New approaches to improve the cord blood unit hematopoietic progenitor cell content

Cord blood is a well-established alternative to bone marrow and peripheral blood stem cell transplantation. To this day, over 400 000 unrelated donor cord blood units have been stored in cord blood banks worldwide. To enable successful cord blood transplantation, recent efforts have been focused on finding ways to increase the hematopoietic progenitor cell content of cord blood units. In this study, factors that may improve the selection and quality of cord blood collections for banking were identified. In 167 consecutive cord blood units collected from healthy full-term neonates and processed at a national cord blood bank, mean platelet volume (MPV) correlated with the numbers of cord blood unit hematopoietic progenitors (CD34+ cells and colony-forming units); this is a novel finding. Mean platelet volume can be thought to represent general hematopoietic activity, as newly formed platelets have been reported to be large. Stress during delivery is hypothesized to lead to the mobilization of hematopoietic progenitor cells through cytokine stimulation. Accordingly, low-normal umbilical arterial pH, thought to be associated with perinatal stress, correlated with high cord blood unit CD34+ cell and colony-forming unit numbers. The associations were closer in vaginal deliveries than in Cesarean sections. Vaginal delivery entails specific physiological changes, which may also affect the hematopoietic system. Thus, different factors may predict cord blood hematopoietic progenitor cell numbers in the two modes of delivery. Theoretical models were created to enable the use of platelet characteristics (mean platelet volume) and perinatal factors (umbilical arterial pH and placental weight) in the selection of cord blood collections with high hematopoietic progenitor cell counts. These observations could thus be implemented as a part of the evaluation of cord blood collections for banking. The quality of cord blood units has been the focus of several recent studies. However, hemostasis activation during cord blood collection is scarcely evaluated in cord blood banks. In this study, hemostasis activation was assessed with prothrombin activation fragment 1+2 (F1+2), a direct indicator of thrombin generation, and platelet factor 4 (PF4), indicating platelet activation. Altogether three sample series were collected during the set-up of the cord blood bank as well as after changes in personnel and collection equipment. The activation decreased from the first to the subsequent series, which were collected with the bank fully in operation and following international standards, and was at a level similar to that previously reported for healthy neonates. As hemostasis activation may have unwanted effects on cord blood cell contents, it should be minimized. The assessment of hemostasis activation could be implemented as a part of process control in cord blood banks. Culture assays provide information about the hematopoietic potential of the cord blood unit. In processed cord blood units prior to freezing, megakaryocytic colony growth was evaluated in semisolid cultures with a novel scoring system. Three investigators analyzed the colony assays, and the scores were highly concordant. With such scoring systems, the growth potential of various cord blood cell lineages can be assessed. In addition, erythroid cells were observed in liquid cultures of cryostored and thawed, unseparated cord blood units without exogenous erythropoietin. This was hypothesized to be due to the erythropoietic effect of thrombopoietin, endogenous erythropoietin production, and diverse cell-cell interactions in the culture. This observation underscores the complex interactions of cytokines and supporting cells in the heterogeneous cell population of the thawed cord blood unit.

Every method counts

In this study we explore the concurrent, combined use of three research methods, statistical corpus analysis and two psycholinguistic experiments (a forced-choice and an acceptability rating task), using verbal synonymy in Finnish as a case in point. In addition to supporting conclusions from earlier studies concerning the relationships between corpus-based and ex- perimental data (e. g., Featherston 2005), we show that each method adds to our understanding of the studied phenomenon, in a way which could not be achieved through any single method by itself. Most importantly, whereas relative rareness in a corpus is associated with dispreference in selection, such infrequency does not categorically always entail substantially lower acceptability. Furthermore, we show that forced-choice and acceptability rating tasks pertain to distinct linguistic processes, with category-wise in- commensurable scales of measurement, and should therefore be merged with caution, if at all.

Functional Identification of Regulatory Elements within the Promoter Region of Platelet-Derived Growth Factor 2

Human platelet-derived growth factor (PDGF) is composed of two polypeptide chains, PDGF-1 and PDGF-2,the human homolog of the v-sis oncogene. Deregulation of PDGF-2 expression can confer a growth advantage to cells possessing the cognate receptor and, thus, may contribute to the malignant phenotype. We investigated the regulation of PDGF-2 mRNA expression during megakaryocytic differentiation of K562 cells. Induction by 12-O-tetradecanoylphorbol-13-acetate (TPA) led to a greater than 200-fold increase in PDGF-2 transcript levels in these cells. Induction was dependent on protein synthesis and was not enhanced by cycloheximide exposure.In our initial investigation of the PDGF-2 promoter, a minimal promoter region, which included sequences extending only 42 base pairs upstream of the TATA signal, was found to be as efficient as 4 kilobase pairs upstream of the TATA signal in driving expression of a reporter gene in uninduced K562 cells. We also functionally identified different regulatory sequence elements of the PDGF-2 promoter in TPA-induced K562 cells. One region acted as a transcriptional silencer, while another region was necessary for maximal activity of the promoter in megakaryoblasts. This region was shown to bind nuclear factors and was the target for trans-activation in normal and tumor cells. In one tumor cell line, which expressed high PDGF-2 mRNA levels, the presence of the positive regulatory region resulted in a 30-fold increase in promoter activity. However, the ability of the minimal PDGF-2 promoter to drive reporter gene expression in uninduced K562 cells and normal fibroblasts, which contained no detectable PDGF-2 transcripts, implies the existence of other negative control mechanisms beyond the regulation of promoter activity.

Comparing type counts

This work is a case study of applying nonparametric statistical methods to corpus data. We show how to use ideas from permutation testing to answer linguistic questions related to morphological productivity and type richness. In particular, we study the use of the suffixes -ity and -ness in the 17th-century part of the Corpus of Early English Correspondence within the framework of historical sociolinguistics. Our hypothesis is that the productivity of -ity, as measured by type counts, is significantly low in letters written by women. To test such hypotheses, and to facilitate exploratory data analysis, we take the approach of computing accumulation curves for types and hapax legomena. We have developed an open source computer program which uses Monte Carlo sampling to compute the upper and lower bounds of these curves for one or more levels of statistical significance. By comparing the type accumulation from women’s letters with the bounds, we are able to confirm our hypothesis.

HIGH-RESOLUTION IMAGING OF THE ATLBS REGIONS: THE RADIO SOURCE COUNTS

The Australia Telescope Low-brightness Survey (ATLBS) regions have been mosaic imaged at a radio frequency of 1.4 GHz with 6 `' angular resolution and 72 mu Jy beam(-1) rms noise. The images (centered at R. A. 00(h)35(m)00(s), decl. -67 degrees 00'00 `' and R. A. 00(h)59(m)17(s), decl. -67.00'00 `', J2000 epoch) cover 8.42 deg(2) sky area and have no artifacts or imaging errors above the image thermal noise. Multi-resolution radio and optical r-band images (made using the 4 m CTIO Blanco telescope) were used to recognize multi-component sources and prepare a source list; the detection threshold was 0.38 mJy in a low-resolution radio image made with beam FWHM of 50 `'. Radio source counts in the flux density range 0.4-8.7 mJy are estimated, with corrections applied for noise bias, effective area correction, and resolution bias. The resolution bias is mitigated using low-resolution radio images, while effects of source confusion are removed by using high-resolution images for identifying blended sources. Below 1 mJy the ATLBS counts are systematically lower than the previous estimates. Showing no evidence for an upturn down to 0.4 mJy, they do not require any changes in the radio source population down to the limit of the survey. The work suggests that automated image analysis for counts may be dependent on the ability of the imaging to reproduce connecting emission with low surface brightness and on the ability of the algorithm to recognize sources, which may require that source finding algorithms effectively work with multi-resolution and multi-wavelength data. The work underscores the importance of using source lists-as opposed to component lists-and correcting for the noise bias in order to precisely estimate counts close to the image noise and determine the upturn at sub-mJy flux density.

Methotrexate Promotes Platelet Apoptosis via JNK-Mediated Mitochondrial Damage: Alleviation by N-Acetylcysteine and N-Acetylcysteine Amide

Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 mu M) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading Delta psi m dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of specific inhibitor for JNK abrogates the MTX-induced activation of pro-apoptotic proteins and downstream events confirming JNK phosphorylation by MTX as a key event. We also demonstrate that platelet mitochondria as prime sources of ROS which plays a central role in MTX-induced apoptosis. Further, MTX induces oxidative stress by altering the levels of ROS and glutathione cycle. In parallel, the clinically approved thiol antioxidant N-acetylcysteine (NAC) and its derivative N-acetylcysteine amide (NACA) proficiently alleviate MTX-induced platelet apoptosis and oxidative damage. These findings underpin the dearth of research on interference of therapeutic drugs with platelets, despite their importance in human health and disease. Therefore, the use of antioxidants as supplementary therapy seems to be a safe bet in pathologies associated with altered platelet functions.

Intranasal Delivery of Plasma and Platelet Growth Factors Using PRGF-Endoret System Enhances Neurogenesis in a Mouse Model of Alzheimer’s Disease

[EN] Neurodegeneration together with a reduction in neurogenesis are cardinal features of Alzheimer’s disease (AD) induced by a combination of toxic amyloid-β peptide (Aβ) and a loss of trophic factor support. Amelioration of these was assessed with diverse neurotrophins in experimental therapeutic approaches. The aim of this study was to investigate whether intranasal delivery of plasma rich in growth factors (PRGF-Endoret), an autologous pool of morphogens and proteins, could enhance hippocampal neurogenesis and reduce neurodegeneration in an amyloid precursor protein/presenilin-1 (APP/PS1) mouse model. Neurotrophic and neuroprotective actions were firstly evident in primary neuronal cultures, where cell proliferation and survival were augmented by Endoret treatment. Translation of these effects in vivo was assessed in wild type and APP/PS1 mice, where neurogenesis was evaluated using 5-bromodeoxyuridine (BdrU), doublecortin (DCX), and NeuN immunostaining 5 weeks after Endoret administration. The number of BrdU, DCX, and NeuN positive cell was increased after chronic treatment. The number of degenerating neurons, detected with fluoro Jade-B staining was reduced in Endoret-treated APP/PS1 mice at 5 week after intranasal administration. In conclusion, Endoret was able to activate neuronal progenitor cells, enhancing hippocampal neurogenesis, and to reduce Aβ-induced neurodegeneration in a mouse model of AD.

An evaluation of geographic and annual variation in morphometric characters and gill-raker counts of yellowfin tuna, Thunnus albacares, from the Pacific Ocean

ENGLISH: Samples of yellowfin tuna, Thunnus albacares, collected from five areas of the Pacific Ocean (Mexico, Ecuador, Australia, Japan, and Hawaii) between January and May of 1988 and 1990 were examined for spatiotemporal variation in morphometric characters and gill-raker counts. 'Iwo-factor analysis of variance, with area and year treated as grouping factors, indicated a significant difference in the means of the total gill-raker counts among fish from different areas, but no significant difference between fish caught in different years. The morphometric data were adjusted by allometric formulae to remove size effects. The correct classification rates for the five groups, using discriminant function analysis, based on adjusted morphometric characters, were 77.60/0 for the samples from 1988 and 74.40/0 for those from 1990. These are 72.00/0 and 68.00/0 (Cohen's kappa statistic) better than would have occurred chance. The pattern of geographic variability, however, is unstable for these two years, thus requiring separate discriminant functions for each year. Although there is annual variability in the morphometric characters, these results demonstrate that the stocks examined are morphometrically distinguishable and that their phenetic relationships reflect their geographic origin. SPANISH: Se examinaron muestras de atún aleta amarilla, Thunnus albacares, tomadas de cinco áreas del Océano Pacífico (México, Ecuador, Australia, Japón, y Hawaii) entre enero y mayo de 1988 y 1990, para descubrir variaciones espaciotemporales en las características morfométricas y los conteos de branquiespinas. Un análisis de varianza de dos factores, con área y año como factores de agrupación, indicó una diferencia significativa en los promedios de los conteos de branquiespinas totales entre peces de distintas áreas, pero ninguna entre peces capturados en distintos años. Se ajustaron los datos morfométricos con fórmulas alométricas para eliminar los efectos de la talla del pez. En un análisis de función discriminante, las tasas de clasificación correcta de los cinco grupos, basadas en características morfométricas ajustadas, fueron 77.60/0 para las muestras de 1988 y 74.40/0 para aquellas de 1990. Estas cifras son 72.00/0 y 68.00/0 (estadístico de kappa de Cohen) mejores de lo que se hubiera obtenido al azar. Sin embargo, la variabilidad geográfica es inestable en estos dos años, requiriendo por lo tanto funciones discriminantes separadas para cada año. Aunque existe variabilidad anual en las características morfométricas, estos resultados demuestran que los stocks examinados son morfamétricamente distinguibles, y que su relación fenética refleja su origen geográfico. (PDF contains 31 pages.)

Efficacy and safety of autologous platelet rich plasma for the treatment of vascular ulcers in primary care: Phase III study

Background: Vascular ulcers are commonly seen in daily practice at all levels of care and have great impact at personal, professional and social levels with a high cost in terms of human and material resources. Given that the application of autologous platelet rich plasma has been shown to decrease healing times in various different studies in the hospital setting, we considered that it would be interesting to assess the efficacy and feasibility of this treatment in primary care. The objectives of this study are to assess the potential efficacy and safety of autologous platelet rich plasma for the treatment of venous ulcers compared to the conventional treatment (moist wound care) in primary care patients with chronic venous insufficiency (C, clinical class, E, aetiology, A, anatomy and P, pathophysiology classification C6). Design: We will conduct a phase III, open-label, parallel-group, multicentre, randomized study. The subjects will be 150 patients aged between 40 and 100 years of age with an at least 2-month history of a vascular venous ulcer assigned to ten primary care centres. For the treatment with autologous platelet rich plasma, all the following tasks will be performed in the primary care setting: blood collection, centrifugation, separation of platelet rich plasma, activation of coagulation adding calcium chloride and application of the PRP topically after gelification. The control group will receive standard moist wound care. The outcome variables to be measured at baseline, and at weeks 5 and 9 later include: reduction in the ulcer area, Chronic Venous Insufficiency Quality of Life Questionnaire score, and percentage of patients who require wound care only once a week. Discussion: The results of this study will be useful to improve the protocol for using platelet rich plasma in chronic vascular ulcers and to favour wider use of this treatment in primary care.

A comparison of phytoplankton and total particle counts

Phytoplankton counts made under the light microscope were compared to counts using an electronic dimensional particle counter. Counts were made on a monthly basis, on water samples taken from one station in the Sanyati Basin. Neither total particle numbers nor total particle volume compare closely with phytoplankton numbers. Total particle numbers were of the order of one and a half to two times greater than the phytoplankton numbers.

Preliminary phytoplankton counts in the Sanyati Basin, Lake Kariba

Phytoplankton counts were made on a monthly basis on water samples, taken from one station in the Sanyati Basin. The results show seasonal fluctuations which are probably nutrient dependant. High phytoplankton numbers occur at times of high nutrient levels as was found with the crustacean zooplankton populations (Marshall 1980). Numbers also decreased with depth down to the thermocline. Below the thermocline there was little or no change in numbers.

Beluga, Delphinapterus leucas, Group Sizes in Cook Inlet, Alaska, Based on Observer Counts and Aerial Video

Belugas, Delphinapterus leucas, groups were videotaped concurrent to observer counts during annual NMFS aerial surveys of Cook Inlet, Alaska, from 1994 to 2000. The videotapes provided permanent records of whale groups that could be examined and compared to group size estimates ade by aerial observers.Examination of the video recordings resulted in 275 counts of 79 whale groups. The McLaren formula was used to account for whales missed while they were underwater (average correction factor 2.03; SD=0.64). A correction for whales missed due to video resolution was developed by using a second, paired video camera that magnified images relative to the standard video. This analysis showed that some whales were missed either because their image size fell below the resolution of hte standard video recording or because two whales surfaced so close to each other that their images appeared to be one large whale. The correction method that resulted depended on knowing the average whale image size in the videotapes. Image sizes were measured for 2,775 whales from 275 different passes over whale groups. Corrected group sizes were calcualted as the product of the original count from video, the correction factor for whales missed underwater, and the correction factor for whales missed due to video resolution (averaged 1.17; SD=0.06). A regression formula was developed to estimate group sizes from aerial observer counts; independent variables were the aerial counts and an interaction term relative to encounter rate (whales per second during the counting of a group), which were regressed against the respective group sizes as calculated from the videotapes. Significant effects of encounter rate, either positive or negative, were found for several observers. This formula was used to estimate group size when video was not available. The estimated group sizes were used in the annual abundance estimates.