Conclusion The potential for contracting a microbial pathogen is highest within a hospital environment and hospital acquired infections are significant contributors to morbidity and mortality. Despite the lack of direct evidence to prove that environmental contaminants are responsible for hospital acquired infections, there is an increasing evidence suggesting that the environment may act as a reservoir for at least some of the pathogens causing nosocomial infections. This

work showed that many different bacterial species can persist on surfaces for months and years. The level of bacterial contamination was related with the this website presence of humidity on selleck inhibitor the surface, and tap water (biofilm) was a point of dispersion of bacterial species, usually involved in nosocomial infections as Pseudomonas

aeruginosa, Stenotrophomonas maltophilia and Enterococcus feacalis. Their presence in the environment, as it seems to be pointed by the analysis of the diversity, increases the risk of transmission to the different materials during hand manipulation. Methods Sampling (ICU, Medicine I, Medicine II and Urology) The study was carried out at the Hospital de Faro, Portugal, which serves a resident population of about 253 thousand people and this value may double or triple the population seasonally (in http://​www.​hdfaro.​min-saude.​pt/​site/​index.​php). Between January 2010 and

September, 2011, the hospital was evaluated 12 times (sampled each two months) and four different wards were investigated for environmental contamination of the following surfaces and equipment: sink, tap (biofilm), surface countertop and workbench of the nurses area, shower (and handrail), bedside table, handrail bed (including bed), serum support, oxygen flask, stethoscope, equipment at bedside, other medical equipment, tray used by nurses, hand gel/soap, table (meal and work). The equipment considered in this study is included in the category of noncritical hospital objects and surfaces. These items have been Tau-protein kinase said to pose no risk to patients, nevertheless, these surfaces and equipment are frequently touched by hand can contribute to the spread of healthcare-associated pathogens as Pseudomonas aeruginosa, Staphilococus aureus, or Acinetobacter baumanii. The evaluation was performed in wards of the Medical Unit I and II, Urology and Intensive Care Unit. Samples were collected in the wards, always in the same period of the day, at the end of the morning and during lunch time, after the medical visits and treatments, and also sometime after a ward cleaning. Swabs were used for collecting the organisms present in an area of 10X10 cm of each surface. Taps were sampled by removing the biofilm.

We thank Mari Nyyssönen for help with the microarray experiments, and thank Jizhong Zhou and Liyou Wu for providing the microarrays. The work was supported by a grant from U.S Department of Energy, Office of Science, DE-FG02-04ER63923 and by the WCU (World Class University) program through the National Research Foundation

of Korea funded by the Ministry of Education, Science and Technology (R33-10076). References 1. Villemur R, Lanthier M, Beaudet R, Lépine F: The Desulfitobacterium genus. FEMS Microbiology Reviews 2006, 30:706–733.PubMedCrossRef 2. Kunapuli U, Jahn MK, Lueders T, Geyer R, Heipieper HJ, Meckenstock RU: Desulfitobacterium aromaticivorans sp. nov. and Geobacter toluenoxydans sp. nov., iron-reducing bacteria capable of anaerobic degradation of monoaromatic hydrocarbons. PI3K inhibitor Int J Syst Evol Microbiol 2010,60(3):686–695.PubMedCrossRef 3. Maymo-Gatell GSK458 in vitro X, Chien Y, Gossett JM, Zinder SH: Isolation of a bacterium that reductively dechlorinates tetrachloroethene to ethene. Science 1997, 276:1568–1571.PubMedCrossRef 4. Madsen T, Licht D: Isolation and characterization of an

anaerobic chlorophenol-transforming bacterium. Appl Environ Microbiol 1992, 58:2874–2878.PubMed 5. Christiansen N, Ahring BK: Desulfitobacterium hafniense sp. nov., an anaerobic, reductively dechlorinating bacterium. Int J Syst Bacteriol 1996, 46:442–448.CrossRef 6. Niggemyer A, Spring S, Stackebrandt E, Rosenzweig RF: Isolation and characterization of a novel As(V)-reducing bacterium: implications for arsenic mobilization and the genus Desulfitobacterium . Appl Environ Microbiol 2001, 67:5568–5580.PubMedCrossRef 7. Lie TJ, Godchaux W, Leadbetter ER: Sulfonates as terminal electron acceptors for growth of sulfite-reducing bacteria ( Desulfitobacterium spp.) and sulfate-reducing bacteria: effects of inhibitors of sulfidogenesis. Pazopanib purchase Appl Environ Microbiol 1999,65(10):4611–4617.PubMed 8. Suyama A, Iwakiri R, Kai K, Tokunaga T, Sera

N, Furukawa K: Isolation and characterization of Desulfitobacterium sp. strain Y51 capable of efficient dechlorination of tetrachloroethene and polychloroethanes. Biosci Biotechnol Biochem 2001, 65:1474–1481.PubMedCrossRef 9. Nonaka H, Keresztes G, Shinoda Y, Ikenaga Y, Abe M, Naito K, Inatomi K, Furukawa K, Inui M, Yukawa H: Complete genome sequence of the dehalorespiring bacterium Desulfitobacterium hafniense Y51 and comparison with Dehalococcoides ethenogenes 195. J Bacteriol 2006,188(6):2262–2274.PubMedCrossRef 10. Suyama A, Yamashita M, Yoshino S, Furukawa K: Molecular characterization of the PceA reductive dehalogenase of Desulfitobacterium sp. Strain Y51. J Bacteriol 2002,184(13):3419–3425.PubMedCrossRef 11. Juhala RJ, Ford ME, Duda RL, Youlton A, Hatfull GF, Hendrix RW: Genomic sequences of bacteriophages HK97 and HK022: pervasive genetic mosaicism in the lambdoid bacteriophages. Journal of Molecular Biology 2000,299(1):27–51.PubMedCrossRef 12.

Free PHB granules, i.e. PHB granules that were not in contact AZD1390 nmr to the nucleoid region were not observed. Apparently, constitutive over-expression of phaM resulted in formation of an increased number of small and nucleoid-attached PHB granules. If PhaM is responsible

for the formation of small granules and for the close contact to the nucleoid region, deletion of phaM should have a phenotype. In fact, R. eutropha ∆phaM cells accumulated only very few (0–2) PHB granules that were significantly larger in diameter than those of the phaM over-expressing mutant or of the wild type (Figure 5). Since the diameters of PHB granules of the ∆phaM strain were considerably larger even at early time points a precise analysis whether or not the granules were attached to the nucleoid region was difficult. In most ∆phaM cells the PHB granules were still located close to the nucleoid; however, BLZ945 ic50 at least in some cells a detachment of PHB granules from the nucleoid region could not be excluded for the wild type or for the phaM over-expressing strain. A clear decision whether the absence of PhaM resulted in detachment from the nucleoid can, however, not be made. Since

R. eutropha expresses at least one other protein with DNA-binding and PHB-binding property (PhaR) [30, 31] it might be that PhaR also contributes to association of PHB with DNA. In summary, our data on mutants with altered expression of PhaM clearly show that number, diameter and subcellular localization of PHB granules depends on the presence and concentration of PhaM. Time course of formation and localization of PHB granules in R. eutropha over-expressing PhaP5 PhaP5 had previously been identified as a phasin in R. eutropha by its in vivo interaction with PhaP2 and other phasins [22]. Remarkably, PhaP5 also interacted

with PhaM. To investigate the influence of PhaP5 on RANTES PHB granule formation the phaP5 gene was cloned in a broad host range plasmid (pBBR1MCS-2) under control of the strong and constitutive phaC1 promotor (PphaC), transferred to R. eutropha H16 and HF39 via conjugation and investigated for PHB granules formation and localization under PHB permissive conditions (Figure 6). In case of strain HF39 a eypf-phaP5 fusion was cloned and used to confirm localization of PhaP5 on the PHB granules by fluorescence microscopy. Controls showed that free eYfp is a soluble protein in R. eutropha (Figure 7). Figure 7 Fluorescence microscopical (FM) investigation of R. eutropha H16 (pBBR1MCS-2-P phaC – eyfp -c1) with over-expression of eYfp (a); R. eutropha H16 (pBBR1MCS-2-P phaC – phaP5 ) with over-expression of PhaP5 (b), and R. eutropha H16 (pBBR1MCS-2-P phaC -eyfp- phaP5 ) with over-expression of eYfp-PhaP5 fusion (c) at various stages of PHB formation.

pneumoniae putative surface protein Orf50 53176-54000 E S. pneumoniae DNA replication protein Orf72 79231-80088 E S. pneumoniae putative bacteriocin Orf51 53993-54478 E S. pneumoniae DUF 3801 Orf73 80162-80773 E S. pneumoniae Predicted transcriptional regulator Orf52 54475-55209 E S. pneumoniae phage antirepressor protein Orf74 80766-81749 E S. pneumoniae Protein with unknown function Orf53 55202-56890 E S. pneumoniae TraG/TraD family protein Orf75 82268-82621 E S. pneumoniae transcriptional regulator, ArsR family Orf54 57454-58486 E – DUF Entospletinib 318 Predicted Permease (HHPred) Orf76 82696-83940 E S. pneumoniae major facilitator superfamily MFS_1 Orf55 59048-59398 D C. fetus glyoxalase

family protein Orf77 83927-84403 E S. pneumoniae toxin-antitoxin system, toxin component, GNAT domain protein Orf56 59411-59938 D C. fetus transcriptional regulator Orf78 84758-86491 E S. pneumoniae DNA topoisomerase III Orf57 59988-61910 D C. fetus tetracycline resistance R406 clinical trial protein Orf79 86484-87449 E S. pneumoniae possible DNA (cytosine-5-)-methyltransferase Orf58

62225-63082 D C. fetus aminoglycoside 6-adenylyltransferase (AAD(6) Orf80 87436-95079 E S. pneumoniae superfamily II DNA and RNA helicase Orf59 63575-64348 E S. pneumoniae replication initiator/phage Orf81 95123-95779 E S. pneumoniae putative single-stranded DNA binding protein Orf60 64345-65172 E S. pneumoniae replicative DNA helicase Orf82 95939-96841 E S. pneumoniae transcriptional regulator, XRE family Orf61 65314-65814 E S. pneumoniae Cyclooxygenase (COX) TnpX site-specific recombinase family protein Orf83 97071-98282 E S. pneumoniae transporter, major facilitator family/multidrug resistance protein 2 Orf62 65938-66399

E S. pneumoniae flavodoxin Orf84 C 99739-98462 E S. pneumoniae relaxase/type IV secretory pathway protein VirD2 Orf63 66817-67302 E S. pneumoniae putative conjugative transposon protein Orf85 C 101169-99795 E S. pneumoniae conjugal transfer relaxosome component TraJ Orf64 67299-68033 E S. pneumoniae phage antirepressor protein Orf86 C 101403-100321 E S. pneumoniae toxin-antitoxin system, toxin component, Fic family Orf65 68026-69816 E S. pneumoniae TraG/TraD family protein/putative conjugal transfer protein Orf87 C 101878-101396 E S. pneumoniae putative membrane protein Orf66 70395-70706 E S. pneumoniae putative single-strand binding protein Orf88 C 102435-101887 E S. pneumoniae putative toxin-antitoxin system, toxin component Orf67 70934-71797 E S. pneumoniae conjugative transposon membrane protein Orf89 C 102845-102444 E S. pneumoniae regulator/toxin-antitoxin system, antitoxin component Orf68 72099-72509 E S. pneumoniae conjugative transposon membrane protein Orf90 103034-103555 E S. pneumoniae conserved hypothetical protein Orf69 72580-74823 E S. pneumoniae type IV conjugative transfer system protein Orf91 103825-104235 E S. pneumoniae sigma-70, region 4 Orf70 74831-77410 E S. pneumoniae conjugative transposon cell wall hydrolase/NlpC/P60 family Orf92 104966-106712 E S.

Conventional photolithography and photoresist stripping processes were employed to construct channels with

the desired depth. A silicon (Si) wafer was cleaned in H2SO4:H2O2 solution LY3009104 (volume ratio of 10:1) at 120°C for 10 min, followed by deionized water (DI) for 4 cycles, then HF:H2O solution (1:50) at 22°C for 1 min and DI water for 4 cycles, and finally spin-dried in hot N2 gas for 15 min. Then, the Si wafer was processed by hexamethyldisiloxane (HMDS) coating and positive photoresist HPR 504 spin-coated at 4,000 rpm for 30 s. The wafer was soft-baked on a hot plate at 110°C for 60 s before exposing to UV via the Mask Aligner (SUSS Microtec MA6-2, Garching Germany) for 5 s. The photoresist was developed using FHD-5 for 60 s and post-baked on a hot plate at 120°C for 60 s. The micropatterns were successfully defined at this stage. The Si wafer was then RG7112 research buy etched by a DRIE machine (Surface Technology Systems, Newport, UK) and followed by photoresist stripping in PS210 Photoresist Asher (PVA Tepla AG, Kirchheim, Germany) for 25 min. After constructing the microchannels, 10 nm of thermal oxide was grown using a diffusion furnace to form silica on the channel wall. After drilling the inlets and outlets on the Si chip by a mechanical driller, the chip has to be sealed to form a closed channel. A thin film of polydimethylsiloxane (PDMS) was applied for such purpose due to the good adhesion between PDMS and the Si chip. PDMS was formulated

from Sylgard 184 silicone elastomer mixture (Dow Corning Corporation, Midland, MI, USA) at a weight ratio of base:curing agent = 10:1. Then, it was poured onto a Si wafer with saline coating on the surface and pressed against a cleaned glass slide. After curing PDMS in an oven at 60°C for 2 h, the microchip was constructed by pressing the Si chip against the glass slide Selleckchem Nutlin-3 with the thin layer of PDMS on its surface. The fabricated microchip is shown in Figure  2a. The microreactor is comprised of two microchannels: channels A and B with a width of 300 μm and a depth of 12 μm and an array (20 channels) of 1D nanochannels that connected the two microchannels

to demonstrate the injection process. It is not necessary to adopt 20 nanochannels. One can increase or decrease the number according to their applications. Fewer nanochannels will result in higher precision, and more nanochannels will give a higher throughput. The inset (a1) in Figure  2a illustrates the multilayer structure showing the PDMS, the silicon chip, and the glass slide. Another inset (a2) shows the structure of the two microchannels connected by the nanochannel array that is highlighted by the green dashed square. When the electric field across channel A and channel B was applied, fluid flowed from channel A to channel B through the nanochannel array as indicated by the green arrow in the same figure. The enlarged scanning electron microscopy (SEM) image of the nanochannel array is shown in Figure  2b. The channel width observed was 10 μm.

Biostatistics 2003, 4:249–64.CrossRefPubMed

72. Tusher VG, Tibshirani R, Chu G: Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA 2001, 98:5116–21.CrossRefPubMed 73. Bioinformatics software for genomic data[http://​bioconductor.​org] 74. Software environment for statistical computing and graphics[http://​www.​r-project.​org] Authors’ contributions IS performed the experiments and helped with the interpretation of the data. ADL designed and developed the probe selection process and performed the bioinformatics CYT387 manufacturer and statistical analyses of microarray data. JAV performed the sequence annotation and revised the manuscript. EMV supervised the study and helped in writing the discussion of the manuscript. MBS designed and coordinated the study, participated in the experiments, the microarray data analysis and the annotation process, and wrote the manuscript. All authors read and approved the final manuscript.”
“Background Francisella tularensis is a highly virulent Gram negative bacterial pathogen and the etiologic

agent of the zoonotic disease tularemia. The bacteria are spread via multiple transmission routes including arthropod bites [1], physical selleck inhibitor contact with infected animal tissues [2], contaminated water [3, 4], and inhalation of aerosolized organisms [5]. Inhalation of as few as 10 colony forming units (CFU) are sufficient to initiate lung colonization [6, 7] and the subsequent development of pulmonary tularemia, which is the most lethal form of Tideglusib the disease exhibiting mortality rates as high as 60% [8]. F. tularensis is a facultative intracellular pathogen that invades, survives and replicates within numerous cell types

including, but not limited to, macrophages [9, 10], dendritic cells [11], and alveolar epithelial cells [12]. Intracellular growth is intricately associated with F. tularensis virulence and pathogenesis, and the intracellular lifestyle of F. tularensis is an active area of investigation. Following uptake or invasion of a host cell wild type F. tularensis cells escape the phagosome and replicate within the cytoplasm [13–15] of infected cells. The phagosome escape mechanism employed by F. tularensis remains essentially unknown, but this property is clearly necessary for F. tularensis intracellular growth since mutants that fail to reach the cytoplasm are essentially unable to replicate within host cells [16, 17]. Following phagosome escape F. tularensis must adapt to the cytoplasmic environment. Purine auxotrophs [18], acid phosphatase [19], clpB protease [20], and ripA mutants [21] reach the cytoplasm but are defective for intracellular growth. RipA is a cytoplasmic membrane protein of unknown function that is conserved among Francisella species [21]. Notably, the majority of attenuating mutations described to date impart intracellular growth defects on the mutant strains.

In the current study, we have used a similar assay to identify chemicals that increase iron uptake into cells and demonstrate that these chemicals are effective in increasing iron transport across Caco2 cells, a model system for studying intestinal iron absorption, and increasing iron uptake into various cancer cell lines, favourably altering several aspects of the malignant phenotype. ubiquitin-Proteasome system Methods

Cell lines and Chemicals All antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA) except for rabbit anti-HIF-1α and -2α which were purchased from Novos Biologicals (Littleton, CO). All analytical chemicals were from Sigma-Aldrich (St. Louis, MO). The chemical libraries were obtained from ChemDiv (San Diego, CA) and TimTec (Newark, DE). CM-H2DCFDA (5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetyl ester) or DCFDA and calcein-AM were from Invitrogen (Carlsbad, CA). The cell lines K562, ITF2357 solubility dmso PC-3, Caco2, MDA-MB231, and 267B1 were

obtained from ATCC (Bethesda, MD). RPMI1640 and DMEM culture media and fetal calf serum (FCS) were obtained from Atlanta Biologicals (Lawrenceville, GA). Screening for chemicals that increase iron uptake K562 cells were loaded with calcein by incubating cells with 0.1 μM of Calcein-AM for 10 min in 0.15 M NaCl-20 mM Hepes buffer, pH 7.4, with 0.1% BSA at 37°C followed by extensive washing with NaCl-Hepes buffer to remove extracellular bound calcein, and aliquoted at 5 × 104 – 1 × 105 cells/well in 96-well plates containing test compounds at 10 μM and incubated for 30 min in a humidified 37°C incubator with 5% CO2 before baseline fluorescence was obtained at 485/520 nm (excitation/emission) with 0.1% DMSO as the vehicle control and DTPA as a strong iron chelator control to block all iron uptake. much The fluorescence was then obtained 30 min after addition of 10 μM ferrous ammonium sulfate in 500 μM ascorbic acid (AA). The percentage of fluorescence quench was calculated relative

to 200 μM DTPA added as a blocking control and DMSO as a vehicle control as follows: (1) where Δ F is the change in fluorescence, or fluorescence quench, observed in any well, F0 represents the fluorescence after 30 min of compound, and Ff represents the fluorescence 30 min after addition of Fe. These results were normalized to the blocking and vehicle controls as follows: (2) where Δ Fn is the normalized quench observed after addition of iron, Fcompound is the Δ F observed with compound, Fmin is the average Δ F of the DMSO control; and Fmax is the average Δ F of the DTPA control. With this normalization 100% indicates that a test compound is as potent as DTPA in blocking iron-induced quenching and 0% indicates no inhibition of iron quenching by a test compound or the same quench as observed with the DMSO vehicle control. Compounds with Δ Fn between 0% and 100% are defined as inhibitors of iron uptake.

Nearly 40% of the starting suspension of yeast cells were recovered when cells were slowly frozen in an 8% DMSO-containing solution and this procedure was selected for long term storage of mutant pools. Although specialized cooling apparatuses can be used to control the freezing rate, we found that simple placement of vials of cells within readily and cheaply obtained styrofoam containers (such as those used for shipments of molecular biology enzymes) was sufficient. Figure 2 Gradual freezing in DMSO maximizes recovery of cryopreserved Histoplasma yeast. selleck products WU15 yeast were frozen in varying concentrations of glycerol (A) or DMSO (B). Histoplasma yeast were grown

to late log/early stationary phase in rich medium and added to the appropriate glycerol- or DMSO-containing solutions before freezing. Final cryoprotectant concentrations

indicated along the x-axis of each graph. Vials were placed immediately mTOR inhibitor at -80°C (rapid freeze) or were placed into a styrofoam container before placement at -80°C (slow freeze). Frozen cell aliquots were thawed after 1 week or 9 weeks and recovery measured as the number of viable cfu relative to the number present before freezing. Generation of mutant pools Insertion mutants were generated in the NAm 2 Histoplasma strain WU15 by co-cultivation of Agrobacterium tumefaciens and Histoplasma yeast cells. Co-cultures were plated onto filters and Histoplasma transformants selected Exoribonuclease by transferring filters to medium containing hygromycin to which resistance is provided by sequences within the T-DNA element [23]. Transformant yeast cells were collected and suspensions from individual plates combined to create pools derived from 100 to 200 independent mutant colonies. Yeast cell suspensions were diluted into fresh medium and allowed to grow for 24-48 hours. Twenty-four pools were prepared representing roughly 4000 insertion mutants. A portion of each culture was reserved for nucleic acid isolation and the remainder frozen in aliquots and stored at -80°C. Nucleic acids were purified from

each pool, diluted to 50 ng/ul, and stored at -20°C until analysis by PCR. With an estimated 9000-10,000 genes encoded by the Histoplasma genome, this collection does not represent the number of insertion mutants required for saturation of the genome. We used two probability functions to estimate the size of the library required for a 95% chance of isolating an insertion in a particular locus in the 40 megabase NAm 2 genome. Both calculations assume no bias in insertion sites. Based on the number of predicted genes, the Poisson approach estimates a library of approximately 30,000 insertions would be required. The single study in which multiple alleles of a single locus were isolated in Histoplasma (five AGS1 alleles isolated in a screen of 50,000 insertions; [23]) supports the Poisson calculation; five alleles would be the most probable number of alleles based on a 9000 or 10,000 target estimate.

Marked changes in blood leukocyte counts resulting from a single bout of high intensity exercise are well known and are due largely to the movement of neutrophils from the marginal pool to the circulating pool as a result of muscular action [44]. It is documented that neutrophilia depends of exercise intensity and duration [7] MLN8237 and also of body temperature attained during exercise [45]. Acute exercise results in a rapid increase in blood neutrophil counts likely due to demargination

caused by shear stress and catecholamines [46], which is followed by a delayed neutrophilia attributed to cortisol-induced release of neutrophils from the bone marrow [46]. An increase in blood neutrophil numbers does not imply better neutrophil function, because neutrophils released as a result of acute exercise are relatively immature and consequently their degranulation and oxidative burst in response to bacterial stimulation may be reduced for many hours after the exercise bout [47–49]. Acute exercise elicits characteristic transient biphasic changes in the numbers of circulating lymphocytes. Typically, a lymphocytosis is observed immediately after exercise, with numbers of cells

falling below pre-exercise levels during the early stages of recovery [50]. Results obtained in this study are in total agreement with this pattern of response, with significant decreases in lymphocyte numbers click here detected at 30 and 150 min after exercise, except for the group supplemented with nucleotides in which a total recovery on the number of lymphocytes was detected at 150 min. Although it has been shown that dietary nucleotides stimulates the maturation of immune cells [17, 51], the rapid recovery in lymphocyte counts registered between 30 and 150 min after the exercise test, suggest a redistribution from other cell compartments. There is considerable evidence demonstrating that

exogenous nucleotides increase the proliferative response to T cell-dependent mitogens (PHA, ConA and PWM) [14, 17]. In the present study, significant differences in lymphocyte proliferation have been detected between treatment groups at 24 h after exercise. On the initial exercise test, lymphoproliferative Methamphetamine activity was higher in the placebo group (P < 0.05), while after supplementation it was higher in the nucleotide group (P < 0.05). Interpretation of the data is hampered by the fact that values are different in the baseline test. This was probably due to the reduced sample size (10 athletes per group) and the randomized nature of the study, which resulted by happenstance (since this result is prior to intervention) in an almost significant effect of exercise in the I group. This may be interpreted to indicate a higher susceptibility of this group to depressed lymphocyte proliferation in the face of intense physical activity. This in turn would be expected to dampen, or hide, a putative effect of the nucleotide supplement in this regard.

Patients and methods Patients This prospective study involved 37 consecutive patients with a median age of 28 years (range: 19-58 years) who underwent an allogeneic hematopoietic stem cell transplantation (HSCT) from June 2009 to February 2011 at the Transplantation Centre of Hematology Department Protein Tyrosine Kinase inhibitor at University Hospital Bratislava. There were 24 males and 13 females. Their diagnosis included acute myeloid leukemia (AML) in 13 patients,

acute lymphoblastic leukemia (ALL) in 14 patients, chronic myeloid leukemia (CML) in 2 patients, Hodgkin’s lymhoma in one patient, myelodysplastic syndrome (MDS) in 3 patients, osteomyelofibrosis in one patient and severe aplastic anemia in 3 patients. Twenty-seven patients were conditioned with myeloablative regimens including cyclophosphamide (CY) 60 mg/kg body weight intravenously on 2 consecutive days in combination with fractionated total

body irradiation (TBI) 12 Gy in six fractions of 2 Gy over 3 days in 12 patiens or in combination with peroral busulphan 4 mg/kg body weight daily for 4 days in 15 patients. The remaining 10 patients were conditioned next with nonmyeloablative Mizoribine chemical structure regimens (cyclophosphamide, busulphan, fludarabine, etoposide, cytosine arabinoside, melphalan, idarubicin, carmustine or with combination of antithymocyte globulin). Fifteen patients received hematopoietic

stem cells from an HLA-matched related donor and 22 patients from an HLA-matched unrelated donor. Cyclosporine A and short-term methotrexate were administered for the prophylaxis of graft-versus-host disease (GVHD). Two patients had arterial hypertension, 2 patients had diabetes mellitus and 14 patients had dyslipidemia before transplantation. One patient had a prior history of a cardiac disease because of leukemic infiltration of the heart (at the time of diagnosis of acute leukemia). The cumulative dose of anthracyclines (ANT) (idarubicin, daunorubicin and mitoxantrone) was calculated as the equivalent dose of doxorubicin. Twenty-nine patients were previously treated with ANT (median 250 mg/m2, range: 100-470). Characteristics of patients are summarized in Table 1.