Expression of cystathionine beta-synthase and histopathological observations in placentas of patients with Down syndrome.

UNLABELLED: Down syndrome is the most frequent aneuploidy in live births, with an overall frequency of 1/600-700 births. The overexpression of cystathionine ß-synthase is thought to participate in the presentation of some phenotypes observed in Down syndrome. OBJECTIVE: The aim of this study was to compare the expression levels of cystathionine ß-synthase and histopathological observations from placentas of infants with Down syndrome and healthy newborns. MATERIALS AND METHODS: Six placentas of fetuses/infants with Down syndrome and sixteen placentas of healthy fetuses were studied. Cystathionine ß-synthase mRNA and protein expression were performed by real-time PCR and immunohistochemistry, respectively. RESULTS: We observed an increase in cystathionine ß-synthase mRNA expression (p = 0.0465) and protein levels (p = 0.009) in placentas of fetus/infants with Down syndrome compared with controls. Significantly more circinate edges (p = 0.0007) and trophoblast inclusions (p = 0.0037) were observed in the group with Down syndrome compared with control group. CONCLUSION: The results demonstrate overexpression of cystathionine ß-synthase mRNA and protein in placentas of fetuses/infants with trisomy 21. Further histological abnormalities were found in placentas of patients with Down syndrome, suggesting an alteration in the development of placenta.

Antioxidant intervention attenuates oxidative stress in children and teenagers with Down syndrome.

We previously demonstrated that systemic oxidative stress is present in Down syndrome (DS) patients. In the present study we investigated the antioxidant status in the peripheral blood of DS children and teenagers comparing such status before and after an antioxidant supplementation. Oxidative stress biomarkers were evaluated in the blood of DS patients (n=21) before and after a daily antioxidant intervention (vitamin E 400mg, C 500 mg) during 6 months. Healthy children (n=18) without DS were recruited as control group. The activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), gamma-glutamyltransferase (GGT), glucose-6-phosphate dehydrogenase (G6PD) and myeloperoxidase (MPO), as well as the contents of reduced glutathione (GSH), uric acid, vitamin E, thiobarbituric acid reactive substances (TBARS), and protein carbonyls (PC) were measured. Before the antioxidant therapy, DS patients presented decreased GST activity and GSH depletion; elevated SOD, CAT, GR, GGT and MPO activities; increased uric acid levels; while GPx and G6PD activities as well as vitamin E and TBARS levels were unaltered. After the antioxidant supplementation, SOD, CAT, GPx, GR, GGT and MPO activities were downregulated, while TBARS contents were strongly decreased in DS. Also, the antioxidant therapy did not change G6PD and GST activities as well as uric acid and PC levels, while it significantly increased GSH and vitamin E levels in DS patients. Our results clearly demonstrate that the antioxidant intervention with vitamins E and C attenuated the systemic oxidative damage present in DS patients.

Alterations of ectonucleotidases and acetylcholinesterase activities in lymphocytes of Down syndrome subjects: relation with inflammatory parameters.

BACKGROUND: Subjects with Down syndrome (DS) have an increased susceptibility to infections and autoimmune disorders. ATP, adenosine, and acetylcholine contribute to the immune response regulation, and NTPDase, adenosine deaminase (ADA) and acetylcholinesterase (AChE) are important enzymes in the control of the extracellular levels of these molecules. We evaluated the activities of these enzymes and the cytokine levels in samples of DS individuals. METHODS: The population consisted of 23 subjects with DS and 23 healthy subjects. Twelve milliliters of blood was obtained from each subject and used for lymphocyte and serum preparation. Lymphocytes were separated on Ficoll density gradients. After isolation, NTPDase and AChE activities were determined. RESULTS: The NTPDase activity using ADP as substrate was increased in lymphocytes of DS patients compared to control (P<0.05); however, no alterations were observed in the ATP hydrolysis. An increase was observed in the AChE activity in lymphocytes and in ADA activity in serum of DS patients when compared to healthy subjects (P<0.05). In DS subjects, an increase in the levels of IL-1ß, IL-6, TNF-α and IFN-γ and a decrease in the IL-10 levels were also observed (P<0.05). CONCLUSIONS: Alterations in the NTPDase, ADA and AChE activities as well changes in the cytokine levels may contribute to immunological alterations observed in DS.

Polymorphism C1420T of Serine hydroxymethyltransferase gene on maternal risk for Down syndrome.

Recent researches have investigated the factors that determine the maternal risk for Down syndrome (DS) in young woman. In this context, some studies have demonstrated the association between polymorphisms in genes involved on folate metabolism and the maternal risk for DS. These polymorphisms may result in abnormal folate metabolism and methyl deficiency, which is associated with aberrant chromosome segregation leading to trisomy 21. In this study, we analyzed the influence of the polymorphism C1420T in Serine hydroxymethyltransferase (SHMT) gene on maternal risk for DS and on metabolites concentrations of the folate pathway (serum folate and plasma homocysteine and methylmalonic acid). The study group was composed by 105 mothers with DS children (case group) and 185 mothers who had no children with DS (control group). The genotype distribution did not show significant statistical difference between case and control mothers (P = 0.24) however a protective effect between genotypes CC (P = 0.0002) and CT (P < 0.0001) and maternal risk for DS was observed. Furthermore, the SHMT C1420T polymorphism (rs1979277) does not affect the concentration of metabolites of folate pathway in our DS mothers. In conclusion, our data showed a protective role for the genotypes SHMT CC and CT on maternal risk for DS. The concentrations of metabolites of folate pathway did not differ significantly between the genotypes SHMT.

Oxidative stress and hematologic and biochemical parameters in individuals with Down syndrome.

OBJECTIVE: To evaluate the levels of thiobarbituric acid reactive substances (TBARS), uric acid, and seric superoxide dismutase and catalase activities, as well as total serum iron, total iron-binding capacity (TIBC), erythrocyte osmotic fragility, and hemograms in people with Down syndrome. PARTICIPANTS AND METHODS: The study sampled (July to December 2003) 50 people with Down syndrome and 50 individuals without Down syndrome (control group) matched by age and sex. The levels of TBARS were measured by the TBARS method. Serum superoxide dismutase and catalase activities and uric acid levels were determined spectrophotometrically. Erythrocyte osmotic fragility was assessed by the percentage of hemolysis. Hemograms, total serum iron level, and TIBC were determined with automated systems. RESULTS: The results showed that levels of TBARS, uric acid, and seric superoxide dismutase and catalase activities were higher in the Down syndrome group compared with the control group. We also observed a slight increase in erythrocyte osmotic fragility in the Down syndrome group, but the total serum iron levels, TIBC, and hemograms for both groups were within the age-related reference values. CONCLUSION: This was the first time, to our knowledge, that increases in seric superoxide dismutase and catalase activities were observed in people with Down syndrome. Although other studies are necessary, our results add to the understanding of the mechanisms responsible for the increased oxidative stress observed in individuals with Down syndrome and may be useful in supporting future antioxidant therapies that will improve the lives of people with Down syndrome.

[A 12-week physical activity program increases glucose-6-phosphate-dehydrogenase activity in Down syndrome adolescents]. / Incremento de la glucosa-6-fosfato-deshidrogenasa eritrocitaria en jóvenes con sindrome de Down tras un programa de actividad física de 12 semanas.

In recent years it has been claimed that trisomic cells are more sensitive to oxidative stress since there is an imbalance in the hydrogen peroxide metabolism. We designed the present study to assess the activity level of antioxidant enzyme glucose-6-phosphate-dehydrogenase (G6PDH) of erythrocytes in 31 male adolescents with Down syndrome (mean age 16.3 +/- 1.1) after performing a 12 week aerobic training program. First of all, a significant increase of 14.9% in the catalytic activity of G6PDH was observed in male adolescents with Down syndrome when compared with age, sex and body mass-matched controls without trisomy. After 12-wk program its activity increased significantly compared to baseline value in Down syndrome individuals. Our data are consistent with previous evidence of the existence of higher oxidative stress in adolescents with Down syndrome when compared to the general population. We may also conclude that G6PDH activity was significantly increased after a 12 week exercise program.

Incremento de la glucosa-6-fosfato-deshidrogenasa eritrocitaria en jóvenes con sindrome de Down tras un programa de actividad física de 12 semanas / A 12-week physical activity program increases glucose-6-phosphate-dehydrogenase activity in Down syndrome adolescents

Recientemente se ha publicado que las células trisómicas presentan una mayor sensibilidad al daño oxidativo, que podría justificar la frecuente asociación de síndrome de Down a aterosclerosis, envejecimiento precoz, etc. Para conocer el posible papel de la actividad física moderada en la mejora de la capacidad antioxidante se estudió el comportamiento de la enzima glucosa-6-fosfato-deshidrogenasa (G6PDH) eritrocitaria en 31 adolescentes varones (16.3 ± 1.1 años) tras desarrollar un programa de 12 semanas con tres sesiones (45-60 minutos) y una intensidad del 60-75% frecuencia cardíaca máxima teórica. Nuestros resultados indican una mayor actividad de G6PDH en individuos con síndrome de Down cuando se compara con controles sin trisomía ajustados a su sexo, edad e índice de masa corporal. Asimismo observamos un incremento significativo de su actividad tras completar nuestro programa de 12 semanas. Podemos concluir que la actividad física moderada mejora la capacidad antioxidante en jóvenes con síndrome de Down.(AU)

In recent years it has been claimed that trisomic cells are more sensitive to oxidative stress since there is an imbalance in the hydrogen peroxide metabolism. We designed the present study to assess the activity level of antioxidant enzyme glucose-6-phosphate-dehydrogenase (G6PDH) of erythrocytes in 31 male adolescents with Down syndrome (mean age 16.3 +/- 1.1) after performing a 12 week aerobic training program. First of all, a significant increase of 14.9% in the catalytic activity of G6PDH was observed in male adolescents with Down syndrome when compared with age, sex and body mass-matched controls without trisomy. After 12-wk program its activity increased significantly compared to baseline value in Down syndrome individuals. Our data are consistent with previous evidence of the existence of higher oxidative stress in adolescents with Down syndrome when compared to the general population. We may also conclude that G6PDH activity was significantly increased after a 12 week exercise program.(AU)

Incremento de la glucosa-6-fosfato-deshidrogenasa eritrocitaria en jóvenes con sindrome de Down tras un programa de actividad física de 12 semanas / A 12-week physical activity program increases glucose-6-phosphate-dehydrogenase activity in Down syndrome adolescents

Recientemente se ha publicado que las células trisómicas presentan una mayor sensibilidad al daño oxidativo, que podría justificar la frecuente asociación de síndrome de Down a aterosclerosis, envejecimiento precoz, etc. Para conocer el posible papel de la actividad física moderada en la mejora de la capacidad antioxidante se estudió el comportamiento de la enzima glucosa-6-fosfato-deshidrogenasa (G6PDH) eritrocitaria en 31 adolescentes varones (16.3 ± 1.1 años) tras desarrollar un programa de 12 semanas con tres sesiones (45-60 minutos) y una intensidad del 60-75% frecuencia cardíaca máxima teórica. Nuestros resultados indican una mayor actividad de G6PDH en individuos con síndrome de Down cuando se compara con controles sin trisomía ajustados a su sexo, edad e índice de masa corporal. Asimismo observamos un incremento significativo de su actividad tras completar nuestro programa de 12 semanas. Podemos concluir que la actividad física moderada mejora la capacidad antioxidante en jóvenes con síndrome de Down.

In recent years it has been claimed that trisomic cells are more sensitive to oxidative stress since there is an imbalance in the hydrogen peroxide metabolism. We designed the present study to assess the activity level of antioxidant enzyme glucose-6-phosphate-dehydrogenase (G6PDH) of erythrocytes in 31 male adolescents with Down syndrome (mean age 16.3 +/- 1.1) after performing a 12 week aerobic training program. First of all, a significant increase of 14.9% in the catalytic activity of G6PDH was observed in male adolescents with Down syndrome when compared with age, sex and body mass-matched controls without trisomy. After 12-wk program its activity increased significantly compared to baseline value in Down syndrome individuals. Our data are consistent with previous evidence of the existence of higher oxidative stress in adolescents with Down syndrome when compared to the general population. We may also conclude that G6PDH activity was significantly increased after a 12 week exercise program.

Glutathione metabolism and antioxidant enzymes in children with Down syndrome.

Oxidative stress has been proposed as a pathogenic mechanism of atherosclerosis, cell aging, and neurologic disorders in Down syndrome. This study demonstrates a systemic decrease of all glutathione forms, including glutathionyl-hemoglobin, in the blood of children with Down syndrome. Furthermore, we obtained a disequilibrium, in vivo, between the antioxidant enzyme activities.

[Mutations in the methylene-tetrahydrofolate reductase gene and Down syndrome]. / Mutações no gene da metilenotetrahidrofolato redutase e síndrome de Down.

Down syndrome (DS) is a complex genetic and metabolic disorder attributed to the presence of three copies of chromosome 21. The extra chromosome derives from the mother in 93% of cases and is due to abnormal chromosome segregation during meiosis (nondisjunction). Except for advanced age at conception, maternal risk factors for meiotic nondisjunction are not well established. A recent preliminary study suggested that abnormal folate metabolism and the 677 (C-->T) mutation in the methylene-tetrahydrofolate reductase (MTHFR) gene may be maternal risk factors for DS. Frequency of the MTHFR 677 (C-->T) and 1298 (A-->C) mutations was evaluated in 36 mothers of children with DS and in 200 controls. The results are consistent with the observation that the MTHFR 677 (C-->T) and 1298 (A-->C) mutations are more prevalent among mothers of children with DS than controls. In addition, the most prevalent genotype was the combination of both mutations. The results suggest that mutations in the MTHFR gene are associated with maternal risk for DS

Cell lines derived from hippocampal neurons of the normal and trisomy 16 mouse fetus (a model for Down syndrome) exhibit neuronal markers, cholinergic function, and functional neurotransmitter receptors.

We have established hippocampal cell lines from normal and trisomy 16 fetal mice, a model of human trisomy 21. Both cell lines, named H1b (derived from a normal animal) and HTk (trisomic) possess neuronal markers by immunohistochemistry (enolase, synaptophysin, microtubule associated protein-2, and choline acetyltransferase) and lack glial markers (glial fibrillary acidic protein and S-100). Also, we evaluated intracellular Ca(2+) levels ([Ca(2+)](i)) in response to neurotransmitter agonists, in cells loaded with the fluorescent Ca(2+) indicators Indo-1 and Fluo-3. Both cell lines responded to glutamatergic stimuli induced by glutamate, N-methyl-D-aspartate, I-amino-2,3-dihydro-5-methyl-3-oxo-4-isoxazole propanoic acid or kainate. Glutamate responses were only partially prevented by addition of 5 mM EGTA and the metabotropic glutamate receptor agonist, trans-(1S,3R)-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD), increased [Ca(2+)](i) in both cell types. These results confirm the presence of glutamatergic metabotropic receptors. In glutamate-induced responses, HTk cells exhibited slower time-dependent decay kinetics than H1b cells. Cholinergic agonists (nicotine and muscarine) induced a rapid, transient increase in [Ca(2+)](i) in both cell types. Furthermore, some cells were sensitive to histamine and norepinephrine. All responses to the aforementioned agonists were prevented by addition of specific antagonists. We also studied incorporation and release of [(3)H]choline in the cells, and observed no differences in uptake parameters. However, release induced by K(+) and nicotine depolarization was greatly reduced in HTk cells. The results show that H1b and HTk cells retain neuronal characteristics and respond to specific neurotransmitter stimuli. The HTk differences could be related to neuronal pathophysiology in Down syndrome.