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Biomedical subjects

M D Alonso

Publications and source records attributed to M D Alonso.

At least 19 recordsLinked to original sources

Genistein-induced cell cycle arrest and apoptosis in a head and neck squamous cell carcinoma cell line.

Epidemiological studies have shown a lower incidence of breast, prostate, and colon cancers in Asian countries, particularly China and Japan, than in the United States. It is believed that genistein, a natural tyrosine kinase inhibitor and a metabolite of soy products, may be responsible for the protection from these cancers. Genistein was shown to inhibit cell proliferation and to induce cell cycle arrest at the G2-M phase in breast, prostate, and jurkat T cell leukemia cell lines. However, such studies have not been reported in squamous cell cancers of the head and neck. In this report, we show that genistein inhibits proliferation of a squamous cell carcinoma cell line HN4. Additionally, genistein caused cell cycle arrest at the S/G2-M phase and induced programmed cell death (apoptosis) in these cells. These effects appear to be dose and time dependent, irreversible, persisting when the cells were recultured in genistein-free medium for up to 72 hours, and specific for tumor cells, because genistein did not affect normal keratinocytes. These results suggest that if genistein shows similar results in clinical trials, it can be a potential chemopreventive/chemotherapeutic agent for cancers of the head and neck.

Anticarcinogenic Agents↗

The human intron-containing gene for glycogenin maps to chromosome 3, band q24.

Glycogenin is the autocatalytic, self-glucosylating primer for glycogen synthesis, providing the anchor on which the macromolecule is constructed. We have sequenced the cDNA coding for human muscle glycogenin and have deduced the corresponding amino acid sequence. By means of the polymerase chain reaction and fluorescence in situ hybridization, we have found the chromosomal location of the gene coding for glycogenin. This is localized to human chromosome 3, band q24.

Amino Acid Sequence↗

Occupational asthma induced by aniseed.

This report deals with clinical and immunologic studies in a butcher with work-related asthma. Both the positive methacholine inhalation test and the significant changes observed in PEFR measurements supported the diagnosis of asthma. The significant fall observed in PEFR measurements when the patient handled aniseed supported the diagnosis of occupational asthma. Skin prick tests carried out with 13 spices showed positive reactions only to aniseed extract. The patient had high levels of specific antianiseed IgE antibodies. The bronchial challenge test with an aniseed extract showed an immediate response without a late response. These findings suggest that the respiratory symptoms in our patient were induced by the inhalation of aniseed dust through an IgE-mediated immunologic mechanism of immediate hypersensitivity. This study established the diagnosis of occupational asthma from aniseed dust sensitization.

Adult↗

Catalytic activities of glycogenin additional to autocatalytic self-glucosylation.

Glycogenin is the autocatalytic, self-glucosylating protein that initiates glycogen synthesis in muscle and other tissues. We have sequenced the cDNA for rabbit muscle glycogenin and expressed and purified the protein in high yield as well as two mutant proteins in which Phe or Thr replaces Tyr-194, the site of glucosylation. While the wild-type protein can self-glucosylate, the mutants cannot, but all three utilize alternative acceptors by intermolecular glucose transfer for which the mutants have altered specificity. Tyr-194 is therefore not essential for the catalytic activity of glycogenin. All three proteins also hydrolyze UDP-glucose to glucose at rates comparable with the rate of self-glucosylation. The hydrolysis is competitive with glucose transfer to p-nitrophenyl alpha-maltoside. Self-glucosylation, glucosylation of other acceptors, and hydrolysis all appear to be catalyzed by the same active center. In the absence of peptidase inhibitors, the homogenous recombinant proteins of M(r) 37,000 break down to equally active species having M(r) 32,000. The kinetics of self-glucosylation catalyzed by the wild-type enzyme suggest that the reaction could be intermolecular rather than, as previously reported, intramolecular. The wild-type recombinant enzyme and native muscle glycogenin, which is phosphorylated, are inhibited quite differently by ATP at physiological concentration.

Amino Acid Sequence↗

New and specific nucleoside diphosphate glucose substrates for glycogenin.

Glycogenin, the autocatalytic, self-glucosylating primer for glycogen synthesis by glycogen synthase, is presumed, in vivo, to use UDP-glucose as the source of the glucose residues it adds to itself. When we tested its ability to utilize other nucleoside diphosphate glucoses, it emerged that purine nucleotides are not utilized but two pyrimidine nucleotides are used, in addition to UDP-glucose. These are CDP-glucose and TDP-glucose. CDP-glucose is utilized at 70% of the rate of UDP-glucose. While there is no evidence that CDP-glucose is a natural substrate for glycogenin, it has the advantage over UDP-glucose in that it can be used specifically to detect and assay glycogenin in the presence of glycogen synthase because CDP-glucose, unlike UDP-glucose, is not a substrate for the synthase.

Glucose↗

A new look at the biogenesis of glycogen.

The discovery of glycogenin as a self-glucosylating protein that primes glycogen synthesis has significantly increased our understanding of the structure and metabolism of this storage polysaccharide. The amount of glycogenin will influence how much glycogen the cell can store. Therefore, the production of active glycogenin primer in the cell has the potential to be the overall rate-limiting process in glycogen formation, capable of overriding the better understood hormonally controlled mechanisms of protein phosphorylation/dephosphorylation that regulate the activities of glycogen synthase and phosphorylase. There are indications that a similar covalent modification control is also being exerted on glycogenin. Glycogenin has the ability to glucosylate molecules other than itself and to hydrolyze UDPglucose. These are independent of self-glucosylation, so that glycogenin, even when it has completed its priming role and become part of the glycogen molecule, retains its catalytic potential. Another new component of glycogen metabolism has been discovered that may have even greater influence on total glycogen stores than does glycogenin. This is proglycogen, a low molecular mass (approximately 400 kDa) form of glycogen that serves as a stable intermediate on the pathways to and from depot glycogen (macroglycogen, mass 10(7) Da, in muscle). It is suggested that glycogen oscillates, according to glucose supply and energy demand, between the macroglycogen and proglycogen, but not usually the glycogenin, forms. The proportion of proglycogen to macroglycogen varies widely between liver, skeletal muscle, and heart, from 3 to 15% to 50% by weight, respectively. On a molar basis, proglycogen is greatly in excess over macroglycogen in muscle and heart, meaning that if the proglycogen in these tissues could be converted into macroglycogen, they could store much more total glycogen. Discovering the factors that regulate the balance between glycogenin, proglycogen, and macroglycogen may have important implications for the understanding and management of noninsulin-dependent diabetes and for exercise physiology.

Animals↗

Properties of carbohydrate-free recombinant glycogenin expressed in an Escherichia coli mutant lacking UDP-glucose pyrophosphorylase activity.

Glycogenin, the self-glucosylating primer for glycogen synthesis, is expressed in wild-type E. coli as a recombinant protein in an already partly glucosylated form, owing to the presence of its substrate, UDP-glucose. By using an E. coli mutant strain lacking in UDP-glucose pyrophosphorylase activity, we have succeeded in expressing carbohydrate-free glycogenin (apo-glycogenin) in good yield. When provided with UDPxylose, it autocatalytically adds 1 xylose residue. With UDP-glucose, an average of 8 glucose residues are added. However, release of the self-synthesized maltosaccharide chains with isoamylase reveals them to be a mixture. Chains as long as 11 glucose residues (maltoundecaose) are present. The ability of recombinant apo-glycogenin to self-glucosylate is further proof that a separate enzyme is not needed for the addition of the first glucose residue to Tyr-194 of the protein.

Escherichia coli↗

Tyrosine-194 of glycogenin undergoes autocatalytic glucosylation but is not essential for catalytic function and activity.

Glycogenin is the protein primer for glycogen synthesis. By autocatalytic transglucosylation from UDPglucose, it creates a malto-octaose chain attached to its Tyr-194. It has been uncertain whether the autocatalysis includes the addition of the first glucose residue to Tyr-194. We now show this to be the case. However, we also demonstrate, contrary to a claim by others, that Tyr-194 is not necessary for the catalytic function and activity of glycogenin.

Animals↗

Asthma and contact urticaria caused by rice in a housewife.

We report the case of an atopic housewife who presented with rhinoconjunctivitis-asthma and contact urticaria from handling rice and other cereals. She tolerated cooke cereals. Both skin prick tests with a rice extract (20% w/v) and a rub test with raw rice gave positive results. Bronchial challenge test with methacholine revealed a PC20 of 0.45 mg/ml. The challenge test with raw rice resulted in immediate and late clinical and spirometric responses; pretreatment with DSCG inhibited both responses. The histamine release test (HRT) with rice was positive, and we detected rice-specific IgE antibodies by REIA in the patient's serum. Skin prick tests, HRT, and RAST with a battery of cereals gave positive results. Finally, the rice REIA was inhibited by rice (75%), rye (63%), corn (64%), and wheat (51%) extracts.

Adult↗

Delayed allergic reaction to cefonicid.

We report a case of delayed cutaneous reaction to intramuscular treatment with cefonicid, a second generation cephalosporin. An isolated late skin test response was observed to cefonicid only and not to other beta-lactams. A patch test with cefonicid 1% was positive after 48 and 96 hours.

Cefonicid↗

Pistachio nut hypersensitivity: identification of pistachio nut allergens.

Type I hypersensitivity to pistachio nut antigens was demonstrated in three patients by means of immediate skin-test reactivity, specific IgE determination by a fluoroimmunoassay (CAP), CAP-inhibition and leucocyte histamine release. Sensitization to other dried fruits and pollens was observed in the patients. The CAP-inhibition studies revealed significant crossreactivity between pistachio and cashew nut belonging to the Anacardiaceae family, and between pistachio nut and other dried fruits belonging to taxonomically unrelated botanical families. No relevant crossallergenicity was observed between pistachio nut and Lolium and Olea pollens. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of a pistachio nut extract followed by immunoblotting analysis identified four IgE-binding bands with molecular weights of 34, 41, 52 and 60 kD.

Allergens↗

Occupational protein contact dermatitis from herring.

We present a case of occupational protein contact dermatitis caused by herring in a dolphinarium worker. The in vivo and in vitro tests results indicated a type I allergic mechanism. In vivo cross-reactivity among fish belonging to the Clupeiformes order was observed.

Adult↗