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Biomedical subjects

R Fischetti

Publications and source records attributed to R Fischetti.

7 recordsLinked to original sources

Comparison of different biochemical and molecular methods for the identification of Vibrio parahaemolyticus.

AIMS: Multicentre evaluation of biochemical and molecular methods for the identification of Vibrio parahaemolyticus. METHODS AND RESULTS: For the biochemical identification methods, API 20E and API 20NE and Alsina's scheme were evaluated in intra- and interlaboratory tests in order to determine the accuracy and concordance of each method. Both in intra- and interlaboratory tests, the Alsina's scheme showed the highest sensitivity (86% of correct identifications in the interlaboratory test). False-positive results were obtained by all methods (specificity was 95% for API 20E, 73% for API 20NE and 84% for Alsina's scheme) and concordance varied from 65% of API 20NE to 84% of API 20E. For the molecular identifications, polymerase chain reaction (PCR) for the detection of toxR gene, tl gene and pR72H fragment were tested on 30 strains by two laboratories. The PCR for toxR showed the highest inclusivity (96%), exclusivity (100%) and concordance (97%). CONCLUSIONS: Among the biochemical identification methods tested, the Alsina's scheme gave more reliable results; however, in order to avoid false-positive results, all the biochemical identifications should be confirmed by means of molecular methods. SIGNIFICANCE AND IMPACT OF THE STUDY: Availability of an efficient identification method of Vibrio parahaemolyticus to use in official control of fisheries products.

Bacterial Proteins↗

The BioCAT undulator beamline 18ID: a facility for biological non-crystalline diffraction and X-ray absorption spectroscopy at the Advanced Photon Source.

The 18ID undulator beamline of the Biophysics Collaborative Access Team at the Advanced Photon Source, Argonne, IL, USA, is a high-performance instrument designed for, and dedicated to, the study of partially ordered and disordered biological materials using the techniques of small-angle X-ray scattering, fiber diffraction, and X-ray absorption spectroscopy. The beamline and associated instrumentation are described in detail and examples of the representative experimental results are presented.

Academies and Institutes↗

Myofilament lattice spacing as a function of sarcomere length in isolated rat myocardium.

The Frank-Starling relationship of the heart has, as its molecular basis, an increase in the activation of myofibrils by calcium as the sarcomere length increases. It has been suggested that this phenomenon may be due to myofilaments moving closer together at longer lengths, thereby enhancing the probability of favorable acto-myosin interaction, resulting in increased calcium sensitivity. Accordingly, we have developed an apparatus so as to obtain accurate measurements of myocardial interfilament spacing (by synchrotron X-ray diffraction) as a function of sarcomere length (by video microscopy) over the working range of the heart, using skinned as well as intact rat trabeculas as model systems. In both these systems, lattice spacing decreased significantly as sarcomere length was increased. Furthermore, lattice spacing in the intact muscle was significantly smaller than that in the skinned muscle at all sarcomere lengths studied. These observations are consistent with the hypothesis that lattice spacing underlies length-dependent activation in the myocardium.

Actin Cytoskeleton↗

Structure and kinetics of the photoproduct of carboxymyoglobin at low temperatures: an X-ray absorption study.

Photolysis and recombination of carboxymyoglobin at low temperatures have been studied by a variety of methods. This paper combines optical and structural studies of carboxymyoglobin photolysis and recombination in the temperature range 4-120 K. The absorbance changes indicate ablation of the characteristic optical transitions of carboxymyoglobin and formation of a photoproduct (Mb*CO) differing from deoxymyoglobin. When the X-ray absorption changes in the 7150-7200-eV region of the X-ray absorption spectrum are used as an indicator of structural change, the photoproduct at 4 K as measured with respect to the unphotolyzed sample is 60% of that observed for the chemically produced deoxy form. Saturation of the change is obtained with repetitive flashes totaling several thousand joules of energy from a xenon flash lamp by using a thin sample (1 mm) at 4 mM concentration as measured by both optical transmission and X-ray absorption criteria. The kinetics of the reaction show the change to occur at 10 K within the resolving time currently available (2 s) in the X-ray absorption measurements. The amplitude of the light-induced change decreases to half its maximal value at 40 K and to zero at 90 K. Steady illumination suggests at least two recombination processes. Analysis of the extended X-ray absorption fine structure (EXAFS) data on Mb*CO indicates small distance changes in the first shell of Fe-N and Fe-C that can be attributed to lengthening of the pyrrole nitrogen bonds and proximal histidine motion, together with a small displacement of the CO molecule on photolysis--a form here designated Mb*CO. This structure of the germinate state, Mb*CO, may elucidate the nature of elementary steps in chemical reactions and in tunneling processes.

Electron Probe Microanalysis↗

Heterogeneity of oxygen delivery in normoxic and hypoxic states: a fluorometer study.

An on-line, real-time histogram display of heterogeneity of oxygen delivery to perfused and in situ organs is afforded by a flying-spot fluorometer that provides excitation for either oxidized flavoprotein of the mitochondrial space or reduced pyridine nucleotide of mitochondrial and cytosolic spaces. Emission from the two fluorochromes is acquired at 10(4) to 10(5) data points/s and histograms of the fluorescence intensity versus the number of occurrences of that intensity are displayed at 1--10 times per second. The histograms show alterations of the intensity and of the degree of heterogeneity of the redox states of perfused heart with model coronary occlusion, of perfused and in situ rat liver, and of rat and gerbil models of stroke. The percentage change of oxygen delivery to the intracellular space can be calculated from the areas under the histogram.

Brain↗