Biomedical subjects

M Skliar

Publications and source records attributed to M Skliar.

5 recordsLinked to original sources

PubMed2001

Effect of culture conditions on the synthesis of vitamin D(3) metabolites in Solanum glaucophyllum grown in vitro.

In cultured Solanum glaucophyllum we have recently described the operation of a light-independent pathway of 1alpha,25-dihydroxy-vitamin D(3) (1alpha,25(OH)(2)D(3)) biosynthesis which involves similar intermediates as in vertebrates. In this work we investigated factors influencing the formation of 1alpha,25(OH)(2)D(3) and related sterols in S. glaucophyllum grown in vitro in darkness. Callus tissue and cells cultured in Murashige and Skoog medium in the absence of light were employed. Lipids were extracted with chloroform-methanol. The remaining water soluble fraction was incubated with beta-glucosidase to release vitamin D(3) compounds from their glycoconjugated derivatives followed by organic solvent extraction. Vitamin D(3) derivatives were isolated by Sephadex LH-20 and high-performance liquid chromatography (HPLC). HPLC or competitive protein binding assays with intestine 1alpha,25(OH)(2)D(3) receptor and serum vitamin D binding protein were used to quantify the metabolites. The levels of 1alpha,25(OH)(2)D(3) in calli varied according to the tissue explant origin, e.g. stem>leaf>fruit. For all organs, the metabolite was mainly present as free sterol (>80% of total). There were larger amounts of 25(OH)D(3) than 1alpha,25(OH)(2)D(3). It was found that Ca(2+), auxin and kinetin are important factors upregulating 1alpha,25(OH)(2)D(3) synthesis in S. glaucophyllum tissue and cells. Irradiation with UV light increased vitamin D(3) but not 1alpha,25(OH)(2)D(3) levels. In agreement with these results, incubation of cells with [3H]25(OH)D(3) revealed a low conversion rate to [3H]1alpha,25(OH)(2)D(3). The operation of a light-dependent pathway formation of vitamin D(3) coupled to higher expression of 25(OH)D(3)-1alpha-hydroxylase may account for the large concentrations of 1alpha,25(OH)(2)D(3) normally found in differentiated field-grown plants.

Calcifediol↗

PubMed2000

Nicotiana glauca: another plant species containing vitamin D(3) metabolites.

Vitamin D(3)-related compounds have been detected in various plant species, mostly belonging to the Solanaceae. In this work we show that Nicotiana glauca, a widespread member of this taxonomic family, contains 7-dehydrocholesterol, vitamin D(3) and hydroxylated derivatives bearing precursor-product metabolic relationships in vertebrates. Leaves collected in the field and callus cultures were used. By means of specific radioreceptor binding assays and mass spectrometry of purified fractions obtained from plant tissue lipid extracts by Sephadex LH-20 and Sep-Pak C18 chromatography followed by HPLC, we established the presence of 7-dehydrocholesterol, vitamin D(3), 25(OH)-vitamin D(3) and 1alpha,25(OH)(2)-vitamin D(3) (1alpha,25(OH)(2)D(3)), the latter being a hormonally relevant metabolite in animals. These results indicate that N. glauca may represent a useful species in which to characterize the biosynthetic pathway and physiological functions of vitamin D(3) compounds in plants. In addition, tissue culture of N. glauca could become a significant tool for biotechnological production of 1alpha,25(OH)(2)D(3).

Journal Article↗

PubMed1998

Identification of 7-dehydrocholesterol, vitamin D3, 25(OH)-vitamin D3 and 1,25(OH)2-vitamin D3 in Solanum glaucophyllum cultures grown in absence of light.

Solanum glaucophyllum contains the calciotropic hormone 1, 25-dihydroxy-vitamin D3 (1,25(OH)2D3). The metabolic pathway leading to the formation of 1,25(OH)2D3 in the plant is largely unknown. Specifically, there is controversy about the participation of a photolytic reaction in the generation of vitamin D3 and its metabolites. To investigate the requirement for light, S. glaucophyllum tissue (callus) and cell suspension cultures grown under strict conditions of darkness were extracted with chloroform/methanol (1:2, v/v) followed by purification of the lipidic fraction by Sephadex LH-20 and high-performance liquid chromatography. HPLC peaks with elution times similar to those of authentic samples of 7-dehydrocholesterol, vitamin D3, 25(OH)D3 and 1,25(OH)2D3 were detected. The presence of 1,25(OH)2D3 was also evidenced by [3H]1,25(OH)2D3 competitive binding analysis using the chick hormone intestinal receptor. Furthermore, 7-dehydrocholesterol, vitamin D3, 25(OH)D3 and 1,25(OH)2D3 were unequivocally identified by mass spectrometry. Incubation of control samples of 7-dehydrocholesterol under the same conditions as S. glaucophyllum cultures did not result in vitamin D3 formation, excluding the influence of light in these experiments. The results suggest that a synthetic route of vitamin D3 compounds independent of light operates in Solanum glaucophyllum cultured in vitro.

Argentina↗

PubMed1997

Implicit Kalman filtering.

For an implicitly defined discrete system, a new algorithm for Kalman filtering is developed and an efficient numerical implementation scheme is proposed. Unlike the traditional explicit approach, the implicit filter can be readily applied to ill-conditioned systems and allows for generalization to descriptor systems. The implementation of the implicit filter depends on the solution of the congruence matrix equation (A1)(Px)(AT1) = Py. We develop a general iterative method for the solution of this equation, and prove necessary and sufficient conditions for convergence. It is shown that when the system matrices of an implicit system are sparse, the implicit Kalman filter requires significantly less computer time and storage to implement as compared to the traditional explicit Kalman filter. Simulation results are presented to illustrate and substantiate the theoretical developments.

Algorithms↗

PubMedDate not supplied

Air-quality monitoring and detection of air contamination in an enclosed environment.

We report on the development of an air-quality monitoring and early detection system for an enclosed environment with specific emphasis on manned spacecraft. The proposed monitoring approach is based on a distributed parameter model of contaminant dispersion and real-time contaminant concentration measurements. Kalman filtering is identified as a suitable method for generating on-line estimation of the spatial contamination profile, and an implicit Kalman filtering algorithm is shown to be preferable for rear-time implementation. The identification of the contaminant concentration profile allows for a straightforward solution of the early detection of an air contamination event and provides information that enables potential automatic diagnosis of an unknown contamination source.

Air Pollutants↗