Enhancement of adenovirus-mediated gene transfer to human bone marrow cells

Adenovirus infection of CD34+ hematopoietic stem/progenitor cells is dependent on the multiplicity of infection (MOI), time of incubation, the volume in which the co-incubation occurs and the presence or absence of growth factors. Studies revealed that a brief co-incubation (1-8 hours), resulted in low levels of transgene expression, suggesting that adenovirus infection of CD34+ cells occurs slowly, and optimal transduction requires a 24 hour exposure to adenovirus. Infection by Ad/beta-gal or Ad/p53 at a MOI of 500:1 provided a high transduction efficiency but inhibited hematopoietic function. However, treatment at a MOI of 50-100 resulted in efficient transduction (10.7-15.7% positive) without detectable toxicity. Secondary proof of adenovirus transgene expression was demonstrated by detection of mRNA for p53 in Ad/p53 infected stem cells. We conclude that a 24 hour exposure to recombinant adenovirus encoding p53 or beta-gal, at a MOI of 50-100 is optimal for in vitro gene transfer to BM cells and has no significant effect on hematopoietic function. Adenovirus-mediated transduction of BM cells can also be modulated by growth factors (IL-3, GM-CSF and G-CSF) with improved gene delivery and maintenance of hematopoietic function. In summary, adenovirus vectors can be used to transiently transduce stem cells, and conditions have been defined to maximize expression and limit inhibitory effects on CD34+ cells. These data support continued investigation of this vector for local cytokine delivery and purging of stem cell products.     

Thrombolytic therapy in Missouri hospital emergency departments: compliance with the National Heart Attack Alert Program guidelines

Adhesive interactions between leukocytes and endothelium are required for subsequent leukocyte extravasation toward inflammatory sites. Understanding the possible kinetic expression of vascular cell adhesion molecule-1 (VCAM-1) in the middle ear cavity during an inflammatory cascade in vivo may be important for clarifying local immunological responses in otitis media. Two inflammatory models were produced in the rat and involved acute middle ear mucosal and cutaneous inflammation induced after inoculation or intradermal injection of lipopolysaccharide (LPS). After intravenous injection of both 125I-labeled anti-VCAM-1 and 131I-labeled control monoclonal antibody (mAb), the kinetic expression of VCAM-1 in the middle ear and skin was assessed by local radionuclide uptake. The biodistribution of an 125I-labeled anti-VCAM-1 mAb as a potential detector of focal inflammation was examined in normal rats. Both inflammatory lesions were characterized by early and sustained (up to 24 h) expression of VCAM-1, with maximal expression at 4 h after LPS stimulation. The kinetics of VCAM-1 expression was similar among the middle ear mucosa or skin specimens studied and different stimulation methods. A similar biodistribution and clearance of radioactivity between 125I-labeled anti-VCAM-1 mAb and 131I- or 99mTc-labeled control mAb were observed. The present result suggest that functional VCAM-1 induced by LPS is expressed in both middle ear tissue and skin lesions and may play a role in the initial stage of inflammatory response produced. Although VCAM-1 upregulation is a very early event in the inflammatory cascade, 125I-labeled anti-VCAM-1 mAb may be useful for the early detection of focal inflammation in the middle ear.     

A glutamate-sensitizing activity in conditioned media derived from rat cerebellar granule cells

When cerebellar granule cells that had been cultured in vitro for 8 days were subjected to a cytotoxic glutamate pulse (100 microM, 30 min incubation), the response varied according to cell density and the volume of medium in which cells were grown. Thus, lowering the cell density by a factor of 4 compared with usual conditions (2.6 x 10(5) cells/cm2) or increasing the volume by an identical 4-fold factor reduced cell death from 90-95% to 20-30%. Addition of a conditioned medium derived from high-density to low-density cultures or to high-volume cultures markedly increased the sensitivity of the cells to glutamate. This glutamate-sensitizing activity, which accelerated by several days the onset of the response of cerebellar cultures to glutamate, was inhibited by actinomycin D and was not detectable in conditioned medium derived from confluent cultures of cerebellar astroglia, or from cell lines such as PC12, GT1-7, 3T3 and CHP 100. Glutamate-sensitizing activity was not mimicked by trilodo-L-thyronine, insulin-like growth factor-I (IGF-I), truncated IGF-I, GPE [a tripeptide (gly-pro-glu) derived from IGF-I], brain-derived neurotrophic factor (BDNF), basic fibroblast growth factor or tumour necrosis factor-alpha. However, IGF-I added to cultures of granule cells plated at high density and grown in basal medium Eagle's without serum or any other constituent of chemically defined media was capable of supporting production of glutamate-sensitizing activity to an extent similar to that shown by whole fetal calf serum. Under the same conditions triiodo-L-thyronine and BDNF did not support the production of glutamate-sensitizing activity. Glutamate-sensitizing activity was not mimicked by glutamate, NMDA, glycine or lactate, and was not inhibited by glucose, haemoglobin or N-omega-nitro-L-arginine methyl ester. At variance with the response of granule cells, the response to glutamate of GABAergic cells present in the same culture was not affected by cell density or by glutamate-sensitizing activity.     

In vivo purging with high-dose cytarabine followed by high-dose chemoradiotherapy and reinfusion of unpurged bone marrow for adult acute myelogenous leukemia in first complete remission

PURPOSE: To evaluate in a prospective study the efficacy of autologous bone marrow transplantation (BMT) in adult patients with acute myelogenous leukemia (AML) in first remission, using a single course of high-dose Cytarabine (HD Ara-C) consolidation therapy as in vivo purging. PATIENTS AND METHODS: Sixty consecutive adult patients with AML in first complete remission (CR) were treated with HD Ara-C consolidation therapy as a method of in vivo purging before marrow collection. High-dose therapy consisted of fractionated total-body irradiation (FTBI) 12 Gy, intravenous etoposide 60 mg/kg, and cyclophosphamide 75 mg/kg, followed by reinfusion of cryopreserved marrow. RESULTS: Sixty patients underwent consolidation treatment with HD Ara-C with the intent to treat with autologous BMT. Sixteen patients were unable to proceed to autologous BMT (10 patients relapsed, one died of sepsis, one developed cerebellar toxicity, two had inadequate blood counts, and two refused). Forty-four patients underwent autologous BMT and have a median follow-up time of 37 months (range, 14.7 to 68.7) for patients who are alive with no relapse. The cumulative probability of disease-free survival (DFS) at 24 months in the intent-to-treat group is 49% (95% confidence interval [CI], 37% to 62%) and in those who actually underwent autologous BMT is 61% (95% CI, 46% to 74%). The probability of relapse was 44% (95% CI, 31% to 58%) and 33% (95% CI, 20% to 49%) for the intent-to-treat and autologous BMT patients, respectively. CONCLUSION: This approach offers a relatively high DFS rate to adult patients with AML in first CR. The results of this study are similar to those achieved with allogeneic BMT.     

A panoramic view and swot analysis of artificial intelligence for achieving the sustainable development goals by 2030: progress and prospects

Applied Intelligence - The17 Sustainable Development Goals (SDGs) established by the United Nations Agenda 2030 constitute a global blueprint agenda and instrument for peace and prosperity...     

Eicosapentaenoic acid (20∶5n-3) from the marine microalgaPhaeodactylum tricornutum

Eicosapentaenoic acid (EPA, 20∶5n-3) was obtained from the marine microalgaePhaeodactylum tricornutum by a three-step process: fatty acid extraction by direct saponification of biomass, polyunsaturated fatty acid (PUFA) concentration by formation of urea inclusion compounds, and EPA isolation by semipreparative high-performance liquid chromatography (HPLC). Alternatively, EPA was obtained by a similar two-step process without the PUFA concentration step by the urea method. Direct saponification of biomass was carried out with two solvents that contained KOH for lipid saponification. An increase in yield was obtained because the problems associated with emulsion formation were avoided by separating the biomass from the soap solution before adding hexane for extraction of insaponifiables. The most efficient solvent, ethanol (96%) at 60°C for 1 h, extracted 98.3% of EPA. PUFA were concentrated by the urea method with a urea/fatty acid ratio of 4∶1 at a crystallization temperature of 28°C and by using methanol and ethanol as urea solvents. An EPA concentration ratio of 1.73 (55.2∶31.9) and a recover yield of 78.6% were obtained with methanol as the urea solvent. This PUFA concentrate was used to obtain 93.4% pure EPA by semipreparative HPLC with a reverse-phase, C₁₈, 10 mm i.d.×25-cm column and methanol/water (1% acetic acid), 80∶20 w/w, as the mobile phase. Eighty-five percent of EPA loaded was recovered, and 65.7% of EPA present inP. tricornutum biomass was recovered in highly pure form by this three-step downstream process. Alternatively, 93.6% pure EPA was isolated from the fatty acid extract (without the PUFA concentration step) with 100% EPA recovery yield. This two-step process increases the overall EPA yield to 98.3%, but it is only possible to obtain 20% as much EPA as that obtained by three-step downstream processing.     

Effect of low‐temperature plasma and chitosan treatment on wool dyeing with Acid Red 27

This study examines in detail the influence of low‐temperature plasma and biopolymer chitosan treatments on wool dyeability. Wool knitted fabrics were treated and characterized by whiteness and shrink‐resistance measurements. Surface modification was assessed by contact‐angle measurements of human hair fibers, which were used as a model to study the wetting properties of the treated wool knitted fabrics. The dyeing behavior was assessed from the diffusion mechanism point of view. The dyeing kinetics were measured at two different pHs (4.2 and 6.5) and three different temperatures (60, 85, and 100°C) to gain information about the contribution of the surface modification treatment to the dyeing mechanism. The exhaustion and reflectance data were compared, and the apparent diffusion coefficients were calculated. On the basis of the obtained results, a model for the dyeing mechanism of the chitosan treated wool was proposed. When treated with chitosan, the polymer sheath spread on the surface of the fibers acted as a predominant dyeing site in very short dyeing times, thus interacting with the dye and in later stages imparting the dye to the wool fiber. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 2204–2214, 2005     

I3WSN: Industrial Intelligent Wireless Sensor Networks for indoor environments

Sensor Web Enablement (SWE) technologies have been successfully applied to a great variety of outdoor scenarios but, in practical terms, little effort has been applied for indoor environments, and even less in the field of industrial applications. This article presents an intelligent SWE application for an indoor and industrial scenario, with the aim of improving and increasing the levels of human safety. The base low-level architecture is built on top of wireless sensor networks (WSN) connected to a Sensor Observation Service (SOS). Higher layers in the architecture include services that make real time decisions based on the collected data. Both simulation and experimental results are presented. The paper shows the viability of our approach in terms of performance, scalability, modularity and safety.     

Iron species incorporated over different silica supports for the heterogeneous photo-Fenton oxidation of phenol

Iron-containing catalysts have been prepared following different synthesis routes and silica supports (amorphous, zeolitic and mesostructured materials). Activity and stability of these materials were assessed on the photo-Fenton degradation of phenolic aqueous solutions using near UV irradiation (higher than 313 nm) at room temperature and initial neutral pH. Their catalytic performance was monitored in terms of phenol and total organic carbon (TOC) conversions. Aromatic compounds and carboxylic acids as by-products coming from incomplete mineralization of phenol as well as the efficiency of each catalytic system in the use of the oxidant were also studied. Stability of the materials throughout the photo-Fenton reaction was evaluated in terms of metal leachibility. Activity and stability depend on the environment of iron species and features of silica support. The evolution of pH with the reaction time and their relationship with TOC degradation and leaching degree has been discussed. A nanocomposite material of crystalline iron oxides supported over mesostructured SBA-15 material is shown the most successful catalyst for degradation of phenolic aqueous solutions by photo-Fenton processes, achieving an outstanding overall catalytic performance accompanied with a noteworthy stability.     

Semiquantitative analysis of thiophenic compounds in light cycle oil (LCO) using C NMR spectroscopy

S-methyltetrafluoroborate salts of the thiophenic compounds (CH₃-S⁺:BF₄⁻) present in LCO petroleum fractions were obtained and analyzed by ¹H and ¹³C NMR spectroscopy. The methylation of the samples was carried out using 99.5% ¹³C enriched methyl iodine, to improve the sensitivity of the technique. The amount of the methylated derivatives was determined by the internal standard method; using dioxane as a reference, 37 sulphur compounds were detected. Among them, benzo[b]thiophene, dibenzo[b,d]thiophene, and several isomers of methyl, dimethyl and trimethyl[b]benzothiophenes were the most abundant. With this research, it was demonstrated that NMR spectroscopy can be used to analyze thiophenic compounds from petroleum medium fractions.