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Biomedical subjects

M Castagnola

Publications and source records attributed to M Castagnola.

At least 91 records · Page 5Linked to original sources

Antigenic characterization of Penicillium camemberti and related common cheese contaminants.

Twenty-four isolates of Penicillium (including a green-spored mutant from a French Brie cheese, Penicillium camemberti) with a proposed relationship to the white cheese mold P. camemberti were investigated by immunological procedures. These penicillia, which are representative of species that have caused considerable taxonomic confusion, had common micromorphology (terverticillate penicilli with rough and smooth stipes and smooth ellipsoidal to subglobose [(3 to 5) X 2 1/2 to 4 1/2 microns] conidia); growth rates; good growth on creatine sucrose agar, cheese, and other products with a high amount of protein and lipid as a primary habitat; production (with the exception of Penicillium solitum) of cyclopiazonic acid; and the ability to grow at low temperatures and water activities. The isolates that were investigated proved to be strictly antigenically related. Absorbed antiserum of the green-spored mutant of P. camemberti showed a specific precipitin band when tested by immunodiffusion either with its homologous reference antigen or with the exoantigens obtained from different isolates. The precipitin band was not present in any P. camemberti starter culture but in many unwanted cheese contaminants. The precipitin band can be used in the purity control of P. camemberti starter culture spore preparations. Analysis of the exoantigens of all the cultures by reversed phase high-performance liquid chromatography allowed us to subdivide these penicillia into nine groups below the species level. The results indicate that P. commune Thom is the wild-type ancestor of P. camemberti.

Antigens, Bacterial↗

[Total nitrogen concentration in human liver tissue. Correlations between morphological and microanalytical data].

The degree of hepatic steatosis and percentage total nitrogen content were evaluated simultaneously in a group of obese patients and after bypass surgery for obesity. The straight line relationship between sample weight and gas chromatographic response integrals shows that samples had the same water content. The higher concentration of total nitrogen in patients with higher levels of steatosis and the inverse correlation between degree of adipose infiltration and degree of total nitrogen, when water content is the same, suggest that the total liver parenchyma nitrogen concentration closely reflects the structural protein content.

Adult↗

Identification and serotyping of Microsporum canis isolates by monoclonal antibodies.

Hybridoma cells were produced by fusing mouse myeloma cells with spleen cells from mice primed with an exoantigen of Microsporum canis. Three clones produced antibodies which were examined by the Western blot technique for their potential usefulness in the identification of M. canis isolates and differentiation of strains within the species. Based on reactions with immunological determinants, all of the M. canis isolates tested presented either species- or strain-specific domains. Monoclonal antibodies proved to be useful reagents for the identification of M. canis isolates and for the differentiation of strains within the species. A purified antigen depleted of common antigenic determinants was obtained in affinity chromatography by using monoclonal antibody.

Animals↗

Changes in Fe site structure from fetal to adult hemoglobin probed by XANES.

Iron X-ray absorption near edge structure (XANES) spectra of human fetal (F) and adult (A) deoxyhemoglobin (deoxyHb) measured at the Frascati synchrotron radiation facility reveal the different geometrical structure of the Fe-porphyrin complexes in the two proteins. By this method, having determined for the first time the variation of atomic positions in fetal and adult hemoglobin in solution (close to the 'in vivo' situation), we give further insight into the structure-function relationship in hemoglobins.

Fetal Hemoglobin↗

Use of killer toxins for computer-aided differentiation of Candida albicans strains.

Killer toxins were isolated from eight selected killer yeasts. Their activity on 100 Candida albicans isolates of human and animal origin was studied. A computer aided system for differentiating C. albicans strains was developed. By using this system, it was possible to differentiate 14 biotypes of C. albicans isolates based on their susceptibility to the killer toxins.

Candida albicans↗

Determination of the non-enzymatic glycation of hemoglobin by isoelectrofocusing of its globin chains.

Micro cation exchange chromatography determination of HbA1c does not provide a complete picture of Hb glycation, for it does not determine all the glycated forms of hemoglobin. For the determination of total glycation, we describe here a rod IEF method, which allows the simultaneous quantitation of glycation on alpha and beta globin chains. The method exhibits good sensitivity; it is not affected by artifacts deriving from temperature, hypertriglyceridemia, Hb variants or labile HbA1 (aldiminic Hb). The results obtained indicate that in a normal population approximately 18% of the beta chain and 8% of the alpha chain are glycated. These mean percentages increase in the diabetic to 28% and 12%, respectively. The beta chain is glycated on both valine and lysine residues, while the alpha chain is glycated only on the latter. HbA1 values from micro cation exchange chromatography are significantly related to both alpha and beta glycation. Thus, valinic or lysinic glycation have roughly the same clinical significance.

Diabetes Mellitus↗

[Rigid phantom dosimetry of brachytherapy sources, with special reference to 125-I].

At present, the brachytherapy dosimetry requires the highest level of accuracy in determining the physical parameters of many different radiation sources. Brachytherapy sources differ not only in the nucleus (widely ranging between 30 keV and 1 MeV), but also in the strength and geometrical and filtration characteristics. This work reports the brachytherapy dosimetry carried out in a rigid perspex phantom which allows measurements by means of solid state dosimeters (TLD, semiconductor), ionization chamber and film. Film densities were measured by means of a computerized Perkin-Elmer 1010G microdensitometer, with a spatial resolution of 50 microns. The isodensity lines were dose calibrated by TLD measurements. The dosimetry of the 125I, 192Ir and 198Au sources is reported. The dose distributions of the low energy 125I source, in two different tissues (muscle and adipose) were studied, by means of laboratory built tissue-equivalent sections of the phantoms. The experimental data suggest that this dosimetric procedure is suitable for intercomparison measurements between different radiotherapy centers using the same nuclides in different sources.

Adipose Tissue↗

Analysis of the globins from fast human haemoglobins by isoelectrofocusing on polyacrylamide gel rods.

The globins from all fast haemoglobin (Hb) components obtainable by Bio-Rex 70 cation-exchange chromatography were examined by isoelectrofocusing on polyacrylamide gel rods with 8.0 mol/l urea. From this analysis HbA1a1 and HbA1a2 seem to be very heterogeneous components. HbA1b is separable into two components, one of which is varied in both the beta chains. Between HbA1b2 and the well-known HbA1c components two chromatographic peaks are separated, one with a noticeable percentage of glucosylated beta chain and one that probably contains HbF. HbA1c has both beta chains glucosylated, while HbA1x seems to be a beta monoglucosylated Hb form. Finally, the early part of the HbAo peak has a large amount of glucosylation on both alpha and beta chains.

Chromatography, Ion Exchange↗

Investigation of the heterogeneity of hemoglobin by cation-exchange chromatography on Bio-REX 70.

The use of Bio-Rex 70 cation-exchange resin for chromatography of normal and diabetic hemoglobin provides a reproducible pattern of the "fast components". Particular attention to the choice of sample preparation, pH of elution, and the increase of ionic strength by sodium chloride linear gradients results in the separation of Hb-A1b into two components and in the isolation of a new component eluting between Hb-A1c and Hb-A0. Experiments with [3H]glucose and the colorimetric test (thiobarbituric acid) normally used to determine the extent of non-enzymatic glycosylation, as well as an increase of this component in diabetic samples compared with normoglycemic ones and a significant linear correlation with Hb-A1c, indicate that this component should be a part of the hemoglobins glucosylated on the epsilon-NH2 group of the lysines of both chains and/or the hemoglobin glucosylated on the alpha-NH2 of the valine of the alpha-chain. We propose to call this component Hb-A1x, pending confirmation of its identity. Normally Hb-A1x accounts for about 3% of Hb-A, but up to 5-7% of glucosylated hemoglobins should be confined to the early part of Hb-A0. In diabetics, the percentage of Hb-A1x rises to 4-5% and that of the other glucosylated hemoglobins increases to 12-16%.

Chromatography, Ion Exchange↗

The chromatographic separation of glycosylated haemoglobins: a comparison between macro- and micromethods.

The microchromatographic determination of "fast haemoglobins" is currently used in long-term control of diabetes: its use is made easier by commercially available kits, that are impaired by some analytical variables. For a good standardization of these kits a comparison with an accurate method of analysis is necessary. The macrochromatographic column allows the determination of each fast component with a good precision, and reveals the great complexity of human haemoglobin. Comparison of the two methods shows that the most appropriate temperature of elution for the kit is 20 degrees C and that the increase in the fraction of "fast haemoglobins" in diabetic samples is only related to the HbA1c component.

Chromatography↗

Can heterologous collagen enhance the granulation tissue growth? An experimental study.

A purified and lyophilized sponge of native bovine tendon collagen (Gelfix) was used to plug gastric, jejunal and colonic surgical holes (phi = 6 mm) on 78 white rats. In a control group (18 rats) colonic holes were plugged with autologous epiploon. The mortality rate due to generalized peritonitis, was higher (Fischer test = 4%) in the control group than in animals treated with the heterologous collagen. The healing process was evaluated by light microscopy at sequential intervals ranging from 4 to 120 days after implantation. The heterologous collagen biological activity was evaluated as follows: 1. The sponge was labelled with Tyrosine 125I or mixed with Collagen Methyl 14C methylated. Then the radioactivity of the isolated granulation tissue cells was evaluated. The electrophoresis of these lysed cells, as well as histoautoradiography were performed. 2. Indirect immunofluorescence studies employing rabbit antibodies against fibronectin were also carried out. The heterologous collagen seemed to act as a mechanical support and as a stimulus to chemotactic migration of fibroblasts. It also appeared to participate as a nutrient substitute in the metabolic activity of the granulation tissue. Finally, heterologous collagen fibers were found to be massively bound by fibronectin.

Animals↗

An experimental study on fluorocarbons as blood substitutes.

A fluorocarbon emulsion (Fluosol DA 20%) was experimented as a blood substitute. Fourty-five rats weighing 320 +/- 30 g. underwent different degrees of hemodilution, by isovolemic exanguino-transfusions. A first group (15 rats) had maximal exchange transfusion (Ht = 0,5%) with Fluosol DA 20% and was followed for 12 hours. A second group (15 rats) (control group) underwent the same procedure, but hydrolyzed gelatin 3% (Emagel) was used instead of Fluosol DA 20%. A third group (15 rats) had partial exchange-transfusion (Ht = 23 +/- 2%) with Fluosol DA 20%, and was followed for 2 months. Cerebral and cardiovascular activity, respiratory exchanges and acid-base balance as well as other basic biological parameters were evaluated in the first and second group. The initial signs of altered cerebral activity were observed with a Ht = 2%, while death occurred at Ht = 0,5% due to respiratory arrest. In the third group, a retention of fluorocarbon particles was demonstrated by gas chromatography in all the organs examined. In this group, lung and spleen lesions were also observed by light microscopy and transmission electron microscopy.

Acid-Base Equilibrium↗

Separation of reduced and oxidized glutathione by micellar electrokinetic capillary chromatography.

The separation of reduced and oxidized glutathione at an absolute sensitivity of about 100 pg by micellar electrokinetic capillary chromatography without derivatization is described. The time required for the separation is less than 10 min (the time between two following injections is about 15 min). The separation is characterized by high efficiency and good reliability. A partition mechanism is responsible for the high resolution observed. The method was utilized for the analysis of commercial preparations of glutathione and a good agreement with the expected results was obtained; the oxidation of the commercial glutathione in solution was easily analysed.

Electrophoresis↗