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Biomedical subjects

M Amado

Publications and source records attributed to M Amado.

9 recordsLinked to original sources

Experimental phytophotodermatitis.

BACKGROUND/AIMS: Phytophotodermatitis (PPD) is defined as a phototoxic reaction of the skin after contact with substances derived from plants and subsequent exposure to sunlight. It is a frequent disease in our outpatient clinics during summer because of contact with Tahitian lemon. Our objectives were to experimentally reproduce PPD in rats, to identify whether PPD is induced by minimal exposure periods to sunlight, to find what kinds of lemons and which parts of the lemon (the fruit juice or the peel juice) may trigger the disease; to know whether the use of sunblock prevents the reaction; and to perform light microscopy of the lesions to describe their histology. METHODS: Adult rats (Rattus norwegicus), three in each experiment, were used. After painting the rats with the fruit juice or the peel juice they were exposed to sunlight for 2.5, 5, 7.5, and 10 min. Tahitian and Sicilian lemons were used in the experiments. Biopsies with 3-mm punches of different times of exposure were performed. RESULTS: The peel juice of both lemons reproduced PPD, which was clinically evident after 48 h. When the peel juice was alone applied there was no reaction; moreover, exposure to sunlight alone triggered no reaction. Two and a half minutes of exposure time was sufficient to induce phototoxic reaction, which was time dependent (the longer the exposure the more intense the reaction). Histopathological studies showed epithelial time-dependent vacuolar degeneration. The use of sunblock diminished the intensity of the reaction but did not prevent it. CONCLUSION: PPD can be reproduced in an animal model. It may be caused by the peel juice of Tahitian and Sicilian lemon. Because of an extremely short time of exposure (2.5 min) is sufficient to induce PPD it is necessary to alert the population, of the need for caution when handling lemons, especially outdoors despite using sunblock.

Animals↗

Characterization of the histo-blood group O(2) gene and its protein product.

BACKGROUND AND OBJECTIVES: This study aimed to show the full sequence and function of the O(2) allele, and investigate whether it accounts for the incompatible expression of A antigens in gastric carcinomas of blood group O persons. MATERIALS AND METHODS: By PCR, we determined the ABO genotype of group O subjects (76 gastric carcinoma patients and 165 blood donors). Two expression constructs, encoding either the putative soluble or full-length O(2) protein, were used to transfect Sf9 cells. The expression and the activity of the O(2) protein were analysed by immunohistochemistry and enzymatic assays, respectively. RESULTS: No significant difference was detectable between the O(2) allele frequency in gastric carcinoma patients (3.9%) and blood donors (4.2%). Sequencing analysis of the O(2) allele revealed an intact reading frame identical to that of A transferase except for four nucleotide substitutions. O(2)-transfected Sf9 cells and gastric carcinomas genotyped as O(1)O(2) both expressed a protein recognized by anti-A/B transferase monoclonal antibodies. In enzymatic assays, the O(2) protein failed to show measurable A transferase activity. CONCLUSION: The O(2) allele has an intact reading frame encoding a protein immunologically related to A/B transferases and enzymatically inactive. Further, our data gave no indication that the O(2) allele is related to the phenomenon of incompatible A antigen expression in gastric cancer.

ABO Blood-Group System↗

Identification and characterization of large galactosyltransferase gene families: galactosyltransferases for all functions.

Enzymatic glycosylation of proteins and lipids is an abundant and important biological process. A great diversity of oligosaccharide structures and types of glycoconjugates is found in nature, and these are synthesized by a large number of glycosyltransferases. Glycosyltransferases have high donor and acceptor substrate specificities and are in general limited to catalysis of one unique glycosidic linkage. Emerging evidence indicates that formation of many glycosidic linkages is covered by large homologous glycosyltransferase gene families, and that the existence of multiple enzyme isoforms provides a degree of redundancy as well as a higher level of regulation of the glycoforms synthesized. Here, we discuss recent cloning strategies enabling the identification of these large glycosyltransferase gene families and exemplify the implication this has for our understanding of regulation of glycosylation by discussing two galactosyltransferase gene families.

Animals↗

A family of human beta3-galactosyltransferases. Characterization of four members of a UDP-galactose:beta-N-acetyl-glucosamine/beta-nacetyl-galactosamine beta-1,3-galactosyltransferase family.

BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of a human UDP-galactose:beta-N-acetyl-glucosamine beta-1, 3-galactosyltransferase, designated beta3Gal-T1, revealed no ESTs with identical sequences but a large number with similarity. Three different sets of overlapping ESTs with sequence similarities to beta3Gal-T1 were compiled, and complete coding regions of these genes were obtained. Expression of two of these genes in the Baculo virus system showed that one represented a UDP-galactose:beta-N-acetyl-glucosamine beta-1, 3-galactosyltransferase (beta3Gal-T2) with similar kinetic properties as beta3Gal-T1. Another gene represented a UDP-galactose:beta-N-acetyl-galactosamine beta-1, 3-galactosyltransferase (beta3Gal-T4) involved in GM1/GD1 ganglioside synthesis, and this gene was highly similar to a recently reported rat GD1 synthase (Miyazaki, H., Fukumoto, S., Okada, M., Hasegawa, T., and Furukawa, K. (1997) J. Biol. Chem. 272, 24794-24799). Northern analysis of mRNA from human organs with the four homologous cDNA revealed different expression patterns. beta3Gal-T1 mRNA was expressed in brain, beta3Gal-T2 was expressed in brain and heart, and beta3Gal-T3 and -T4 were more widely expressed. The coding regions for each of the four genes were contained in single exons. beta3Gal-T2, -T3, and -T4 were localized to 1q31, 3q25, and 6p21.3, respectively, by EST mapping. The results demonstrate the existence of a family of homologous beta3-galactosyltransferase genes.

Amino Acid Sequence↗

Dimeric sialyl-Le(x) expression in gastric carcinoma correlates with venous invasion and poor outcome.

BACKGROUND & AIMS: High expression of sialyl-Le(x) in tumors of different organs correlates with hematogenous metastasis and adverse outcome. Dimeric sialyl-Le(x) expression in gastric carcinoma was evaluated, and its prognostic significance within this setting was determined. METHODS: Dimeric sialyl-Le(x) immunohistochemical expression in 97 gastric carcinomas was analyzed using the FH6 monoclonal antibody. Scoring was based on the percentage of immunoreactive cells: negative, low expression (< or = 25%), and high expression (> 25%). RESULTS: Immunoreactivity was observed in 45 cases (46.4%), encompassing 27 and 18 cases with low and high expression, respectively. Significant relationships were found between dimeric sialyl-Le(x) expression and venous invasion (P = 0.0025) and histological classification (P = 0.05). No correlation was observed with other clinicopathologic features. Patients with tumors showing high expression of dimeric sialyl-Le(x) had a significantly shorter survival time than those with low or no expression (P = 0.03). By multivariate analysis, pathological TNM (pTNM) staging and venous invasion emerged as independent prognostic factors in the whole series. Within the group of patients with tumors in pTNM stages II and III, dimeric sialyl-Le(x) was the only independent prognostic factor. CONCLUSIONS: High expression of dimeric sialyl-Le(x) correlates with venous invasion and poor outcome in gastric carcinoma.

Adult↗

A family of human beta4-galactosyltransferases. Cloning and expression of two novel UDP-galactose:beta-n-acetylglucosamine beta1, 4-galactosyltransferases, beta4Gal-T2 and beta4Gal-T3.

BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of the human UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferase, designated beta4Gal-T1, revealed a large number of ESTs with identical as well as similar sequences. ESTs with sequences similar to that of beta4Gal-T1 could be grouped into at least two non-identical sequence sets. Analysis of the predicted amino acid sequence of the novel ESTs with beta4Gal-T1 revealed conservation of short sequence motifs as well as cysteine residues previously shown to be important for the function of beta4Gal-T1. The likelihood that the identified ESTs represented novel galactosyltransferase genes was tested by cloning and sequencing of the full coding region of two distinct genes, followed by expression. Expression of soluble secreted constructs in the baculovirus system showed that these genes represented genuine UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferases, thus designated beta4Gal-T2 and beta4Gal-T3. Genomic cloning of the genes revealed that they have identical genomic organizations compared with beta4Gal-T1. The two novel genes were located on 1p32-33 and 1q23. The results demonstrate the existence of a family of homologous galactosyltransferases with related functions. The existence of multiple beta4-galactosyltransferases with the same or overlapping functions may be relevant for interpretation of biological functions previously assigned to beta4Gal-T1.

Amino Acid Sequence↗

Continuous nebulization for status asthmaticus.

Continuous nebulization of beta 2 agonists is now recognized as a useful treatment for severe exacerbations of asthma. This mode of administration has been described both for adults and children in the emergency room and in the intensive care unit. It has been suggested that early use of continuous inhalation therapy may reduce or prevent the need for intensive care unit admissions and potentially toxic treatments such as intravenous beta agonists and mechanical ventilation. A number of methods have been proposed for continuous administration of beta agonists, however, there is no concensus as to the best one. The lack of controlled studies clearly demonstrating superior outcomes in patients who receive this treatment leaves many questions unanswered. In addition, differences in efficacy and safety between frequent and continuous nebulization, if any, are unclear. Adverse effects primarily consist of muscle cramps, hypokalemia, and hyperglycemia. It is suggested that use of this treatment should be considered for the asthmatic patient who demonstrates progression towards significant obstruction before institution of the more toxic treatments listed above. While continuous nebulization has not been proven to prevent the need for them, it is clearly safer. Should population studies eventually fail to demonstrate its superior efficacy, continuously nebulized beta 2 agonist therapy may still be beneficial for individual patients.

Adolescent↗