Early embryonic death of glutamate carboxypeptidase II (NAALADase) homozygous mutants.

Glutamate carboxypeptidase II (EC 3.4.17.21) catalyzes the hydrolysis (Km = 0.2 microM) of the neuropeptide N-acetylaspartylglutamate to yield N-acetylaspartate and glutamate and also serves as a high-affinity folate hydrolase in the gut, cleaving the polyglutamate chain to permit the absorption of folate. N-acetylaspartylglutamate is an agonist at the mGluR3 metabotropic receptor and a source of extracellular glutamate through hydrolysis by glutamate carboxypeptidase II. Given the important role of glutamate in brain development and function, we were interested in the effects of a null mutation of glutamate carboxypeptidase II that would potentiate the effects of N-acetylaspartylglutamate. The PGK-Neomycin cassette was inserted to delete exons 9 and 10, which we previously demonstrated encode for the zinc ligand domain essential for enzyme activity. Successful germline transmission was obtained from chimeras derived from embryonic stem cells with the targeted mutation of glutamate carboxypeptidase II. Homozygous null mutants did not survive beyond embryonic day 8. Folate supplementation of the heterozygous mothers did not rescue the homozygous embryos. Mice heterozygous for the null mutation appeared grossly normal and expressed both mutated and wild-type mRNA but the activity of glutamate carboxypeptidase II is comparable to the wild-type mice. The results indicate that the expression of glutamate carboxypeptidase II is upregulated when one allele is inactivated and that its activity is essential for early embryogenesis.

[Utilization of polymerase chain reaction for detection and genotyping of Helicobacter pylori in gastric biopsy using paraffin: a retrospective study]. / Utilización de la reacción en cadena de la polimerasa para la detección y genotipificación de Helicobacter pylori en biopsias gástricas incluídas en parafina: un estudio retrospectivo.

BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.

[A simple method to detect disaccharides deficiency]. / Métodos simples de detección de deficiencias de disacaridasas.

A simple method, easy to perform during an endoscopic procedure, fast and inexpensive, that allows detecting deficiencies in lactase, sucrase or maltase activities is presented. Briefly, method consists in placing a duodenal biopsy sample in an adequate vial containing lactose, sucrose or maltose solution during a few minutes, and then, adding a few drops of a glucose reactive from commercial origin. Presence of any enzymatic activity is demonstrated when released glucose from any of the disaccharides chosen reacts with the second reactive, turning solution to a red colour. Its utility is discussed and compared with other diagnostic methods.

Métodos simples de detección de deficiencias de disacaridases / A simple method to detect disaccharides deficiency

A simple method, easy to perform during an endoscopic procedure, fast and inexpensive, that allows detecting deficiencies in lactase, sucrase or maltase activities is presented. Briefly, method consists in placing a duodenal biopsy sample in an adequate vial containing lactose, sucrose or maltose solution during a few minutes, and then, adding a few drops of a glucose reactive from commercial origin. Presence of any enzymatic activity is demonstrated when released glucose from any of the disaccharides chosen reacts with the second reactive, turning solution to a red colour. Its utility is discussed and compared with other diagnostic methods.

Utilización de la reacción en cadena de la polimerasa para le detección y genotificación de Helicobacter pylori gástricas incluidas en parafina: un estudio retrospectivo / Utilization of polymerase chain reaction for detection and genotyping of Helicobacter pylori in gastric biopsy using paraffin: a retrospective study

BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.

Utilización de la reacción en cadena de la polimerasa para la detección y genotipificación de Helicobacter pylori en biopsias gástricas incluídas en parafina: un estudio retrospectivo. / [Utilization of polymerase chain reaction for detection and genotyping of Helicobacter pylori in gastric biopsy using paraffin: a retrospective study]

BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.

Métodos simples de detección de deficiencias de disacaridasas. / [A simple method to detect disaccharides deficiency]

A simple method, easy to perform during an endoscopic procedure, fast and inexpensive, that allows detecting deficiencies in lactase, sucrase or maltase activities is presented. Briefly, method consists in placing a duodenal biopsy sample in an adequate vial containing lactose, sucrose or maltose solution during a few minutes, and then, adding a few drops of a glucose reactive from commercial origin. Presence of any enzymatic activity is demonstrated when released glucose from any of the disaccharides chosen reacts with the second reactive, turning solution to a red colour. Its utility is discussed and compared with other diagnostic methods.

Métodos simples de detección de deficiencias de disacaridases / A simple method to detect disaccharides deficiency

A simple method, easy to perform during an endoscopic procedure, fast and inexpensive, that allows detecting deficiencies in lactase, sucrase or maltase activities is presented. Briefly, method consists in placing a duodenal biopsy sample in an adequate vial containing lactose, sucrose or maltose solution during a few minutes, and then, adding a few drops of a glucose reactive from commercial origin. Presence of any enzymatic activity is demonstrated when released glucose from any of the disaccharides chosen reacts with the second reactive, turning solution to a red colour. Its utility is discussed and compared with other diagnostic methods. (AU)

Utilización de la reacción en cadena de la polimerasa para le detección y genotificación de Helicobacter pylori gástricas incluidas en parafina: un estudio retrospectivo / Utilization of polymerase chain reaction for detection and genotyping of Helicobacter pylori in gastric biopsy using paraffin: a retrospective study

BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.(AU)