Adeno-associated virus 2-mediated high efficiency gene transfer into immature and mature subsets of hematopoietic progenitor cells in human umbilical cord blood

Recombinant adeno-associated virus 2 (AAV) virions were constructed containing a gene for resistance to neomycin (neoR), under the control of either the herpesvirus thymidine kinase (TK) gene promoter (vTK-Neo), or the human parvovirus B19 p6 promoter (vB19-Neo), as well as those containing an upstream erythroid cell-specific enhancer (HS-2) from the locus control region of the human beta-globin gene cluster (vHS2-TK-Neo; vHS2-B19-Neo). These recombinant virions were used to infect either low density or highly enriched populations of CD34+ cells isolated from human umbilical cord blood. In clonogenic assays initiated with cells infected with the different recombinant AAV-Neo virions, equivalent high frequency transduction of the neoR gene into slow-cycling multipotential, erythroid, and granulocyte/macrophage (GM) progenitor cells, including those with high proliferative potential, was obtained without prestimulation with growth factors, indicating that these immature and mature hematopoietic progenitor cells were susceptible to infection by the recombinant AAV virions. Successful transduction did not require and was not enhanced by prestimulation of these cell populations with cytokines. The functional activity of the transduced neo gene was evident by the development of resistance to the drug G418, a neomycin analogue. Individual high and low proliferative colony-forming unit (CFU)-GM, burst-forming unit-erythroid, and CFU-granulocyte erythroid macrophage megakaryocyte colonies from mock-infected, or the recombinant virus-infected cultures were subjected to polymerase chain reaction analysis using a neo-specific synthetic oligonucleotide primer pair. A 276-bp DNA fragment that hybridized with a neo-specific DNA probe on Southern blots was only detected in those colonies cloned from the recombinant virus-infected cells, indicating stable integration of the transduced neo gene. These studies suggest that parvovirus-based vectors may prove to be a useful alternative to the more commonly used retroviral vectors for high efficiency gene transfer into slow or noncycling primitive hematopoietic progenitor cells, without the need for growth factor stimulation, which could potentially lead to differentiation of these cells before transplantation.     

Establishment of human parotid pleomorphic adenoma cells in culture: morphological and biochemical characterization

This study reports the establishment of alpha-amylase-producing human parotid pleomorphic adenoma cell lines (2HP and 2HP1) which have been maintained in culture for over 1 yr. The procedures required preparation of cellular clumps from tumor tissue and plating them on plasma clot or precoated dishes. During the initial phase of growth they required modified MCDB-153 medium without serum. When cells showed signs of degeneration they were changed to MCDB-153 medium containing first 2% and then 10% heat inactivated fetal bovine serum. Although cells grew well in MCDB-153 containing 10% serum, the epithelial cell morphology was not distinct. Therefore, the growth and morphology of cells grown in MCDB-10% serum were compared with those in RPMI growth medium containing 10% fetal bovine serum and F12 containing 10% agammaglobulin newborn bovine serum. Although the growth of cells was a little slower in F12 medium than those in MCDB and RPMI, the epithelial cell morphology was maintained better than in other growth media. The cells of 2HP and 2HP1 produce low levels of alpha-amylase and relatively high levels of alpha-amylase mRNAs of 1176 and 702 bp and contain neurofilament-160, a neuronal-specific marker. The cells of 2HP1 are tumorigenic when tested in athymic mice, but the cells of 2HP are not. The establishment of amylase-producing human parotid adenoma cell lines of different characteristics in culture provides a new opportunity to study the mechanisms of differentiation and transformation, and regulation of alpha-amylase in these cells.     

Integrating statistical and nonstatistical audit evidence using belief functions: A case of variable sampling

The main purpose of this article is to show how one can integrate statistical and nonstatistical items of evidence in the belief function framework. First, we use the properties of consonant belief functions to define the belief that the true mean of a variable lies in a given interval when a statistical test is performed for the variable. Second, we use the above definition to determine the sample size for a statistical test when a desired level of belief is needed from the sample. Third, we determine the level of belief that the true mean lies in a given interval when a statistical test yields certain values for the sample mean and the standard deviation of the mean for the variable. Finally, we use the auditing situation to illustrate the process of integrating statistical and nonstatistical items evidence. © 1994 John Wiley & Sons, Inc.     

Hierarchical Fuzzy Systems Integrated with Particle Swarm Optimization for Daily Reference Evapotranspiration Prediction: a Novel Approach

Water Resources Management - Reference evapotranspiration (ET0) is a crucial element for deriving irrigation scheduling of major crops. Thus, precise projection of ET0 is essential for better...     

Dielectric behaviour of hot pressed AlN ceramic exposed to inorganic acid vapours

Dielectric behaviour of hot pressed AIN ceramic is studied before and after exposing the samples to inorganic acid (HCl and HNO₃) vapours with a specific aim to study the effect of these vapours on the dielectric constant () and dissipation factor (tan ). Four samples having different volume percentage of porosity (0.2 to 15%) are selected for this study. Dielectric dispersion increases after exposing the samples to the above acid vapours. Tan also increases quite appreciably; the increase being more at higher porosity. Recovery studies show that the exposure effect is reversible. The exposure time dependence of and tan indicates that these parameters show a maxima at a particular exposure time. However, no such maxima is observed in the gravimetric measurements. The increase in dielectric parameters after exposure to acid vapours is explained in terms of the ionic conduction due to the dissociation of these vapours in the presence of moisture. The porosity dependence of this effect is discussed in terms of closed and open porosity reported by other workers.     

Interaction behavior of nitrogen in liquid niobium

The kinetics and mechanism of absorption/desorption of nitrogen in liquid Nb were investigated in the temperature range of 2470 °C to 2670 °C in samples levitated in a N₂/Ar stream with various nitrogen partial pressures. The nitrogen solution reaction in liquid Nb was found to be exothermic, with the standard enthalpy and entropy of solution of −236.4 ± 23.3 kJ/mol and −-5.3 ± 8.3 J/K · mol, respectively. Above the threshold flow rate of the N₂/Ar stream, the absorption process was determined to be second order with respect to nitrogen concentration, indicating that the rate-controlling step is either the adsorption of nitrogen molecules on the liquid surface or dissociation of adsorbed nitrogen molecules into surface-adsorbed atoms. The desorption process was found to be second order as well. At lower flow rates, however, the absorption rate was found to depend on the gas-phase mass transfer rate. The rate equation for nitrogen absorption in the range of 2470 °C to 2670 °C is given by with the value ofQ calculated to be −327.2 ± 20.6 kJ/mol, while nitrogen desorption at 2670 °C follows the relation      

Data stream classification with novel class detection: a review,comparison and challenges

Knowledge and Information Systems - Developing effective and efficient data stream classifiers is challenging for the machine learning community because of the dynamic nature of data streams. As a...     

Study of automatic text summarization approaches in different languages

Artificial Intelligence Review - Nowadays we see huge amount of information is available on both, online and offline sources. For single topic we see hundreds of articles are available, containing...     

Consensus-based data replication protocol for distributed cloud

The Journal of Supercomputing - Data availability ensures efficient data accessibility by the readers anytime and from anywhere. It can be addressed by creating multiple copies of each data file...     

Correction to: EDVWDD: Event‑Driven Virtual Wheel‑based Data Dissemination for Mobile Sink‑Enabled Wireless Sensor Networks

The Journal of Supercomputing - A correction to this paper has been published: https://doi.org/10.1007/s11227-021-03790-9