Population dynamics of the long-snouted seahorse (Hippocampus guttulatus Cuvier, 1829) in the Mar Menor coastal lagoon.

A 10-year monitoring program was developed to quantify the population dynamics of the long-snouted seahorse population in the Mar Menor coastal lagoon. Based on 985 underwater visual censuses, we estimated the long-snouted seahorse (Hippocampus guttulatus Cuvier, 1829) population size in the Mar Menor lagoon and its reduction in size in the last decades, as well as the effect of eutrophication crises in 2016 and 2019 on the species. The annual recruitment for the 2013-2020 period was estimated by comparing the relative abundance of early seahorse life stages in the ichthyoplankton. The density ranged from 0.0458 specimens/m3 at the beginning of the sampling period to 0.0004 at the end, showing a statistically significant difference between the three analyzed periods (Hgl=2 = 14.0, p = 0.001). The long-snouted seahorse population from the Mar Menor lagoon exemplifies the impact of fishing activities and human pressure, especially euxinic episodes and habitat destruction. As a result of this, the Mar Menor population has decreased from several million specimens to a few thousand, in only three decades. This species showed considerable resilience, the seahorse population began to recover once fishing activity stopped. In contrast, the long-snouted seahorse showed high vulnerability to habitat loss and an episodic flooding event. Adult seahorses showed preferences for highly complex habitats, especially Caulerpa prolifera-Cymodocea nodosa mixed meadows and habitats of high complexity and anthropogenic origin, such as harbors, jetties, or breakwaters. In contrast, juvenile seahorses preferred monotonous seabeds with low complexity, such as the sandy beds that are characteristic of the Mar Menor lagoon littoral.

Dynamic modulation of inflammatory pain-related affective and sensory symptoms by optical control of amygdala metabotropic glutamate receptor 4.

Contrary to acute pain, chronic pain does not serve as a warning signal and must be considered as a disease per se. This pathology presents a sensory and psychological dimension at the origin of affective and cognitive disorders. Being largely refractory to current pharmacotherapies, identification of endogenous systems involved in persistent and chronic pain is crucial. The amygdala is a key brain region linking pain sensation with negative emotions. Here, we show that activation of a specific intrinsic neuromodulatory system within the amygdala associated with type 4 metabotropic glutamate receptors (mGlu4) abolishes sensory and affective symptoms of persistent pain such as hypersensitivity to pain, anxiety- and depression-related behaviors, and fear extinction impairment. Interestingly, neuroanatomical and synaptic analysis of the amygdala circuitry suggests that the effects of mGlu4 activation occur outside the central nucleus via modulation of multisensory thalamic inputs to lateral amygdala principal neurons and dorso-medial intercalated cells. Furthermore, we developed optogluram, a small diffusible photoswitchable positive allosteric modulator of mGlu4. This ligand allows the control of endogenous mGlu4 activity with light. Using this photopharmacological approach, we rapidly and reversibly inhibited behavioral symptoms associated with persistent pain through optical control of optogluram in the amygdala of freely behaving animals. Altogether, our data identify amygdala mGlu4 signaling as a mechanism that bypasses central sensitization processes to dynamically modulate persistent pain symptoms. Our findings help to define novel and more precise therapeutic interventions for chronic pain, and exemplify the potential of optopharmacology to study the dynamic activity of endogenous neuromodulatory mechanisms in vivo.

Nonconvex prior image constrained compressed sensing (NCPICCS): theory and simulations on perfusion CT.

PURPOSE: To present and evaluate a new image reconstruction method for dynamic CT based on a nonconvex prior image constrained compressed sensing (NCPICCS) algorithm. The authors systematically compared the undersampling potential, functional information recovery, and solution convergence speed of four compressed sensing (CS) based image reconstruction methods using perfusion CT data: Standard l1-based CS, nonconvex CS (NCCS), and l1-based and nonconvex CS, including an additional constraint based on a prior image (PICCS and NCPICCS, respectively). METHODS: The Shepp-Logan phantom was modified such that its uppermost ellipses changed attenuation through time, simulating both an arterial input function (AIF) and a homogeneous tissue perfusion region. Data were simulated with and without Poisson noise added to the projection data and subsequently reconstructed with all four CS-based methods at four levels of undersampling: 20, 12, 6, and 4 projections. Root mean squared (RMS) error of reconstructed images and recovered time attenuation curves (TACs) were assessed as well as convergence speed. The performance of both PICCS and NCPICCS methods were also evaluated using a kidney perfusion animal experiment data set. RESULTS: All four CS-based methods were able to reconstruct the phantoms with 20 projections, with similar results on the RMS error of the recovered TACs. NCCS allowed accurate reconstructions with as few as 12 projections, PICCS with as few as six projections, and NCPICCS with as few as four projections. These results were consistent for noise-free and noisy data. NCPICCS required the fewest iterations to converge across all simulation conditions, followed by PICCS, NCCS, and then CS. On animal data, at the lowest level of undersampling tested (16 projections), the image quality of NCPICCS was better than PICCS with fewer streaking artifacts, while the TAC accuracy on the selected region of interest was comparable. CONCLUSIONS: The authors have presented a novel method for image reconstruction using highly undersampled dynamic CT data. The NCPICCS method takes advantage of the information provided by a prior image, as in PICCS, but employs a more general nonconvex sparsity measure [such as the l(p)-norm (0 < p < or = 1)] rather than the conventional convex l1-norm. Despite the lack of guarantees of a globally optimal solution, the proposed nonconvex extension of PICCS consistently allowed for image reconstruction from fewer samples than the analogous l1-based PICCS method. Both nonconvex sparsity measures as well as prior image information (when available) significantly reduced the number of iterations required for convergence, potentially providing computational advantages for practical implementation of CS-based image reconstruction techniques.

Analysis of dynamic networks based on the Ising model for the case of study of co-authorship of scientific articles.

Two computational methods based on the Ising model were implemented for studying temporal dynamic in co-authorship networks: an interpretative for real networks and another for simulation via Monte Carlo. The objective of simulation networks is to evaluate if the Ising model describes in similar way the dynamic of the network and of the magnetic system, so that it can be found a generalized explanation to the behaviours observed in real networks. The scientific papers used for building the real networks were acquired from WoS core collection. The variables for each record took into account bibliographic references. The search equation for each network considered specific topics trying to obtain an advanced temporal evolution in terms of the addition of new nodes; that means 3 steps, a time to reach the interest of the scientific community, a gradual increase until reaching a peak and finally, a decreasing trend by losing of novelty. It is possible to conclude that both methods are consistent with each other, showing that the Ising model can predict behaviours such as the number and size of communities (or domains) according to the temporal distribution of new nodes.

[Factors associated to high-flow nasal cannula treatment failure in pediatric patients with respiratory failure in two pediatric intensive care units at high altitude]. / Factores asociados al fracaso de la terapia con cánulas nasales de alto flujo en pacientes pediátricos con insuficiencia respiratoria en dos unidades de cuidados críticos pediátricos a gran altitud.

INTRODUCTION: Acute respiratory failure is the leading cause of hospitalization in pediatrics. High-flow nasal cannulas (HFNCs) offer a new alternative, but the evidence and indications are still debated. The performance of HFNCs at high altitude has not been described to date. OBJECTIVE: To describe the use of HFNCs in pediatric patients admitted with respiratory failure and explore the factors associated with treatment failure. METHODOLOGY: A prospective cohort study was carried out in patients between 1 month and 18 years of age managed with HFNCs. The demographic and treatment response data were recorded at baseline and after 1, 6 and 24hours. The number of failures was determined, as well as the length of stay, complications and mortality. Patients with treatment failure were compared with the rest. RESULTS: A total of 539 patients were enrolled. Infants (70.9%) of male sex (58.4%) and airway diseases such as asthma and bronchiolitis (61.2%) were more frequent. There were 53 failures (9.8%), with 21 occurring in the first 24hours. The median length of stay was 4 days (IQR 4); there were 5 deaths (0.9%) and 13 adverse events (epistaxis) (2.2%). Improvement was observed in vital signs and severity over time, with differences in the group that failed, but without interactions. The final logistic model established an independent relationship of failure between the hospital (OR 2.78, 95%CI 1.48-5.21) and the initial respiratory rate (OR 1.56, 95%CI 1.21-2.01). CONCLUSIONS: HFNCs afford good clinical response, with few complications and a low failure rate. The differences found between institutions suggest a subjective relationship in the decision of therapy failure.

Effects of feed withdrawal times prior to slaughter on some animal welfare indicators and meat quality traits in commercial pigs.

The aim of this study was to determinate the effects of on-farm fasting periods and lairage times on the physiological behavioral indicators, and on the killing-out and meat quality traits in commercial pigs. Two-hundred and forty pigs were submitted to a 2X4X3 experimental design, with two on-farm periods (0 or 8 h), four lairage times (1, 8, 12, 24 h) of fasting and three replicates. The blood values of cortisol, urea, and non-esterified fatty acids were above the baseline values for the species Fasting on the farm, lairage time, and total fasting (on farm plus lairage time) had a significant effect on carcass yield and on the hot and cold carcass yields (P < 0.01), but not on live weight losses (P > 0.01). The total fast had significant effects on the pH values, color L * and on water-holding capacity of the meat (P < 0.01). Therefore, in favor of animal welfare and better meat quality we suggest a total fast no >12 h.

Inhibiting diacylglycerol acyltransferase-1 reduces lipid biosynthesis in bovine blastocysts produced in vitro.

Diacylglycerol acyltransferase-1 (DGAT1) is one of the DGAT enzymes that catalyzes the final step in the synthesis of triacylglycerol, which is a major component of the lipid droplets in embryos. Intracellular lipids accumulated in embryos produced in vitro have been associated with reduced cryotolerance and quality. The objective of the present study was to investigate the influence of DGAT1 inhibition on embryo development, quality, and post-vitrification survival, in addition to expression profiles of selected lipid metabolism-regulating and oxidative stress genes. Bovine cumulus-oocyte complexes were matured and fertilized in vitro and were cultured in synthetic oviduct fluid (SOF) supplemented with 5% fetal calf serum (FCS) alone (Control) or with 1, 5, 10 or 50 µM DGAT1 inhibitor (A922500®; D1, D5, D10, and D50, respectively) or 0.1% dimethyl sulfoxide (CDMSO: vehicle for DGAT1 inhibitor dilution) from 54 h post-insemination until Day 8 post insemination. No differences were found in blastocyst yield on days 7 and 8 in Control, CDMSO, D10, and D50 groups. Embryos cultured with 10 or 50 µM DGAT1 inhibitor had greater mitochondrial activity (P < 0.01), and increased number of cells (P < 0.05), while the cytoplasmic lipid content was reduced (P < 0.01), the latter associated with altered expression profiles of selected genes regulating lipid metabolism or genes related with oxidative stress (transcript abundance increased for SLC2A1 and SLC2A5 and decreased for DGAT1 and GPX1). Importantly, the survival rate of blastocysts produced with 10 µM DGAT1 was higher than that of Control, CDMSO and D50 groups at 72 h after vitrification and warming (73.8 vs 57.1, 55.9 and 56.1%, respectively, P < 0.001). In conclusion, inhibition of DGAT1 synthesis in bovine embryos produced in vitro abrogates the negative effect of FCS by decreasing their lipid content, increasing mitochondria activity and improving embryo cryotolerance, as well as favoring the expression of lipid metabolism regulating and oxidative stress-related transcripts.

Improved dual-energy material discrimination for dual-source CT by means of additional spectral filtration.

The use of additional spectral filtration for dual-energy (DE) imaging using a dual-source CT (DSCT) system was investigated and its effect on the material-specific DE(ratio) was evaluated for several clinically relevant materials. The x-ray spectra, data acquisition, and reconstruction processes for a DSCT system (Siemens Definition) were simulated using information provided by the system manufacturer, resulting in virtual DE images. The factory-installed filtration for the 80 kV spectrum was left unchanged to avoid any further reductions in tube output, and only the filtration for the high-energy spectrum was modified. Only practical single-element filter materials within the atomic number range of 40 < or = Z < or = 83 were evaluated, with the aim of maximizing the separation between the two spectra, while maintaining similar noise levels for high- and low-energy images acquired at the same tube current. The differences between mean energies and the ratio of the 140 and 80 kV detector signals, each integrated below 80 keV, were evaluated. The simulations were performed for three attenuation scenarios: Head, body, and large body. The large body scenario was evaluated for the DE acquisition mode using the 100 and 140 kV spectra. The DE(ratio) for calcium hydroxyapatite (simulating bone or calcifications), iodine, and iron were determined for CT images simulated using the modified and factory-installed filtration. Several filter materials were found to perform well at proper thicknesses, with tin being a good practical choice. When image noise was matched between the low- and high-energy images, the spectral difference in mean absorbed energy using tin was increased from 25.7 to 42.7 keV (head), from 28.6 to 44.1 keV (body), and from 20.2 to 30.2 keV (large body). The overlap of the signal spectra for energies below 80 keV was reduced from 78% to 31% (head), from 93% to 27% (body), and from 106% to 79% (large body). The DE(ratio) for the body attenuation scenario increased from 1.45 to 1.91 (calcium), from 1.84 to 3.39 (iodine), and from 1.73 to 2.93 (iron) with the additional tin filtration compared to the factory filtration. This use of additional filtration for one of the x-ray tubes used in dual-source DECT dramatically increased the difference between material-specific DE ratios, e.g., from 0.39 to 1.48 for calcium and iodine or from 0.28 to 1.02 for calcium and iron. Because the ability to discriminate between different materials in DE imaging depends primarily on the differences in DE ratios, this increase is expected to improve the performance of any material-specific DECT imaging task. Furthermore, for the large patient size and in conjunction with a 100/140 kV acquisition, the use of additional filtration decreased noise in the low-energy images and increased contrast in the DE image relative to that obtained with 80/140 kV and no additional filtration.

On the fitting of binding data when receptor dimerization is suspected.

Mechanistic and empirical modelling are compared in context of dimeric receptors. In particular, the supposed advantages of the two-state dimer model for fitting of binding data with respect to classical approaches such as the two-independent sites model are investigated. The two models are revisited from both the mechanistic and empirical point of views. The problem of overparameterized models and the benefits of the concurrent use of mechanistic and empirical models for mechanism analysis are discussed. The pros and cons of mathematical models are examined with special emphasis given to the interpretation of the connection between the shapes of the curves and receptor cooperativity. It is shown that a given pharmacological phenotype (curve shape) can be obtained from different receptor genotypes (as, for instance, non-interconvertible monomeric receptor species, receptor-G protein interactions and dimeric receptors), though values of the Hill coefficient greater than one are indicative of receptor oligomerization. The existence of a relationship between the recently defined dimer cooperativity index and the more familiar Hill coefficient is proven.

Iso-Kikuchi Syndrome: Report of 3 Pediatric Cases. / Síndrome de Iso-Kikuchi: 3 casos en la edad pediátrica.

Iso-Kikuchi syndrome, or congenital onychodysplasia of the index finger, is an uncommon condition characterized by total anonychia or dysplasia of the nail of the index finger. It is occasionally accompanied by underlying bone abnormalities and is rarely associated with other conditions. Although various hypotheses have been put forward to explain the pathophysiology of the syndrome, its etiology remains unknown. We report the cases of 3 pediatric patients (2 boys and 1 girl) with nail changes and bone abnormalities consistent with Iso-Kikuchi syndrome. We highlight the importance of recognizing this entity early to avoid the need for additional tests and unnecessary treatment.

Bandgap and pseudohelicity effects over conductance in gapped graphene junctures.

We study the conductance in gapped single-layer graphene junctures as a function of bangap, pseudohelicity and charge carriers density. To do it, we first calculate the transmission coefficients of massive charge carries for p-n and n-p-n junctures of gapped single-layer graphene. Next, we calculate the conductance for these two systems using the Landauer formula. Only for the p-n juncture case and non-zero bandgap values, we find the existence of a contribution to the conductance from pseudohelicity inversion states, which is small compared to the contribution from pseudohelicity conservation states. Also, we find for both type of junctures that there exists a window of charge carriers densities values where the conductance is zero (conductance gap), in such a way that the size of this window depends on the squared of the bandgap. We observe that the existence of a bandgap in the system leads to valley mixing and this fact could be useful for the future design of devices based on single-layer graphene.

Manual MGIT™ system for the detection of Mycobacterium tuberculosis: insights from a high TB burden setting.

OBJECTIVE: To evaluate the diagnostic performance of manual MGIT™ (MMGIT) compared to the gold standard, Löwenstein-Jensen (LJ), in the diagnosis of pulmonary tuberculosis (TB) in a high-burden setting. METHODS: Individuals with suspected TB enrolled in parallel diagnostic trials during 2007-2011 were included. Two samples were obtained from each patient and inoculated into MMGIT and LJ medium. Diagnostic tests were performed, and the incremental yield of a second test and time to detection (TTD) were calculated. Analyses were performed per patient and per sample. Gold standard was based on LJ culture. RESULTS: In the per patient and per sample analysis, we evaluated 1436 patients and 4142 samples. The sensitivity and specificity for smear and MMGIT per sample were respectively 89.9%/92.2% and 97.1%/98.9%. Contamination was observed in 1.4% of samples on MMGIT. The mean TTD (days) was 11.8 for MMGIT and 22.9 for LJ. The sensitivity and specificity for smear and MMGIT per patient were respectively 89.9% and 92.2% and 97.1% and 98.3%. A second MMGIT culture had an incremental yield of 1.6%. CONCLUSIONS: MMGIT has high sensitivity and specificity, regardless of smear result, with a 50% reduction in TTD compared to LJ. These features make MMGIT an acceptable TB diagnostic method for use in resource-limited settings.

Factores asociados al fracaso de la terapia con cánulas nasales de alto flujo en pacientes pediátricos con insuficiencia respiratoria en dos unidades de cuidados críticos pediátricos a gran altitud / Factors associated to high-flow nasal cannula treatment failure in pediatric patients with respiratory failure in two pediatric intensive care units at high altitude

Introducción: El fallo respiratorio agudo es la principal causa de hospitalización en pediatría. Las cánulas nasales de alto flujo (CNAF) ofrecen una nueva alternativa, pero sigue existiendo debate en torno a la evidencia e indicaciones. No se ha descrito su comportamiento en gran altitud. Objetivo Describir el uso de CNAF en pacientes pediátricos que ingresan con insuficiencia respiratoria y explorar los factores asociados al fracaso de la terapia. Metodología Estudio de cohortes prospectivo. Pacientes entre un mes y 18 años manejados con CNAF. Se describieron datos demográficos y se evaluó la respuesta al inicio, 1.a, 6.a y 24.a horas. Se determinó el número de fracasos, así como estancia, complicaciones y mortalidad. Se compararon los pacientes con fracaso al tratamiento. Resultados Ingresaron 539 pacientes. Fueron más frecuentes los lactantes (70,9%) de sexo masculino (58,4%) con afecciones respiratorias como asma y bronquiolitis (61,2%). Se presentaron 53 fracasos (9,8%), 21 en las primeras 24 horas. La mediana de estancia fue de 4 días (RIQ 4), hubo 5 éxitus (0,9%) y 13 eventos adversos –epistaxis– (2,2%). Se observó mejoría de signos vitales y gravedad en el tiempo con diferencias en el grupo que fracasó, pero sin interacciones. El modelo logístico final estimó una relación independiente del fracaso, entre el hospital (OR 2,78; IC95% 1,48-5,21) y la frecuencia respiratoria inicial (OR 1,56; IC95% 1,21-2,01). Conclusión La CNAF es un sistema con buena respuesta clínica, pocas complicaciones y una baja tasa de fracasos. Las diferencias entre las instituciones sugieren una relación subjetiva de la decisión del fracaso. (AU)

Introduction: Acute respiratory failure is the leading cause of hospitalization in pediatrics. High-flow nasal cannulas (HFNCs) offer a new alternative, but the evidence and indications are still debated. The performance of HFNCs at high altitude has not been described to date. Objective To describe the use of HFNCs in pediatric patients admitted with respiratory failure and explore the factors associated with treatment failure. Methodology A prospective cohort study was carried out in patients between 1 month and 18 years of age managed with HFNCs. The demographic and treatment response data were recorded at baseline and after 1, 6 and 24hours. The number of failures was determined, as well as the length of stay, complications and mortality. Patients with treatment failure were compared with the rest. Results A total of 539 patients were enrolled. Infants (70.9%) of male sex (58.4%) and airway diseases such as asthma and bronchiolitis (61.2%) were more frequent. There were 53 failures (9.8%), with 21 occurring in the first 24hours. The median length of stay was 4 days (IQR 4); there were 5 deaths (0.9%) and 13 adverse events (epistaxis) (2.2%). Improvement was observed in vital signs and severity over time, with differences in the group that failed, but without interactions. The final logistic model established an independent relationship of failure between the hospital (OR 2.78, 95%CI 1.48-5.21) and the initial respiratory rate (OR 1.56, 95%CI 1.21-2.01). Conclusions HFNCs afford good clinical response, with few complications and a low failure rate. The differences found between institutions suggest a subjective relationship in the decision of therapy failure. (AU)

Understanding the salinity effect on cationic polymers in inducing flocculation of the microalga Neochloris oleoabundans.

A mechanistic study was performed to evaluate the effect of salinity on cationic polymeric flocculants, that are used for the harvesting of microalgae. The polyacrylamide Synthofloc 5080H and the polysaccharide Chitosan were employed for the flocculation of Neochloris oleoabundans. In seawater conditions, a maximum biomass recovery of 66% was obtained with a dosage of 90mg/L Chitosan. This recovery was approximately 25% lower compared to Synthofloc 5080H reaching recoveries greater than 90% with dosages of 30mg/L. Although different recoveries were obtained with both flocculants, the polymers exhibit a similar apparent polymer length, as was evaluated from viscosity measurements. While both flocculants exhibit similar polymer lengths in increasing salinity, the zeta potential differs. This indicates that polymeric charge dominates flocculation. With increased salinity, the effectivity of cationic polymeric flocculants decreases due to a reduction in cationic charge. This mechanism was confirmed through a SEM analysis and additional experiments using flocculants with various charge densities.

Conformational analysis of GpA and GpAp in aqueous solution by molecular dynamics and statistical methods.

Barnase, an extracellular endoribonuclease from Bacillus amyloliquefaciens, hydrolyses single-stranded RNA. Its very low catalytic activity toward GpN dinucleotides, where N stands for any nucleoside, is markedly increased when a phosphate is added to the 3'-end, as in GpNp. Here we investigate the conformational properties of GpA and GpAp in solution, in order to determine whether differences in these properties may be related to the changes in enzymatic activity. Two independent 1.3 ns molecular dynamics trajectories are generated for each dinucleotide in the presence of explicit water molecules and counter ions. These trajectories are analysed by monitoring molecular properties, such as the solvent accessible surface area, the distance and orientation between the bases, the behaviour of torsion angles and formation of intramolecular H-bonds. To identify relevant correlations between these parameters, statistical techniques, comprising multiple regression, clustering and discriminant analysis are used. Results show that GpA has a significant propensity to form folded conformations (approximately 50%), fostered by a small number of intramolecular H-bonds, whereas GpAp remains essentially extended. The latter behaviour seems to be due to an H-bond between the terminal phosphate and adenosine ribose group, which restricts rotation about the adenine Agamma angle. We also find that GpA folding is induced by a concerted motion of specific torsion angles, which is closely coupled to the formation of a network of flexible hydrogen bonds. Finally, on the basis of an expression for barnase KM, which incorporates the folded/extended conformational equilibria of the dinucleotide substrates, it is argued that our findings on the differences between these equilibria, can qualitatively rationalize the experimentally measured differences in enzymatic properties.

Ageing affects nitric oxide synthase, cyclooxygenase and oxidative stress enzymes expression differently in mesenteric resistance arteries.

1 Our aim was to study the role of nitric oxide (NO) and arachidonic acid pathways in the alpha(1)-adrenoceptor-mediated vasoconstriction in mesenteric resistance arteries from 3--4 and 22 to 23-month-old Sprague-Dawley rats. 2 The expression of NO synthase (NOS), cyclooxygenase (COX) isoforms, soluble guanylate cyclase, superoxide dismutase and the NAD(P)H oxidase subunits p22(phox) and p 47(phox) were determined. 3 The N(G)-nitro-l-arginine methyl ester, a non-selective NOS inhibitor, shifted to the left but indomethacin and NS 398, non-selective and selective COX-2 inhibitors, shifted to the right the concentration-response curve for the vasoconstriction by phenylephrine in both age groups. 4 Ageing up-regulated endothelial NOS and p22(phox) expression but did not modify COX, soluble guanylate cyclase, superoxide dismutase and p 47(phox) expression. 5 These data suggest that the observed enhancement of eNOS protein expression could constitute a compensatory mechanism to counter-regulate a chronic loss of NO possibly through increased superoxide anion production from NAD(P)H oxidase induced by age.

Effects of L-NG-nitro-arginine on noradrenaline induced contraction in the rat anococcygeus muscle.

1. The influence of L-NG-nitro-arginine (L-NOARG, 30 microM) on contractile responses to exogenous noradrenaline was studied in the rat anococcygeus muscle. 2. Noradrenaline (0.1-100 microM) contracted the muscle in a concentration-dependent manner. L-NOARG (30 microM) had no effect on noradrenaline responses. 3. Phenoxybenzamine (Phz 0.1 microM) depressed by 46% (P < 0.001) the maximum response and shifted to the right (P < 0.001) the E/[A] curve to noradrenaline (pEC50 control: 6.92 +/- 0.09; pEC50 Pbz: 5.30 +/- 0.10; n = 20). 4. The nested hyperbolic null method of analysing noradrenaline responses after phenoxybenzamine showed that only 0.61% of the receptors need to be occupied to elicit 50% of the maximum response, indicating a very high functional receptor reserve. 5. Contractile responses to noradrenaline after partial alpha 1-adrenoceptor alkylation with phenoxybenzamine (0.1 microM) were clearly enhanced by L-NOARG. 6. The potentiating effect of L-NOARG on noradrenaline responses after phenoxybenzamine was reversed by (100 microM) L-arginine but not by (100 microM) D-arginine. 7. These results indicate that spontaneous release of NO by nitrergic nerves can influence the alpha -adrenoceptor-mediated response to exogenous noradrenaline.

Effect of NG-nitro-L-arginine methylester (L-NAME) on functional and biochemical alpha 1-adrenoceptor-mediated responses in rat blood vessels.

1. The modulation by NG-nitro-L-arginine methylester (L-NAME) of alpha 1-adrenoceptor-mediated contraction was investigated on isolated segments of rat tail artery and aorta. The influence of L-NAME on inositol phosphates accumulation by alpha 1-adrenoceptor agonists was also investigated to elucidate the intracellular mechanism responsible for this modulation. 2. In aorta but not in tail artery L-NAME (30 microM) enhanced the sensitivity (3.3 times) and the maximum contraction (Emax) induced by the full agonist, phenylephrine. 3. St-587, a partial alpha 1-adrenoceptor agonist, behaved as a weak agonist in the aorta (22.2% of phenylephrine Emax). However, when the same agonist was studied in tail artery rings a maximum contraction that was 78.4% of the phenylephrine induced Emax was reached. 4. L-NAME increased (3.3 times) the Emax for St-587 contraction in the aorta but not in the tail artery. Sensitivity to St-587 was slightly but significantly (P < 0.001) enhanced (1.9 times) by L-NAME in tail artery segments. 5. Contractile responses to phenylephrine after partial alkylation with phenoxybenzamine were analyzed by the nested hyperbolic null method. To elicit 50% of Emax for contraction only 1.1% of the receptors in the tail artery and 21% of the receptors in the aorta need to be occupied. These results indicate a higher receptor reserve for the tail artery than the aorta. 6. In the tail artery but not in the aorta, St-587 activates phosphoinositide turnover. The presence of L-NAME was without effect on inositol phosphates accumulation induced by this partial alpha 1-adrenoceptor agonist. 7. The maximum contraction induced by phenylephrine, after partial alpha-adrenoceptor alkylation, was enhanced by L-NAME in tail artery rings. However, the NO synthase inhibitor was unable to modify the phenylephrine-induced accumulation of inositol phosphates in the presence of phenoxybenzamine. 8. These results indicate that the differences in St-587-induced contraction and the modulation by L-NAME of alpha 1-adrenoceptor-mediated contraction observed between the tail artery and aorta are associated with differences in receptor reserve. In addition, our biochemical studies indicate that the potentiating effect of L-NAME is independent of intracellular calcium release via phosphatidylinositol turnover.

Modelling the changes induced by chronic desipramine treatment on the factors governing the agonism at prejunctional alpha 2-adrenoceptors.

1. The adaptational changes induced after chronic desipramine treatment on functional responsiveness of alpha 2-adrenoceptor activation were investigated in prostatic portions of the rat vas deferens. 2. For this purpose, clonidine and xylazine were studied for their effects on twitch contractions elicited by electrical field stimulation of prostatic portions removed 48 h after the last injection to the animals of vehicle or desipramine (10 mg kg-1, i.p.; 14 days). Operational model-fitting and the nested hyperbolic method were used to analyse the effects of irreversible receptor alkylation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ, 300 nM) on the alpha 2-adrenoceptor-mediated effects of clonidine, either in vehicle- or in desipramine-treated animals. 3. Treatment with desipramine decreased the potency (increased the EC50) of clonidine and xylazine by about 12 and 9 fold respectively. However, the treatment did not modify the maximal effect (alpha) elicited by either agonist. The estimates of apparent affinity for clonidine did not depend on the method of calculation as the 'null' method and the 'operational' method gave similar answers. Estimates of tau values for both agonists revealed that chronic desipramine treatment resulted in significant decreases in the efficacy of agonists. However, desipramine treatment was not associated with significant changes in the affinity constant for clonidine while for xylazine, the operational model provided a higher estimate of KA (lower affinity) after desipramine treatment. 4. The results indicate a large receptor reserve at prejunctional alpha 2-adrenoceptors which is modulated by chronic desipramine treatment. 5. The comparison of results obtained after chronic desipramine exposure with those by using EEDQ suggests that chronic desipramine treatment is not a useful experimental intervention for the purpose of estimating agonist affinities and efficacies.

Endothelial modulation of alpha 1-adrenoceptor contractile responses in the tail artery of spontaneously hypertensive rats.

1. Vascular contraction induced by phenylephrine was studied in tail artery rings from spontaneously hypertensive (SHR) and Wistar Kyoto rats (WKY) with particular focus on the role of endothelium. The influence of receptor reserve and the density of alpha 1-adrenoceptors on the possible differences observed were also analysed. 2. Phenylephrine (0.01-100 microM) induced concentration-dependent vasoconstrictions. The maximum response (alpha, P < 0.001) was greater but the pEC50 (P < 0.05) smaller in rings from SHR than from WKY rats irrespective of the presence or absence of endothelium. 3. Removal of endothelial cells resulted in a decrease of the maximum contraction with no modification in the pEC50 in arteries from both WKY and SHR. 4. The density of alpha 1-adrenoceptors (Bmax) and the dissociation constant (KD) were found to be the same for preparations from SHR and WKY rats in [3H]-prazosin binding experiments. 5. The apparent affinity (pKA) determined by the nested hyperbolic method and the operational model was similar in tail arteries from the two rat strains, irrespective of the presence or absence of endothelium. However, in endothelium-denuded rings, the pKA value was enhanced when compared with intact rings, in both SHR and WKY rats. 6. In rings from hypertensive rats, the operational parameter maximum possible effect (Em) was greater and the agonist efficacy (tau) was smaller than in rings from normotensive rats. When the endothelium was removed log tau and Em diminished in preparations from both rat strains. 7. In summary, the increased maximum responsiveness to phenylephrine in rings from SHR could be due to enhancement in Em. The log tau values indicate a deterioration in the transduction of the stimulus provided by the agonist in tail arteries from hypertensive animals. This study also suggests that the absence of endothelium modifies the alpha 1-adrenoceptor-mediated vasoconstriction probably by altering the transduction signalling mechanisms. The importance of analysing the degree of endothelium functionality when comparing results from different groups of rats is stated.