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Despite the increasing role of molecular markers, differential counts and morphology of hematopoietic cells in the bone marrow (BM) remain essential diagnostic criteria in hematological diseases. However, the respective reference values for BM myelogram commonly used came from small series with limited numbers of healthy individuals. We evaluated the myelograms of 236 healthy individuals who underwent unrelated bone marrow donation. Health check-ups were performed 4 weeks prior to harvest. Samples for this study, taken from the first aspiration, were stained according to the standard Pappenheim method. Three experienced investigators assessed cellularity, megakaryopoiesis, and differential counts independently. The median donor age was 31 (range 18–51) years. Predonation tests did not reveal any relevant morbidity. Thirty-seven out of 42 hypocellular marrow samples were from younger donors up to 39 years. Content of megakaryocytes was normal in 210 specimens (89%). Gender and body mass index had significant impact on hematopoiesis, whereas age had not. The number of erythroblasts was higher (about 32%) and the proportion granulopoiesis slightly lower (about 50%) compared with previous studies. Differential counts showed also some differences with respect to individual maturation stages in these lines. Interrater comparisons showed greater reliability for the assignment of cells to the different hematopoietic cell lines than for single-cell diagnoses. This study largely confirms the results for cell counts in normal human bone marrow available from previous reports and provides some insights into factors that affect individual cell populations. It also reveals substantial variability among even experienced investigators in cytological diagnoses.

This study aimed to evaluate the changes in the erythropoietin level and hematological variables in wrestlers after intermittent hypoxic exposure (IHE). Twelve wrestlers were assigned into two groups: hypoxia (sports training combined with IHE, n=6) and control (sports training, n=6). An IHE was performed for 10 days, with one day off after 6 days, once a day for about an hour. The concentrations of hydrogen peroxide (H(2)O(2)), nitric oxide (NO), vascular endothelial growth factor (VEGF) and erythropoietin (EPO), as well as total creatine kinase activity (CK) were measured. Also, the hematological markers (Hb -hemoglobin, Ht - hematocrit, RBC - red blood cell, WBC - white blood cell, Ret - reticulocytes) were analyzed. The 6-day IHE caused an increase in the levels of H(2)O(2), NO and VEGF. Similarly, the EPO level and WBC count reached the highest value after 6 days of IHE. The total Ret number increase constantly during 10 days of IHE. The hypoxia group showed a higher CK activity compared to the control. In conclusion, 10-day IHE in combination with wrestling training elevates levels of H(2)O(2), NO and VEGF, and improves the oxygen transport capacity by the release of EPO and Ret in circulation.

Background Fewer circulating endothelial progenitor cells (EPCs) and increased plasma (C-term) stromal cell-derived factor 1α (SDF-1α), a substrate of DPP-4, are biomarkers, and perhaps mediators, of cardiovascular risk and mortality. Short-term/acute treatment with DPP-4 inhibitors improve EPC bioavailability; however, long-term effects of DPP-4i on EPCs bioavailability/plasma (C-term) SDF-1α are unknown. Methods Randomized (2:1) open-label trial to compare the effects of vildagliptin (V) (100 mg/day) vs glibenclamide (G) (2.5 mg bid to a maximal dose of 5 mg bid) on circulating EPC levels at 4 and 12 months of treatment in 64 patients with type 2 diabetes in metformin failure. At baseline, and after 4 and 12 months, main clinical/biohumoral parameters, inflammatory biomarkers, concomitant therapies, EPC number (CD34 + /CD133 + /KDR + /10 6 cytometric events) and plasma (C-term) SDF-1α (R&D system) were assessed. Results Baseline characteristics were comparable in the two groups. V and G similarly and significantly (p < 0.0001) improved glucose control. At 12 months, V significantly increased EPC number (p < 0.05) and significantly reduced (C-term) SDF-1α plasma levels (p < 0.01) compared to G, with no differences in inflammatory biomarkers. Conclusions V exerts a long-term favorable effect on EPC and (C-term) SDF-1α levels at glucose equipoise, thereby implying a putative beneficial effect on vascular integrity. Trial registration Clinical Trials number: NCT01822548; name: Effect of Vildagliptin vs. Glibenclamide on Circulating Endothelial Progenitor Cell Number Type 2 Diabetes. Registered 28 March, 2013

Besides stimulating hematopoiesis, erythropoietin (EPO) protects against experimental ischemic injury in the heart. The present study evaluated the safety and tolerability of EPO treatment in non-anemic patients with acute myocardial infarction (MI). In this single-center, investigator-initiated, prospective study, patients with a first acute MI were randomized to one bolus of 300 microg darbepoetin alfa or no additional medication before primary coronary intervention. Twenty-two patients (mean age 59 +/- 2 years) were included. In the darbepoetin group, serum EPO-levels increased to 130-270 times that of controls, within the first 24 h. After darbepoetin administration, only small and non-significant changes in hematocrit levels were observed, while endothelial progenitor cells (EPCs, CD34+/CD45-) were increased at 72 h (2.8 vs. 1.0 cells/microl in control group, p < 0.01). No adverse events were recorded during the 30-day follow-up. After 4 months, left ventricular ejection fraction was similar in the two groups (52 +/- 3% in darbepoetin vs. 48 +/- 5% in control group, p = NS). Intravenous single high-dose darbepoetin alfa in acute MI is both safe and well tolerated. Darbepoetin treatment after MI stimulates EPCs mobilization. The results of this first pilot study support a larger scale clinical trial to establish efficacy of EPO administration in patients after acute MI.

Podocyte Number in Normotensive Type 1 Diabetic Patients With Albuminuria Kathryn E. White 1 , Rudolf W. Bilous 1 , Sally M. Marshall 1 , Meguid El Nahas 2 , Giuseppe Remuzzi 3 , Giampiero Piras 4 , Salvatore De Cosmo 5 , GianCarlo Viberti 6 and on behalf of the European Study for the Prevention of Renal Disease in Type 1 Diabetes (ESPRIT) 1 Department of Medicine, Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, U.K. 2 Sheffield Kidney Institute, Northern General Hospital Trust, Sheffield, U.K. 3 Mario Negri Institute for Pharmacological Research, Bergamo, Italy 4 Diabetes Unit, Hospital Brotzu, Cagliari, Italy 5 Division and Research Unit of Endocrinology, “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Italy 6 Department of Diabetes, Endocrinology & Internal Medicine, Guy’s Hospital, King’s College London, London, U.K. Abstract We estimated glomerular cell number in 50 normotensive type 1 diabetic patients with raised albumin excretion rate (AER) and investigated any change after 3 years in a subgroup of 16 placebo-treated patients. Biopsies from 10 normal kidney donors were used as controls. Mesangial and endothelial cell number was increased in the 50 diabetic patients at the start of the study compared with control subjects. There was no difference in podocyte number. Glomerular volume was increased in diabetic patients, but surface area of glomerular basement membrane (GBM) underlying the podocytes did not differ between groups. AER correlated positively with mesangial cell number in microalbuminuric patients ( r = 0.44, P = 0.012) and negatively with podocyte number in proteinuric patients ( r = −0.48, P = 0.040). In the 16 placebo-treated patients, glomerular volume increased after 3 years owing to matrix accumulation and increased GBM surface area. Although overall cell number did not differ significantly from baseline, the decrease in podocyte number during follow-up correlated with AER at follow-up ( r = −0.72, P = 0.002). In conclusion, cross-sectional analysis of podocyte number in type 1 diabetic patients with raised AER but normal blood pressure shows no significant reduction compared with nondiabetic control subjects. Longitudinal data provide evidence for an association between podocyte loss and AER, but whether cellular changes are a response to, a cause of, or concomitant with the progression of nephropathy remains uncertain. Footnotes Address correspondence and reprint requests to Dr. K.E. White, Department of Medicine, Medical School, Framlington Place, Newcastle upon Tyne, NE2 4HH U.K. E-mail: k.e.white{at}ncl.ac.uk . Received for publication 20 February 2002 and accepted in revised form 12 July 2002. AER, albumin excretion rate; BP, blood pressure; ESPRIT, European Study for the Prevention of Renal Disease in Type 1 Diabetes; GBM, glomerular basement membrane; GFR, glomerular filtration rate. DIABETES

This popular pocket book has been updated and expanded throughout, providing a concise view of diagnostic haematology, in a convenient and practical format. A Beginner's Guide to Blood Cells is an ideal for; Trainee laboratory technicians and scientists Students studying the physiology or pathology of the blood Those preparing for haematology examinations Why Buy This Book? Unique pocket guide, written by Barbara Bain - a world-renowned expert in the field of blood morphology Outlines the basics of diagnostic haematology Includes an invaluable self-assessment section NEW EDITION - now includes more information on clinical aspects and further tests.

Background Endothelial progenitor cells (EPC) are bone marrow-derived cells which can undergo differentiation into endothelial cells and participate in endothelial repair and angiogenesis. Insulin facilitates this in vitro mediated by the IGF-1 receptor. Clinical trials showed that the number of circulating EPCs is influenced by glucose control and EPC are a predictor of cardiovascular death. To study direct effects of insulin treatment on EPCs in type 2 diabetes patients, add-on basal insulin treatment was compared to an escalation of oral medication aiming at similar glucose control between the groups. Methods 55 patients with type 2 diabetes (61.6±5.9 years) on oral diabetes medication were randomized in a 2:2:1 ratio in 3 groups. Patients were treated additionally with insulin glargine (n=20), NPH insulin (n=22) or escalated with oral medication (n=13). Number of circulating EPC, EPC-outgrowth, intima media thickness, skin microvascular function and HbA1c were documented at baseline and/or after 4 weeks and 4 months. Results HbA1c at baseline was, 7.3+/-0.7% in the oral group, 7.3+/-0.9% and 7.5+/-0.7% in the glargine and NPH insulin respectively (p=0.713). HbA1c after 4 months decreased to 6.8+/-0.8%, 6.6+/-0.7% and 6.7+/-0.6%, in the oral, glargine and NPH insulin group respectively (p=0.61). FACS analysis showed no difference in number of circulating EPC between the groups after 4 weeks and 4 months. However, the outgrowth of EPCs as detected by colony forming assay was increased in the NPH insulin and glargine groups (29.2+/-6.4 and 29.4+/- 6.7 units respectively) compared to the group on oral medication (23.2+/-6.3, p=0.013) after 4 months of treatment. A significant decrease of IMT from 0.80mm (+/-0.14) at baseline to 0.76mm (+/-0.12) after 4 months could be observed in all patients only (p=0.03) with a trend towards a reduction of IMT after 4 months when all patients on insulin treatment were compared to the oral treatment group (p=0.06). Skin microvascular function revealed no differences between the groups (p=0.74). Conclusion The study shows that a 4-month treatment with add-on insulin significantly increases the outgrowth of EPC in patients with type 2 diabetes mellitus. Trial registration (Clinical Trials Identifier: NCT00523393).

Different optical coherence tomography （OCT）, both tirne-domain OCT and spectral-domain OCT, has been used to evaluate anterior chamber cells in previous studies. Recently, commercial swept-source OCT was available. The comparison among three kinds of OCTs have not been done before. Here, we compared their feasibility in discerning aqueous cells in vitro. The peripheral blood was diluted to eight different cell concentrations and was packed into plastic-film bag. Each sample was test- ed using different scan modes of three OCTs. It was easy to discern aqueous cells from background noise in both Visante AS-OCT and RTVue XR Avanti OCT, but difficult in Casia SS-1000 OCT. The measured blood cell densities in high-resolution corneal cross-sectional scan of Visante AS-OCT, cornea line scan and 3D cornea scan of RTVue XR Avanti OCT were highly correlated with the actual blood cell concentration （Pearson correlation=0.991, 0.989, and 0.993 respective- ly）. The mean adjusted cell densities of cornea line scan （10.46±2.87 spots mm-2） and 3D cornea scan （11.01±2.47 spots mm-2） of RTVue XR Avanti OCT were significantly larger than that of Visante AS-OCT （1.55±1.75 spots mm-2） （P〈0.01）. Our study demonstrated that RTVue XR Avanti OCT, a spectral-domain OCT, is superior to other two OCTs in evaluating aqueous cells.

Although the elementary unit of biology is the cell, high-throughput methods for the microscale manipulation of cells and reagents are limited. The existing options either are slow, lack single-cell specificity, or use fluid volumes out of scale with those of cells. Here we present printed droplet microfluidics, a technology to dispense picoliter droplets and cells with deterministic control. The core technology is a fluorescence-activated droplet sorter coupled to a specialized substrate that together act as a picoliter droplet and single-cell printer, enabling high-throughput generation of intricate arrays of droplets, cells, and microparticles. Printed droplet microfluidics provides a programmable and robust technology to construct arrays of defined cell and reagent combinations and to integrate multiple measurement modalities together in a single assay.

Abstract Background The clinical utility of bronchoalveolar lavage fluid (BAL) cell analysis for the diagnosis and management of patients with interstitial lung disease (ILD) has been a subject of debate and controversy. The American Thoracic Society (ATS) sponsored a committee of international experts to examine all relevant literature on BAL in ILD and provide recommendations concerning the use of BAL in the diagnosis and management of patients with suspected ILD. Purpose To provide recommendations for (1) the performance and processing of BAL and (2) the interpretation of BAL nucleated immune cell patterns and other BAL characteristics in patients with suspected ILD. Methods A pragmatic systematic review was performed to identify unique citations related to BAL in patients with ILD that were published between 1970 and 2006. The search was updated during the guideline development process to include published literature through March 2011. This is the evidence upon which the committee's conclusions and recommendations are based. Results Recommendations for the performance and processing of BAL, as well as the interpretation of BAL findings, were formulated by the committee. Conclusions When used in conjunction with comprehensive clinical information and adequate thoracic imaging such as high-resolution computed tomography of the thorax, BAL cell patterns and other characteristics frequently provide useful information for the diagnostic evaluation of patients with suspected ILD.