Biomedical subjects

P Reschiglian

Publications and source records attributed to P Reschiglian.

10 recordsLinked to original sources

PubMed2002

Improved performance of gravitational field-flow fractionation for screening wine-making yeast varieties.

Performance of gravitational field-flow fractionation (GFFF) is improved here with respect to the ability to fractionate and distinguish different varieties of wine-making yeast from Saccharomyces cerevisiae. A new GFFF channel with non-polar walls has been employed to enhance fractionation selectivity and reproducibility. Since GFFF retention depends from first principles on particle size, Coulter counter measurements were performed in order to compare size distribution profiles with GFFF profiles. From such a comparison, GFFF was shown to be able to reveal differences in yeast cells other than size. This could make use of GFFF for screening different varieties of wine-making yeast towards future quality assessment procedures based on a possible correlation between yeast cell morphology indexes and quality indexes.

Fractionation, Field Flow↗

PubMed2001

Gravitational field-flow fractionation for the characterisation of active dry wine yeast.

Gravitational field-flow fractionation (GrFFF) is applied to the fractionation of active dry wine yeast. An experimental approach to the analysis of the effects that field variation by changing mobile phase composition and flow-rate have on the fractionation process of standard particles (polystyrene) was first developed to further obtain effective fractionation of wine yeast by GrFFF. Scanning electron microscopy and Coulter counter particle size measurements were used to monitor the fractionation extent and capabilities of GrFFF to describe the distribution of yeast cells populations.

Chemical Fractionation↗

PubMed2000

Working without accumulation membrane in flow field-flow fractionation.

Nonideal interaction of sample with the separation device is a difficulty found in chromatographic methods as well as in field-flow fractionation. However, in field-flow fractionation (FFF), greater flexibility in the choice of carrier solution composition is possible, thus reducing the need of a wide choice of surface chemistry when nonideal sample interaction is to be minimized. The use of an ultrafiltration membrane as the surface for the accumulation wall is common practice in flow field-flow fractionation. Typical membranes in use are laminates of a skin membrane onto a backing material such as woven polyester. At this point, only a limited choice of membrane chemistries is available. Many membranes have been developed for protein applications as membranes are widely used in the pharmaceutical industries. While these membranes work well for protein applications, flow field-flow fractionation is applicable to polymeric particulate as well as protein samples. Thus, sample interaction with the membrane surface is possible with nonprotein applications and these interactions can induce significant secondary effects on retention ratio and affect instrumental reliability. Also, the woven texture of membranes may detrimentally affect the FFF separation. For these reasons, the study of flow field-flow fractionation using a flat, smooth surface of controlled chemistry is of relevance. We present here the results of a new, membraneless channel that uses a bare frit as the accumulation wall and that is intended for analysis of micrometer-sized particles only. Selectivity results are comparable to those obtained with the membrane, while relative sample recovery indicates that the best quantitative performance can be obtained without the membrane. Moreover, neither sample immobilization nor losses through the frit occur when operating membraneless. On the other hand, first experimental evidence of a certain level of frit surface activity suggests that optimization of experimental conditions is required.

Journal Article↗

PubMed2000

Continuous split-flow thin cell and gravitational field-flow fractionation of wheat starch particles.

The combined employment of the SPLITT (split-flow thin) cell--a relatively new system for fast, continuous binary separation--and of gravitational field-flow fractionation (GrFFF)--a fractionation technique suitable for micron particle size distribution determination--was investigated for starch separation and characterization. Emphasis is placed on the main advantages of both techniques: operating under gentle earth gravity field, low cost and ease of maintenance. The reproducibility of GrFFF is demonstrated. Both the SPLITT separation and GrFFF fractionation results were checked by optical microscopy. Application examples of typical starch fractionation experiments are reported and discussed.

Chemical Fractionation↗

PubMed2000

Quantitative analysis in field-flow fractionation using ultraviolet-visible detectors: an experimental design for absolute measurements

In previous works, it has been shown that a standard ultraviolet-visible detection system can be used for quantitative analysis of heterogeneous systems (dispersed supermicron particles) in field-flow fractionation (FFF) by single peak area measurements. Such an analysis method was shown to require either experimental measurements (standardless analysis) or an accurate model (absolute analysis) to determine the extinction efficiency of the particulate samples. In this work, an experimental design to assess absolute analysis in FFF through prediction of particles' optical extinction is presented. Prediction derives from the semiempirical approach by van de Hulst and Walstra. Special emphasis is given to the restriction of the experimental domain of instrumental conditions within which absolute analysis is allowed. Validation by statistical analysis and a practical application to real sample recovery studies are also given.

Journal Article↗

PubMed1997

A new fast and standardless method for direct determination of metals associated with particulate matter in air: avoiding errors in the determination of Pb in an urban environment.

We describe a new method and the relevant instrumentation necessary for its implementation in the analysis of metals associated with particulate matter in air. The procedure can be divided into two steps: in the first step the sample is accumulated in a device through electrostatic precipitation whose center is a graphite tube; in the second step the graphite tube itself is used as an atomization device for the determination of the metals present in the sample through the electrothermal atomic absorption technique. The method is simple, fast, accurate, and inexpensive. Moreover, if the experimental conditions are well chosen, there is no need for calibration, which is very convenient in the case of samples such as particulate matter in the air. The elements that can be determined with the present apparatus are Hg, Cd, Tl, Ag, Mg, and Mn. These are highly or medium volatile because the materials used cannot reach very high temperatures for long periods. The experiments are confined to air, but other gases, in which a corona discharge is possible, would give the same results. With the method proposed, it was possible to show that the official method for Pb determination in the urban environment of Bologna presents a negative systematic error of about 25%.

Lead↗

PubMed1995

Simulation of fractograms of fat emulsions in power-programmed sedimentation field-flow fractionation (SdFFF).

A quasi-empirical approach to the simulation of fractograms was examined to verify that the elution behavior of emulsions in power-based field programmed sedimentation field-flow fractionation (SdFFF) is consistent and predictable. The approach was applied to Intralipid, a commercial soybean emulsion and to an investigational medium chain triglyceride emulsion. The simulations predicted the fractograms that were obtained under various conditions of field strength, field decay and velocity of the suspending fluid, using distribution parameters obtained from one preliminary measurement of size distribution profile. Predicted fractograms were compared to experimental ones, under various fractionating powers. Good agreement was observed in most cases, in which interference of the secondary relaxation effects was not effective. The agreement confirmed the applicability of the approach to emulsions and that the simulations can be used instead of actual experiments for the optimization of their characterization by power-programmed SdFFF.

Chemical Fractionation↗

PubMed1988

Expression of hepatitis B surface antigen in human cells by a recombinant BK virus DNA vector.

The construction of the first stable human cell lines that express and secrete authentic hepatitis B virus surface antigen (HBsAg), using a BK virus (BKV) episomal plasmid vector, is described. The amount of HBsAg produced by BKV vectors (up to 600 ng/10(7) cells) was comparable to other eukaryotic vector systems. The level of HBsAg expression remained the same regardless of the orientation of the HBsAg gene, substitution of the HBsAg gene promoter with the mouse metallothionein I gene promoter or the tissue origin of the human cell lines used to establish stable cellular transformants. Northern blot analysis also indicated synthesis of normal HBsAg transcripts. Surprisingly, however, the vectors were maintained at far lower than expected copy number (one to five copies/cell). Reasons for this are discussed.

BK Virus↗

PubMed1987

Induction of malignant subcutaneous sarcomas in hamsters by a recombinant DNA containing BK virus early region and the activated human c-Harvey-ras oncogene.

Malignant undifferentiated sarcomas were induced in 11 of 15 (73.3%) newborn Syrian hamsters by s.c. inoculation of a recombinant DNA (pBK/c-rasA) containing BK virus (BKV) early region gene and the activated human c-Harvey-ras(c-Ha-ras) oncogene derived from T24 bladder carcinoma. The two genes inoculated independently as well as a recombinant DNA of BKV early region gene and normal human c-Ha-ras proto-oncogene were not tumorigenic. Tumor-derived cell lines propagated in culture were immortalized and had growth characteristics consistent with a fully transformed phenotype. Tumors and tumor cell lines showed tandem insertions of pBK/c-rasA in high copy number and expressed BKV- and c-Ha-ras-specific transcripts as well as BKV T-antigen and c-Ha-ras protein with a molecular weight of 21,000. We conclude that BKV DNA requires interaction with other oncogenic functions for tumorigenicity. These findings may be relevant to the role of BKV in human neoplasia, where cooperation or synergism between BKV and cellular oncogenes could occur as an aspect of the multifactorial process of carcinogenesis.

Animals↗

PubMed1986

Molecular and biological properties of BK virus-IR, a BK virus variant isolated from a human tumor.

We describe the molecular and biological properties of BK virus (BKV)-IR, a new BKV variant isolated from a human tumor of pancreatic islets. BKV-IR bears a 253-base-pair (bp) deletion and an 80-bp insertion in the early region of the genome. The deletion abolishes the expression of small-t antigen. The inserted sequences, grouped in four clusters, produce rearrangements in the first and second enhancer elements. They are bound by 12-bp direct repeats and could form a 217-base stem-loop structure suggestive of an insertion sequence. As compared with wild-type BKV, BKV-IR transformed hamster cells with a reduced efficiency and induced ependymomas in hamsters at a lower frequency and with a longer latency period. Tumors induced by BKV-IR, however, showed features of higher malignancy. The possible role of the insertion sequence-like element in transformation by BKV-IR is discussed.

Adenoma, Islet Cell↗