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Se midió el desarrollo corporal de terneros destetados a los tres meses de edad en pasturas de Andropogon gayanus, A. gayanus + Centrosema acutifolium y A. gayanus + Pueraria phaseoloides. Se utilizaron 10 terneros/tratamiento (70.3 ± 1.73 kg). El ensayo empezó en plena temporada de lluvias, que en este año fueron excesivas y afectaron negativamente la recuperación de las praderas. El desarrollo de los animales se redujo seriamente por la falta de forraje. Se hicieron algunas comparaciones subjetivas con un grupo de terneros que permaneció con sus madres en sabana nativa, hasta los nueve meses de edad, en la cual se realiza el destete tradicional. La cantidad de lluvia no permitió hacer comparaciones entre las pasturas. Los terneros destetados precozmente mostraron un desempeño muy pobre con respecto a los que permanecieron con sus madres; sin embargo, el grupo que pastoreó A. gayanus presentó los peores resultados en ganancia de peso vivo, porcentaje de proteína y minerales en suero, hematocrito, etc., muriendo tres animales por efecto del tratamiento. El destete precoz redujo el anestro posparto de las vacas y mejoró la recuperación de peso de las mismas.The development of early-weaned calves (three months old) in Andropogon gayanus, A. gayanus, Centrosema acutifolium and A. gayanus + Pueraria phaseoloides was investigated. Ten animals were used per treatment; the experiment beginning at the start of the rainy season. In this year 100 % and 45 % more rain fell in may and june respectively than the yearly average for these months. Several subjective comparison were made between the calves in the improved and group maintaned with their mothers in the native savannah until traditional early weaning occurs. The abnormally high rain-fall adverselty affected pasture recovery to the extent that it was impossible to make comparisons between the three pasture treatments. In general, however, all early-weaned calves did not perform as well as those remaining with their mothers. Results from the group in the pure stand of A. gayanus were the worst overall (live weight gains, protein and minerals in serum red blood cell), with three animals dying during the course of the trial. Early-weaning reduced anestrum pospartum and improved weight recovery of the cows.

Thanks to the asset management regulations, modern maintenance practices are rapidly getting a more managerial role, and is used to achieve savings and optimize energy use. Therefore, knowing at all times the status of the equipment allows for timely and necessary interventions that generate value to the company, and recover the initial conditions of them. Heat exchanger networks, rather than a productive asset, constitute an energysaving strategy, to have lower fuel costs, emission control, rational use of energy, etc. in the atmospheric furnaces of the crude distillation units, and other units that perform similar processes. Thus, keeping them in their best conditions, most of the time is necessary. In this paper, we propose a methodology for the diagnosis of the equipment of the network, and the maintenance planning justified by the energy efficiency of them and the economic impact of the intervention. In addition, an indicator is presented to provide an economic justification for maintenance interventions, as well as for briefly showing the results of the application of maintenance mainly on efficiency and the use of the KPI J proposed for programming the maintenance schedule of some of the equipment (pilot test) of the heat exchanger network under study. The methodology developed uses real operation values, and its results provided savings up to USD 150,000. [ABSTRACT FROM AUTHOR]

Five beta-adrenoceptor blockers, propranolol, acebutolol, diacebutolol, atenolol and bisoprolol, were tested for their antagonistic effect against isoprenaline on human isolated bronchi. The results showed (1) that only propranolol exerted a competitive antagonistic effect against isoprenaline (pA2 = 9.40 +/− 0.22, n = 7) whereas the other drugs did not, and (2) that, in the presence of beta-adrenoceptor blockers in the plasma concentrations reported after a single usual therapeutic dose, the doses of isoprenaline giving the same bronchodilator effect must be multiplied by 32.6, 5.51, 4.63, 2.82 and 1.95 respectively with propranolol, diacetolol, acebutolol, atenolol, and bisoprolol. It was concluded that (1) atenolol and bisoprolol were the least potent drugs at bronchial level in therapeutic plasma concentrations and (2) that tests performed on the human isolated bronchus might be a useful screening procedure for new drugs with potential activity on the airways. [ABSTRACT FROM AUTHOR]

Abstract Background Dietary fibre is both preventive and therapeutic for bowel functional diseases. Soluble cocoa products are good sources of dietary fibre that may be supplemented with this dietary component. This study assessed the effects of regularly consuming two soluble cocoa products (A and B) with different non-starch polysaccharides levels (NSP, 15.1 and 22.0% w/w, respectively) on bowel habits using subjective intestinal function and symptom questionnaires, a daily diary and a faecal marker in healthy individuals. Methods A free-living, two-stage, randomized, crossover, single-blind intervention was carried out in 44 healthy men and women, between 18-55 y old, who had not taken dietary supplements, laxatives, or antibiotics six months before the start of the study. In the four-week-long intervention stages, separated by a three-week-wash-out stage, two servings of A and B, that provided 2.26 vs. 6.60 g/day of NSP respectively, were taken. In each stage, volunteers' diet was recorded using a 72-h food intake report. Results Regularly consuming cocoa A and B increased fibre intake, although only cocoa B significantly increased fibre intake (p < 0.001) with respect to the non-cocoa stage. No changes in body weight were observed in either of the 4 week interventions. With cocoa product B, the number of daily bowel movements increased (p = 0.002), the frequency of having a bowel movement once a day increased (p = 0.009), the time to have a bowel movement was lower (p = 0.016) as well as the feeling of constipation (p = 0.046) without inducing adverse gastrointestinal symptoms, only flatulence increased (p = 0.019). Conclusions Regular consumption of the cocoa products increases dietary fibre intake to recommended levels and product B improves bowel habits. The use of both objective and subjective assessments to evaluate the effects of food on bowel habits is recommended.

Abstract: The effects of virgin olive oil phenols, hydroxytyrosyl acetate (HTy-Ac) and hydroxytyrosol (HTy), on cell integrity and steady-state values of cellular redox status were assessed in HepG2 cells, as well as their potential protective effects against oxidative stress induced by tert-butyl hydroperoxide (t-BOOH). Direct treatment for 20h with 0.5–10μM HTy or HTy-Ac reduced ROS generation and increased glutathione peroxidase activity at the higher concentrations. Furthermore, after t-BOOH exposure, pretreatment with HTy-Ac and HTy for 2 or 20h counteracted cell viability damage from 1μM, counterbalanced reduced glutathione levels from 0.5μM, protected against lipid peroxidation from 0.5μM, decreased ROS generation from 1μM as well as antioxidant enzyme activities from 1μM. All these changes were statistically significant. HTy-Ac presents antioxidative stress protective effects at physiological concentrations similar to or even slightly higher than that of HTy, thus contributing to the protective role of virgin olive oil. [Copyright &y& Elsevier]

Síntesis de las investigaciones llevadas a cabo en "Cova Remigia", Ares del Maestre (Castellón). Se ha realizado una revisión a partir de una base de datos, codificando los diferentes elementos y características de los pictogramas: tipología, conservación, técnica, tamaño y color, etc., que han permitido la identificación de seis momentos diferentes en la realización del panel.

Problem: Recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF) represent distinct clinical conditions with established definitions, both of which have been linked to an underlying pro-inflammatory state. This study aimed to explore the levels of monocytic-myeloid-derived suppressor cells (M-MDSCs) and regulatory T cells (T Reg ) in a cohort of RPL and RIF women and their potential contribution to RPL and RIF., Method of Study: One hundred and eight non-pregnant women were evaluated: 40 RPL, 41 RIF, and 27 fertile healthy controls (HC). A multiparametric flow cytometry approach was utilized to measure and quantify the frequency of M-MDSCs and T Reg cells. Cytokine levels in plasma samples were evaluated through a multiplex assay. M-MDSCs levels were significantly higher in RPL and RIF patients compared to HC., Results: M-MDSCs levels were significantly higher in RPL (9.4% [7-11.6]) and RIF (8.1% [5.9-11.6]) patients compared to HC (6% [4.2-7.6]). An optimal cut-off of 6.1% for M-MDSCs disclosed a sensitivity of 75.6% and 89.7% and a specificity of 57.7% and 57.7% in RIF and RPL groups, respectively. A significant negative correlation was observed between M-MDSCs and T Reg (p = .002, r = -.51)., Conclusions: Our preliminary data allowed us to build a predictive model that may aid as a potential diagnostic tool in the clinic. These findings could provide a better understanding of these pathologies and a better definition of patients that could benefit from personalized treatments to promote pregnancy. Additional exploration and confirmation in distinct study groups are needed to fully assess the diagnostic capabilities of this biomarker., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This review collects and critically examines data on the levels of tumour necrosis factor-alpha (TNF-α) in lean, overweight and obese subjects, and the effects of intervention with different foods and food products containing bioactive constituents in overweight/obese individuals. We additionally explore the influence of different single nucleotide polymorphisms (SNPs) on TNF-α levels and compare the response to food products with that to some anti-obesity drugs. Our aim was to provide an overview of the variability, consistency, and magnitude of the reported effects of dietary factors on TNF-α, and to envisage the reliability of measuring changes in the levels of this cytokine as a biomarker responsive to food intervention in association with the reduction in body weight. Regarding the circulating levels of TNF-α, we report: (i) a large intra-group variability, with most coefficients of variation (CV%) values being ≥30% and, in many cases, >100%; (ii) a large between-studies variability, with baseline TNF-α values ranging from <1.0 up to several hundred pg/mL; (iii) highly variable effects of the different dietary approaches with both statistically significant and not significant decreases or increases of the protein, and the absolute effect size varying from <0.1 pg/mL up to ≈50 pg/mL. Within this scenario of variability, it was not possible to discern clear differentiating limits in TNF-α between lean, overweight, and obese individuals or a distinct downregulatory effect on this cytokine by any of the different dietary approaches reviewed, i.e., polyunsaturated fatty acids (PUFAs), Vitamin-D (VitD), mixed (micro)nutrients, (poly)phenols or other phytochemicals. Further, there was not a clear relationship between the TNF-α responses and body weight changes. We found similarities between dietary and pharmacological treatments in terms of variability and limited evidence of the TNF-α response. Different factors that contribute to this variability are discussed and some specific recommendations are proposed to reinforce the need to improve future studies looking at this cytokine as a potential biomarker of response to dietary approaches.

Olive-pomace oil is rich in oleic acid, and thus it can be an interesting dietary fat alternative as it can allow reaching the recommendation of consuming 20% of total diet energy in the form of monounsaturated fatty acids. In addition, olive-pomace oil also contains a wide range of minor components that may contribute to its healthy properties. The major components identified with healthy properties are triterpenic dialcohols and acids, squalene, tocopherols, sterols, fatty alcohols and phenolic compounds. The refining process, that the crude pomace-oil must undergo for commercial purposes, significantly reduces the content of phenolic compounds, while the other minor components remain at concentrations which can induce positive health effects, especially on cardiovascular health, outstanding pentacyclic triterpenes and aliphatic fatty alcohols in olive-pomace oil. Numerous in vitro and preclinical studies support that mainly the pure compounds, or extracts isolated from plant sources, play an important role in preventing cardiovascular disease and risk factors. Likewise, tocopherols, squalene and phytosterols, in addition to the minor fraction of phenolic compounds, have shown high biological activity with particular association to the cardiovascular function. In the light of the foregoing, and taking into consideration the absence of clinical studies with olive-pomace oil, it would be of great interest to develop randomized, crossover, controlled, double-blind studies to extend the knowledge and understanding on the health effects of olive-pomace olive.

Cocoa has beneficial health effects partly due to its high flavanol content. This study was aimed at assessing the absorption and metabolism of polyphenols in two soluble cocoa products: a conventional (CC) and a flavanol-rich product (CC-PP). A crossover, randomized, blind study was performed in 13 healthy men and women. On two different days, after an overnight fast, volunteers consumed one serving of CC (15 g) or CC-PP (25 g) in 200 mL of semi-skimmed milk containing 19.80 mg and 68.25 mg of flavanols, respectively. Blood and urine samples were taken, before and after CC and CC-PP consumption, and analyzed by high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometry (HPLC-ESI-QToF-MS). Up to 10 and 30 metabolites were identified in plasma and urine, respectively. Phase II derivatives of epicatechin were identified with kinetics compatible with small intestine absorption, although the most abundant groups of metabolites were phase II derivatives of phenyl-γ-valerolactone and phenylvaleric acid, formed at colonic level. 5-(4'-Hydroxyphenyl)-γ-valerolactone-sulfate could be a sensitive biomarker of cocoa flavanol intake. CC and CC-PP flavanols showed a dose-dependent absorption with a recovery of 35%. In conclusion, cocoa flavanols are moderately bioavailable and extensively metabolized, mainly by the colonic microbiota.

Bioavailability of yerba mate phenolic compounds was assessed in healthy humans. More than 34 metabolites were identified in biological fluids, mainly sulfated conjugates of caffeic and ferulic/isoferulic acids, in addition to non-metabolized caffeoyl-, feruloyl- and p-coumaroilquinic acids, with rapid appearance and clearance in plasma indicative of small intestinal absorption. These compounds amounted to 13.1% of the urinary metabolites. Delayed absorption of dihydrocaffeic, dihydroferulic and dihydrocoumaric acids and their phase II metabolites, in addition to feruloylglycine, pointed to their microbial origin and colonic absorption, accounting for 81.0% of excreted metabolites. Phase II flavonol metabolites (0.2%) derived mainly from rutin after colonic transformation and absorption were also detected. Additionally, dihydroferuloyl-, dihydrocaffeoyl- and dihydrocoumaroylquinic acids (5.7%) were identified, showing the most delayed kinetics. Total phenolic excretion (147.6μmol) corresponded to 13.2% of ingested phenols. In conclusion, yerba mate polyphenols are partially bioavailable and extensively metabolized, mainly by the colonic microbiota., (Copyright © 2017. Published by Elsevier Ltd.)

Roasting greatly reduces the phenolic content in green coffee beans. Considering the beneficial effects of coffee polyphenols, blends containing green coffee beans are being consumed as a healthier alternative to roasted coffee. This study was aimed at assessing the absorption and metabolism of hydroxycinnamates in an instant green/roasted (35/65) coffee blend in healthy humans. Twelve fasting men and women consumed a cup of coffee containing 269.5 mg (760.6 μmol) of chlorogenic acids. Blood and urine samples were taken before and after coffee consumption at different times and analyzed by LC-MS-QToF. Up to 25 and 42 metabolites were identified in plasma and urine, respectively, mainly in the form of sulfate and methyl derivatives, and to a lower extent as glucuronides. Un-metabolized hydroxycinnamate esters (caffeoyl-, feruloyl-, and coumaroylquinic acids), hydroxycinnamic acids (caffeic, ferulic and coumaric acids) and their phase II metabolites, in addition to phase II derivatives of lactones, represented a minor group of metabolites (16.3% of the metabolites excreted in urine) with kinetics compatible with small intestine absorption. Dihydrohydroxycinnamic acids and their phase II derivatives, in addition to feruloylglycine, showed delayed kinetics due to their colonic origin and represented the most abundant group of metabolites (75.7% of total urinary metabolites). Dihydrohydroxycinnamate esters (dihydroferuloyl-, dihydrocaffeoyl- and dihydrocoumaroylquinic acids) have been identified for the first time in both plasma and urine, with microbial origin (excreted 8-12 h after coffee intake) amounting to 8% of total urinary metabolites. In conclusion, coffee polyphenols are partially bioavailable and extensively metabolized, mainly by the colonic microbiota.

Considering that nitrocatechols present putative effects against Parkinson's disease, the absorption and metabolism of nitroderivatives of hydroxytyrosol (HT) were assessed using human cell model systems. The test compounds nitrohydroxytyrosol (NO2HT), nitrohydroxytyrosyl acetate (NO2HT-A), and ethyl nitrohydroxytyrosyl ether (NO2HT-E) were efficiently transferred across human Caco-2 cell monolayers as an intestinal barrier model, NO2HT-A and NO2HT-E being better (p < 0.05) absorbed (absorption rate (AR) = 1.4 ± 0.1 and 1.5 ± 0.2, respectively) than their precursor, NO2HT (AR = 1.1 ± 0.1). A significant amount of the absorbed compounds remained unconjugated (81, 70, and 33% for NO2HT, NO2HT-A, and NO2HT-E, respectively) after incubation in Caco-2 cells, being available for hepatic metabolism. Nitrocatechols were extensively taken up and metabolized by human hepatoma HepG2 cells as a model of the human liver. Both studies revealed extensive hydrolysis of NO2HT-A into NO2HT, whereas NO2HT-E was not hydrolyzed. Glucuronide (75-55%), methylglucuronide (25-33%), and methyl derivatives (0-12%) were the main nitrocatechol metabolites detected after metabolism in Caco-2 and HepG2 cells. In conclusion, NO2HT, NO2HT-A, and NO2HT-E show high in vitro bioavailability and are extensively metabolized by hepatic cells.

Background: In community-acquired pneumonia host inflammatory response against the causative microorganism is necessary for infection resolution. However an excessive response can have deleterious effects. In addition to antimicrobial effects, macrolide antibiotics are known to possess immunomodulatory properties., Methods: A prospective study was performed on 52 admitted patients who developed an inadequate response after 72 hours of antibiotic treatment - non-responders community-acquired pneumonia - (blood and bronchoalveolar lavage), and two control groups: 1) community-acquired pneumonia control (blood) and 2) non-infection control (blood and bronchoalveolar lavage). Cytokine profiles (interleukin (IL)-6, IL-8, IL-10), tumour necrosis factor α and clinical outcomes were assessed., Results: Non-responders patients treated with macrolide containing regimens showed significantly lower levels of IL-6 and TNF-α in bronchoalveolar lavage fluid and lower IL-8 and IL-10 in blood than those patients treated with non-macrolide regimens. Clinical outcomes showed that patients treated with macrolide regimens required fewer days to reach clinical stability (p < 0.01) and shorter hospitalization periods (p < 0.01)., Conclusions: After 72 hours of antibiotic effect, patients who received macrolide containing regimens exhibited lower inflammatory cytokine levels in pulmonary and systemic compartments along with faster stabilization of infectious parameters.

Neutrophil activation state and its relationship with an inflammatory environment in community-acquired pneumonia (CAP) remain insufficiently elucidated. We aimed to evaluate the neutrophil apoptosis and cytokine pattern in CAP patients after 72 h of treatment, and their impact on infection resolution. Apoptosis of blood and bronchoalveolar lavage (BAL) neutrophils was measured in nonresponding CAP (NCAP), in responding CAP (blood only) and in patients without infection (control). Pro-inflammatory (interleukin (IL)-6, IL-8) and anti-inflammatory (IL-10) cytokines were measured. Main outcomes were clinical stability and days of hospitalisation. Basal neutrophil apoptosis was higher in the BAL and blood of NCAP, whereas spontaneous apoptosis (after 24 h culture) was lower. Cytokines in NCAP were higher than in responding CAP and control: IL-6 was increased in BAL and blood, IL-8 in BAL and IL-10 in blood. An increased basal apoptosis (≥20%) in BAL of NCAP was associated with lower systemic IL-10 (p<0.01), earlier clinical stability (p=0.05) and shorter hospital stay (p=0.02). A significant correlation was found for systemic IL-6 and IL-10 with days to reach stability and length of stay. After 72 h of treatment, an increased basal alveolar neutrophil apoptosis might contribute to downregulation of inflammation and to faster clinical stability.

The objective of the study was to compare faecal monitoring and area under the curve (AUC) techniques for measuring iron absorption from a stable isotope label. Nine healthy female subjects (age: 33+/-8 yr) were recruited to take part in the study. After an overnight fast, each subject received a 5 mg dose of highly enriched Fe-57 and all faecal samples were collected for 10 days post-dose to allow for estimation of iron absorption by the faecal monitoring method (geometric mean: 25%). Serial blood samples (12 in total) were also collected from each volunteer for 6h post-dose to estimate iron absorption by the well-validated area under the plasma iron concentration curve (AUC) method (geometric mean: 18%). The faecal monitoring method yields higher iron absorption results compared to AUC. This discrepancy may be due to the steps involved in faecal processing where systematic errors are likely to take place and also to the lack of complete faecal recovery associated with issues of cleanliness and hygiene when passing stools. Both of these lead to losses of the unabsorbed isotope and to an overestimate of iron absorption., (Copyright 2010 Elsevier GmbH. All rights reserved.)

As part of an iron absorption study, we needed to accurately count reticulocytes in the peripheral blood of healthy human volunteers before measuring their enrichment with stable iron isotopes given in an oral dose. Recent studies have suggested the usefulness of reticulocyte counting by flow cytometry, through a combination of differential light scatter and measurement of the stoichiometric binding of thiazole orange (TO) to RNA within the maturing erythrocyte. Using this method we set out to improve the precision of our quantitative analysis by counting more cells, as reticulocytes normally comprise <2% of the red cell population. To ensure exclusion of other cell types, we identified WBCs and platelets with CD16+CD45- allophycocyanin and CD61- phycoerythrin, respectively. After removal of CD16(+) CD45(+) TO(+) WBCs and CD61(+) TO(-) platelets from analysis, the remaining cells were a combination of CD61(-) TO(-) erythrocytes, CD61(-) TO(+) reticulocytes and CD61(+) TO(+) reticulated platelets. Reticulocyte counts were lower after exclusion of CD61(+) TO(+) cells from analysis. They were similarly lower when erythrocyte precursors were positively identified through their glycophorin A expression and TO uptake. We conclude that it is necessary to exclude reticulated platelets from flow cytometric reticulocyte analysis.

Background: Iron deficiency anaemia is one of the most important nutritional diseases, with high prevalence worldwide. The G277S transferrin mutation has been implicated as a risk factor for iron deficiency in menstruating women. However, the subject is controversial and there are no data concerning the possible influence of this polymorphism on iron absorption., Aim of the Study: To undertake a pilot study to investigate the effect of carrying the G277S transferrin mutation on non-haem iron absorption from a meal in young menstruating women compared to wild-type controls., Methods: Menstruating women with low iron stores (serum ferritin < 30 microg/l) or who had suffered from iron deficiency anaemia or had a family history of anaemia were recruited (n = 162). Haematological parameters were analysed, including haemoglobin, ferritin, total-iron binding capacity and transferrin saturation. Non-haem iron absorption from a meal was measured in 25 non-anaemic women either with the G277S/G277G (n = 10) or the wild type G277G/G277G (n = 15) genotype. The incorporation of stable isotopes of iron into erythrocytes was used to measure absorption., Results and Conclusions: There were no significant differences in iron status indices or non-haem iron absorption between genotypes. However, G277S carriers did not show the usual inverse association between iron stores and non-haem iron absorption. Further studies should focus on the effects of a combination of polymorphisms in iron metabolism genes on iron absorption.

Objective: The Maillard reaction and lactose isomerization may be induced during the manufacture of infant formulas. We studied the effects of dehydration and sterilization on iron bioavailability from an infant formula in suckling and weanling rats., Methods: In experiment 1, a previously reconstituted powdered infant formula and an in-bottle-sterilized liquid infant formula from the same manufacturer were fed from drinking bottles to 2-wk-old suckling rat pups for 7 d. In experiment 2, the same formulas were complemented with AIN-76 and fed to weanling rats for 7 d after a 4-d adaptation period. In both experiments, intake, body weight, and fecal and urinary excretions were monitored, and the following iron indexes were calculated: apparent absorption and retention and the coefficients percentage of absorption versus intake, percentage of retention versus absorption, and percentage of retention versus intake., Results: The liquid infant formula resulted in lower body weights on day 4, particularly among the younger rats that had significantly lower food intakes (P = 0.045). In weanling rats fed powdered and liquid infant formulas, food intake and body weight were not significantly different. The pups showed significantly higher absorption (percentage of absorption versus intake) and retention (percentages of retention versus absorption and retention versus intake) efficiencies than did the weanling rats (P < 0.001 for the three indexes). Hemoglobin values (P = 0.001) and liver iron concentrations (P = 0.009) were significantly higher in the weanling rats than in the pups. In contrast, erythrocyte iron concentrations and hematocrit were higher in the pups (P = 0.016 and 0.053, respectively)., Conclusions: In rat pups, iron bioavailability is negatively affected by the consumption of in-bottle-sterilized infant formula, possibly as the result of the content of Maillard reaction products, altered proteins, and lactulose. However, when this formula is included in a mixed diet and given to weanling rats, this detrimental effect no longer occurs.

The effects of ouabain, an inhibitor of the plasmalemmal Na(+)/K(+)-ATPase activity, were examined in human isolated bronchus. Ouabain produced concentration-dependent contraction with -logEC(50)=7.16+/-0.11 and maximal effect of 67+/-4% of the response to acetylcholine (1 mM). Ouabain (10 microM)-induced contraction was epithelium-independent and was not depressed by inhibitors of cyclooxygenase and lipoxygenase, antagonists of muscarinic, histamine H(1)-receptors and alpha-adrenoceptors, or neuronal Na(+) channel blockade. The inhibition of ouabain contraction in tissues bathed in K(+)-free medium, and the inhibition by ouabain of the K(+)-induced relaxation confirm that the contractile action of ouabain is mediated by inhibition of Na(+)/K(+)-ATPase. Furthermore, depolarization (16.4+/-0.9 mV) was observed in human isolated bronchus by intracellular microelectrode recording. Ouabain (10 microM)-induced contractions were abolished by a Ca(2+)-free solution but not by blockers of L-type Ca(2+) channels. In human cultured bronchial smooth muscle cells, ouabain (10 microM) produced a sustained increase in [Ca(2+)](i) (116+/-26 nM) abolished in Ca(2+)-free medium. Incubation with a Na(+)-free medium or amiloride (0.1 mM) markedly inhibited the spasmogenic effect of ouabain thus suggesting the role of Na(+)/Ca(2+) exchange in ouabain contraction while selective inhibitors of Na(+)/H(+)-antiport, Na(+)/K(+)/Cl(-)-antiport, or protein kinase C had no effect. Ouabain (10 microM) failed to increase inositol phosphate accumulation in human bronchus. Ouabain (10 microM) did not alter bronchial responsiveness to acetylcholine or histamine but inhibited the relaxant effects of isoprenaline, forskolin, levcromakalim, or sodium nitroprusside. These results indicate that ouabain acts directly to produce contraction of human airway smooth muscle that depends on extracellular Ca(2+) entry unrelated to L-type channels and involving the Na(+)/Ca(2+)-antiporter.

The muscarinic functional antagonism of isoproterenol relaxation and the contribution of muscarinic M2 receptors were examined in human isolated bronchus. In intact tissues, acetylcholine (ACh) precontraction decreased isoproterenol potency and maximal relaxation (-log EC50 shift = -1.49 +/- 0.16 and E(max) inhibition for 100 microM ACh = 30%) more than the same levels of histamine contraction. The M2 receptor-selective antagonist methoctramine (1 microM) reduced this antagonism in ACh- but not histamine-contracted tissues. Similar results were obtained for forskolin-induced relaxation. After selective inactivation of M3 receptors with 4-diphenylacetoxy-N-(2-chloroethyl)piperadine hydrochloric acid (30 nM), demonstrated by abolition of contractile and inositol phosphate responses to ACh, muscarinic recontractile responses were obtained in U-46619-precontracted tissues fully relaxed with isoproterenol. Methoctramine antagonized recontraction, with pK(B) (6.9) higher than in intact tissues (5.4), suggesting participation of M2 receptors. In M3-inactivated tissues, methoctramine augmented the isoproterenol relaxant potency in U-46619-contracted bronchus and reversed the ACh-induced inhibition of isoproterenol cAMP accumulation. These results indicate that M2 receptors cause indirect contraction of human bronchus by reversing sympathetically mediated relaxation and contribute to cholinergic functional antagonism.

1. Cysteinyl-leukotrienes cause contractions and/or relaxations of human isolated pulmonary vascular preparations. Although, the localization and nature of the receptors through which these effects are mediated have not been fully characterized, some effects are indirect and not mediated via the well-described LT1 receptor. 2. In human pulmonary veins (HPV) with an intact endothelium, leukotriene D4 (LTD4) induced contraction above basal tone. This response was observed at lower concentrations of LTD4 in the presence of nitric oxide synthase inhibitor N omega-nitro-L-arginine (L-NOARG). Contractions (in the absence and presence of L-NOARG) were partially blocked by the LT1 antagonists (MK 571 and ICI 198615). 3. LTD4 relaxed HPV previously contracted with noradrenaline. This relaxation was potentiated by LT1 antagonists, but was abolished by removal of the endothelium. LTD4 also relaxed human pulmonary arteries (HPA) precontracted with noradrenaline but this effect was not modified by LT1 antagonists. 4. The results suggest that contraction of endothelium-intact HPV by LTD4 is partially mediated via LT1 receptors. Further, in endothelium-intact HPV, this contraction was opposed by a relaxation induced by LTD4, dependent on the release of nitric oxide, which was mediated, at least in part, via a non-LT1 receptor. In addition, LTD4 relaxation on contracted HPA was not mediated by LT1 receptors. 5. The mechanical effects of LTD4 on human pulmonary vasculature are complex and involve both direct and indirect mechanisms mediated via at least two types of cysteinyl-leukotriene receptors.

The three endothelins ET-1, ET-2 and ET-3 induced a potent contractile response in the human isolated bronchus with an intact epithelium, which proceeded on two different stages. The first stage was observed at low concentrations (high potency) (10(-12) to 10(-9) M) but corresponded to a low intrinsic activity (Emax maximal effect induced by ACh 10(-3) M), the second stage appeared at higher concentrations and corresponded to higher intrinsic activity. The rank order of potency for the two stages of contractile activity was ET-1 greater than ET-2 = ET-3. Removal of the epithelium significantly enhanced the two stages of the contractile responses to the three endothelins and abolished the differences in potency efficacy that were observed between ET-1, ET-2 and ET-3 when the epithelium was present. Phosphoramidon (10(-5) M), an enkephalinase inhibitor, was as potent as epithelium removal in enhancing the contractile responses to these agonists at low concentrations (first stage of contraction, 10(-16) to 10(-9) M). However, with high concentrations of endothelins (greater than 10(-9) M, second stage of contraction), phosphoramidon was less potent than epithelium removal in enhancing the contractile responses. In epithelium-denuded strips, preincubation with phosphoramidon did not further increase the maximal contractions induced by/or the potencies of ET-1, ET-2 or ET-3. After epithelium removal, responses to low doses of endothelins were attenuated by nicardipine (10(-6) M) whereas responses to high doses of the endothelins were not affected, as was also observed when the epithelium was present. (ABSTRACT TRUNCATED AT 250 WORDS)

1. The effects of three endothelins: (i) the classical or human/porcine endothelin (ET-1); (ii) [Trp6, Leu7] endothelin (ET-2) and (iii) [Thr2, Phe4, Thr5, Tyr6, Lys7, Tyr14] endothelin or rat endothelin (ET-3) were tested on the human isolated bronchus. 2. ET-1 produced a concentration-dependent contraction of the human isolated bronchus that proceeded in two different steps. The first step was observed at very low concentrations (pD2 = 11.01 +/- 0.17, n = 10) but corresponded to a low intrinsic activity (Emax = 15.6 +/- 1.8% of Emax = 26.1 +/- 2.9% of ACh 3 x 10(-3) M, n = 5, P less than 0.05), reduced by nicardipine 10(-6) M (Emax = 6.0 +/- 2.6% of ACh 3 x 10(-3) M, n = 5, P less than 0.05) and strongly inhibited in calcium-free medium. The second step of the action of ET-1 corresponded to a lesser potency (pD2 = 7.90 +/- 0.17, n = 9) but a higher intrinsic activity (Emax = 82.5 +/- 4.7% of ACh 3 x 10(-3) M). This effect was not significantly modified by nicardipine 10(-6) M or by Bay K 8644 10(-7) M. Neither of the two effects was modified by indomethacin 3 x 10(-6) M. 3. The effects of ET-2 and ET-3 were qualitatively similar to those of ET-1 but quantitatively different; for these two steps of contracting activity and for potency and efficacy the ranking was: ET-1 greater than ET-2 = ET-3. 4. Thus, ET-1 appears to be the most potent of these three substances in its effect on the human isolated bronchus. Its activity seems to involve the action of voltage-dependent calcium channels at low concentrations (10-12 to 10-9M), whereas other mechanisms are involved at higher concentrations (10-8 to 3 x 10-7M).

Zinc sulphate (50 mg kg-1 p.o.) did not modify basal gastric mucosal blood flow, as measured by [3H]aniline clearance, but inhibited its reduction by noradrenaline (3.5 micrograms kg-1 min-1). Zinc sulphate also influenced gastric emptying of phenol red but its effects depended upon the dose; 30 mg kg-1 caused no variation whereas 80 mg kg-1 induced a significant delay. The nature of both actions is discussed and their implications in the development and prevention of gastric ulceration have been analysed.

Verapamil (3, 10, 20 mg/kg-1) increases the necrotizing effects of oral 25% NaCl or 100% ethanol. Damage by 0.6 N HCl was not equally affected since 1 mg/kg-1 of verapamil decreased the ulcer index whereas the higher doses augmented it. Pharmacologically induced gastric lesions were also differently affected by verapamil, ulcers produced by histamine being greatly enhanced and those of reserpine inhibited. Neither indomethacin nor compound 48/80 ulcers were modified. These results suggest that verapamil modifies the susceptibility of the gastric mucosa to damage.

The antiulcer effects of two classical antisecretory agents, cimetidine and pirenzepine, were compared with those of prostaglandin E2 and carbenoxolone sodium. The acid inhibitory action of drugs was not important in our study since the ulcer model employed consisted of perfusing the stomach continuously, at a high intraluminal pressure (120 mm H2O), with a simulated gastric juice (0.1 M HCl plus 600 mg pepsin/l). Cimetidine and pirenzepine failed to inhibit gastric erosions, whereas prostaglandin E2 and carbenoxolone greatly reduced the severity of the mucosal lesions. Long-term treatment with cimetidine did not prevent gastric damage. These findings indicate that pure antisecretory agents do not directly increase the capacity of the mucosa to resist damage.