Effects of ascorbic acid addition on the oxidative stress response of Oryza sativa L. plants to As(V) exposure.

Accumulation of noxious elements in the edible part of crops and its impact on food safety is of increasing concern. Rice is one of the major staple food crops worldwide, including arsenic (As)-polluted areas, in which dietary As exposure is becoming a widespread health threat. Plant chemical priming has been shown to be an effective strategy to enhance tolerance to environmental stresses, including metal(loid) exposure. The priming effect of ascorbic acid (AsA) was assessed in rice seedlings exposed to As(V) in a hydroponics experiment. AsA treatment (co-addition to the growing media concomitantly (t0) or 24 h in advance (t24)) prevented an excessive accumulation of As in the roots (that decreased âˆ¼ 60%) and stimulated the activities of photosynthetic and antioxidant attributes (∼1.2-fold) in the aerial part of the plants. The increase in proline levels in both shoots (∼2.1-fold) and roots (∼2.4-fold) was found to be the most sensitive stress parameter, and was able to reflect the AsA-induced reduction of As toxic effects (concentrations back to Control levels, both simultaneously added or added as a pretreatment) in the aerial part of the plants. However, the phytotoxic effects related to As exposure were not fully prevented by priming with AsA, and further research is needed to find alternative priming approaches.

Sigma-Lognormal Modeling of Speech.

Human movement studies and analyses have been fundamental in many scientific domains, ranging from neuroscience to education, pattern recognition to robotics, health care to sports, and beyond. Previous speech motor models were proposed to understand how speech movement is produced and how the resulting speech varies when some parameters are changed. However, the inverse approach, in which the muscular response parameters and the subject's age are derived from real continuous speech, is not possible with such models. Instead, in the handwriting field, the kinematic theory of rapid human movements and its associated Sigma-lognormal model have been applied successfully to obtain the muscular response parameters. This work presents a speech kinematics-based model that can be used to study, analyze, and reconstruct complex speech kinematics in a simplified manner. A method based on the kinematic theory of rapid human movements and its associated Sigma-lognormal model are applied to describe and to parameterize the asymptotic impulse response of the neuromuscular networks involved in speech as a response to a neuromotor command. The method used to carry out transformations from formants to a movement observation is also presented. Experiments carried out with the (English) VTR-TIMIT database and the (German) Saarbrucken Voice Database, including people of different ages, with and without laryngeal pathologies, corroborate the link between the extracted parameters and aging, on the one hand, and the proportion between the first and second formants required in applying the kinematic theory of rapid human movements, on the other. The results should drive innovative developments in the modeling and understanding of speech kinematics.

The aroma of La Mancha Chelva wines: Chemical and sensory characterization.

The aim of this study is the chemical and sensory description of the aroma of wines white Chelva through five successive vintages. Solid Phase Extraction (SPE) was used for the isolation of volatile compounds and Gas Chromatography Mass Spectrometry (GC-MS) was employed by their analysis. Aroma Sensory Profile of wines was defined using the Quantitative Descriptive Sensory Analysis (QDA). A total of 77 volatile compounds were identified in Chelva wines. Chelva wines present a great quantity of aromatic compounds of very different chemical nature and with different aromatic descriptors, which will increase its aromatic complexity of these wines. Aroma sensory profile of Chelva wines was defined by higher intensity of fresh and peach aromas with notes citric, green apple, tropical fruit and sweet. This study showed that the Chelva grapes variety cultivated in La Mancha region present a great aroma potential and a complex sensory profile and can be considered a viable alternative to traditional grape varieties cultivated in this region for increasing the offer to the consumer.

Replication of PTPRC as genetic biomarker of response to TNF inhibitors in patients with rheumatoid arthritis.

Genetic biomarkers could be useful for orienting treatment of patients with rheumatoid arthritis (RA), but none has been convincingly validated yet. Putative biomarkers include 14 single nucleotide polymorphisms that have shown association with response to TNF inhibitors (TNFi) in candidate gene studies and that we assayed here in 755 RA patients. Three of them, in the PTPRC, IL10 and CHUK genes, were significantly associated with response to TNFi. The most significant result was obtained with rs10919563 in PTPRC, which is a confirmed RA susceptibility locus. Its RA risk allele was associated with improved response (B=0.33, P=0.006). This is the second independent replication of this biomarker (P=9.08 × 10(-8) in the combined 3003 RA patients). In this way, PTPRC has become the most replicated genetic biomarker of response to TNFi. In addition, the positive but weaker replication of IL10 and CHUK should stimulate further validation studies.

Nonsuppressed ion chromatographic determination of total calcium in milk.

A novel approach for measuring calcium in milk by the use of high performance nonsuppressed ion chromatography is proposed. Total calcium as well as the calcium present in the colloidal and salt state in milk can be quantified. Because the presence of citrate in the milk serum interferes with the measurement of calcium by the conductivity detector, the free and chelated calcium can also be distinguished. By using a sample preparation procedure using Amberlite resin (Fluka, Steinheim, Germany), the amount of soluble calcium in milk can be determined. Because the quantification of calcium in milk and milk derivatives is becoming increasingly important in order to understand the functionality of the milk proteins, better methods need to be developed to determine, with a high level of accuracy, the amount of calcium present in milk in various forms.

Rheological properties of rennet gels containing milk protein concentrates.

Different milk protein concentrates (MPC), with protein concentrations of 56, 70, and 90%, were dispersed in water under different treatments (hydration, shear, heat, and overnight storage at 4 degrees C), as well as in a combination of all the treatments in a factorial design. The particle size distribution of the dispersions was then measured to determine the optimal conditions for the dispersion. Heating at 60 degrees C for 30 min with 5 min of shear was chosen as the best condition to dissolve MPC powders. The samples were also characterized for composition, presence of protein aggregates, and ratio of calcium to protein. The total calcium present in MPC increased with increasing concentration of protein; however, the total calcium-to-protein ratio was lower in MPC90 than in MPC56 and MPC70. The level of whey protein denaturation, the presence of kappa-casein-whey protein aggregates in the supernatant after centrifugation, and the amount of caseins dissociated from the micelle increased as the protein concentration in the powder increased. The total amount of casein macropeptide released was lower in samples from powders with a higher protein concentration than for MPC56 or the skim milk control. The gelation behavior of reconstituted MPC was tested in systems dispersed in water (5% protein) as well as in systems dispersed in skim milk (6% protein). The gelation time of MPC dispersions was considerably lower and the gel modulus was higher than those of reconstituted skim milk with the same protein concentration. When MPC dispersions were dialyzed against skim milk, a significant decrease in the gelation time and modulus were shown, with a complete loss of gelling functionality in MPC90 dispersed in water. This demonstrated that the ionic equilibrium was key to the functionality of MPC.

[Serologic diagnosis of Helicobacter pylori in endoscopy personnel. Serology in endoscopists]. / Diagnóstico serológico de Helicobacter pylori en endoscopistas. Serología en endoscopistas.

OBJECTIVE: the Helicobacter pylori entered into the medical world, particularly in the field of the Gastroenterology, revolutionizing it, and changing partially our physiopathological concepts and treatment of the gastroduodenal ulcer. This infestation results in an immune response, that is measured by means of serologic tests, which are very useful in the massive screening studies, mainly in population at risk like those who are institutionalized and the personnel of health, particularly workers of the departments of Gastroenterology. METHOD: motivated for this, we carry out a prospective study in 38 endoscopists using as controls 38 workers not related with a Gastroenterology department. Thus, we analyzed the presence of antibodies against Helicobacter pylori among the personnel of the departments of Gastroenterology of three hospitals of Havana City. These personnel filled a form designed for this purpose containing following data: age, sex, time of work as endoscopist, use of protection means, and history of gastroduodenal pathology. RESULTS: there were 24 women in each group (63,2%). Whereas the seroprevalence among endoscopists was 39,47%, in the control group, only three people were seropositives (7,89%). We found a positive correlation between the time working as endoscopists and the rate of seropositivity. CONCLUSIONS: our study demonstrated that endoscopists have a significantly higher risk than the controls of contracting the infection by the Helicobacter pylori, and that the seropositive endoscopists have sintomatology more frequently than the seronegative endoscopists, although these differences were not statistically significant.

[Lines of action for the insertion mounting and manipulation of a system of valves for the central versus catheters]. / Líneas de actuación para la inserción, montaje y manipulación de un sistema de válvulas para los catéteres venosos centrales.

In order to reduce the incidence of catheter related sepsis and the factors associated to it, our unit established some performance lines for the insertion, assembly and handling of a valvular system for the central venous catheters.

A poly(ADP-ribose) polymerase haplotype spanning the promoter region confers susceptibility to rheumatoid arthritis.

OBJECTIVE: To investigate the association of the poly(ADP-ribose) polymerase 1 (PARP-1) gene promoter polymorphism with rheumatoid arthritis (RA) predisposition. METHODS: An association study with 213 Spanish RA patients and 242 healthy subjects was carried out to investigate the association of all known PARP-1 gene promoter polymorphisms, i.e., a CA microsatellite repeat, a poly(A)(n), and 3 single point mutations (C410T, C1362T, and G1672A), with disease susceptibility. Additionally, we analyzed the distribution of PARP-1 polymorphisms in 58 Spanish families with 1 or more affected members. RESULTS: Upon complete genotyping of the panel of 455 samples, strong linkage disequilibrium was observed among the 5 PARP-1 polymorphisms. Only 2 PARP-1 haplotypes were detected: haplotype A (410T-[A](10)-[CA](10-12)-1362C, which includes short PARP-1 CA alleles) and haplotype B (410C-[A](11)-[CA](13-20)-1362T, always paired with long PARP-1 CA variants). Regarding the G1672A variation, although linkage disequilibrium was detected, it did not seem to be part of the conserved haplotypes described. Haplotype B was statistically overrepresented in the RA patient group compared with the healthy subjects (odds ratio 1.42, 95% confidence interval 1.06-1.91, P = 0.019). In addition, a significant dose effect of PARP-1 haplotype carriage on disease predisposition was observed. Of note, within haplotype B, the PARP-1 CA 97-bp allele was found to be the RA-predisposing marker (odds ratio 2.17, 95% confidence interval 1.27-3.72, P = 0.003, corrected P < 0.05). CONCLUSION: Our results demonstrate the existence of 2 unique PARP-1 haplotypes in the Spanish population and provide the first evidence that PARP-1 haplotypes play a role in susceptibility to RA.

Antioxidant systems and O(2)(.-)/H(2)O(2) production in the apoplast of pea leaves. Its relation with salt-induced necrotic lesions in minor veins.

The present work describes, for the first time, the changes that take place in the leaf apoplastic antioxidant defenses in response to NaCl stress in two pea (Pisum sativum) cultivars (cv Lincoln and cv Puget) showing different degrees of sensitivity to high NaCl concentrations. The results showed that only superoxide dismutase, and probably dehydroascorbate reductase (DHAR), were present in the leaf apoplastic space, whereas ascorbate (ASC) peroxidase, monodehydroascorbate reductase (MDHAR), and glutathione (GSH) reductase (GR) seemed to be absent. Both ASC and GSH were detected in the leaf apoplastic space and although their absolute levels did not change in response to salt stress, the ASC/dehydroascorbate and GSH to GSH oxidized form ratios decreased progressively with the severity of the stress. Apoplastic superoxide dismutase activity was induced in NaCl-treated pea cv Puget but decreased in NaCl-treated pea cv Lincoln. An increase in DHAR and GR and a decrease in ASC peroxidase, MDHAR, ASC, and GSH levels was observed in the symplast from NaCl-treated pea cv Lincoln, whereas in pea cv Puget an increase in DHAR, GR, and MDHAR occurred. The results suggest a strong interaction between both cell compartments in the control of the apoplastic ASC content in pea leaves. However, this anti-oxidative response does not seem to be sufficient to remove the harmful effects of high salinity. This finding is more evident in pea cv Lincoln, which is characterized by a greater inhibition of the growth response and by a higher rise in the apoplastic hydrogen peroxide content, O(2)(.-) production and thiobarbituric acid-reactive substances, and CO protein levels. This NaCl-induced oxidative stress in the apoplasts might be related to the appearance of highly localized O(2)(.-)/H(2)O(2)-induced necrotic lesions in the minor veins in NaCl-treated pea plants. It is possible that both the different anti-oxidative capacity and the NaCl-induced response in the apoplast and in the symplast from pea cv Puget in comparison with pea cv Lincoln contributes to a better protection of pea cv Puget against salt stress.

[Patterns of drug resistance in Staphylococcus aureus from human clinical isolates]. / Patrones de drogorresistencia de cepas de Staphylococcus aureus de origen clínico humano.

Fifty circulating strains of Staphylococcus aureus of clinical origin were characterized by their drug susceptibility to 15 antimicrobials through the method of radial diffusion in Mueller Hinton medium. Also, beta-lactam production was determined by acidimetric and chromogenic methods as well as the presence of methicillin-resistant strains. It was confirmed that 32% of strain was susceptible to tested antimicrobials, the most effective of which were imipenem, norfloxacyn, and amikacyn for 98, 96 and 92% susceptibility respectively. Twenty-seven different drug resistance patterns were found in the studied strains. 22% of the total strains was beta-lactam producers whereas 27% of the latter turned out to be methicilline-resistant.

Does diphenylene iodonium chloride have any effect on the O2- -generating step of plant peroxidases?

The O2*- -generating step of plant peroxidases during their catalytic cycle is represented by the decay of compound III (CoIII) into ferriperoxidase, which most likely involves the dissociation of a ferric-O2*- complex to yield the ferric form of the enzyme and O2*-. Diphenylene iodonium chloride (DPI), at 50-100 microM, does not significantly enhance the stability of CoIII of peroxidase, as judged by the values of k(decay), and therefore, DPI appears to have no effect on the O2*- -generating step of plant peroxidases. From these results, it is concluded that caution should be exercised when considering peroxidase as a possible enzyme target of O2*- -mediated plant physiological processes sensitive to DPI inhibition.

Expression and characterization of three tomato 1-aminocyclopropane-1-carboxylate oxidase cDNAs in yeast.

Heterologous expression in yeast has previously shown that the tomato cDNA LE-ACO1 encodes a functional 1-aminocyclopropane-1-carboxylate (ACC) oxidase (ACO) protein [Hamilton, A. J., Bouzayen, M. & Grierson, D. (1991) Proc. Natl Acad. Sci. USA 88, 7434-7437]. In the present work, full-length cDNAs encoding the two other members of the tomato ACO family (LE-ACO2 and LE-ACO3) were isolated and expressed in Saccharomyces cerevisiae. Analysis of the predicted amino acid sequences showed that the ACO1 and ACO3 proteins are highly similar (95%) while ACO2 is more divergent (89%). Yeast strains transformed with each of the three cDNAs were able to convert exogenous ACC to ethylene, the ACO1 strain exhibiting the highest activity in vivo and the ACO3 and ACO2 strains reaching 65% and 45% of ACO1 maximum activity, respectively. None of the ACO activities expressed in yeast required addition of ascorbate in vivo. ACO activities assayed in vitro revealed no significant differences between the three isoforms with regards to optimum temperature (29 degrees C), optimum pH (6.8-7.2), absolute dependence for ascorbate, Fe2+ and carbon dioxide, and inhibition by iron-chelating agents (1,10-phenanthroline and EDTA), Co2+ and free-radical scavengers (n-propyl gallate). However, differences were detected in the apparent Km values for ACC, the pI and the specific activity. The biochemical features that might explain the differences between the isoenzyme activities are discussed.

Effect of fosetyl-A1 on peroxidase from grapevine (Vitis vinifera) cells.

Grapevine (Vitis vinifera cv. Monastrell) suspension cell cultures were treated with fosetyl-A1, a widely used systemic fungicide for grapevine diseases caused by oomycetes, and examined at the electron microscope level for peroxidase cytochemistry. The results showed that treatment with fosetyl-A1 provokes an activation of both vacuolar sap and tonoplast-located peroxidase, already described as due to the basic peroxidase isoenzyme, B5, which was previously characterized as a constitutive marker of disease resistance against Plasmopara viticola in axillary bud cultures of Vitis spp. This activation of peroxidase isoenzyme B5, as seen at the electron microscope level, was confirmed by cytophotometric methods, but is in contrast with the unchanged enzyme level determined by biochemical methods. These results suggest a metabolic activation of peroxidase isoenzyme B5 as a consequence of fosetyl-A1 treatment, probably due to an acidification of the vacuole. This response was accompanied by the appearance of myelin-like structures inside the cytoplasm and osmiophylic-bodies inside the mitochondria. However, the latter structural changes cannot easily be related to the above described specific peroxidase response.

Purification of a basic peroxidase isoenzyme from Capsicum fruits and the immunoinhibition of its capsaicin oxidation capacity by antibodies raised against horseradish peroxidase.

Pepper fruits contain a peroxidase isoenzyme of basic pI, the peroxidase isoenzyme B6, located in vacuoles and the principal component of peroxidase polymorphism in the whole fruit. This isoenzyme was purified by preparative isoelectric focusing in glycerol-stabilized 3.0-10.0 pH gradients and characterized for its ability to oxidize capsaicin (8-methyl-N-vanillyl-6-nonenamide). Spectrophotometric studies illustrated that the capsaicin oxidation by pepper peroxidase isoenzyme B6 was H2O2-dependent and was totally abolished by antibodies raised against horseradish peroxidase. From these studies, it can be concluded that capsaicin is oxidized by pepper peroxidase isoenzyme B6, thus confirming a role for this peroxidase isoenzyme in capsaicin turnover and degradation.

Hammett rho sigma correlation for the inhibition by indoles of coniferyl alcohol oxidation catalyzed by cell wall peroxidases.

The inhibitory effect of indole-3-acetic acid, and of its peroxidase-mediated degradation products of an indole nature, on the oxidation rate of coniferyl alcohol catalyzed by cell wall peroxidases has been studied. The results show that the inhibitory effect of indole-3-acetic acid and indole-3-carbinol may be explained, in part, by their properties as peroxidase substrates. However, I50 values for a series of indole compounds not regarded as peroxidase substrates show a good correlation with the electron-donating or electron-withdrawing nature of the 3-substituents, as judged by the linearity of the Hammett rho sigma plot. These results suggest that although the properties of indole compounds as peroxidase substrates may be responsible, in part, for their inhibitory effects on the peroxidase-mediated oxidation of coniferyl alcohol, the inhibitory effect appears to be mainly determined by the acidity of the imino group of the indole nucleus.

A comparative study of a progestin-only oral contraceptive versus non-hormonal methods in lactating women in Buenos Aires, Argentina.

A non-randomized comparative clinical trial of the progestin-only oral contraceptive (POC), Ovrette (75 mcg norgestrel) (Wyeth), versus non-hormonal methods was conducted at two clinics in Buenos Aires, Argentina. The trial was designed to assess the breast-feeding patterns of women choosing progestin-only oral contraception and non-hormonal methods of contraception, and to study the relationship between lactation and the clinical performance of a POC. Five-hundred women were allocated to either the progestin-only pill group (n = 250) or to the non-hormonal group (n = 250) and were followed up monthly for six months after admission. Measurements in mean infant weight, mean infant length, and mean head circumference were similar throughout the follow-up period. Non-hormonal users reported significantly more self-perceived decreases in milk production at the 5th and 6th month follow-up intervals. Acceptance and continued use of the pill were excellent, with only one woman discontinuing because of a pregnancy which was attributed to user failure. The principal side effect reported by women in both groups was intermenstrual bleeding.

PIP: A nonrandomized, comparative, clinical trial of the progestin only oral contraceptive (OC) Ovrette (75 mcg norgestrel) vs. nonhormonal methods was conducted at 2 clinics in Buenos Aires, Argentina. The trial was designed to assess breast-feeding patterns of women choosing progestin- only OCs and nonhormonal methods of contraception and to study the relationship between lactation and the clinical performance of the OC. 500 women were allocated to either the progestin-only pill group (n=250) or t o the nonhormonal group (n=250) and were followed monthly for 6 months after admission. Measurements in mean infant weight, mean infant length, and mean head circumference were similar throughout the follow- up period. Nonhormonal users reported significantly more self-perceived decreases in milk production at the 5th an 6th month follow-up intervals. Acceptance and continued use of the pill were excellent, with only 1 women discontinuing because of a pregnancy which was the result of user failure. The principal side effect reported by women in both groups was intermenstrual bleeding.

Oxidation of coniferyl alcohol by cell wall peroxidases at the expense of indole-3-acetic acid and O2. A model for the lignification of plant cell walls in the absence of H2O2.

The oxidation of coniferyl alcohol (CA), a lignin precursor, by cell wall peroxidases may take place at the expense of indole-3-acetic acid (IAA) and O2, and in the absence of H2O2. The peroxidase-catalyzed oxidation of CA shows an optimum at an IAA concentration of 0.33 mM, while higher IAA concentrations are inhibitory. The observation that the oxidation of CA by cell wall peroxidase at the expense of IAA and O2 is inhibited by genistein, a putative endogenous inhibitor of lignification in lupin hypocotyls, supports the view that the H2O2-generating system coexists with cell wall peroxidase activities involved in lignification, and that it takes place at the expense of IAA and O2.

Distribution of isoflavones in lupin hypocotyls. Possible control of cell wall peroxidase activity involved in lignification.

The distribution of 4 key isoflavones (luteone, genistein, 2'-hydroxygenistein and wighteone) in lupin (Lupinus albus L. cv. multolupa) hypocotyls shows a gradient that diminishes from young to old tissues. A spatial gradient occurs within the hypocotyl, and a temporal gradient in both the outermost vascular and epidermal tissues. Not only does a gradient exist in respect to the quantity of isoflavones, but there is also a gradient in respect to the type of isoflavone. Thus, wighteone is mainly associated with the non-meristematic zones of the lupin hypocotyl. A close relationship was found between the distribution and the localization in the walls of phloem cells of both peroxidase (EC 1.11.1.7) and isoflavones. This observation suggests an in vivo peroxidase-isoflavone interconnection. In fact, lupin isoflavones are able to inhibit the peroxidase-catalyzed oxidation of the lignin precursor coniferyl alcohol, probably due to the co-oxidation of isoflavones in the reaction media. The results are discussed on the basis of a possible role for isoflavones in controlling cell wallperoxidase activity involved in the lignification of phloem cells.

Indole-3-methanol is the main product of the oxidation of indole-3-acetic acid catalyzed by two cytosolic basic isoperoxidases from Lupinus.

The nature of the products of the auxin catabolism mediated by both basic and acidic isoperoxidases has been studied. While indole-3-methanol is only a minor product of the oxidation of indole-3-acetic acid catalyzed by extracellular acidic isoperoxidases, it is the only product of the oxidation of indole-3-acetic acid catalyzed by two cytosolic basic isoperoxidases (EC 1.11.1.7) from lupin (Lupinus albus L.) hypocotyls. The putative indole-3-methanol formed by these latter isoperoxidases was isolated and then characterized by mass spectrometry and (1)H-nuclear magnetic resonance spectrometry. These results are discussed with respect to the diversity and compartmentation of the catabolism of indole-3-acetic acid in plant tissues.