Engraftment after autologous hematopoietic stem cell transplantation in patients mobilized with Plerixafor: A retrospective, multicenter study of a large series of patients.

Plerixafor (PLX) appears to effectively enhance hematopoietic stem-cell mobilization prior to autologous hematopoietic stem cell transplantation (auto-HCT). However, the quality of engraftment following auto-HCT has been little explored. Here, engraftment following auto-HCT was assessed in patients mobilized with PLX through a retrospective, multicenter study of 285 consecutive patients. Information on early and 100-day post-transplant engraftment was gathered from the 245 patients that underwent auto-HCT. The median number of PLX days to reach the stem cell collection goal (≥2 × 106 CD34+ cells/kg) was 1 (range 1-4) and the median PLX administration time before apheresis was 11 h (range 1-18). The median number of apheresis sessions to achieve the collection goal was 2 (range 1-5) and the mean number of CD34+ cells collected was 2.95 × 106/kg (range 0-30.5). PLX administration was safe, with only 2 mild and transient gastrointestinal adverse events reported. The median time to achieve an absolute neutrophil count (ANC) >500/µL was 11 days (range 3-31) and the median time to platelet recovery >20 × 103/µL was 13 days (range 5-69). At 100 days after auto-HCT, the platelet count was 137 × 109/L (range 7-340), the ANC was 2.3 × 109/L (range 0.1-13.0), and the hemoglobin concentration was 123 g/L (range 79-165). PLX use allowed auto-HCT to be performed in a high percentage of poorly mobilized patients, resulting in optimal medium-term engraftment in the majority of patients in whom mobilization failed, in this case mainly due to suboptimal peripheral blood CD34+ cell concentration on day +4 or low CD34+ cell yield on apheresis.

Early synergistic interactions between the HPV16­E7 oncoprotein and 17ß-oestradiol for repressing the expression of Granzyme B in a cervical cancer model.

Although high-risk human papillomavirus (HR­HPV) infection has a prominent role in the aetiology of cervical cancer (CC), sex steroid hormones may also be involved in this process; however, the cooperation between oestrogen and HR­HPV in the early stages of cervical carcinogenesis is poorly understood. Since 17ß-oestradiol (E2) and the HPV type 16­E7 oncoprotein induce CC in transgenic mice, a microarray analysis was performed in the present study to generate global gene expression profiles from 2­month­old FVB (non­transgenic) and K14E7 (transgenic) mice who were left untreated or were treated for 1 month with E2. Upregulation of cancer-related genes that have not been previously reported in the context of CC, including glycerophosphodiester phosphodiesterase domain containing 3, interleukin 1 receptor type II, natriuretic peptide type C, MGAT4 family member C, lecithin-retinol acyltransferase (phosphatidylcholine-retinol-O-acyltransferase) and glucoside xylosyltransferase 2, was observed. Notably, upregulation of the serine (or cysteine) peptidase inhibitor clade B member 9 gene and downregulation of the Granzyme gene family were observed; the repression of the Granzyme B pathway may be a novel mechanism of immune evasion by cancer cells. The present results provide the basis for further studies on early biomarkers of CC risk and synergistic interactions between HR­HPV and oestrogen.

Reduced proliferation of endothelial colony-forming cells in unprovoked venous thromboembolic disease as a consequence of endothelial dysfunction.

BACKGROUND: Venous thromboembolic disease (VTD) is a public health problem. We recently reported that endothelial colony-forming cells (ECFCs) derived from endothelial cells (EC) (ECFC-ECs) from patients with VTD have a dysfunctional state. For this study, we proposed that a dysfunctional status of these cells generates a reduction of its proliferative ability, which is also associated with senescence and reactive oxygen species (ROS). METHODS AND RESULTS: Human mononuclear cells (MNCs) were obtained from peripheral blood from 40 healthy human volunteers (controls) and 50 patients with VTD matched by age (20-50 years) and sex to obtain ECFCs. We assayed their proliferative ability with plasma of patients and controls and supernatants of cultures from ECFC-ECs, senescence-associated ß-galactosidase (SA-ß-gal), ROS, and expression of ephrin-B2/Eph-B4 receptor. Compared with cells from controls, cells from VTD patients showed an 8-fold increase of ECFCs that emerged 1 week earlier, reduced proliferation at long term (39%) and, in passages 4 and 10, a highly senescent rate (30±1.05% vs. 91.3±15.07%, respectively) with an increase of ROS and impaired expression of ephrin-B2/Eph-4 genes. Proliferation potential of cells from VTD patients was reduced in endothelial medium [1.4±0.22 doubling population (DP)], control plasma (1.18±0.31 DP), or plasma from VTD patients (1.65±0.27 DP). CONCLUSIONS: As compared with controls, ECFC-ECs from individuals with VTD have higher oxidative stress, proliferation stress, cellular senescence, and low proliferative potential. These findings suggest that patients with a history of VTD are ECFC-ECs dysfunctional that could be associated to permanent risk for new thrombotic events.

Protocolo de actuación ante valores críticos en pruebas de laboratorio en el lugar de asistencia al paciente en una unidad neonatal / Critical values protocol for point of care testing in a neonatal unit

Introducción. En las unidades asistenciales que realizan pruebas de laboratorio en el lugar de asistencia al paciente (POCT) sería recomendable establecer las mismas políticas de calidad que en el laboratorio central, entre ellas disponer de un protocolo de actuación ante valores críticos. El objetivo del estudio consistió en elaborar el listado de valores críticos para pruebas POCT realizadas en una unidad neonatal y establecer el protocolo de actuación. Material y métodos. Las magnitudes estudiadas fueron pH, pCO2, pO2, saturación de oxígeno, hemoglobina, sodio, potasio, calcio ionizado, glucosa, bilirrubina y lactato. Estas magnitudes fueron determinadas en un analizador ABL90 FLEX. El listado de valores críticos se elaboró mediante revisión de la bibliografía y consenso con los neonatólogos. El protocolo de actuación se adaptó a partir del protocolo del laboratorio de urgencias. La revisión del listado preliminar se basó en la práctica clínica y en los datos de frecuencia de aparición de valores críticos. Resultados. Se expone nuestra experiencia en la elaboración de un listado de valores críticos para POCT y en la implantación del protocolo de actuación. Conclusiones. Para establecer el listado de valores críticos y protocolo de actuación resultó fundamental la experiencia del laboratorio central. Para conseguir la implantación de un protocolo de actuación ante valores críticos en POCT es necesaria una estrecha colaboración entre la unidad asistencial y el personal del laboratorio. La frecuencia de aparición de valores críticos y la experiencia de los clínicos son herramientas que se complementan en la revisión del listado de valores críticos (AU)

Introduction. Care units performing point of care testing (POCT) should have the same quality policies as the central laboratory, including having a protocol for critical values. The aim of the study was to develop a list of critical values for POCT in a neonatal unit, and set the protocol performance. Material and methods. The tests included in the protocol were pH, pCO2, pO2, oxygen saturation, haemoglobin, sodium, potassium, ionized calcium, glucose, bilirubin, and lactate. These tests were determined using a POCT ABL90FLEX analyser. To prepare the list of critical values, a bibliography review was performed, as well as meetings with the neonatologists. The revision of the preliminary list was based on clinical practice and data frequency of critical values. To set the protocol, an adaptation of our emergency laboratory protocol was performed. Results. We show our experience in the preparation of a list of critical values for POCT and the implementation of a protocol. Conclusions. Central laboratory experience was a key element in establishing the list of critical values and action protocol. A close collaboration between health care unit and laboratory was required to achieve the implementation of a POCT critical values protocol. The frequency of critical values and clinician experience were complementary tools to revise the list of critical values (AU)

Susceptibility to infectious diseases based on antimicrobial peptide production.

In the last few years, the great impact of antimicrobial peptides on infectious disease susceptibility and natural resistance has been reported. In some cases, susceptibility to diseases is related to antimicrobial peptide polymorphisms and gene copy numbers, but for the vast majority of infectious diseases, these phenomena need to be elucidated. This review is focused on the current knowledge about susceptibility and resistance conferred by genetic variations in antimicrobial peptide expression in infectious diseases.

Genetic homogeneity of axenic isolates of Giardia intestinalis derived from acute and chronically infected individuals in Mexico.

Twenty-six axenic isolates of Giardia intestinalis, established in Mexico City over an 11-year period from symptomatic and asymptomatic individuals with acute or chronic infections, were typed genetically. A segment of the glutamate dehydrogenase gene was amplified by PCR and examined by restriction analysis using BspH1 and ApaI to determine the major genetic assemblages to which the isolates belonged. This was coupled with the amplification and analysis of segments of variant-specific surface protein genes to determine genetic subgroupings. Despite their heterogeneous clinical backgrounds, the isolates were found to be genetically homogeneous-all belonging to genetic group I of assemblage A. The results show that type A-I G. intestinalis is ubiquitous in Mexico City and that host factors play an important, if not dominant, role in determining the clinical outcome of Giardia infections in humans.

Biology of human hematopoietic stem and progenitor cells present in circulation.

Circulating hematopoietic stem and progenitor cells play important roles in the physiology and homeostasis of the hematopoietic system. The frequency of these cells varies throughout development, being more abundant during gestation. In the adult, the numbers of such cells are extremely low; however, they can be increased by intravenous administration of chemotherapy and/or recombinant cytokines to individuals. This mechanism--known as mobilization--involves the disruption of the interactions between primitive hematopoietic cells and microenvironment elements (stromal cells and extracellular matrix molecules), which are mediated by a group of molecules known as cell adhesion molecules. During the last two decades, circulating cells of newborns (those present in umbilical cord blood) and adults (mobilized peripheral blood) have gained relevance not only because of their biology, but also because of their clinical application. Indeed, at present the number of mobilized peripheral blood-derived hematopoietic cell transplants performed worldwide is clearly superior to the number of bone marrow transplants being done annually. On the other hand, the number of cord blood transplants has significantly increased during the last few years, and cord blood banking has expanded in a significant manner over the last decade. Circulating stem and progenitor cells are being manipulated ex vivo, both in cellular and molecular terms, and the clinical use of these manipulated cells is just beginning. Undoubtedly, hematopoietic cells present in circulation will play a key role in the development of both gene and cellular therapies for a variety of diseases.

Comparative analysis of the in vitro proliferation and expansion of hematopoietic progenitors from patients with aplastic anemia and myelodysplasia.

Aplastic anemia (AA) and myelodysplasia (MDS) show great similarities in their biology. To date, however, it is still unclear to what extent hematopoietic progenitor cells (HPCs) from AA and MDS share biological properties and what the functional differences are between them. In trying to address this issue, in the present study we have analyzed, in a comparative manner, the proliferation and expansion capacities of bone marrow (BM) progenitor cells from AA and MDS in response to recombinant cytokines. BM samples from normal subjects (NBM) and patients with AA and MDS were enriched for HPC by immunomagnetic-based negative selection. Selected cells were cultured in the absence (control) or in the presence of early-acting cytokines (Mix I), or early-, intermediate- and late-acting cytokines (Mix II). Proliferation and expansion were assessed periodically. In NBM and MDS cultures apoptosis was also determined. In NBM cultures, Mix I induced a nine-fold increase in total cell numbers and a 3.6-fold increase in colony-forming cell (CFC) numbers. In Mix II-supplemented cultures, total cells were increased 643-fold, and CFC 12.4-fold. In AA cultures, no proliferation or expansion were observed in Mix I-supplemented cultures, whereas only a four-fold increase in total cell numbers was observed in the presence of Mix II. In MDS cultures, a 12-fold increase in total cells and a 2.9-fold increase in CFC were observed in the presence of Mix I; on the other hand, Mix II induced a 224-fold increase in total cells and a 5.9-fold increase in CFC. Apoptosis was reduced in cytokine-supplemented cultures from NBM. In contrast, Mix II induced a significant increase in the rate of apoptosis in MDS cultures. Our results demonstrate that, as compared to their normal counterparts, AA and MDS progenitors are deficient in their proliferation and expansion potentials. Such a deficiency is clearly more pronounced in AA cells, which seem to be unable to respond to several cytokines. MDS progenitors, on the other hand, are capable to proliferate and expand in response to cytokines; however, their rate of apoptosis is increased by intermediate- and late-acting cytokines, so that the overall proliferation and expansion are significantly lower than those of normal progenitor cells.