The quality of the antioxidant defence system in term and preterm twin neonates

Abstract

Objective: Multiple pregnancy is associated with an enhanced metabolism and demand for O2, which may lead to the overproduction of reactive oxygen species and the development of oxidative stress. The degree of oxidative damage depends on the level of the antioxidant protection system of the foetus. The objective of the study was to identify the relationship between the state of the maturity and the antioxidant status of twin neonates. Investigations of the umbilical cord blood were carried out to detect differences in the antioxidant defence system between mature and premature twin neonates.

Methods: The activities of the superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) enzymes, the levels of reduced glutathione (GSH), protein carbonyls and oxidized lipids and the total antioxidant capacity of the plasma were determined.

Results: The level of lipid peroxidation was significantly higher in the premature neonates. An increase in the total antioxidant capacity was accompanied by a decrease in the damaged protein concentration. Significantly elevated activities of GPx alone were observed in the premature twins, though the GSH content too tended to be increased. The activity of SOD was decreased in the premature neonates.

Discussion: The antioxidant status of twin neonates are mainly influenced by maturity. We suggest that the level of lipid peroxidation might be of clinical value as a marker of pre- and perinatal distress in twins.     

The size of differences as a quality criterion for characters in phylogenetics

One important aspect regarding the suitability of a character for phylogenetic reconstruction is the separability of compared groups with regard to different character states. Therefore, it is recommendable to standardize mean differences with respect to the dispersion of individual data.     

Conductance of the cytoplasm of erythrocytes as a criterion for evaluating the quality of blood preservation.

It has been established that enzymic systems become exhausted during the preservation of blood, which causes disturbances in the transport of ions across the cell membrane. These changes occurring in the cell may be recorded by measuring the electrical conductance of cytoplasm. The conductance has been determined for three selected preserving liquids in relation to the blood storage time. Results of this study showed that the dynamics of changes in the electrical conductance of the cytoplasm is characteristic for a given type of the preserving liquid. Hence, the determination of changes in the electrical conductance of the cytoplasm may provide a new important criterion for evaluating various methods of blood preservation.     

The secretor locus as a marker for prenatal prediction of myotonic dystrophy (DM)

Summary To verify the reliability of secretor status for prenatal diagnosis of myotonic dystrophy (DM), 179 amniotic fluid samples were compared with saliva or urine samples of the infants by hemagglutination inhibition. While no discrepancies were observed, problems could arise with intermediate results. Additionally, secretor typing is only informative in 8.4% of patients.     

Competitive inhibition of cytochrome P-450scc by (22R)- and (22S)-22-aminocholesterol. Side-chain stereochemical requirements for C-22 amine coordination to the active-site heme

Two diastereomeric aminocholesterols, (22R)-22-aminocholesterol and (22S)-22-aminocholesterol, are both found to be potent inhibitors of the biosynthesis of pregnenolone from cholesterol by purified bovine mitochondrial P-450scc. Both steroids are competitive versus cholesterol, but the stereochemically correct analog (22R)-22-aminocholesterol is bound approximately 1000 times more tightly than (22S)-22-aminocholesterol. The dissociation constants are 25 nM and 13 microM, respectively. Direct comparisons between spectroscopic and enzymatic properties of the two enzymesterol complexes and the 22-amino-23,24-bisnor-5-cholen-3 beta-ol complex are made, underlining the importance of the stereochemistry at the C-22 position.     

Parental consanguinity as a cause for increased incidence of births defects in a study of 238,942 consecutive births.

The risk for birth defects in the offspring of first-cousin matings has been estimated to increase sharply compared to non consanguineous marriages. As a general decline in the frequency of consanguineous marriages was observed in this century, one wonders whether consanguinity is still a factor in the appearance of birth defects in developed countries. Based on our registry of congenital anomalies we tried to answer to this question. In the population studied in North-Eastern France a consanguineous mating was known in 1.21% of the cases with congenital anomalies, vs. 0.27% in controls, (p < 0.001). The frequency of the malformations recorded paralleled the degree of consanguinity: out of 89 malformed children, 51 were seen in first-cousins mating (10.3 times more frequent than in offspring of non consanguineous couples), 17 in second-cousins marriages and 18 in more distant relatives mating. Three were uncle-niece marriage. Excluding known mendelian conditions these numbers were 73, 36, 17 and 17 respectively and the corresponding relative risk were 3.68, 3.01, 3.41 and 4.89 respectively. Therefore there is a negative dose-response effect between level of inbreeding and risk of congenital malformations. Consanguineous mothers were more often pregnant than non consanguineous mothers (p < 0.01) and they had more stillbirths than non consanguineous mothers. These results show that consanguinity is still a factor of birth defects and they must be taken into account for genetic counseling of inbred marriages, in developed countries.     

The equivalent dose as a dosimetric criterion for acute radiation injury by radiations of various quality. The generalized equivalent dose

Using different quality coefficients as functions of LET, generalized equivalent doses were calculated, and the survival of dogs after gamma/neutron irradiation was determined using the concept of an equivalent dose. LET functions of the generalized quality coefficients providing a good agreement between theoretical and experimental results were chosen. Various functions of the generalized quality coefficients for different ingredients of bone-marrow haemopoiesis activity should be used to estimate the severity of radiation injury to red bone marrow.     

The sequence dependence of fiber organization. A comparative molecular dynamics study of the islet amyloid polypeptide segments 22-27 and 22-29

Amyloid fiber formation and the possible polymorphism of molecular arrangements depend on the polypeptide length and composition. Here, we seek the chemical clues underlying these processes. Our starting point is based on the experimental observation that some short peptide segments are able to develop fibers that are very similar to those of their original parent proteins. We focus our study on the NFGAILSS peptide, derived from the human islet amyloid polypeptide (residues 22-29). This peptide turned out to be a perfect example, illustrating the fact that the amyloid microscopic organization is highly complex, rather than simply involving hydrogen bond formation. Furthermore, obtaining a reliable molecular model has allowed us to analyze the differences between the amyloid structure we have obtained for this peptide and that obtained for the previously studied, two residues shorter, segment (residues 22-27, NFGAIL). This comparative study yields some clues about chemical events that govern the aggregation of proteins into oriented fibers, such as molecular packing between sheets and the degree of interaction specificity. We characterize the important role played by the hydrophobic and aromatic residues in the inter-sheet association and present new approaches toward the understanding of the nature of events that are likely to take place during fibril formation. These include analysis of interaction patterns derived from specific sheet-associated packing.     

Take-off capacity as a criterion for quality control in mass-produced predators,Rhizophagus grandis (Col.: Rhizophagidae) for the biocontrol of bark beetles,Dendroctonus micans (Col.: Scolytidae)

Time spent by adult beetles in cold storage at 3–7°C accounted for 81% of the loss of take-off capacity inRhizophagus grandis Gyllenhal in windtunnel experiments. At the age of three weeks, the insects were at their highest take-off capacity at about 80%. This was followed by a steady decrease, 7% of the insects failing to take-off each month. Changes in the fat reserves during cold storage could explain at least partly this reduction of flight capacity although there was no significant difference in fresh weight between insects that were able/unable to take-off. Sex had a significant influence on take-off rates, with an 8.7% higher take-off rate in females. Take-off capacity was further reduced when the insects were mass-produced in cultures using parent beetles submitted to a blend of synthetic oviposition stimulants instead of live prey larvae. Response to synthetic attractants by those insects which took-off, however, was not influenced by cold storage or by the use of synthetic stimulants in the culture medium.

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Résumé Le temps passé en conservation à basse température (3–7°C) depuis la métamorphose est intervenu pour 81% dans la perte de la capacité d'envol chez le coléoptère prédateurRhizophagus grandis Gyll. lors d'expériences en tunnel de vol. A l'age de trois semaines, les insectes sont à leur plus haut niveau d'envol (envol d'environ 80% des insectes). Par la suite, il y a un déclin constant de la capacité d'envol, à raison de 7% des insectes chaque mois. Des changements dans les réserves lipidiques peuvent partiellement expliquer cette réduction, bien qu'il n'y ait pas eu de différence entre le poids frais d'insectes capables de s'envoler et celui d'individus qui en étaient incapables. Le sexe a une influence sur le taux d'envol, avec un taux d'envol significativement plus élevé de 8.7% chez les femelles. La capacité d'envol est encore réduite chez des insectes qui ont été produits dans des élevages de masse où les parents étaient soumis à un mélange de stimuli de ponte de synthèse au lieu d'être mis en présence de larves deD. micans vivantes. Chez les insectes qui prennent leur vol, cependant, la réponse aux attractifs de synthèse est indépendante de l'age ainsi que des conditions d'élevage.

     

Ammonia toxicity as a criterion for the evaluation of larval quality in the prawn Macrobrachium rosenbergii

The feasibility of a short-term ammonia toxicity test as an evaluation criterion for larval quality was assessed in three trials. In each one, Macrobrachium rosenbergii larvae originating from the same spawn were nutritionally differentiated in two groups by feeding them either a nutrient-rich (Artemia nauplii enriched for 24 h with n-3 highly unsaturated fatty acids (HUFA) and ascorbic acid (AA)) or a nutrient-poor diet (Artemia nauplii starved for 24 h). Throughout their development, larvae from both treatments were exposed during 24 h to six concentrations of total ammonia (NH₄⁺+NH₃) and a control (no ammonia added). Based on mortality rates, the median lethal concentration for 50% of the population (LC₅₀) was estimated. As expected from earlier work, larvae fed the optimal diet presented higher n-3 HUFA and AA contents as well as higher growth and metamorphosis rates. From the moment the effect of diet quality was analytically detectable in the tissues of the larvae, the ammonia test was able to distinguish both groups of larvae. Differences in ammonia tolerance were observed as early as larval stage 4 and remained evident throughout larval development. The short-term ammonia toxicity test proved to be a valuable, sensitive and reproducible criterion for the establishment of larval quality.     

Odonata (Insecta) as a tool for the biomonitoring of environmental quality

Despite the fundamental dependence of human populations on water resources, a range of anthropogenic impacts, in particular the removal of riparian vegetation, threaten freshwater environments. One of the most effective means of evaluating the effects of anthropogenic disturbance in aquatic ecosystems is the use of bioindicators, and the insects of the order Odonata are among the most efficient indicators, due to their enormous sensitivity to environmental changes. In this context, the present study aimed to verify which parameters of the odonate community (species richness, abundance/biomass, composition, taxonomic diversity and taxonomic/phylogenetic distinctness) are most effective for the evaluation of the loss of environmental integrity. The study focused on 50 streams in the northeast of the Brazilian state of Pará. The streams were sampled during the dry season, between June and August 2011. The physical characteristics of each stream were evaluated using a Habitat Integrity Index (HII). The species composition provided the best parameter for the evaluation of ecological integrity, providing a relatively accurate assessment at a lower mean research cost than other parameters. Taxonomic diversity and distinctness also provided relatively reliable results, contributed additional information on the evolutionary relationships among the odonate taxa, and also provided a low-cost approach. Deconstructing communities is necessary to detect impacts, considering the considerable variation in the environmental requirements of the different species. Overall, the parameter that best responded to gradients of disturbance was species composition, followed by diversity and taxonomic distinctness. Given these findings, odonate-based biomonitoring should focus on these parameters to guarantee the optimal detection and evaluation of habitat alterations.     

Heat-shock protein 27 (Hsp27) as a target of methylglyoxal in gastrointestinal cancer

The molecular mechanisms underlying the posttranslational modification of proteins in gastrointestinal cancer are still unknown. Here, we investigated the role of methylglyoxal modifications in gastrointestinal tumors. Methylglyoxal is a reactive dicarbonyl compound produced from cellular glycolytic intermediates that reacts non-enzymatically with proteins. By using a monoclonal antibody to methylglyoxal-modified proteins, we found that murine heat-shock protein 25 and human heat-shock protein 27 were the major adducted proteins in rat gastric carcinoma mucosal cell line and human colon cancer cell line, respectively. Furthermore, we found that heat-shock protein 27 was modified by methylglyoxal in ascending colon and rectum of patients with cancer. However, methylglyoxal-modified heat-shock protein 25/heat-shock protein 27 was not detected in non cancerous cell lines or in normal subject. Matrix-associated laser desorption/ionization mass spectrometry/mass spectrometry analysis of peptide fragments identified Arg-75, Arg-79, Arg-89, Arg-94, Arg-127, Arg-136, Arg-140, Arg-188, and Lys-123 as methylglyoxal modification sites in heat-shock protein 27 and in phosphorylated heat-shock protein 27. The transfer of methylglyoxal-modified heat-shock protein 27 into rat intestinal epithelial cell line RIE was even more effective in preventing apoptotic cell death than that of native control heat-shock protein 27. Furthermore, methylglyoxal modification of heat-shock protein 27 protected the cells against both the hydrogen peroxide- and cytochrome c-mediated caspase activation, and the hydrogen peroxide-induced production of intracellular reactive oxygen species. The levels of lactate converted from methylglyoxal were increased in carcinoma mucosal cell lines. Our results suggest that posttranslational modification of heat-shock protein 27 by methylglyoxal may have important implications for epithelial cell injury in gastrointestinal cancer.     

Mass spectrometric study of the enzymatic conversion of cholesterol to (22R)-22-hydroxycholesterol, (20R,22R)-20,22-dihydroxycholesterol, and pregnenolone, and of (22R)-22-hydroxycholesterol to the lgycol and pregnenolone in bovine adrenocortical preparations. Mode of oxygen incorporation.

Incubation of cholesterol with a bovine adrenocortical mitochondrial acetone-dried powder preparation yielded (22R)-22-hydroxycholesterol (I), (20R,22R)-20,22-dihydroxycholesterol(II), and pregnenolone (III) which were conclusively identified by combined gas chromatography-mass spectrometry. Incubations with [4-14C]cholesterol yielded I, II, and III with specific activities (determined from partial mass-spectral scans) not significantly different from those of the used substrate or the cholesterol reisolated after the incubation, demonstrating that the isolated compounds arose mostly, if not entirely, from the substrate cholesterol. Incubations in an 18O-enriched atmosphere yielded I, II, and III with 18O at position C-22, C-20 and C-22, and C-20, respectively, providing evidence that the hydroxyl groups of the side chain of I and II and the C-20 oxygen atom of III originated from molecular oxygen. The distribution of the oxygen atoms in II after incubation with 18O2 and 16O2 (devoid of 16O18O) proved that the hydroxyl groups of the side chain of II were introduced from two different molecules of oxygen, consistent with a sequential hydroxylation of cholesterol. No (20S)-20-hydroxycholesterol was found. Incubation of I in an 18O-enriched atmosphere afforded II and III with 18O at C-20.     

The generation of big-endothelin (1-22) (endothelin-valine) from big-endothelin in the soluble fraction of porcine lung.

The degradation of big-endothelin (big-ET) in the soluble fraction of porcine lung was investigated. The degradation in the presence of p-chloromercuribenzoate (PCMB), pepstatin A, and EDTA resulted in the accumulation of two newly-formed fragments, big-ET (23-39) and big-ET (1-22), the latter called endothelin-valine (ET-Val). The generation of the two fragments was inhibited by diisopropylfluorophosphate (DFP). The enzyme responsible, called ET-Val-generating endopeptidase, was isolated from porcine lung by a procedure including chromatographies on columns of DEAE-cellulose, hydroxylapatite, Mono Q, p-mercuribenzoate-Sepharose, and Superose 6. The molecular weight of the enzyme was 140,000 and the pH optimum of the activity was 7.0. The activity was strongly inhibited by DFP, but scarcely inhibited by PCMB, EDTA, and pepstatin A. Thus, the isolated enzyme was classified as a serine protease cleaving big-ET at the Val22-Asn23 bond.