Fluorescence in situ hybridization (FISH): DNA probe production and hybridization criteria

We describe methods for the production of fluorescence in situ hybridization (FISH) probes and the utilization of these probes for the detection of complementary DNA sequences with accuracy and sensitivity for application in both basic research and clinical diagnosis. Due to the frequent use of FISH in many laboratories, it is important to apply the most convenient and reproducible approach. This review describes some of the most recent techniques, and includes versatile, effective and simple methods of probe production and fluorescence in situ hybridization. We also describe methods for the production of region-specific and chromosome-specific DNA probes and hybridization techniques for the visualization of these probes.     

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.     

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.     

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.     

An intelligent hybrid approach for hepatitis disease diagnosis: Combining enhanced k‐means clustering and improved ensemble learning

In real world, the automatic detection of liver disease is a challenging problem among medical practitioners. The intent of this work is to propose an intelligent hybrid approach for the diagnosis of hepatitis disease. The diagnosis is performed with the combination of k‐means clustering and improved ensemble‐driven learning. To avoid clinical experience and to reduce the evaluation time, ensemble learning is deployed, which constructs a set of hypotheses by using multiple learners to solve a liver disease problem. The performance analysis of the proposed integrated hybrid system is compared in terms of accuracy, true positive rate, precision, f‐measure, kappa statistic, mean absolute error, and root mean squared error. Simulation results showed that the enhanced k‐means clustering and improved ensemble learning with enhanced adaptive boosting, bagged decision tree, and J48 decision tree‐based intelligent hybrid approach achieved better prediction outcomes than other existing individual and integrated methods.     

Investigation of multiple heterogeneous relationships using a q-rung orthopair fuzzy multi-criteria decision algorithm

Neural Computing and Applications - Q-rung orthopair fuzzy (q-ROF) set is one of the powerful tools for handling the uncertain multi-criteria decision-making (MCDM) problems, various MCDM methods...     

I-convergence in probabilistic n-normed space

In this article we introduce the notion of I-Cauchy sequence and I-convergent sequence in probabilistic n-normed space. The concept of I*-Cauchy sequence and I*-convergence in probabilistic n-normed space are also introduced and some of their properties related to these notions have been established.     

Use of AgInTe2 for optoelectronic switching at higher frequencies

A possible application of AgInTe₂ for optoelectronic switching is reported. Alternating and direct current (a.c. and d.c.) response of the material has been studied over a wide frequency range (5 Hz to 1 MHz) through measurements of conductance, capacitance and photocurrent at different temperature and illumination levels. The switching/recovery time has been analysed in terms of time constant and found to be of the order of microseconds for this material. Higher frequencies of operation (>40 kHz) are preferable because, at such frequencies, becomes almost independent of illumination level and temperature. Furthermore, the switching speed can be conveniently controlled with d.c. bias applied across the sample. Elliott's CBH model has been satisfactorily used to explain the switching behaviour for AgInTe₂.     

Correlation of burst pressure of GRP cylinders with the stress wave factor

Glass fibre-reinforced plastic (GRP) cylinders are increasingly used for highly stressed structural elements. The higher the demands on the materials, the higher are the fault detection requirements to be met by non-destructive materials testing methods. Acousto-ultrasonics is a valuable aid for the non-destructive evaluation of GRP composite materials, because it may be the answer to evaluating effects of subtle defects in composites. The aim of the research is to evaluate the burst pressure of GRP cylinders by acousto-ultrasonics techniques. The theoretical results have been found to be in good agreement with the experimental values. Hence the results strongly suggest that stress wave factor measurements can be exploited successfully to predict burst pressure of GRP cylinders.Nomenclature P Internal pressure, kgf cm^–2 - d Internal diameter, cm - t Thickness of cylinder, cm - (N ,N ,N ) Resultant forces, kgf - (M ,M ,M ) Moments, kg cm - [A] Extensional stiffness matrix - [B] Bending stretching coupling matrix - [D] Flexural stiffness matrix - ( ⁰ , ⁰ , ⁰ ) Midplane strains - (k ,k ,k ) Curvatures - n Number of laminae - Z Distance from midplane, cm - _u Ultimate tensile strength of GRP composite, kg cm^–2 - S _W Stress wave factor - m Material parameter - Filament winding angle     

Effect of laser irradiation on the electrical properties of amorphous germanium films

Amorphous films of germanium were grown using a vacuum evaporation technique, on glass substrates kept at room temperature. As-grown films were irradiated with Q-Switched Nd-YAG laser pulses (=1.06 m, 20nsec, 10 to 50Jcm^–2). The d.c. conductivity measurements were made in the temperature range 77 to 300 K. It was observed that the effect of laser irradiation was similar to the effect caused by the thermal annealing of the films. The d.c. conductivity data were analysed in the light of Mott's theory of a variable range hopping conduction process.