Digestive enzyme activities during early ontogeny in Common snook (Centropomus undecimalis).

Common snook (Centropomus undecimalis) is one of the most important marine species under commercial exploitation in the Gulf of Mexico; for this reason, interest in developing its culture is a priority. However, larviculture remains as the main bottleneck for massive production. In this sense, our objective was to determine the changes of digestive enzymes activities using biochemical and electrophoretic techniques during 36 days of Common snook larviculture fed with live preys (microalgae, rotifers, and Artemia). During larviculture, all digestive enzymatic activities were detected with low values since yolk absorption, 2 days after hatching (dah) onwards. However, the maximum values for alkaline protease (6,500 U mg protein(-1)), trypsin (0.053 mU × 10(-3) mg protein(-1)), and Leucine aminopeptidase (1.4 × 10(-3) mU mg protein(-1)) were detected at 12 dah; for chymotrypsin at 25 dah (3.8 × 10(-3) mU mg protein(-1)), for carboxypeptidase A (280 mU mg protein(-1)) and lipase at 36 dah (480 U mg protein(-1)), for α-amylase at 7 dah (1.5 U mg protein(-1)), for acid phosphatases at 34 dah (5.5 U mg protein(-1)), and finally for alkaline phosphatase at 25 dah (70 U mg protein(-1)). The alkaline protease zymogram showed two active bands, the first (26.3 kDa) at 25 dah onwards, and the second (51.6 kDa) at 36 dah. The acid protease zymogram showed two bands (RF = 0.32 and 0.51, respectively) at 34 dah. The digestive enzymatic ontogeny of C. undecimalis is very similar to other strictly marine carnivorous fish, and we suggest that weaning process should be started at 34 dah.

On-line arsenic co-precipitation on ethyl vinyl acetate turning-packed mini-column followed by hydride generation-ICP OES determination.

An alternative and new system for on-line preconcentration of arsenic by sorption on a mini-column associated to hydride generation--inductively coupled plasma--optical emission spectrometry determination was studied. It is based on the sorption of arsenic on a column packed with ethyl vinyl acetate (EVA) turnings and the use of La(III) as co-precipitant reagent. This polymeric material was employed here for the first time as filling material for column preconcentration. It could work both as adsorbent and as sieve material. Sample and co-precipitant agent (lanthanum nitrate) were off-line mixed and merged with ammonium buffer solution (pH 10.0), which promoted precipitation and quantitative collection on the small EVA turnings. The arsenic preconcentrated by co-precipitation with lanthanum hydroxide precipitate was subsequently eluted with hydrochloric acid, which was the medium used for hydride generation. Considering a flow rate of 5 ml/min, three enrichment factors were obtained, 28-, 38- and 45-fold at three different sampling times, 60, 120 and 180s; respectively. The detection limits (3s) obtained for each case were 0.013, 0.009 and 0.007 microg/l. Additionally, the calculated precisions expressed as relatively standard deviation (R.S.D.) were 0.9, 1.3 and 1.1%. Satisfactory results were obtained for the determination of arsenic in standard reference material NIST 1643e Trace Elements in Water and drinking water samples.

On-line preconcentration and determination of mercury in biological and environmental samples by cold vapor-atomic absorption spectrometry.

An on-line procedure for the determination of traces of total mercury in environmental and biological samples is described. The present methodology combines cold vapor generation associated to atomic absorption spectrometry (CV-AAS) with preconcentration of the analyte on a minicolumn packed with activated carbon. The retained analyte was quantitatively eluted from the minicolumn with nitric acid. After that, volatile specie of mercury was generated by merging the acidified sample and sodium tetrahydroborate(III) in a continuous flow system. The gaseous analyte was subsequently introduced via a stream of Ar carrier into the atomizer device. Optimizations of both, preconcentration and mercury volatile specie generation variables were carried out using two level full factorial design (2(3)) with 3 replicates of the central point. Considering a sample consumption of 25mL, an enrichment factor of 13-fold was obtained. The detection limit (3sigma) was 10ngL(-1) and the precision (relative standard deviation) was 3.1% (n=10) at the 5microgL(-1) level. The calibration curve using the preconcentration system for mercury was linear with a correlation coefficient of 0.9995 at levels near the detection limit up to at least 1000microgL(-1). Satisfactory results were obtained for the analysis of mercury in tap water and hair samples.

Determination of the binding constant of indomethacin-beta-cyclodextrin complex by capillary electrophoresis: experimental optimization and temperature study.

The apparent electrophoretic mobilities of indomethacin in beta-cyclodextrin at a range of concentrations were measured directly by capillary electrophoresis. Three different linear plots and a non linear plot are proposed for the apparent binding constant calculations, based on the fact that the molar ratio of the inclusion complex was 1:1. K values obtained at 298 K were 421 M(-1) (double reciprocal fit), 488 M(-1) (x-reciprocal fit), 428 M(-1) (y-reciprocal fit) and 490 M(-1) (non linear fit). The corresponding K values at 313 K were 380 M(-1) (double reciprocal fit), 355 M(-1) (x-reciprocal fit), 366 M(-1) (y-reciprocal fit) and 339 M(-1) (non linear fit). Using the proposed methods, the binding constant of the indomethacin-beta-cyclodextrin inclusion complex can be obtained easily. The methods have been applied to obtain the values of the constant K under different experimental conditions. Under optimized conditions the K constant is temperature dependent and non-arrhenian behaviour was observed.

Fingerprint analysis and synthetic adulterant search in Hedera helix formulations by capillary electrophoresis.

A high-performance capillary electrophoretic (CE) method has been developed for obtaining electropherograms of various extracts and the commercial formulation (fingerprints) of Hedera helix L used in Argentina as a cough's treatment. Also, a capillary zone electrophoresis (CZE) method was developed for the search, identification and determination of some possible adulterants. These likely adulterants are common synthetic drugs used in respiratory diseases (antitussive, decongestant and bronchodilator agents). Under optimum conditions, the analytes (ephedrine, codeine, diphenhydramine and constituents of H. helix formulations) were separated within less than 10 min in 20 mM sodium tetraborate buffer (pH 9.0). The present procedure was validated with respect to selectivity, linearity range, limits of detection (LOD) and quantification (LOQ), precision (repeatability and intermediate precision), solution stability and accuracy; the results obtained were satisfactory. Good linearity was obtained over two orders of magnitude and detection limits (S/N = 3) were better than 1.2 microg ml(-1) for all analytes. The CE methodology was successfully applied to the search and subsequent determination of ephedrine, codeine and diphenhydramine in H. helix extracts and its phytopharmaceutical products.

Trace element determination of Argentine wines using ETAAS and USN-ICP-OES.

The objective of this work was to develop a method to determine the metal content in wine samples from the province of Mendoza in Argentina. Ten samples of white wine and 10 samples of red wine available in the supermarket were analyzed for the metals aluminium, cadmium, calcium, chromium, copper, iron, nickel, lead and zinc by electrothermal atomic spectrometry (ETAAS) and ultrasonic nebulization was coupled to inductively coupled plasma optical emission spectrometry (USN-ICP-OES). The aluminium, cadmium, calcium, copper, iron, lead, zinc, chromium concentrations were between 17.0-18.0 microg l(-1), 1.0-4.7microg l(-1), 10.0-15.0 mg l(-1), 23.0-28.0 microg l(-1), 480-790 microg l(-1), 50-90 microg l(-1), 24-130 microg l(-1), and <0.2-6.25 microg l(-1), respectively. The levels compare well with those reported for similar wines from some other parts of the world. A significant aspect in this paper is the samples mineralization step, which allowed the direct determination of the metals. Concerning to the Cd determination, a refluxing digestion system was used for the pretreatment of the samples.

On-line preconcentration/determination of copper in parenteral solutions using activated carbon by inductively coupled plasma optical emission spectrometry.

A method for the on-line preconcentration of copper using a minicolumn packed with activated carbon and its subsequent determination by inductively coupled plasma optical emission spectrometry (ICP-OES) coupled with flow injection (FI) was studied. In order to determinate the copper concentration present in parenteral solutions; it was retained on activated carbon (AC) at pH 9.5. A sensitivity enrichment factor of 30-fold was obtained with respect to the copper determination by ICP-OES without preconcentration. The detection limit for the preconcentration of 25 ml of sample was 0.1 microgl(-1). The precision for the ten replicate determinations at the 2.5 microgl(-1) Cu level was 3.0% relative standard deviation (R.S.D.), calculated with the peak heights. The calibration graph using the preconcentration method for cooper species was linear with a correlation coefficient of 0.9996 at levels near the detection limits up to at least 200 microgl(-1). The method was successfully applied to the determination of copper in parenteral solutions.

On-line preconcentration/determination of lead in Ilex paraguariensis samples (mate tea) using polyurethane foam as filter and USN-ICP-OES.

The present paper proposes an on-line preconcentration procedure for lead determination in Ilex paraguariensis (St. Hilaire) samples by ultrasonic nebulization associated to inductively coupled plasma optical emission spectrometry (USN-ICP-OES). It is based on the precipitation of lead(II) ion on a minicolumn packed with polyurethane foam using 2-(5-bromo-2-pyridilazo)-5-diethylaminophenol (5-Br-PADAP) as precipitating reagent. The collected analyte precipitate was quantitatively eluted from the minicolumn with 20% (v/v) nitric acid. An enhancement factor of 225-fold was obtained (15 for USN and 15 for preconcentration). The detection limit (DL) value for the preconcentration of 10.0 ml of sample was 40.0 ng/l. The relative standard deviation (R.S.D.) was 3.0% for a Pb concentration of 1 microg/l, calculated from the peak heights obtained. The calibration graph using the preconcentration system for lead was linear with a correlation coefficient of 0.9997, at levels near the detection limits up to at least 100 microg/l. The preconcentration procedure was successfully applied to the determination of lead in mate tea samples.

Enhancement of glycine receptor function by ethanol: role of phosphorylation.

1. The effects of several kinase inhibitors (staurosporine, GF 109203X, H89, KN62, genistein) and of the phosphatase inhibitor calyculin A were studied on the ethanol potentiation and on the function of homomeric alpha1 glycine receptor expressed in Xenopus oocytes using a two electrode voltage clamp recording technique. 2. The function of the homomeric alpha1 glycine receptor was not modified in Xenopus oocytes pretreated with kinase inhibitors or with the phosphatase inhibitor calyculin A. 3. The potentiation of the glycine receptor function induced by ethanol (10-200 mM) was significantly reduced in Xenopus oocytes pretreated with the PKC inhibitors staurosporine or GF 109203X. 4. No differences in propofol (2.5 microM) or halothane (250 microM) actions were found after exposure of Xenopus oocytes to staurosporine. 5. No differences in ethanol sensitivity were found after exposure of Xenopus oocytes expressing glycine alpha1 receptors to H89, KN62, genistein or to the phosphatase inhibitor calyculin A. 6. The mutant alpha1 (S391A), in which the PKC phosphorylation site at serine 391 was mutated to alanine, was less sensitive to the effects of ethanol than was the alpha1 wild type receptor. Moreover, the ethanol potentiation of the glycine receptor function was not affected by treatment with staurosporine in oocytes expressing alpha1 (S391A). 7. The splice variant of the alpha1 glycine receptor subunit, alpha1ins, containing eight additional amino acids and a potential phosphorylation site for PKA, did not differ from wild type for sensitivity to ethanol. 8. These results indicate that phosphorylation by PKC of the homomeric alpha1 glycine receptor subunit modulates ethanol potentiation, but not the function of the glycine receptor.

Biphasic effect of cadmium in non-cytotoxic conditions on the secretion of nitric oxide from peritoneal macrophages.

The effects of cadmium (Cd) in non-cytotoxic conditions on the nitric oxide (NO) production in peritoneal macrophages (pM) were studied. Peritoneal macrophages from Balb/c mice were incubated over 18 h with 5, 10, 20, or 25 microM Cd2+ (as CdCl2 21:2 H2O) in the culture medium. Concentrations of 20 microM Cd2+ and over had cytotoxic effects, measured by MTT assay. Cell viability with 10 microM Cd2+ in the medium was above 90% after 18 h of incubation, and above 80% after 72 h. At this same Cd2+ concentration, NO production increased from 6 to 18 h. At 24 h production decreased but was still above control levels. At 48 h production NO was near control levels, and continued to decrease until the end of the experiment (72 h). NO levels produced with Cd2+ concentrations of 5, 10 and 20 microM in the medium were above the control at 18 h. NO production and lipoperoxidation increased simultaneously after 18 h with 10 microM of Cd in the medium. Amounts of inducible nitric oxide synthase (iNOS) protein and iNOS activity also increased. At a concentration of 10 microM Cd has a biphasic effect on NO production over time.

Spectrophotometric determination of gadolinium(III) with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP).

Reaction between gadolinium(III) and 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5- Br-PADAP) was studied for delineating optimal conditions for complexation. This reagent can be used for the spectrophotometric determination of Gd(III) in concentrations ranging from 0.04 to 1.2 ppm (a = 1.76(+/- 0.03) x 10(5) (1.(-1) mole(-1). cm). The reaction takes place at a pH between 9.2 and 11.6. In the presence of Triton X-100 this complex is soluble in water. In order to overcome difficulties caused by the presence of other lanthanides, an ion exchange chromatographic technique was used.

Optimization of the preconcentration system of cadmium with 1(2-thiazolylazo)-p-cresol using a knotted reactor and flame atomic absorption spectrometric detection.

The present paper proposes an on-line preconcentration procedure for cadmium determination in drinking water samples. It is based on the precipitation of cadmium(II) ions on a knotted reactor (KR) using 1(2-thiazolylazo)-p-cresol (TAC) as complexing reagent. The optimization step was performed using a full factorial design involving the variables: pH, eluent concentration (nitric acid) and TAC concentration. The results of this experiment demonstrated that these variables at chosen levels are not statistically significant. Under optimized experimental established conditions, analytical parameters for the preconcentration method were: a detection limit of 40.0 ng/l, precision as relative standard deviation (RSD) of 1.2 and 1.0%, for cadmium concentration of 2.5 and 20.0 microg/l, respectively. The preconcentration factor considering the slopes of the analytical curves with and without preconcentration is 23 for a sample volume of 10 ml. This system shows a sampling frequency of 25 h(-1). In order to check the accuracy, the standard reference material, NIST SRM 1643d trace elements in water was analyzed. A comparison, using t-test demonstrates that there is not significant difference among the achieved results with proposed method and the certified values. The addition/recovery experiments in the samples analyzed demonstrated the accuracy and applicability of the system developed for cadmium determination in water samples.

On-line complexation/preconcentration system for the determination of lead in wine by inductively coupled plasma-atomic emission spectrometry with ultrasonic nebulization.

An on-line lead preconcentration and determination system implemented with inductively coupled plasma-atomic emission spectrometry (ICP-AES) with ultrasonic nebulization (USN) in association with flow injection was studied. For the preconcentration of lead, a Pb-quinolin-8-ol complex was formed on-line at pH 6.8 and retained on Amberlite XAD-16 resin. The lead was removed from the microcolumn by countercurrent elution with nitric acid. A total enhancement factor of 225 was obtained with respect to ICP-AES with pneumatic nebulization (15.0 for USN and 15.0 for the column). The detection limit for Pb for the preconcentration of a 10 mL wine sample was 0.15 microg/L. The precision for 10 replicate determinations at a Pb level of 25 microg/L was a relative standard deviation of 2.5%, calculated from the peak heights obtained. The calibration graph obtained by using the preconcentration system for lead was linear with a correlation coefficient of 0.9995 for levels near the detection limit up to > or = 1000 microg/L. The method was successfully applied to the determination of lead in wine samples.

Morphogenesis in short-term and long-term anther derived calli of Oenothera hookeri de vries.

Anther culture of O. hookeri on Murashige and Skoog (1962) medium supplemented with 2 mg l-1 2,4-dichlorophenoxyacetic acid and 2 mg-1 1-naphthaleneacetic acid produced callus formation. When subcultured onto medium lacking auxin, the callus regenerated through the organogenic pathway. Non-organogenic and organogenic callus was observed using histological methods after 2, 3 and 24 weeks in culture. Three types of calli were recognized: non-organogenic friable calli, organogenic friable calli with roots and organogenic hard calli with shoots. The microscopical sections showed striking differences in tissue organization among friable and compact calli. Vascular bundles were prominent in compact calli, but were not found in friable calli. Calli sections showed at light microscopy cells at two developmental stages; differentiated highly vacuolated cells and meristematic small isodiametric cells with densely stained cytoplasm. At electron microscopy level abnormal chloroplasts were present in non-organogenic calli, while chloroplasts were well developed in organogenic hard calli. Peroxisomes with paracrystalline protein bodies were abundant in both types of calli.

Preconcentration and speciation of chromium in drinking water samples by coupling of on-line sorption on activated carbon to ETAAS determination.

An on-line flow injection (FI) preconcentration-electrothermal atomic absorption spectrometry (ETAAS) method is developed for trace determination of chromium in drinking water samples by sorption on a conical minicolumn packed with activated carbon (AC) at pH 5.0. The chromium was removed from the minicolumn with 1.0% (v/v) nitric acid. An enrichment factor (EF) of 35-fold for a sample volume of 10ml was obtained. The detection limit (DL) value for the preconcentration method proposed was 3.0ngl(-1). The precision for 10 replicate determinations at the 0.5mugl(-1) Cr level was 4.0% relative standard deviation (R.S.D.), calculate with the peak heights obtained. The calibration graph using the preconcentration system for chromium was linear with a correlation coefficient of 0.9992 at levels near the detection limits up to at least 50mugl(-1). The method was successfully applied to the determination of Cr(III) and Cr(VI) in drinking water samples.

[Analysis of cesarean section indication at a center for active management of labor]. / Análisis de la indicación de cesárea en un centro de manejo activo del parto.

Since 1985, when active management of labor was introduced at Hospital Dr. Gustavo Fricke, a significative decrease of cesarean section delivery has been observed. In 918 women operated on between October 1987 and March 1989, the main indication was as follow: elective cesarean section, repeated section, fetal distress in labor and abnormal lying of the fetus with an incidence of 31.2%, 18.4%, 17.9% and 14.3% respectively. Dystocia, which appears to be the main cause in international literature represented only 12.3% of the total cesarean section in our service.

On-line preconcentration and determination of nickel in natural water samples by flow injection-inductively coupled plasma optical emission spectrometry (FI-ICP-OES).

An on-line nickel preconcentration and determination system implemented with inductively coupled plasma optical emission spectrometry (ICP-OES) associated to flow injection (FI) was studied. Trace amounts of nickel were preconcentrated by sorption on a conical minicolumn packed with activated carbon (AC) at pH 5.0. The nickel was removed from the minicolumn with 20% nitric acid. An enrichment factor of 80-fold for a sample volume of 50 ml was obtained. The detection limit (DL) value for the preconcentration method proposed was 82 ng l(-1). The precision for ten replicate determinations at the 0.5 microg l(-1) Ni level was 3.0% relative standard deviation (R.S.D.), calculated from the peak heights obtained. The calibration graph preconcentration method for nickel was linear with a correlation coefficient of 0.9997 at levels near the detection limits (DL) up to at least 100 microg l(-1). The method was successfully applied to the determination of nickel in natural water samples.

Speciation and preconcentration of vanadium(V) and vanadium(IV) in water samples by flow injection-inductively coupled plasma optical emission spectrometry and ultrasonic nebulization.

An on-line separation, preconcentration and determination system for vanadium(IV) and vanadium(V) comprising inductively coupled plasma optical emission spectrometry (ICP-OES) coupled to a flow injection (FI) method with an ultrasonic nebulization (USN) system was studied. The vanadium species were retained on an Amberlite XAD-7 resin as a vanadium-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (V-5-Br-PADAP) complex at pH 3.7. Enhanced selectivity was obtained with the combined use of the formation on-line of the complexes and 1,2-cyclohexanediaminetetraacetic acid (CDTA) as masking agent. The vanadium complexes were removed from the microcolumn with 25% v/v nitric acid. A sensitivity enhancement factor of 225 was obtained with respect to ICP-OES using pneumatic nebulization (15-fold for USN and 15-fold for the microcolumn). The detection limit for the preconcentration of 10 mL of aqueous solution was 19 ng L-1. The precision for 10 replicate determinations at the 5 micrograms L-1 V level was 2.3% relative standard deviation (RSD), calculated from the peak heights obtained. The calibration graph using the separation and preconcentration system for vanadium species was linear with a correlation coefficient of 0.9992 at levels from near the detection limits up to at least 100 micrograms L-1. The method was successfully applied to the speciation of vanadium in river water samples.