Leucaféresis , Mieloma Múltiple/sangre , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Terapia Combinada , Ciclofosfamida/administración & dosificación , Ciclofosfamida/farmacología , Doxorrubicina/administración & dosificación , Doxorrubicina/farmacología , Esquema de Medicación , Estudios de Evaluación como Asunto , Femenino , Movilización de Célula Madre Hematopoyética/economía , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Mieloma Múltiple/tratamiento farmacológico , Mieloma Múltiple/terapia , Prednisona/administración & dosificación , Prednisona/farmacología , Estudios Retrospectivos , Vincristina/administración & dosificación , Vincristina/farmacologíaRESUMEN
SUMMARY: Hematological inherited diseases can be cured by hematopoietic stem cell transplantation (HSCT) from an human leukocyte antigen (HLA)-identical sibling donor (MSD), but the outcome of unrelated donors (URD) or haploidentical donors (HMD) has been a cause of concern. In all, 94 children affected with inherited diseases underwent HSCT at a single center using MSD (group A, n=31), URD (group B, n=23) or HMD (group C, n=40). There was no difference in the rate of engraftment or in the incidence of grades III-IV acute graft-versus-host disease (GVHD) between the groups. Survival rate was 80.6% in group A, 62.5% in group B and 47.5% in group C (P=0.023). In group B, survival rate was 73.7% in the subgroup with zero or one class I mismatch, and 25% in the subgroup with two or more class I mismatches (P=0.04). In group C, survival rate was 83.3% in the 9/10-identical subgroup, 64.3% in the seven or 8/10 subgroup, and 25% in the five or 6/10 subgroup (P=0.0007). Thus, engraftment, incidence of GVHD and survival are similar in recipients of grafts from MSD, URD with 0-1 class I-mismatch, or HMD with at least 7/10 HLA matches. The low success of HSCT using more disparate donors suggests reserving them for patients with very poor prognosis.
Asunto(s)
Enfermedades Genéticas Congénitas/terapia , Trasplante de Células Madre Hematopoyéticas , Prueba de Histocompatibilidad , Histocompatibilidad/genética , Adolescente , Niño , Preescolar , Enfermedades Genéticas Congénitas/complicaciones , Enfermedades Genéticas Congénitas/mortalidad , Genotipo , Supervivencia de Injerto/inmunología , Enfermedad Injerto contra Huésped/inmunología , Haplotipos , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Trasplante de Células Madre Hematopoyéticas/mortalidad , Trasplante de Células Madre Hematopoyéticas/estadística & datos numéricos , Humanos , Lactante , Infecciones Oportunistas/inmunología , Análisis de Supervivencia , Donantes de Tejidos , Trasplante Homólogo , Trasplante Isogénico , Resultado del TratamientoRESUMEN
In order to study the influence of bone marrow CD34(+) cell dose on the outcome of allogeneic bone marrow transplantation (BMT), we analysed the results of BMT from HLA-identical siblings donors in 50 patients with chronic myeloid leukaemia (CML). The median numbers of nucleated cells (NC) and CD34(+) cells infused were 2.18 x 10(8)/kg (0.05-4.14 x 10(8)/kg) and 3.12 x 10(6)/kg (0.35-8.52 x 10(6)/kg), respectively. All patients engrafted. In univariate analysis, there was no correlation between the number of CD34(+) cells infused and the time to neutrophil recovery (P = 0.17). The Kaplan-Meier estimate of grade II-IV acute graft-versus-host disease (GVHD) at day 100 was 53 +/- 14% and 2-year survival was 46 +/- 15%. A number of CD34(+) cells infused greater than the median was the main factor increasing survival (P = 0.0006) and decreasing 100 day transplant-related mortality (P = 0.009). Patient-, disease- and transplant-related characteristics were not statistically different among patients receiving more or less than the median number of CD34(+) cells. The rate of infectious deaths was significantly higher in patients receiving less than 3.12 x 10(6) CD34/kg (48% vs 16%, P = 0.01). In a multivariable analysis, two factors associated with increased risk of death were advanced disease status at transplant (HR: 2.5 (95% CI: 1.09-5.75), P = 0.03) and a lower number of marrow CD34(+) cells infused/kg (HR: 4.55 (95% CI: 1.87-10.90), P = 0.0008).
Asunto(s)
Antígenos CD34/análisis , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Adolescente , Adulto , Antígenos CD34/farmacología , Células de la Médula Ósea/citología , Células de la Médula Ósea/inmunología , Trasplante de Médula Ósea/inmunología , Causas de Muerte , Recuento de Células , Femenino , Citometría de Flujo , Estudios de Seguimiento , Supervivencia de Injerto , Enfermedad Injerto contra Huésped , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/complicaciones , Leucemia Mielógena Crónica BCR-ABL Positiva/mortalidad , Masculino , Persona de Mediana Edad , Núcleo Familiar , Tasa de Supervivencia , Trasplante Isogénico/inmunología , Resultado del TratamientoRESUMEN
Blood, brain, lung, and gastric contents of a drowning victim recovered from an automobile in which there was a strong odor of gasoline were examined for the presence of gasoline. Heated headspace, capillary column gas chromatography (GC) was employed for the analysis of the samples with simple pattern recognition being used to complete the determination. A mathematical discussion of the probability of artifact components or other organic compounds in the samples causing results which could falsely appear to be gasoline is discussed.
Asunto(s)
Gasolina/análisis , Petróleo/análisis , Adulto , Cromatografía de Gases/métodos , Medicina Legal , Gasolina/envenenamiento , Humanos , Masculino , Reconocimiento de Normas Patrones AutomatizadasRESUMEN
Concomitant findings of isopropanol and acetone in biospecimens of decedents known not to have been exposed to the alcohol prompted a study to explain its origins. Mixtures of acetone, ADH, and NADH at pHs 7.3 and 8.8 were incubated at 37 degrees C for varying intervals. Reaction products were then analyzed by headspace GC and assured identification made by GC/MS. It was found that isopropanol is produced by reduction of acetone at pH 7.3 (to a lesser extent at pH 8.8), providing evidence for an alternate metabolic route for acetone. A mechanism for this reduction is proposed. Ranges for isopropanol (in mg/dL or mg/100 g) found in unexposed decedents were: blood 1-29; liver 7-59; brain 2-12; kidney 6-26. Thus, the forensic investigator must interpret isopropanol results cautiously, particularly when low concentrations are found.
Asunto(s)
1-Propanol/análisis , Medicina Legal , Acetona/análisis , Acetona/metabolismo , Alcohol Deshidrogenasa , Oxidorreductasas de Alcohol/farmacología , Cromatografía de Gases , Humanos , Concentración de Iones de HidrógenoRESUMEN
Qualitative and quantitative analysis of tissues and body fluids for multiple volatile organic compounds were performed by a combination of packed and open tubular capillary GC and GC/MS. This paper describes methods for such analyses in a case involving the exposure of two persons to methyl ethyl ketone, methyl isobutyl ketone, toluene and the three isomeric xylenes. Tissue and body fluid concentrations of these substances in the two decedents are presented and discussed briefly.
Asunto(s)
Butanonas/análisis , Cetonas/análisis , Metil n-Butil Cetona/análisis , Tolueno/análisis , Xilenos/análisis , Cromatografía de Gases , Medicina Legal , Cromatografía de Gases y Espectrometría de Masas , HumanosRESUMEN
The stability of nortriptyline in aqueous solutions containing various concentrations of formaldehyde was investigated. Amitriptyline, as a reaction product, was determined by gas chromatography/mass spectrometry (GC/MS) in these experiments. Factors that may contribute to this phenomenon, including pH, formaldehyde concentration, and incubation time were evaluated. At 40% (v/v) formaldehyde concentration and pH 4, there was a 68% decrease in nortriptyline concentration along with a concomitant formation of amitriptyline after 24 h. The N-methylated product was responsible for 48% of the total tricyclic drug present. The data also clearly indicate that the formation of amitriptyline is favored at elevated pH.
Asunto(s)
Amitriptilina/química , Formaldehído/química , Nortriptilina/química , Amitriptilina/análisis , Embalsamiento , Cromatografía de Gases y Espectrometría de Masas , Concentración de Iones de Hidrógeno , Nortriptilina/análisisRESUMEN
Hematopoietic progenitor cells are present in umbilical cord blood; placental blood (PB) previously considered as waste product now constitutes an alternative source of hematopoietic stem cells for bone marrow reconstitution. This has promoted the establishment of cord blood banks for use in unrelated transplants. The banking of PB offers many advantages: the donors do not require anesthesia, stored PB can be a valuable source of stem cells for patients from ethnic minorities underrepresented in volunteer registers, and stored PB can be made available much faster than bone marrow from unrelated donors. Preliminary clinical experience suggests that, due to the immunological immaturity of PB cells, graft versus host disease might be lower than when using bone marrow from adult donors and HLA restrictions might be less stringent. If the number of nucleated cells in PB often appears low for patients weighing more than 40 kg, clinical data suggest that the number of stem cells may be sufficient for adult transplantation. The number of cord blood banks throughout the world is increasing rapidly. In the USA and Europe, more than 10,500 PB units are stored and available for transplantation. In the next 5 years, a total of 50,000 PB will be reached which may be sufficient to provide for the majority of candidates for unrelated BM transplantation. The practices of umbilical cord blood collection, mother selection, infectious disease screening, cell manipulation and storage must be standardized. Some accreditation process should be mandatory for assessing operating procedures and the quality assurance programs of the banks, and for allowing the international exchange of placental blood between transplant centers.
Asunto(s)
Bancos de Sangre/normas , Sangre Fetal , Trasplante de Células Madre Hematopoyéticas , Adulto , Bancos de Sangre/economía , Recolección de Muestras de Sangre/normas , Enfermedades Genéticas Congénitas/diagnóstico , Prueba de Histocompatibilidad , Humanos , Recién Nacido , Padres , Control de CalidadRESUMEN
Ethanol was determined by gas chromatography in a variety of tissues and body fluids secured at autopsy in 61 cases. The specimens tested included right and left heart blood, femoral blood, pericardial fluid, cerebrospinal fluid, vitreous humor, urine, stomach contents, and brain. Statistical analysis of the cases revealed no significant differences among the various blood sites tested. However, the variations in blood ethanol concentrations among the various sampling sites within each case were as follows: 40 cases showed differences of less than 25%; 16 cases revealed variability between 25% and 50%, 4 cases had differences exceeding 50%. In one case, satisfactory blood analyses could not be accomplished. The larger variances occurred especially in those instances in which stomach alcohol concentration was 0.50% or greater. In one case, the variability amongst the different blood sites exceeded 400% (femoral blood--0.043%, right atrium--0.070%, root of aorta--0.156%); the brain was 0.050%, and the stomach contents was 1.2%. For all 61 cases, variances in blood alcohol content among the different sampling sites in a single cadaver ranged from 1.8 to 428%.
Asunto(s)
Autopsia/métodos , Recolección de Muestras de Sangre , Etanol/sangre , Medicina Legal/métodos , Cambios Post Mortem , Absorción , Análisis de Varianza , Líquidos Corporales/química , Química Encefálica , Cromatografía de Gases , Etanol/análisis , Etanol/farmacocinética , Contenido Digestivo/química , HumanosRESUMEN
A nurse was accused of attempting to murder her anesthesiologist husband on two occasions by administering to him a neuromuscular blocking agent. In both episodes, urine specimens were obtained from the victim shortly after the suspected assaults. The samples were initially tested fluorometrically using Rose Bengal dye as a pairing agent. Both were presumptively positive for pancuronium. Confirmation of these results was achieved by pairing the drug with potassium iodide, extracting the complex, and submitting the extract to thin-layer chromatography (TLC) cleanup, elution at the appropriate retardation factor (Rf), and, finally, gas chromatography/mass spectrometry (GC/MS) analysis in the selected-ion monitoring mode. The two quaternary amines of pancuronium appear to undergo pyrolytic N-demethylation in the injection port to yield an entity amenable to capillary column gas chromatography. The mass spectrum of the compound consists of a base peak of m/z 322, with additional fragments of 292, 323, 338, and 397 m/z, each of which was monitored. The confirmed positive findings were instrumental in adjudicating the case.
Asunto(s)
Homicidio , Pancuronio/envenenamiento , Pancuronio/orina , Cromatografía de Gases y Espectrometría de Masas , Humanos , MasculinoRESUMEN
PURPOSE: Today, haematopoietic stem cell graft from placental blood concerns more than 15 % of allogeneic grafts. An inter-laboratory study of the quality control of defrosted cord blood units has been coordinated by the French society for cell and tissue bioengineering (SFBCT), with the cord blood bank of Bourgogne Franche-Comté and controlled by the French health products safety agency (Afssaps). The aim of this study is to ensure the inter-laboratory reproducibility of the quality controls practised by the banks during defrosting. The cellular outputs were analyzed according to the defrosting techniques, according to the method used in flow cytometry: single-platform (SP) versus double-platform (DP), or the product nature, i.e. in total blood or miniaturized. METHODS: Forty-two units of placental blood (USP), which were out of range were provided for defrosting to 14 participating sites. USP were defrosted and controlled according to the procedures of each bank. Once the USP is defrosted, a part of the product was controlled by the site and the other part by Afssaps. Following controls were carried out: numeration of the total nucleated cells (TNC) and of CD34+ cells (made by a SP method in Afssaps) and functional assay. RESULTS: Concerning TNC, the defrosting sites obtained a cellular output of 94 %+/-28 in day 0 compared with an output of 72 %+/-24 in Afssaps showing a rather good stability of the USP transmitted with an average deviation of 23 %+/-22. The freezing process with or without reduction of volume does not affect this variation. Concerning the numeration of CD34+ cells, the average deviation between the participating sites and Afssaps was 29 %+/-23 compared with 21 %+/-16 for the sites using a SP method against 47 %+/-25 for those using a DP method. The CD34+ outputs are equal to 82 % +/- 60 in day 0 for the participating sites against 52 %+/-20 for Afssaps. For the sites using a DP method, it is stressed that this output is particularly high with a rate of 126 %+/-90 (n=15) whereas it is 62 %+/-20 (n=32) for the sites using a SP method. CONCLUSION: These results underline a good stability of viable CD34+ cells and a greater reliability of the SP methods for the CD34+ cell numeration for these defrosted USP. Lastly, the results of the functional assay regarding the average clonogenicities (equal to 15 %) reinforce the conclusions on the quality of the defrosted products.