Biomedical subjects

M J Alonso

Publications and source records attributed to M J Alonso.

At least 55 records · Page 3Linked to original sources

PubMed1999

Enhancement of nasal absorption of insulin using chitosan nanoparticles.

PURPOSE: To investigate the potential of chitosan nanoparticles as a system for improving the systemic absorption of insulin following nasal instillation. METHODS: Insulin-loaded chitosan nanoparticles were prepared by ionotropic gelation of chitosan with tripolyphosphate anions. They were characterized for their size and zeta potential by photon correlation spectroscopy and laser Doppler anemometry, respectively. Insulin loading and release was determined by the microBCA protein assay. The ability of chitosan nanoparticles to enhance the nasal absorption of insulin was investigated in a conscious rabbit model by monitoring the plasma glucose levels. RESULTS: Chitosan nanoparticles had a size in the range of 300-400 nm, a positive surface charge and their insulin loading can be modulated reaching values up to 55% [insulin/nanoparticles (w/w): 55/100]. Insulin association was found to be highly mediated by an ionic interaction mechanism and its release in vitro occurred rapidly in sink conditions. Chitosan nanoparticles enhanced the nasal absorption of insulin to a greater extent than an aqueous solution of chitosan. The amount and molecular weight of chitosan did not have a significant effect on insulin response. CONCLUSIONS: Chitosan nanoparticles are efficient vehicles for the transport of insulin through the nasal mucosa.

Absorption↗

PubMed1999

Role of iNOS in the vasodilator responses induced by L-arginine in the middle cerebral artery from normotensive and hypertensive rats.

1. The substrate of nitric oxide synthase (NOS), L-arginine (L-Arg, 0.01 microM - 1 mM), induced endothelium-independent relaxations in segments of middle cerebral arteries (MCAs) from normotensive Wistar-Kyoto (WKY) and hypertensive rats (SHR) precontracted with prostaglandin F2alpha (PGF2alpha). These relaxations were higher in SHR than WKY arteries. 2. L-N(G)-nitroarginine methyl ester (L-NAME) and 2-amine-5,6-dihydro-6-methyl-4H-1,3-tiazine (AMT), unspecific and inducible NOS (iNOS) inhibitors, respectively, reduced those relaxations, specially in SHR. 3. Four- and seven-hours incubation with dexamethasone reduced the relaxations in MCAs from WKY and SHR, respectively. 4. Polymyxin B and calphostin C, protein kinase C (PKC) inhibitors, reduced the L-Arg-induced relaxation. 5. Lipopolysaccharide (LPS, 7 h incubation) unaltered and inhibited these relaxations in WKY and SHR segments, respectively. LPS antagonized the effect polymyxin B in WKY and potentiated L-Arg-induced relaxations in SHR in the presence of polymyxin B. 6. The contraction induced by PGF2alpha was greater in SHR than WKY arteries. This contraction was potentiated by dexamethasone and polymyxin B although the effect of polymyxin B was higher in SHR segments. LPS reduced that contraction and antagonized dexamethasone- and polymyxin B-induced potentiation, these effects being greater in arteries from SHR. 7. These results suggest that in MCAs: (1) the induction of iNOS participates in the L-Arg relaxation and modulates the contraction to PGF2alpha; (2) that induction is partially mediated by a PKC-dependent mechanism; and (3) the involvement of iNOS in such responses is greater in the hypertensive strain.

Animals↗

PubMed1999

Mechanisms involved in relaxation induced by exogenous nitric oxide in pig coronary arteries.

The aim of the present study was to analyze the mechanisms involved in the vasodilator responses elicited by nitric oxide (NO) in segments of porcine posterior descending coronary artery. Exogenous NO (0.1-30 microM) induced concentration-dependent relaxations in segments precontracted with a concentration of the thromboxane A2 mimetic, U-46619 (30-300 nM) that produced a contraction 70% (submaximal contraction) of that elicited by 75 mM K+ (maximal contraction). The relaxations were almost abolished by 6-anilinoquinoline-5,8-quinone (LY-83583, 10 microM), an inhibitor of guanylate cyclase, and with precontraction with 40 or 60 mM K+. Relaxations were reduced by 5 mM tetraethylammonium (TEA), a blocker of Ca(2+)-activated K+ channels (Kca channels) and unaltered by ouabain (500 microM), 4-aminopyridine (1 mM), glibenclamide (10 microM), apamin (1 microM) and charybdotoxin (0.3 microM), inhibitors of sodium pump, voltage-sensitive, ATP-sensitive, small-conductance Kca and large-conductance KcaK+ channels, respectively. These results suggest that the relaxation caused by exogenous NO is mediated by guanylate cyclase activation, with only a slight participation of a hyperpolarizing mechanism mediated by activation of Kca channels.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed1999

Mechanisms of bradykinin-induced relaxation in pig coronary arteries.

Bradykinin (BK) induced endothelium- and concentration-dependent relaxations in segments of porcine posterior descending coronary arteries submaximally precontracted with the thromboxane A2 mimetic, U-46619. The effects of BK were reduced by L-NG-monomethylarginine (L-NMMA) and 6-anilinoquinoline-5,8-quinone (LY-83583), respective inhibitors of nitric oxide (NO) synthase and guanylate cyclase, but were unaffected by the cytochrome P450 monoxygenase blocker, thiopentone sodium; however, BK effects were slightly reduced by dimethyl sulfoxide (DMSO), an hydroxyl radical scavenger. Relaxant responses were reduced markedly by ouabain, a sodium pump inhibitor but only slightly by tetraethylammonium (TEA) and charybdotoxin, respective blockers of Ca-activated (KCa) and large-conductance (BKCa) K+ channels. However, BK responses were practically abolished by TEA + L-NMMA + ouabain while unaffected by apamin, 4-aminopyridine and glibenclamide, blockers of small-conductance KCa voltage-sensitive and ATP-sensitive K+ channels, respectively. In segments submaximally precontracted with K+, BK-induced relaxation was lower than that of those precontracted with U-46619, and was further reduced by L-NMMA, LY-83583 and especially, ouabain; L-NMMA + ouabain + TEA abolished the effect. Precontraction of segments with higher K+ concentrations almost abolished the relaxation. These results suggest that the relaxation to BK is mediated: 1) by endothelial NO release which activates guanylate cyclase of smooth muscle cells; 2) by hydroxyl radicals; and 3) by an endothelial hyperpolarizing factor, that does not seem to be a metabolite derived from cytochrome P450 monoxygenases and that relaxes activating K+ channels (mainly BKCa), and especially, the sodium pump.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed1998

Design and evaluation of chitosan/ethylcellulose mucoadhesive bilayered devices for buccal drug delivery.

This paper describes the preparation of new buccal bilayered devices comprising a drug-containing mucoadhesive layer and a drug-free backing layer, by two different methods. Bilaminated films were produced by a casting/solvent evaporation technique and bilayered tablets were obtained by direct compression. The mucoadhesive layer was composed of a mixture of drug and chitosan, with or without an anionic crosslinking polymer (polycarbophil, sodium alginate, gellan gum), and the backing layer was made of ethylcellulose. The double-layered structure design was expected to provide drug delivery in a unidirectional fashion to the mucosa and avoid loss of drug due to wash-out with saliva. Using nifedipine and propranolol hydrochloride as slightly and highly water-soluble model drugs, respectively, it was demonstrated that these new devices show promising potential for use in controlled delivery of drugs to the oral cavity. The uncrosslinked chitosan-containing devices absorbed a large quantity of water, gelled and then eroded, allowing drug release. The bilaminated films showed a sustained drug release in a phosphate buffer (pH 6.4). Furthermore, tablets that displayed controlled swelling and drug release and adequate adhesivity were produced by in situ crosslinking the chitosan with polycarbophil.

Adhesives↗

PubMed1998

Formulation of L-asparaginase-loaded poly(lactide-co-glycolide) nanoparticles: influence of polymer properties on enzyme loading, activity and in vitro release.

This paper describes the preparation and characterisation of poly(lactide-co-glycolide) (PLG) nanoparticles containing the enzyme L-asparaginase. L-Asparaginase was encapsulated in PLG nanospheres using a water-in-oil-in-water solvent evaporation technique. The effect of the copolymer molecular weight and the presence of carboxyl-end groups in the copolymer chain on the physicochemical and in vitro release properties of the nanoparticles was investigated. Results indicated that size, encapsulation efficiency and in vitro release properties (enzymatic activity retention and protein quantification) of the nanoparticles were affected by the PLG molecular weight. As expected, nanoparticles made of high-molecular-weight PLG had a larger size, a higher loading and la slower release rate than those made od a low-molecular-weight PLG. Nevertheless, the most relevant factor affecting the entrapment and release of L-asparaginase from PLG nanoparticles was the presence of free carboxyl-end groups in the PLG chain. The nanoparticles made of PLG with free carboxyl-end groups had a high protein loading (4.86%, w/w) and provided a continuous delivery of the active enzyme for 20 days. However, the enzyme loading was lower (2.65%, w/v) and no active enzyme was detected in the release medium after a 14-day incubation period when nanoparticles were made of PLG with carboxyl-end groups esterified. These results give evidence of the potential of PLG nanospheres for the continuous delivery of L-asparaginase for extended periods of time and show the effect of the PLG chain end-groups in the amount and activity of the enzyme loaded into the nanospheres.

Asparaginase↗

PubMed1998

Design of microencapsulated chitosan microspheres for colonic drug delivery.

Among the different approaches to achieve colon-selective drug delivery, the use of polymers, specifically biodegraded by colonic bacteria, holds great promise. In this work a new system which combines specific biodegradability and pH-dependent release is presented. The system consists of chitosan (CS) microcores entrapped within acrylic microspheres. Sodium diclofenac (SD), used as a model drug, was efficiently entrapped within CS microcores using spray-drying and then microencapsulated into Eudragit L-100 and Eudragit S-100 using an oil-in-oil solvent evaporation method. The size of the CS microcores was small (1.8-2.9 microns) and they were encapsulated within Eudragit microspheres (size between 152 and 233 microns) forming a multireservoir system. Even though CS dissolves very fast in acidic media, at pH 7.4, SD release from CS microcores was delayed, the release rate being adjustable (50% dissolved within 30-120 min) by changing the CS molecular weight (MW) or the type of CS salt. Furthermore, by coating the CS microcores with Eudragit, perfect pH-dependent release profiles were attained. No release was observed at acidic pHs, however, when reaching the Eudragit pH solubility, a continuous release for a variable time (8-12 h) was achieved. A combined mechanism of release is proposed, which considers the dissolution of the Eudragit coating, the swelling of the CS microcores and the dissolution of SD and its further diffusion through the CS gel cores. In addition, infrared (IR) spectra revealed that there was an ionic interaction between the amine groups of CS and the carboxyl groups of Eudragit, which provided the system with a new element for controlling the release. In conclusion, this work presents new approaches for the modification of CS as well as a new system with a great potential for colonic drug delivery.

Chitin↗

PubMed1998

Synaptic immunolocalization of glyoxylate-complex molecules in the striate areas of the rat cerebral cortex: light and electron microscopic studies.

The location of glyoxylate-complex molecules has been investigated in several areas of the rat cerebral cortex using the immunohistochemical peroxidase-antiperoxidase (PAP) method. Antibodies against glyoxylate-complex molecules have been developed in the rabbit after immunization with a glyoxylate-bovine serum albumin conjugate. Observations carried out with the light microscope demonstrated positive immunostaining at the membrane level of scattered neurons located in all cortical areas, mainly in cortical layer IV. The striate areas (17, 18, 18a) had both the greatest number of immunopositive neurons and the most intense ones. At the electron microscopic level, it was observed that in the striate areas an immunopositive reaction was located mainly in the periphery of synaptic vesicles of some nerve endings, and in both pre- and postsynaptic membranes of these synaptic structures. The presence of glyoxylic acid and glyoxylate-complex molecules in such areas leads us to suggest that these substances could play an important role in selected synaptic contacts in which some pyramidal and non-pyramidal neurons are involved.

Animals↗

PubMed1998

Influence of experimental parameters on the characteristics of poly(lactic acid) nanoparticles prepared by a double emulsion method.

Nanoparticles were prepared by the double emulsion method (w/o/w), using methylene chloride as an organic solvent and polyvinyl alcohol (PVA) or human serum albumin (HSA) as a surfactant. Experimental parameters such as the preparation temperature, the solvent evaporation methods, the internal aqueous phase volume, the surfactant concentration and the polymer molecular weight were investigated for particle size, the zeta potential, the residual surfactant percentage and the polydispersity index. Preparation parameters leading to particles with well-defined characteristics such as an average size around 200 nm and a polydispersity index lower than 0.1 were identified. The conditions were optimized to ensure protein encapsulation: a cool temperature, a short processing time, a sufficient internal aqueous phase and careful washing. It appeared that the higher the surfactant concentration in the external aqueous phase was, the smaller the particles, the lower the polydispersity index and the higher the residual amount of surfactant were. For PVA or HSA, the agreement between the convenient surfactant concentration and its critical aggregation concentration could be emphasized. Otherwise, an increased polymer molecular weight led both to a slightly decreased particle size and to a lower polydispersity index. Moreover, multilayer absorption of PVA which does not depend on Poly(lactic-acid) molecular weight was exhibited. Finally, the zeta potential resulted from the polymer molecular weight and the residual PVA.

Drug Delivery Systems↗

PubMed1998

Protein encapsulation within polyethylene glycol-coated nanospheres. I. Physicochemical characterization.

The development of injectable nanoparticulate "stealth" carriers for protein delivery is a major challenge. We have shown the possibility of entrapping human serum albumin (HSA) in polyethylene glycol (PEG)-coated monodisperse biodegradable nanospheres with a mean diameter of about 200 nm, prepared from amphiphilic diblock PEG-polylactic acid (PLA) copolymers, with loadings up to 9% (w/w). Microscopic techniques and surface analysis studies enabled us to prove that the protein was well entrapped and not adsorbed onto the particle surface. Zeta potential and water uptake studies corroborated that part of the PEG chains are located in the nanosphere matrix. Water uptake in the nanospheres was related to their chemical composition, i.e., the respective wt% of PEG and PLA in the matrix, and not on their fabrication procedure. The hydrophilic PEG blocks absorbed up to 130% (w/w) water, whereas PLA absorbed only about 10% (w/w). However, the rate of swelling at the beginning of the process was related to the structure of the matrix, more particularly to the manner in which PEG was disposed at the surface. Furthermore, it was shown that the PEG "brush" at the nanosphere surface drastically reduces HSA adsorption on the PEG-PLA nanospheres compared to the PLA ones.

Adsorption↗

PubMed1998

Development of new chitosan-cellulose multicore microparticles for controlled drug delivery.

Chitosan (CS) is a very interesting biomaterial for drug delivery; however its use in oral administration is restricted by its fast dissolution in the stomach and limited capacity for controlling the release of drugs. To address this limitation, a new microparticulate CS controlled release system, consisting of hydrophilic CS microcores entrapped in a hydrophobic cellulosic polymer, such as cellulose acetate butyrate (CAB) or ethyl cellulose (EC) was proposed. These microparticles were obtained with different types of CS and various core/coat ratios, with the particle size in all cases being smaller that 70 microns. Using sodium diclofenac (SD) and fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) as model compounds, the properties of these new microparticles for the entrapment and controlled release of drugs and proteins were investigated. Results showed that the entrapment efficiency of SD was very high irrespective of the processing conditions. Furthermore, for both model compounds (SD and FITC-BSA) it was possible to modulate the in vitro release of the encapsulated molecules by changing the core properties (CS salt, Mw, core/coat ratio) or the coating polymer. The microparticles were stable at low pH and thus, suitable for oral delivery without requiring any harmful cross-linkage treatment.

Animals↗

PubMed1998

Protein encapsulation and release from poly(lactide-co-glycolide) microspheres: effect of the protein and polymer properties and of the co-encapsulation of surfactants.

Despite the recognised role of the poly(lactide-co-glycolide) (PLGA) in the encapsulation and release of proteins from PLGA microspheres, the importance that the characteristics of the protein have in these processes has not yet been sufficiently investigated. The aim of this work was to study the simultaneous effect of the protein and PLGA properties and of the microencapsulation process on the physicochemical and in vitro release characteristics of protein-loaded PLGA microspheres. Two model proteins of different isoelectric points (pI), bovine serum albumin (BSA) (pI = 4.6) and lysozyme (LZM) (pI = 11.2), and two different molecular weights (Mw) of PLGA were selected. Microspheres were prepared using the water-in-oil-in-water (w/o/w) solvent extraction and the oil-in-oil (o/o) solvent evaporation techniques. Results showed that BSA was efficiently encapsulated independent of the PLGA Mw, whereas the encapsulation of LZM was favoured with the low Mw PLGA. The co-encapsulation of a surfactant (poloxamer 188 or 331) reduced the protein encapsulation efficiency, especially of BSA. These results suggested that the tensoactive properties of the protein and its affinity for the PLGA are major determinants of the protein encapsulation. Both proteins released faster from the microspheres prepared by the o/o solvent evaporation procedure, with respect to those prepared by the w/o/w solvent extraction technique. In addition, both polymer Mw and protein type had an effect on the protein release rate. The release rate of both proteins, in the absence of a surfactant, was faster from the low Mw PLGA microspheres. However, the release rate constant was higher for BSA than for LZM irrespective of the PLGA Mw. In addition, the co-encapsulation of a surfactant led, in most cases, to a faster release of the encapsulated protein. To conclude, these results suggest that protein release from PLGA microspheres is not only governed by the PLGA erosion rate and protein diffusion through the water-filled channels, but is highly affected by the protein properties and its possible interaction with PLGA and its degradation products.

Chemical Phenomena↗

PubMed1998

Stealth PLA-PEG nanoparticles as protein carriers for nasal administration.

PURPOSE: The aim of the study was to encapsulate a model protein antigen, tetanus toxoid (TT), within hydrophobic (PLA) and surface hydrophilic (PLA-PEG) nanoparticles and to evaluate the potential of these colloidal carriers for the transport of proteins through the nasal mucosa. METHODS: TT-loaded nanoparticles, prepared by a modified water-in-oil-in-water solvent evaporation technique, were characterized in their size, zeta potential and hydrophobicity. Nanoparticles were also assayed in vitro for their ability to deliver active antigen for extended periods of time. Finally, 125I-TT-loaded nanoparticles were administered intranasally to rats and the amount of radioactivity recovered in the blood compartment, lymph nodes and other relevant tissues was monitored for up to 48 h. RESULTS: PLA and PLA-PEG nanoparticles had a similar particle size (137-156 nm) and negative surface charge, but differed in their surface hydrophobicity: PLA were more hydrophobic than PLA-PEG nanoparticles. PLA-PEG nanoparticles, especially those containing gelatine as an stabilizer, provided extended delivery of the active protein. The transport of the radiolabeled protein through the rat nasal mucosa was highly affected by the surface properties of the nanoparticles: PLA-PEG nanoparticles led to a much greater penetration of TT into the blood circulation and the lymph nodes than PLA nanoparticles. Furthermore, after administration of 125I-TT-loaded PLA-PEG nanoparticles, it was found that a high amount of radioactivity persisted in the blood compartment for at least 48 h. CONCLUSIONS: A novel nanoparticulate system has been developed with excellent characteristics for the transport of proteins through the nasal mucosa.

Administration, Intranasal↗

PubMed1998

Role of lipid peroxidation and the glutathione-dependent antioxidant system in the impairment of endothelium-dependent relaxations with age.

1. Age-related changes in the blood prooxidant-antioxidant state, as well as its influence on the relaxant responses to acetylcholine (ACh) were studied in the tail artery from 6-, 24- and 30-month-old Sprague-Dawley (SD) rats. 2. Malondialdehyde (MDA) plasma levels increased 2 and 3 times in 24- and 30-month-old rats, respectively, when compared with 6-month-old rats (0.43+/-0.09 microM). This increase was accompanied by an induction of 6-phosphogluconate dehydrogenase (6PG-DH) and glutathione reductase (GR) activities in red blood cells from 24-month-old rats. In 30-month-old rats, a further induction of these enzymatic activities, as well as glucose-6-phosphate dehydrogenase (G6P-DH) and glutathione peroxidase (GPx) activities was observed. 3. No differences with age were found in the concentration-response curves to ACh in isolated tail artery segments from 6- and 24-month-old rats precontracted with 0.3 microM noradrenaline (NA). However, a decrease in sensitivity to ACh-induced relaxation was observed in 30-month-old rats; EC30 values were 3.5 (1.3-8.0) x 10(-7) M and 18.1 (8.9-30.1) x 10(-7) M for 6- and 30-month-old rats, respectively. Moreover, a decrease in maximum ACh relaxation (10 microM) was found in 30-month-old rats in comparison with that obtained in 6-month-old rats (58.5+/-3.9% and 42.5+/-3.4% of previous NA contraction, respectively). 4. Incubation of tail artery segments with MDA (0.5, 1 or 10 microM) caused a reduction of ACh-induced relaxations that was different in the three ages. Thus, the reduction of ACh-induced relaxations became significant with 0.5 microM MDA in 6-, with 1 microM MDA in 24-, and with 10 microM MDA in 30-month-old rats. In addition, MDA did not cause a shift in the concentration-response curve to ACh, but a decrease in the maximum response. 5. Superoxide dismutase (SOD; 150 u ml(-1), a superoxide anion scavenger) reversed the inhibitory effect of MDA on ACh-induced relaxations at all ages studied. 6. We conclude that: (1) ageing produces an increase in lipid peroxidation process, as indicated by the increase in MDA plasma levels, that is accompanied by an induction of lipid peroxide detoxification enzymes: (2) the changes in prooxidant-antioxidant equilibrium with age contribute, at least partially, to the impairment of the relaxant responses evoked by ACh; and (3) the effect of MDA appears to be mediated by superoxide anion at all ages studied.

Acetylcholine↗

PubMed1998

The L-arginine inhibition of rat middle cerebral artery contractile responses is mediated by inducible nitric oxide synthase.

1. The effect of L-arginine (L-Arg), the nitric oxide synthase (NOS) substrate, on the responses to prostaglandin F2alpha (PGF2alpha, 10 microM) and K+ (120 mM) in rat middle cerebral artery (MCA) segments was analysed. 2. PGF2alpha induced a stable contraction of 0.35+/-0.06 mN mm(-1); the subsequent addition of bradykinin (BK, 1 microM) produced a relaxation of 42+/-9% of the PGF2alpha-induced tone. K+ induced a response consisting of a rapid basal tone increase (1.42+/-0.16 mN mm(-1)) followed by a decrease to a stable phase (1.24+/-0.15 mN mm(-1)). 3. L-Arg (0.1 mM), but not D-Arg, decreased the basal tone and reduced the contraction to PGF2alpha in segments with and without endothelium. The contractile response to K+ was also reduced and not maintained in the presence of L-Arg. 4. The inhibitory effect of L-Arg on the PGF2alpha- and K+-induced contractions was completely reversed by the NOS inhibitor, NG-monomethyl-L-arginine (L-NMMA, 0.1 mM). 5. Pre-incubation of segments with dexamethasone (1 microM), to inhibit inducible NOS (iNOS), or with the antibiotic polymyxin B (10 microg ml(-1)) reduced the L-Arg inhibition, whereas it was increased by lipopolysaccharide (LPS, 100 ng ml(-1)), an inductor of iNOS. L-NMMA antagonized the effects of dexamethasone and LPS. 6. The present results suggest that L-Arg inhibition of the PGF2alpha- and K+-induced contractions in rat MCA is the result of NO synthesis by iNOS stimulation.

Animals↗

PubMed1998

Efficacy of topical cyclosporine-loaded nanocapsules on keratoplasty rejection in the rat.

PURPOSE: To evaluate the efficacy of a topical formulation of nanocapsules loaded with 1% cyclosporine A (CsA), which has been previously demonstrated to enhance CsA corneal penetration, compared to 1% CsA in migliol oil on a penetrating keratoplasty rejection model in the rat. METHODS: Lewis rats received orthotopic corneal allografts from inbred Fisher donors. Rats were treated with 10 microliters of the following topical solutions four times daily for 30 days, starting one day before surgery: Group 1 (n = 9), 1% CsA-loaded nanocapsules; group 2 (n = 13), 1% CsA dissolved in migliol oil; group 3 (n = 12), migliol oil; group 4 (n = 13), no treatment. Rejection index, mean survival time and rejection percentage were calculated for each group, and CsA levels in blood were measured. RESULTS: The rejection percentage was 84.6% for group 2, 91.7% for group 3, and 100% for groups 1 and 4, with no significant differences among groups. Mean graft survival time was 7.3 days for group 1, 15.5 days for group 2, 8.36 days for group 3, and 7.69 days for group 4, with significant differences between group 2 and the other groups. Systemic CsA levels were only detectable in group 2. CONCLUSIONS: CsA formulated in migliol oil delayed corneal rejection onset, but blood levels were evident in this group. CsA loaded-nanocapsules showed no effect on rejection and the drug was not detectable in blood. These data, along with the current concepts on corneal graft rejection immunology, suggest that the immunomodulatory effect of topical CsA in the prevention of corneal graft rejection may be systemically-mediated.

Administration, Topical↗

PubMed1998

Endothelial factors and autoregulation during pressure changes in isolated newborn piglet cerebral arteries.

To analyze newborn cerebrovascular autoregulation, middle cerebral arteries from 3-4-d-old piglets were cannulated, and diameter changes after transmural pressure variation were measured. After an equilibration period at 30 mm Hg, pressure was modified from 10 to 70 mm Hg in 20-mm Hg steps. Segments with endothelium showed vasodilation during pressure decrease and vasoconstriction during pressure increase. In each case the maximum response was about 5% that of the resting diameter. Segments without endothelium responded passively to pressure change. Vasodilation during pressure decrease was reduced by the preferential calcium-activated potassium (KCa) channel blocker, tetraethylammonium (1 mM), and was absent with the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methylester (L-NAME, 10 microM). The NO synthase substrate, L-arginine (10 microM), counteracted the dilation blockade caused by L-NAME. The cyclooxygenase inhibitor indomethacin (10 microM) and the endothelin A receptor antagonist BQ-123 (10O microM) eliminated the pressure increase-induced vasoconstriction. The ATP-sensitive potassium channel blocker, glibenclamide (1 microM), and the endothelin B receptor antagonist, BQ-788 (10 nM), did not modify the autoregulatory response. None of these drugs modified the passive changes produced by pressure variations in segments without endothelium. These results suggest that: 1) piglet middle cerebral artery autoregulation is endothelium-dependent; 2) NO and KCa channels are involved in vasodilation during transmural pressure decrease, and 3) endothelin-1, through endothelin A receptors, and prostanoids mediate vasoconstriction during pressure increase.

Animals↗

PubMed1998

Comparative study of microbiological and histopathological techniques used for the detection of Helicobacter pylori.

This study evaluates the sensitivity, specificity and predictive values of several techniques commonly used for the detection of Helicobacter pylori in an analysis of 105 biopsy specimens (gastric and duodenal). For comparative purposes, the techniques investigated were divided into 2 groups: histopathological and microbiological. The former included hematoxylin-eosin and Giemsa stains, a Gram stain modified for use in tissue, and immunohistochemical techniques. Microbiological analysis was performed using culture, the urease test and the conventional Gram stain. The immunohistochemical techniques proved to be the most sensitive (93%). The modified Gram stain was sufficiently sensitive (92%) and specific (97%) for the detection of the bacterium. When combined with a microbiological technique such as the urease test, this stain showed increased sensitivity (96%) but its specificity was reduced to 94%. This combination of tests is recommended for the detection of H. pylori in biopsy specimens since it is easily performed at low cost and gives excellent results. For economical reasons, it is suggested that the use of immunohistochemical techniques should be restricted to specific cases.

Bacteriological Techniques↗