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J A Hernández

Publications and source records attributed to J A Hernández.

At least 19 recordsLinked to original sources
PubMed2007

Expression of furA is modulated by NtcA and strongly enhanced in heterocysts of Anabaena sp. PCC 7120.

Fur (ferric uptake regulator) proteins are principally responsible for maintaining iron homeostasis in prokaryotes. Iron is usually a scarce resource. Its limitation reduces photosynthetic rates and cell growth in cyanobacteria in general and especially in cyanobacteria that are fixing dinitrogen, a process that requires the synthesis of numerous proteins with a high content of iron. This paper shows that in the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120, levels of furA mRNA and FurA protein increase significantly in response to nitrogen deprivation, and that furA up-regulation takes place specifically in proheterocysts and mature heterocysts. Great differences in a Northern blot, probed with furA, of RNA from an ntcA mutant relative to wild-type Anabaena sp. were attributable to binding of NtcA, a global regulator of nitrogen metabolism, to the promoter of furA and to the promoter of the furA antisense transcript alr1690-alpha-furA.

Anabaena↗
PubMed2006

A single dose of bovine somatotropin 5 days before the end of progestin-based estrous synchronization increases prolificacy in sheep.

Bovine somatotropin (bST) enhances ovarian follicular and embryonic development in sheep and cattle. In the present study, the objective was to assess whether bST given 5 days before the end of progestin-based estrous synchronization improves prolificacy and lambing rate in sheep. Pelibuey ewes (n=92) exhibiting estrous cycles at regular intervals received an intravaginal sponge containing 45mg of FGA for 12 days. Five days before sponge withdrawal, ewes were treated with either 125mg of bST sc (bST group; n=47) or saline solution (control; n=45). After the sponge was removed, ewes were observed for estrus and subsequently mated twice. Lambing rate and prolificacy was determined at birth. Blood samples were taken from the time of treatment until day 15 after estrus in eight ewes from the bST group and nine from the control group. Concentrations of IGF-I were determined by immunoradiometric assay and progesterone by RIA. Treatment with bST increased (P<0.01) the proportion of ewes with more than one lamb (bST, 56% compared with control, 26%) and prolificacy (bST, 1.6 compared with control, 1.3). Treatment with bST increased (P<0.05) the lambing rate of multiparous (bST, 92% compared with control, 67%) but not in ewes at the first time they were mated (bST, 71% compared with control, 87%; P>0.05). IGF-I concentrations were greater (P<0.01) in ewes treated with bST than in control ewes from 2 days after treatment. Progesterone concentrations did not vary (P>0.05) between groups. It is concluded that a single dose of bST 5 days before progestin withdrawal increases lambing rate and prolificacy in sheep. These effects are associated with an increase in circulating concentrations of IGF-I.

Animals↗
PubMed2006

The apoplastic antioxidant system in Prunus: response to long-term plum pox virus infection.

This work describes, for the first time, the changes taking place in the antioxidative system of the leaf apoplast in response to plum pox virus (PPV) in different Prunus species showing different susceptibilities to PPV. The presence of p-hydroxymercuribenzoic acid (pHMB)-sensitive ascorbate peroxidase (APX) (class I APX) and pHMB-insensitive APX (class III APX), superoxide dismutase (SOD), peroxidase (POX), NADH-POX, and polyphenoloxidase (PPO) was described in the apoplast from both peach and apricot leaves. PPV infection produced different changes in the antioxidant system of the leaf apoplast from the Prunus species, depending on their susceptibility to the virus. In leaves of the very susceptible peach cultivar GF305, PPV brought about an increase in class I APX, POX, NADH-POX, and PPO activities. In the susceptible apricot cultivar Real Fino, PPV infection produced a decrease in apoplastic POX and SOD activities, whereas a strong increase in PPO was observed. However, in the resistant apricot cultivar Stark Early Orange, a rise in class I APX as well as a strong increase in POX and SOD activities was noticed in the apoplastic compartment. Long-term PPV infection produced an oxidative stress in the apoplastic space from apricot and peach plants, as observed by the increase in H2O2 contents in this compartment. However, this increase was much higher in the PPV-susceptible plants than in the resistant apricot cultivar. Only in the PPV-susceptible apricot and peach plants was the increase in apoplastic H2O2 levels accompanied by an increase in electrolyte leakage. No changes in the electrolyte leakage were observed in the PPV-inoculated resistant apricot leaves, although a 42% increase in the apoplastic H2O2 levels was produced. Two-dimensional electrophoresis analyses revealed that the majority of the polypeptides in the apoplastic fluid had isoelectric points in the range of pI 4-6. The identification of proteins using MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) and peptide mass fingerprinting analyses showed the induction of a thaumatin-like protein as well as the decrease of mandelonitrile lyase in peach apoplast due to PPV infection. However, most of the selected polypeptides showed no homology with known proteins. This fact emphasizes that, at least in Prunus, most of the functions of the apoplastic space remain unknown. It is concluded that long-term PPV infection produced an oxidative stress in the leaf apoplast, contributing to the deleterious effects produced by PPV infection in leaves of inoculated, susceptible Prunus plants.

Antioxidants↗
PubMed2006

Epithelial fluid transport: protruding macromolecules and space charges can bring about electro-osmotic coupling at the tight junctions.

The purpose of the present work is to investigate whether the idea of epithelial fluid transport based on electro-osmotic coupling at the level of the leaky tight junction (TJ) can be further supported by a plausible theoretical model. We develop a model for fluid transport across epithelial layers based on electro-osmotic coupling at leaky tight junctions (TJ) possessing protruding macromolecules and fixed electrical charges. The model embodies systems of electro-hydrodynamic equations for the intercellular pathway, namely the Brinkman and the Poisson-Boltzmann differential equations applied to the TJ. We obtain analytical solutions for a system of these two equations, and are able to derive expressions for the fluid velocity profile and the electrostatic potential. We illustrate the model by employing geometrical parameters and experimental data from the corneal endothelium, for which we have previously reported evidence for a central role for electro-osmosis in translayer fluid transport. Our results suggest that electro-osmotic coupling at the TJ can account for fluid transport by the corneal endothelium. We conclude that electro-osmotic coupling at the tight junctions could represent one of the basic mechanisms driving fluid transport across some leaky epithelia, a process that remains unexplained.

Animals↗
PubMed2006

Interaction of FurA from Anabaena sp. PCC 7120 with DNA: a reducing environment and the presence of Mn(2+) are positive effectors in the binding to isiB and furA promoters.

The Fur (ferric uptake regulator) protein is a global regulator in most prokaryotes that controls a large number of genes. Fur is a classical repressor that uses ferrous iron as co-repressor and binds to specific DNA sequences (iron boxes) as a dimer. Three different genes coding for Fur homologues have been identified in Anabaena sp. PCC 7120. FurA controls the transcription of flavodoxin, the product of the isiB gene, and is moderately autoregulated. In this work, the promoter of the furA gene was defined and the FurA protected regions in the furA and isiB promoters were identified, showing that the binding sites for Anabaena FurA contain A/T-rich sequences with a variable arrangement compared to the conventional 19-base pair Fur consensus. The influence of different factors on the interaction between FurA and the promoters was evaluated in vitro. The affinity of FurA for the DNA targets was significantly affected by the redox status of this regulator and the presence of Mn(2+). The optimal binding conditions were observed in the presence of both Mn(2+) and DTT. Those results suggest that, in addition to iron availability, FurA-DNA interaction is modulated by redox conditions.

Anabaena↗
PubMed2006

The cellular resistance against oxidative stress (H2O2) is independent of neutral trehalase (Ntc1p) activity in Candida albicans.

The protective role of trehalose against oxidative stress caused by hydrogen peroxide in Candida albicans has been investigated in the homozygous mutant ntc1Delta/ntc1Delta, disrupted in the NTC1 gene, which encodes the neutral (cytosolic) trehalase (Ntc1p). After a severe oxidative exposure (50 mM H(2)O(2)), both parental (CAI-4) and ntc1Delta/ntc1Delta exponential-phase cells stored large amounts of intracellular trehalose. In turn, the degree of cell survival was roughly equivalent in both strains, although slightly higher in ntc1Delta/ntc1Delta cultures. The mechanism of 'adaptive tolerance' was functional in the two strains. Thus, a gently oxidative pretreatment (5 mM H(2)O(2)) increased the recovery of cellular viability when it was followed by a severe challenge (50 mM H(2)O(2)); this phenomenon was accompanied by a significant elevation of the endogenous trehalose content. Oxidative stress also induced specific activation of the antioxidant enzymes catalase and glutathione reductase upon gentle oxidative treatment (5 mM H(2)O(2)), whereas superoxide dismutase activity was only activated upon prolonged exposure. Taken together, these results strongly suggest that in C. albicans neutral trehalase activity does not play an essential role in the protective response against oxidative stress. They also suggest that a diminished Ntc1p activity might favour the growth of C. albicans cells subjected to a strong oxidative exposure.

Candida albicans↗
PubMed2005

A general channel model accounts for channel, carrier, counter-transport and co-transport kinetics.

In this work we propose a unifying model of mediated membrane transport, based upon the idea that the integral membrane proteins involved in these processes operate via complex channel mechanisms. In the first part, we briefly review literature about the structural aspects of membrane transporters. We conclude that there is a substantial amount of evidence suggesting that most membrane proteins performing transport are embodied with channel-like structures that may constitute the translocation paths. This includes cases where the phenomenological transport kinetics do not correspond to the classical channel behavior. In the second part of this article we introduce the general channel model of mediated transport and employ it to derive specific examples, like simple one- or two-ligand channels, water-ligand channels, simple carriers, co- and counter-transport systems and more complex water-ligand carriers. We show that, for the most part, these particular cases can be obtained by the application of the techniques of diagram reduction to the full model. The necessary conditions for diagram reduction reflect physical properties of the protein and its surroundings.

Biological Transport, Active↗
PubMed2002

Thermoluminescence properties of KCl(1-X)KBrX:Pb2+ mixed crystals.

A study is presented of the thermoluminescence (TL) of phosphors based on potassium halides doped with divalent lead, such as KCl:Pb2+, KBr:Pb2+ and the crystalline series KCl(1-X)Br(X):Pb2+. The defects in the crystals generated by irradiation have been investigated as well as the trapping and room temperature recombination mechanisms. The samples were gamma irradiated to a dose of 10 kGy. The TL spectral response during the recombination stage was obtained through simultaneous measurements of temperature, emitted light intensity and emission wavelength by using an automated thermoluminescence system with optical fibre couplings and a diode array as a detection device. The TL emission is a broad band in the 350-600 nm range with a maximum strongly dependent upon the mixed composition.

Bromides↗
PubMed2001

Open channel-like behavior of reduced carrier models.

Previous theoretical analyses have demonstrated that some extended channel models can exhibit carrier kinetics, even for the case of models where the transporter does not experience any conformational change. The purpose of this work is to explore the inverse situation, that is, the finding of channel-like transport properties in well-defined carrier models. This approximate behavior has been previously encountered for the case that rapid equilibrium is established between the empty carrier states. This study explores another situation in which a typical carrier can exhibit open channel-like kinetic properties, namely, the reduction of a classical four-state simple carrier model for the case that one of the empty states is a transient intermediate. For this case I analyze here if, for some conditions, the kinetics of the three-state reduced model can be satisfactorily approximated by a channel description. The analysis performed here derives conditions under which this reduced model exhibits channel-like transport kinetics. The work is illustrated by the numerical studies of two comparative examples: one corresponding to a carrier satisfying the necessary conditions for the channel-like behavior, and another for a carrier not meeting those conditions. The finding that a well-defined carrier model may exhibit channel-like transport properties represents a further argument in favor of the concept that typical channels and carriers are particular cases of more general mechanisms of facilitative transport.

Biological Transport, Active↗
PubMed2001

Cloning, overexpression and interaction of recombinant Fur from the cyanobacterium Anabaena PCC 7119 with isiB and its own promoter.

A gene coding for a Fur (ferric uptake regulation) protein from the cyanobacterium Anabaena PCC 7119 has been cloned and overexpressed in Escherichia coli. DNA sequence analysis confirmed the presence of a 151-amino-acid open reading frame that showed homology with the Fur proteins reported for the unicellular cyanobacteria Synechococcus 7942 and Synechocystis PCC 6803. Two putative Fur-binding sites were detected in the promoter regions of the fur gene from Anabaena. Partially purified recombinant Fur binds to the flavodoxin promoter as well as its own promoter. This suggests that the Fur gene is autoregulated in Anabaena.

Amino Acid Sequence↗
PubMed2001

Identification and characterization by high-performance liquid chromatography/electrospray ionization mass spectrometry of a new variant hemoglobin, Mataro [beta134(H12) Val > Ala.

This work illustrates the practical use of combined microbore reversed-phase high-performance liquid chromatography (RP-HPLC) with electrospray ionization mass spectrometry (ESI-MS) in protein identification. The approach consisted of the detection of the abnormal beta-globin chain by ESI-MS analysis of mixtures of intact globins, which simultaneously provided their molecular masses. Separation of the polypeptide globin chains was carried out using microbore C4 RP-HPLC on-line with ESI-MS. Direct peptide-mapping ESI-MS without previous chromatographic separation was performed in order to identify tryptic peptides from whole blood. For the sequence confirmation of the abnormal peptide containing the mutation point, C18 RP-HPLC tryptic separation of the globin mixture on-line with collision-induced dissociation (CID) fragmentation was done. The y series ions allowed the identification of the hemoglobin (Hb) variant as [beta134(H12) Val > Ala]. This new Hb was named Hb Mataró, after the city where it was detected.

Alanine↗
PubMed2001

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.

Adaptation, Physiological↗
PubMed2000

An elementary kinetic model of energy coupling in biological membranes.

The purpose of this work is to contribute to the understanding of the fundamental kinetic properties of the processes of energy coupling in biological membranes. For this, we consider a model of a microorganism that, in its plasma membrane, expresses two electrogenic enzymes (E(1) and E(2)) transporting the same monovalent cation C and electrodiffusive paths for C and for a monovalent anion A. E(1) (E(2)) couples transport C to the reaction S(1)<-->P(1) (S(2)<-->P(2)). We developed a mathematical model that describes the rate of change of the electrical potential difference across the membrane, of the internal concentrations of C and A, and of the concentrations of S(2) and P(2). The enzymes are incorporated via two-state kinetic models; the passive ionic fluxes are represented by classical formulations of electrodiffusion. The microorganism volume is maintained constant by accessory regulatory devices. The model is utilized for stationary and dynamic studies for the case of bacteria employing the electrochemical gradient of Na(+) as energetic intermediate. Among other conclusions, the results show that the membrane potential represents the relevant kinetic intermediate for the overall coupling between the energy donor reaction S(1)<-->P(1) and the synthesis of S(2).

Bacteria↗
PubMed2000

Electrogenic properties of the sodium pump in a dynamic model of membrane transport.

The general purpose of this theoretical work is to contribute to understand the physiological role of the electrogenic properties of the sodium pump, by studying a dynamic model that integrates diverse processes of ionic and water transport across the plasma membrane. For this purpose, we employ a mathematical model that describes the rate of change of the intracellular concentrations of Na+, K+ and Cl-, of the cell volume, and of the plasma membrane potential (Vm). We consider the case of a nonexcitable, nonpolarized cell expressing the sodium pump; Na+, K+, Cl- and water channels, and cotransporters of KCl and NaCl in its plasma membrane. We particularly analyze here the conditions under which the physiological Vm can be generated in a predominantly electrogenic fashion, as a result of the activity of the sodium pump. A major conclusion of this study is that, for the cell model considered, a low potassium permeability is not a sufficient condition for a predominantly electrogenic generation of the Vm by the sodium pump. The presence of an electroneutral exchange of Na+ and K+ represents a necessary additional requirement.

Biological Transport↗