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1.
Ann Oncol ; 31(9): 1251-1259, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32464282

RESUMEN

BACKGROUND: Outcomes for patients with high-risk diffuse large B-cell lymphoma (DLBCL) treated with R-CHOP chemotherapy are suboptimal but, to date, no alternative regimen has been shown to improve survival rates. This phase 2 trial aimed to assess the efficacy of a Burkitt-like approach for high-risk DLBCL using the dose-intense R-CODOX-M/R-IVAC regimen. PATIENTS AND METHODS: Eligible patients were aged 18-65 years with stage II-IV untreated DLBCL and an International Prognostic Index (IPI) score of 3-5. Patients received alternating cycles of CODOX-M (cyclophosphamide, vincristine, doxorubicin and high-dose methotrexate) alternating with IVAC chemotherapy (ifosfamide, etoposide and high-dose cytarabine) plus eight doses of rituximab. Response was assessed by computed tomography after completing all four cycles of chemotherapy. The primary end point was 2-year progression-free survival (PFS). RESULTS: A total of 111 eligible patients were registered; median age was 50 years, IPI score was 3 (60.4%) or 4/5 (39.6%), 54% had a performance status ≥2 and 9% had central nervous system involvement. A total of 85 patients (76.6%) completed all four cycles of chemotherapy. There were five treatment-related deaths (4.3%), all in patients with performance status of 3 and aged >50 years. Two-year PFS for the whole cohort was 67.9% [90% confidence interval (CI) 59.9-74.6] and 2-year overall survival was 76.0% (90% CI 68.5-82.0). The ability to tolerate and complete treatment was lower in patients with performance status ≥2 who were aged >50 years, where 2-year PFS was 43.5% (90% CI 27.9-58.0). CONCLUSIONS: This trial demonstrates that R-CODOX-M/R-IVAC is a feasible and effective regimen for the treatment of younger and/or fit patients with high-risk DLBCL. These encouraging survival rates demonstrate that this regimen warrants further investigation against standard of care. TRIAL REGISTRATION: ClinicalTrials.gov (NCT00974792) and EudraCT (2005-003479-19).


Asunto(s)
Linfoma de Burkitt , Linfoma de Células B Grandes Difuso , Adolescente , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfoma de Burkitt/tratamiento farmacológico , Ciclofosfamida/uso terapéutico , Supervivencia sin Enfermedad , Doxorrubicina/uso terapéutico , Humanos , Ifosfamida/uso terapéutico , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Persona de Mediana Edad , Prednisona/uso terapéutico , Rituximab/uso terapéutico , Reino Unido , Vincristina/uso terapéutico , Adulto Joven
2.
Ann Bot ; 121(2): 321-334, 2018 02 12.
Artículo en Inglés | MEDLINE | ID: mdl-29300863

RESUMEN

Background and Aims: Wildfires are common in seasonally dry parts of the world with a Mediterranean climate. Prescribed burning is used to reduce fuel load and fire risk, but often without reliable information on its effects. This study investigated the effects of prescribed burns in different seasons on Pterostylis revoluta, an autumn-flowering Australian terrestrial orchid, and its orchid mycorrhizal fungi (OMFs) to find the least damaging season for a prescribed burn. Methods: Burns were conducted mid-season in spring and summer 2011 and autumn and winter 2012. Orchids were enumerated and measured during their flowering season in autumn 2011-2014 and mycorrhizal fungi were isolated before and after the burns in autumn 2011, 2012 and 2014. Micro-organisms isolated were characterized. DNA was extracted from the OMFs, and the internal transcribed spacer region was amplified by PCR. Amplicons were clustered by restriction fragment length polymorphism (RFLP), and representative amplicons were sequenced. OMF were tested for sensitivity to smoke water. Key Results: The number of plants increased up to 4-fold and 90 % of plants became vegetative during this study. Isolation of mycorrhizal fungi increased and isolation of bacteria decreased. Before the burns, the main OMF isolated was unexpectedly Tulasnella calospora (Boud.) Juel. By 2014, after the burns, the expected Ceratobasidium sp. D.P. Rogers was the only OMF isolated in most burnt quadrats, whereas T. calospora was confined to a minority of unburnt 'control' and the 'spring burn' quadrats, which were also the only ones with flowering plants. Conclusions: The decline in rainfall during 2010-2012 probably caused the switch from mainly flowering to mainly vegetative plants and the change in OMFs. Burning in spring to summer was less damaging to this orchid than burning in autumn to winter, which should be noted by authorities in fire management plans for fire-prone areas in which this orchid occurs.


Asunto(s)
Incendios , Micorrizas/fisiología , Orchidaceae/fisiología , Lluvia , Conservación de los Recursos Naturales/métodos , ADN de Hongos/genética , Micorrizas/genética , Orchidaceae/microbiología , Dinámica Poblacional , Estaciones del Año , Victoria
3.
Ann Oncol ; 28(10): 2511-2516, 2017 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-28961838

RESUMEN

BACKGROUND: Central nervous system (CNS) relapse of diffuse large B-cell lymphoma (DLBCL) is associated with a dismal prognosis. Here, we report an analysis of CNS relapse for patients treated within the UK NCRI phase III R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone) 14 versus 21 randomised trial. PATIENTS AND METHODS: The R-CHOP 14 versus 21 trial compared R-CHOP administered two- versus three weekly in previously untreated patients aged ≥18 years with bulky stage I-IV DLBCL (n = 1080). Details of CNS prophylaxis were retrospectively collected from participating sites. The incidence and risk factors for CNS relapse including application of the CNS-IPI were evaluated. RESULTS: 177/984 patients (18.0%) received prophylaxis (intrathecal (IT) methotrexate (MTX) n = 163, intravenous (IV) MTX n = 2, prophylaxis type unknown n = 11 and IT MTX and cytarabine n = 1). At a median follow-up of 6.5 years, 21 cases of CNS relapse (isolated n = 11, with systemic relapse n = 10) were observed, with a cumulative incidence of 1.9%. For patients selected to receive prophylaxis, the incidence was 2.8%. Relapses predominantly involved the brain parenchyma (81.0%) and isolated leptomeningeal involvement was rare (14.3%). Univariable analysis demonstrated the following risk factors for CNS relapse: performance status 2, elevated lactate dehydrogenase, IPI, >1 extranodal site of disease and presence of a 'high-risk' extranodal site. Due to the low number of events no factor remained significant in multivariate analysis. Application of the CNS-IPI revealed a high-risk group (4-6 risk factors) with a 2- and 5-year incidence of CNS relapse of 5.2% and 6.8%, respectively. CONCLUSION: Despite very limited use of IV MTX as prophylaxis, the incidence of CNS relapse following R-CHOP was very low (1.9%) confirming the reduced incidence in the rituximab era. The CNS-IPI identified patients at highest risk for CNS recurrence. CLINICALTRIALS.GOV: ISCRTN number 16017947 (R-CHOP14v21); EudraCT number 2004-002197-34.


Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Neoplasias del Sistema Nervioso Central/patología , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Anticuerpos Monoclonales de Origen Murino/administración & dosificación , Ciclofosfamida/administración & dosificación , Doxorrubicina/administración & dosificación , Esquema de Medicación , Femenino , Humanos , Linfoma de Células B Grandes Difuso/patología , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/patología , Prednisona/administración & dosificación , Estudios Prospectivos , Rituximab/administración & dosificación , Vincristina/administración & dosificación
4.
Ann Oncol ; 28(7): 1540-1546, 2017 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-28398499

RESUMEN

BACKGROUND: There is an on-going debate whether 2- or 3-weekly administration of R-CHOP is the preferred first-line treatment for elderly patients with diffuse large B-cell lymphoma (DLBCL). The UK NCRI R-CHOP14v21 randomized phase 3 trial did not demonstrate a difference in outcomes between R-CHOP-14 and R-CHOP-21 in newly diagnosed DLBCL patients aged 19-88 years, but data on elderly patients have not been reported in detail so far. Here, we provide a subgroup analysis of patients ≥60 years treated on the R-CHOP14v21 trial with extended follow-up. PATIENTS AND METHODS: Six hundred and four R-CHOP14v21 patients ≥60 years were included in this subgroup analysis, with a median follow-up of 77.7 months. To assess the impact of MYC rearrangements (MYC-R) and double-hit-lymphoma (DHL) on outcome in elderly patients, we performed a joint analysis of cases with available molecular data from the R-CHOP14v21 (N = 217) and RICOVER-60 (N = 204) trials. RESULTS: Elderly DLBCL patients received high dose intensities with median total doses of ≥98% for all agents. Toxicities were similar in both arms with the exception of more grade ≥3 neutropenia (P < 0.0001) and fewer grade ≥3 thrombocytopenia (P = 0.05) in R-CHOP-21 versus R-CHOP-14. The elderly patient population had a favorable 5-year overall survival (OS) of 69% (95% CI: 65-73). We did not identify any subgroup of patients that showed differential response to either regimen. In multivariable analysis including individual factors of the IPI, gender, bulk, B2M and albumin levels, only age and B2M were of independent prognostic significance for OS. Molecular analyses demonstrated a significant impact of MYC-R (HR = 1.96; 95% CI: 1.22-3.16; P = 0.01) and DHL (HR = 2.21; 95% CI: 1.18-4.11; P = 0.01) on OS in the combined trial cohorts, independent of other prognostic factors. CONCLUSIONS: Our data support equivalence of both R-CHOP application forms in elderly DLBCL patients. Elderly MYC-R and DHL patients have inferior prognosis and should be considered for alternative treatment approaches. TRIAL NUMBERS: ISCRTN 16017947 (R-CHOP14v21); NCT00052936 (RICOVER-60).


Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Biomarcadores de Tumor/genética , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-6/genética , Proteínas Proto-Oncogénicas c-myc/genética , Factores de Edad , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales de Origen Murino/administración & dosificación , Anticuerpos Monoclonales de Origen Murino/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Ciclofosfamida/administración & dosificación , Ciclofosfamida/efectos adversos , Doxorrubicina/administración & dosificación , Doxorrubicina/efectos adversos , Esquema de Medicación , Femenino , Reordenamiento Génico , Humanos , Estimación de Kaplan-Meier , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/mortalidad , Linfoma de Células B Grandes Difuso/patología , Masculino , Persona de Mediana Edad , Análisis Multivariante , Selección de Paciente , Medicina de Precisión , Prednisona/administración & dosificación , Prednisona/efectos adversos , Factores de Riesgo , Rituximab , Factores de Tiempo , Resultado del Tratamiento , Reino Unido , Vincristina/administración & dosificación , Vincristina/efectos adversos
5.
Mycorrhiza ; 27(2): 95-108, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27639577

RESUMEN

Terrestrial orchids depend on orchid mycorrhizal fungi (OMF) as symbionts for their survival, growth and nutrition. The ability of OMF from endangered orchid species to compete for available resources with OMF from common species may affect the distribution, abundance and therefore conservation status of their orchid hosts. Eight symbiotically effective OMF from endangered and more common Caladenia species were tested for their ability to utilise complex insoluble and simple soluble carbon sources produced during litter degradation by growth with different carbon sources in liquid medium to measure the degree of OMF variation with host conservation status or taxonomy. On simple carbon sources, fungal growth was assessed by biomass. On insoluble substrates, ergosterol content was assessed using ultra-performance liquid chromatography (UPLC). The OMF grew on all natural materials and complex carbon sources, but produced the greatest biomass on xylan and starch and the least on bark and chitin. On simple carbon sources, the greatest OMF biomass was measured on most hexoses and disaccharides and the least on galactose and arabinose. Only some OMF used sucrose, the most common sugar in green plants, with possible implications for symbiosis. OMF from common orchids produced more ergosterol and biomass than those from endangered orchids in the Dilatata and Reticulata groups but not in the Patersonii and Finger orchids. This suggests that differences in carbon source utilisation may contribute to differences in the distribution of some orchids, if these differences are retained on site.


Asunto(s)
Carbono/metabolismo , Especies en Peligro de Extinción , Micorrizas/fisiología , Orchidaceae/microbiología , Cromatografía Liquida , Ergosterol/química , Ergosterol/metabolismo , Micorrizas/clasificación
6.
Vox Sang ; 108(2): 160-8, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25469449

RESUMEN

BACKGROUND: We assessed the haemostatic capacity of thawed plasma produced after ambient storage of whole blood for 24 h (RTFP24), and the supernatant of buffy-coat derived platelet concentrates (PC). METHODS: Platelet concentrates (n = 20) were tested on days 1, 5 and 7 of storage at 22°C and RTFP24 (n = 10) immediately following thawing and after 4 and 6 days storage at 4°C. Coagulation factor activity, thrombin generation ± an activator of protein C (PROTAC) and rotational thromboelastometry (ROTEM) were assessed. RESULTS: In plasma and buffy-coat derived PC, there was a < 10% loss of factors II, IX and FX, but much higher loss of factors FV, FVII and FVIII. In plasma, the total or peak amount of thrombin generated was unaffected by storage for 6 days, with or without Protac, but there was an increase in lag time and decreased rate of clot formation by ROTEM. In PC, but not plasma, there was a 16% increase in FXII activity and increase in resistance to activated protein C, co-incidental to 30% loss of free protein S. CONCLUSIONS: These data suggest thrombin generation is relatively unaltered when RTFP24 is thawed and stored for 6 days, and that the supernatant of PC has significant haemostatic capacity.


Asunto(s)
Factores de Coagulación Sanguínea/metabolismo , Plaquetas/metabolismo , Congelación , Plasma/metabolismo , Trombina/metabolismo , Coagulación Sanguínea , Humanos
7.
Int J Qual Health Care ; 27(6): 528-35, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26487508

RESUMEN

OBJECTIVE: In the context of increasing over-testing and the implications for patient safety, to establish the prevalence and nature of pathology test-ordering of GP trainees, and to describe the associations of this test-ordering. DESIGN: A cross-sectional analysis of data from the Registrar Clinical Encounters in Training (ReCEnT) cohort study. SETTING: Five of Australia's 17 general practice regional training providers, encompassing urban-to-very remote practices. PARTICIPANTS: GP trainees. MAIN OUTCOME MEASURES: The number of pathology tests ordered per problem/diagnosis managed. RESULTS: A total of 856 individual trainees (response rate 95.2%) contributed data from 1832 trainee-terms, 108 759 encounters and 169 304 problems. Pathology test-ordering prevalence was 79.3 tests (95% CI: 78.8-79.8) per 100 encounters, 50.9 (95% CI: 50.6-51.3) per 100 problems, and at least 1 test was requested in 22.4% of consultations. Most commonly ordered was full blood count (6.1 per 100 problems). The commonest problem prompting test-ordering was 'check-up' (18.6%). Test-ordering was significantly associated, on multivariable analysis, with the trainee having worked at the practice previously; the patient being adult, male and new to both trainee and practice; the practice being urban; the problem/diagnosis being new; imaging being ordered; referral being made and follow-up being arranged. Trainees were significantly less likely to order tests for problems/diagnoses for which they had sought in-consultation information or advice. CONCLUSIONS: Compared with the established GPs, trainees order more pathology tests per consultation and per problem managed, and in a higher proportion of consultations. Our findings will inform educational policy to enhance quality and safety in general practice training.


Asunto(s)
Pruebas Diagnósticas de Rutina , Medicina General , Médicos Generales/educación , Pautas de la Práctica en Medicina , Australia , Estudios Transversales
8.
Haemophilia ; 20(2): e144-8, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24329777

RESUMEN

Pregnancy is associated with significant haemostatic changes, with a progressive rise in most clotting factors. There is limited data on the changes of factor XIII (FXIII) level during pregnancy. This study assesses changes in FXIII activity during normal pregnancy and establish FXIII reference range during each trimester of pregnancy and immediate postnatal period. This is a cross sectional study of 376 women with normal uneventful pregnancies. Plasma FXIII activity was measured during first (weeks 0-12, n = 116), second (weeks 13-28, n = 132), third trimester (weeks 29-42, n = 128) and postnatal (day 0-3; n = 30). Samples were also collected from non-pregnant women (n = 25) as a control group. FXIII was assayed on CS-5100 analyser using chromogenic reagent. The mean ± SD FXIII activity was 112 ± 29 IU dL(-1) during first trimester, 96 ± 26 IU dL(-1) during second trimester, 83 ± 21 IU dL(-1) during third trimester, 90 ± 19 IU dL(-1) during postnatal period, and 113 ± 26 IU dL(-1) in the control. The reference range was calculated during the first (55-169 IU dL(-1)), second (45-147 IU dL(-1)), third trimester (42-125 IU dL(-1)) and postnatal period (61-137 IU dL(-1)). There was a significant reduction in the mean FXIII activity during the second and third trimester compared to the first trimester and control group (P < 0.0001). During the immediate postnatal period, the mean FXIII activity was not statistically different compared to the third and second trimester levels but was significantly lower compared to the first trimester (P < 0.0001) level and the control group (P = 0.0002). This study establishes the reference range for FXIII activity during the three trimesters of normal pregnancy and immediate postnatal period. Women have a significantly decreased level of FXIII activity during a normal uneventful pregnancy.


Asunto(s)
Factor XIII/metabolismo , Trimestres del Embarazo/sangre , Adulto , Estudios Transversales , Femenino , Humanos , Persona de Mediana Edad , Embarazo , Resultado del Embarazo , Valores de Referencia , Factores de Riesgo , Adulto Joven
9.
Haemophilia ; 20(1): 147-53, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24028703

RESUMEN

von Willebrand disease (VWD) is caused by a quantitative and/or qualitative deficiency of the von Willebrand factor (VWF). The laboratory diagnosis of VWD is dependent on the measurement of VWF antigen (VWF:Ag) and ristocetin cofactor activity (VWF:RCo). The aim of this study was to undertake a two-centre evaluation of two new automated VWF:Ag and VWF:RCo assays systems from Instrumentation Laboratory (Bedford, USA). Using the two new analytical systems that operated with different detection principles: immunoturbidimetric (TOP500 analyser) and chemiluminescent (AcuStar analyser), VWF:Ag and VWF:RCo levels were determined in samples from 171 healthy normal subjects, 80 VWD patients (16 type 1, 58 type 2 and 6 type 3) and 7 acquired von Willebrand syndrome patients. With commercial lyophilized normal and pathological plasmas VWF: Ag and VWF:RCo assays performed on both analysers exhibited low levels of inter-assay imprecision (AcuStar: CV% range 3.3-6.9; TOP500: CV% range 2.6-6.3). Samples from normal healthy subjects (range: VWF:Ag 44.6-173.9 IU dL(-1) ; VWF:RCo 43.1-191.5 IU dL(-1)) and patients (range: VWF:Ag <0.3-115.1 IU dL(-1) ; VWF:RCo <0.5-57.2 IU dL(-1)) showed a good correlation between the two VWF:Ag and VWF:RCo methods (rs = 0.92 and 0.82 respectively), with only a few inconsistent cases among the patients' samples evaluated. The chemiluminescent assays had a lower limit of detection for both VWF:Ag and VWF:RCo compared to immunoturbidimetric tests (0.3 IU dL(-1) vs. 2.2 IU dL(-1) and 0.5 IU dL(-1) vs. 4.4 IU dL(-1) respectively). The TOP500 and AcuStar VWF:Ag and VWF:RCo assays were precise and compare well between centres, making these systems suitable for the diagnosis of VWD in non-specialized and reference laboratories.


Asunto(s)
Pruebas de Coagulación Sanguínea/métodos , Ristocetina/metabolismo , Factor de von Willebrand/metabolismo , Pruebas de Coagulación Sanguínea/instrumentación , Humanos , Reproducibilidad de los Resultados , Ristocetina/sangre , Sensibilidad y Especificidad , Enfermedades de von Willebrand/sangre , Enfermedades de von Willebrand/diagnóstico , Factor de von Willebrand/inmunología
10.
Haemophilia ; 19(2): 338-42, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23205618

RESUMEN

The ristocetin cofactor assay (VWF:RCo) is the reference method for assessing von Willebrand factor (VWF) activity in the diagnosis of von Willebrand's Disease (VWD). However, the assay suffers from poor reproducibility and sensitivity at low levels of VWF and is labour intensive. We have undertaken an evaluation of a new immunoturbidimetric VWF activity (VWF:Ac) assay (INNOVANCE(®) VWF Ac. Siemens Healthcare Diagnostics, Marburg, Germany) relative to an established platelet-based VWF:RCo method. Samples from 50 healthy normal subjects, 80 patients with VWD and 50 samples that exhibited 'HIL' (i.e. Haemolysis, Icterus or Lipaemia) were studied. VWF:Ac, VWF:RCo and VWF:Ag were performed on a CS-analyser (Sysmex UK Ltd, Milton Keynes, UK), all reagents were from Siemens Healthcare Diagnostics. The VWF:Ac assay, gave low intra- and inter-assay imprecision (over a 31-day period, n = 200 replicate readings) using commercial normal (Mean 96.2 IU dL(-1), CV < 3.0%) and pathological (Mean 36.1 IU dL(-1), CV < 3.5%) control plasmas. The normal and clinical samples exhibited good correlation between VWF:RCo (range 3-753 IU dL(-1)) and VWF:Ac (rs = 0.97, P < 0.0001), with a mean bias of 5.6 IU dL(-1). Ratios of VWF:Ac and VWF:RCo to VWF:Ag in the VWD samples were comparable, although VWF:Ac had a superior lower level of detection to that of VWF:RCo (3% and 5% respectively). A subset (n = 97) of VWD and HIL samples were analysed for VWF:Ac at two different dilutions to assess the effect on relative potency, no significant difference was observed (P = 0.111). The INNOVANCE(®) VWF Ac assay was shown to be reliable and precise.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Enfermedades de von Willebrand/diagnóstico , Factor de von Willebrand/análisis , Anticuerpos Monoclonales , Humanos , Receptores de GABA-B/metabolismo , Reproducibilidad de los Resultados
11.
Eur Respir J ; 39(4): 945-55, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-21885399

RESUMEN

Pulmonary hypertension (PH) is a heterogeneous condition. To date, no registry data exists reflecting the spectrum of disease across the five diagnostic groups encountered in a specialist referral centre. Data was retrieved for consecutive, treatment-naïve cases diagnosed between 2001 and 2010 using a catheter-based approach. 1,344 patients were enrolled, with a mean follow-up of 2.9 yrs. The 3-yr survival was 68% for pulmonary arterial hypertension (PAH), 73% for PH associated with left heart disease, 44% for PH associated with lung disease (PH-lung), 71% for chronic thromboembolic PH (CTEPH) and 59% for miscellaneous PH. Compared with PAH, survival was inferior in PH-lung and superior in CTEPH (p<0.05). Multivariate analysis demonstrated that diagnostic group independently predicted survival. Within PAH, Eisenmenger's survival was superior to idiopathic PAH, which was superior to PAH associated with systemic sclerosis (p<0.005). Within PH-lung, 3-yr survival in sleep disorders/alveolar hypoventilation (90%) was superior to PH-lung with chronic obstructive pulmonary disease (41%) and interstitial lung disease (16%) (p<0.05). In CTEPH, long-term survival was best in patients with surgically accessible disease undergoing pulmonary endarterectomy. In this large registry of consecutive, treatment-naïve patients identified at a specialist PH centre, outcomes and characteristics differed between and within PH groups. The current system of classification of PH has prognostic value even when adjusted for age and disease severity, emphasising the importance of systematic evaluation and precise classification.


Asunto(s)
Grupos Diagnósticos Relacionados/clasificación , Hipertensión Pulmonar/clasificación , Hipertensión Pulmonar/diagnóstico , Derivación y Consulta/estadística & datos numéricos , Sistema de Registros/estadística & datos numéricos , Adulto , Anciano , Grupos Diagnósticos Relacionados/estadística & datos numéricos , Endarterectomía/mortalidad , Femenino , Estudios de Seguimiento , Cardiopatías Congénitas/clasificación , Cardiopatías Congénitas/diagnóstico , Cardiopatías Congénitas/mortalidad , Humanos , Hipertensión Pulmonar/mortalidad , Hipertensión Pulmonar/cirugía , Masculino , Persona de Mediana Edad , Análisis Multivariante , Pronóstico , Enfermedad Pulmonar Obstructiva Crónica/clasificación , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Enfermedad Pulmonar Obstructiva Crónica/mortalidad , Enfermedad Pulmonar Obstructiva Crónica/cirugía , Índice de Severidad de la Enfermedad , Trastornos del Sueño-Vigilia/clasificación , Trastornos del Sueño-Vigilia/diagnóstico , Trastornos del Sueño-Vigilia/mortalidad , Análisis de Supervivencia , Tromboembolia/clasificación , Tromboembolia/diagnóstico , Tromboembolia/mortalidad
12.
Nat Cell Biol ; 1(1): 60-7, 1999 May.
Artículo en Inglés | MEDLINE | ID: mdl-10559866

RESUMEN

Indirubin is the active ingredient of Danggui Longhui Wan, a mixture of plants that is used in traditional Chinese medicine to treat chronic diseases. Here we identify indirubin and its analogues as potent inhibitors of cyclin-dependent kinases (CDKs). The crystal structure of CDK2 in complex with indirubin derivatives shows that indirubin interacts with the kinase's ATP-binding site through van der Waals interactions and three hydrogen bonds. Indirubin-3'-monoxime inhibits the proliferation of a large range of cells, mainly through arresting the cells in the G2/M phase of the cell cycle. These results have implications for therapeutic optimization of indigoids.


Asunto(s)
Antibióticos Antineoplásicos/farmacología , Quinasas CDC2-CDC28 , Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Animales , Antibióticos Antineoplásicos/química , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular , Quinasa 2 Dependiente de la Ciclina , Quinasas Ciclina-Dependientes/química , Células HL-60 , Humanos , Carmin de Índigo , Indoles/química , Indoles/farmacocinética , Indoles/farmacología , Isatina/química , Isatina/farmacocinética , Isatina/farmacología , Células Jurkat , Células K562 , Leucemia L1210 , Medicina Tradicional China , Ratones , Modelos Moleculares , Conformación Molecular , Conformación Proteica , Proteínas Serina-Treonina Quinasas/química , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/química , Spodoptera , Transfección
13.
Haemophilia ; 17(2): 252-6, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21070498

RESUMEN

von Willebrand's disease (VWD) is regarded as the most common congenital bleeding disorder, and although not available in all laboratories von Willebrand factor (VWF) activity is most frequently assessed as ristocetin cofactor (VWF:RCo). This test can be technically challenging, is subject to poor sensitivity (∼20 IU dL(-1) VWF:RCo) and has a high degree of inter- and intra-assay imprecision [coefficient of variation (cv) > 25%]. We studied an automated assay using a combined fixed platelet/ristocetin reagent (BC von Willebrand reagent, Siemens Healthcare Diagnostics) on the CS-2000i analyser (Sysmex UK Ltd). Initially inter- and intra-assay imprecision was assessed. The automated method showed good day-to-day reproducibility and linearity of standard curves. This technique, also gave low intra- and inter-assay imprecision using commercial normal (cv < 4.5%) and pathological (cv < 8.1%) control plasmas. We then compared automated technique results from 30 healthy normal subjects and 39 VWD patients to those obtained using standard aggregometry (Bio/Data, PAP4) with lyophilised fixed platelets (Helena BioSciences) and ristocetin (American Biochemical and Pharmaceutical Ltd). The automated method had a sensitivity limit of approximately 10 IU dL(-1) vs. 20 IU dL(-1) for aggregometry. Samples giving results within the aggregometry measurable range (n = 50) exhibited good correlation with the automated technique (median 70 IU dL(-1), range 7-184 IU dL(-1); and 64 IU dL(-1), 6-138 IU dL(-1) respectively; R(2) = 0.85). We subsequently compared 3 different batches of BC von Willebrand reagent, using a second group of normal subjects and VWD patients (n = 35, 55-139 IU dL(-1) and n = 30, <10-50 IU dL(-1)). The CS-2000i results exhibited no clinically significant variation between batches (mean cv = 7%). The automated VWF:RCo assay offers a more sensitive, reproducible, robust and less laborious alternative to standard aggregometry.


Asunto(s)
Pruebas de Coagulación Sanguínea/métodos , Enfermedades de von Willebrand/diagnóstico , Factor de von Willebrand/análisis , Humanos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Enfermedades de von Willebrand/sangre
14.
Vox Sang ; 99(3): 232-8, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-20840338

RESUMEN

BACKGROUND: Octapharma PPGmbH has recently modified its manufacturing process for solvent/detergent-treated plasma to incorporate a prion reduction step, in which a 3 log reduction has been demonstrated. The current study was undertaken to assess the impact of this procedure on haemostatic variables in the new product OctaplasLG in comparison with standard Octaplas. METHODS: Production batches of standard Octaplas (n=4) and OctaplasLG (n=16) were assessed for levels of coagulation factors, physiological protease inhibitors, markers of activation and procoagulant microparticles. Global haemostasis was assessed by a thrombin generation test (TGT) and rotational thromboelastometry (ROTEM). RESULTS: Mean levels of factors: II, V, VII, IX, X, XI, XII and XIII, VWF:Ag, antithrombin, protein C and free protein S were all >75 u/dl. ADAMTS-13 activity levels were normal. Factor VIII and VWF:RCo were >55 u/dl. TGT and ROTEM were similar in both preparations, and microparticles were present at negligible levels. Two units of OctaplasLG had slightly elevated levels of Prothrombin Fragments 1+2, but D-Dimer and thrombin-antithrombin complexes were normal in all batches. CONCLUSION: These studies indicate that the affinity chromatography procedure used in OctaplasLG does not appear to adversely affect the proven haemostatic quality of Octaplas, while offering a selective reduction in the concentration of pathological prion proteins.


Asunto(s)
Proteínas Sanguíneas/análisis , Hemostáticos/análisis , Plasma/química , Priones , Cromatografía de Afinidad/métodos , Humanos
15.
Nat Protoc ; 15(2): 540-574, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31915391

RESUMEN

The number of people aged over 65 is expected to double in the next 30 years. For many, living longer will mean spending more years with the burdens of chronic diseases such as Alzheimer's disease, cardiovascular disease, and diabetes. Although researchers have made rapid progress in developing geroprotective interventions that target mechanisms of aging and delay or prevent the onset of multiple concurrent age-related diseases, a lack of standardized techniques to assess healthspan in preclinical murine studies has resulted in reduced reproducibility and slow progress. To overcome this, major centers in Europe and the United States skilled in healthspan analysis came together to agree on a toolbox of techniques that can be used to consistently assess the healthspan of mice. Here, we describe the agreed toolbox, which contains protocols for echocardiography, novel object recognition, grip strength, rotarod, glucose tolerance test (GTT) and insulin tolerance test (ITT), body composition, and energy expenditure. The protocols can be performed longitudinally in the same mouse over a period of 4-6 weeks to test how candidate geroprotectors affect cardiac, cognitive, neuromuscular, and metabolic health.


Asunto(s)
Envejecimiento/fisiología , Salud , Envejecimiento/metabolismo , Animales , Composición Corporal , Electrocardiografía , Metabolismo Energético , Prueba de Tolerancia a la Glucosa , Fuerza de la Mano , Resistencia a la Insulina , Estudios Longitudinales , Ratones , Ratones Endogámicos C57BL , Reconocimiento en Psicología
16.
Vox Sang ; 96(3): 206-12, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19175566

RESUMEN

BACKGROUND: We have previously shown that fresh-frozen plasma (FFP) contains red blood cell-derived procoagulant microparticles (MPs) that are removable by 0.2 microm filtration. Given the limitations of current methods for accurately sizing MPs, we have applied the novel approach of dynamic light scattering (DLS) to characterize the size distributions of these MPs within FFP. METHODS: Fresh-frozen plasma was prepared from blood Group A and O donations (n = 10 of each) after an overnight hold of whole blood at 4 degrees C. On the day of analysis, plasma was thawed to 37 degrees C and daughter aliquots were studied pre- and post-filtration (0.2 microm filtration device, Ceveron MFU-500, Technoclone). MP size and dispersity was assessed using a Zetasizer Nano S (Malvern Instruments Ltd), which employs a 173 degrees backscatter detector and an N5 Submicron Particle Size Analyser (Beckman Coulter) using multi-angle measurements (30.1 degrees , 62.6 degrees and 90 degrees ). The analysers presented MP size distribution graphically as intensity plots, mean size, standard deviation and polydispersity index. RESULTS: Of the instruments used, only the N5 utilizing a 30.1 degrees angle of measurement could detect MPs of the expected size distribution and demonstrate their removal by filtration. MPs (range of mean particle diameters: pre, 101-464 nm; post, 21-182 nm filtration) were significantly smaller post-filtration (P < 0.0001), but polydispersity index (median: pre, 0.746, post, 0.769) exhibited no significant change. There was no significant difference between the size of MPs from blood Group O (pre, 247 nm) and Group A (pre, 289 nm) samples (P = 0.44). CONCLUSION: Our data demonstrates that DLS offers a novel approach to assessing MP size and distribution, a technique that could be easily adopted as a means of assessing MPs within either FFP or other blood products.


Asunto(s)
Micropartículas Derivadas de Células/química , Luz , Tamaño de la Partícula , Plasma/química , Dispersión de Radiación , Humanos
17.
Br J Haematol ; 141(4): 536-44, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-18341632

RESUMEN

Autoantibodies to ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type I motif, member 13) play an important role in the development of microthrombosis in thrombotic thrombocytopenic purpura (TTP). In severe cases of antiphospholipid syndrome (APS), microthrombosis can occur similar to that seen in TTP, suggesting possible mutual pathogenic factors. However, the role of ADAMTS13 in APS is unknown. We hypothesised that aberrations in ADAMTS13 may occur in APS and evaluated ADAMTS13 and von Willebrand factor (VWF) in 68 patients with antiphospholipid antibodies (aPA) including 52 with APS. Thirty-three (49%) had IgG anti-ADAMTS13 with 12 of these patients having reduced ADAMTS13 activity, suggesting neutralising antibodies. Low ADAMTS13 activity (median 34%) was demonstrated in 22/68 (33%), all with normal ADAMTS13 antigen levels consistent with dysfunctional ADAMTS13. Reduced ADAMTS13 activity was not secondary to elevated von Willebrand factor (VWF), or increased VWF secretion (normal VWF propeptide), although a reduced VWF clearance was noted in APS. Analysis found no associations between the ADAMTS13 abnormalities and any aPA profile or thrombotic/obstetric complications, although this study was not adequately powered to address clinical associations. Nevertheless, these findings highlight that ADAMTS13 autoantibodies and ADAMTS13 dysfunction can occur in APS, and although the clinical significance remains undetermined, ADAMTS13 dysfunction may be contributory to thrombogenesis in autoimmune conditions other than TTP.


Asunto(s)
Proteínas ADAM/inmunología , Síndrome Antifosfolípido/inmunología , Autoanticuerpos/sangre , Factor de von Willebrand/metabolismo , Proteínas ADAM/sangre , Proteínas ADAM/fisiología , Proteína ADAMTS13 , Adulto , Anciano , Femenino , Semivida , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad
18.
Vox Sang ; 95(3): 197-204, 2008 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-19121184

RESUMEN

BACKGROUND: We have previously demonstrated that clot formation in fresh-frozen plasma (FFP) is influenced by the presence of microparticles (MP). In this study, the cellular source(s), properties and influence of MPs on clot formation within FFP were further characterized. METHODS: Fresh-frozen plasma was prepared after an overnight hold of whole blood at 4 degrees C. We examined the effect of a 0.2 microm filtration device designed to remove cellular MPs on thrombin generation test (TGT) and Thrombelastography (TEG(R)) as well as clotting factors and physiological inhibitors: prothrombin time (PT); activated partial thromboplastin time (APTT), fibrinogen (Fg), factor VIII (FVIII), von Willebrand factor antigen (VWF:Ag), antithrombin III (AT-III) and protein C (PC). MPs were measured using a functional assay and also by flow cytometry. RESULTS: Microparticle levels by functional assay were reduced by filtration (pre- 5.11 vs. post- 4.43 nmol/l phosphatidylserine equivalent, P < 0.0001). Flow cytometry showed that the most numerous MPs were derived from red blood cells, with ~87% binding annexin V, most of which (94%) were removed by filtration. MP removal had minimal effect on the PT, APTT, Fg, VWF:Ag, AT-III or PC or FVIII, but a major effect on TGT (endogenous thrombin potential: pre- 1722 vs. post- 990 nM thrombin, P < 0.0001; peak thrombin: pre- 91 vs. post- 44 nM thrombin, P < 0.0001), which in turn reflected the changes seen in TEG(R), where post-filtration clots started forming more slowly and the rate of clot formation was reduced. CONCLUSION: These data suggest that MPs contribute towards clot formation in FFP.


Asunto(s)
Coagulación Sanguínea , Micropartículas Derivadas de Células/metabolismo , Plasma/metabolismo , Pruebas de Coagulación Sanguínea/métodos , Transfusión de Componentes Sanguíneos/métodos , Citometría de Flujo/métodos , Humanos
19.
Vox Sang ; 94(4): 306-14, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-18266780

RESUMEN

BACKGROUND: Factor VIII (FVIII) levels are used as a quality marker of fresh-frozen plasma (FFP); however, other clotting factors are not routinely measured. METHODS: We assessed additional haemostatic parameters and the dynamics of coagulation using Thrombelastography (TEG) and a thrombin generation test (TGT). FFP was prepared on the day of donation (Day 0) or after overnight hold at 4 degrees C (Day 1). RESULTS: Factor VIII in Day 1 FFP was 18% lower than in Day 0. TEG parameters in Day 1 FFP were consistent with increased coagulability and did not correlate with altered levels of clotting factors, but were consistent with the increased levels of microparticles seen in the Day 1 samples. TGT studies exhibited increased lag time, time to peak and reduced peak thrombin generation, but no change in endogenous thrombin potential (ETP) on Day 1. There was a weak association between FVIII level and both ETP and peak thrombin (ETP r(s)> or = 0.22, P< or = 0.003; peak thrombin r(s)> or = 0.48, P< or = 0.0001), which was influenced by ABO group, with the lowest levels in group O. CONCLUSION: We conclude that levels of FVIII do not predict the haemostatic potential of FFP and that there may be a role for alternative technologies in monitoring the quality of FFP.


Asunto(s)
Coagulación Sanguínea/fisiología , Factor VIII/análisis , Plasma/fisiología , Humanos , Plasma/química , Control de Calidad , Tromboelastografía
20.
J Thromb Haemost ; 16(8): 1604-1613, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29897666

RESUMEN

Essentials New VWF activity assays are increasingly used but information on their comparability is limited. This is an ISTH SSC-organized study (expert labs, 5 countries) to compare all available assays. VWF activity by six assays correlated well with each other. The new assays show improved characteristics - minor differences are noted. SUMMARY: Background Several new assays have become available to measure von Willebrand factor (VWF) activity. The new assays appear to have improved performance characteristics compared with the old reference standard, ristocetin cofactor activity (VWF:RCo), but information is limited about how they compare with VWF:RCo and each other. Methods The von Willebrand factor Subcommittee of the International Society for Thrombosis and Haemostasis (ISTH) Scientific and Standardization Committee (SSC) designed a collaborative study involving expert laboratories from several countries to compare available tests with each other and with VWF:RCo. Eight laboratories from five countries were provided with blinded samples from normal healthy individuals and well-characterized clinical cases. Laboratories measured VWF activity using all tests available to them; data from six laboratories, not affected by thawing during transportation, are included in this study. Results All tests correlated well with VWF:RCo activity (r-values ranged from 0.963 to 0.989). Slightly steeper regression lines for VWF:Ab and VWF:GPIbM were clinically insignificant. The new assays showed improved performance characteristics. Of the commercially available assays, the VWF:GPIbR using the AcuStar system was the most sensitive and could reliably detect VWF activity below 1 IU dL-1 . The lower limit of the measuring interval for the VWF:GPIbM and the VWF:GPIbR assays was in the 3-4 and 3-6 IU dL-1 range, respectively. Inter-laboratory variation was also improved for most new assays. Conclusion All VWF activity assays correlated well with each other and the VWF:RCo assay. The slight differences in characteristics found in the COMPASS-VWF study will assist the VWF community in interpreting and comparing activity results.

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