Biomedical subjects

M J Alonso

Publications and source records attributed to M J Alonso.

At least 73 records · Page 4Linked to original sources

PubMed1997

[Eosinophilic gastroenteritis caused by bee pollen sensitization].

A 34-year-old Spanish woman with a lifelong history of seasonal rhinoconjunctivitis and honey intolerance (pyrosis and abdominal pain) developed, 3 weeks after starting ingestion of bee pollen, astenia, anorexia, abdominal pain, diarrhoea, peripheral blood hypereosinophilia and elevated serum total IgE levels. A duodenal biopsy showed eosinophilic infiltration of the mucosal layer. Other causes of hypereosinophilia were not found. Repeated parasitological stool studies, as well as a duodenal aspirate showed negative results. Symptoms, hypereosinophilia and elevated IgE levels resolved after bee pollen ingestion was stopped. This is a typical case of eosinophilic gastroenteritis by ingestion of bee pollen in a woman with intolerance to honey bee, because the patient fulfilled the usual diagnostic criteria: gastrointestinal symptoms were present, eosinophilic infiltration of the digestive tract was demonstrated by biopsy, no eosinophilic infiltration of other organs was found and the presence of parasites was excluded. Honey intolerance and/or bee pollen administration should be considered as a cause of eosinophilic gastroenteritis.

Adult↗

PubMed1997

Effect of lysozyme on the stability of polyester nanocapsules and nanoparticles: stabilization approaches.

The efficacy of colloidal particles as drug carriers is closely related to their interaction with proteins and enzymes in different body fluids. In the present work, we analysed the interaction phenomenon between lysozyme (LZM), a positively charged enzyme that is highly concentrated in mucosas, and two different drug carriers: nanocapsules made of an oily core coated by the polymer poly-epsilon-caprolactone (PECL) and nanoparticles made solely of PECL. Results showed that the interaction of LZM with these colloidal drug carriers is highly affected by their surface charge. Nanocapsules, because of their important negative charge (-40 mV), adsorbed a great amount of LZM, which is positively charged. This adsorption process, which was also evidenced by the significant reduction of the nanocapsules' negative surface charge, led to the degradation of the polymer coating and the aggregation of the nanocapsules. In contrast, nanoparticles had a low negative surface charge (-8 mV) and adsorbed only a small amount of LZM, which did not cause the destabilization of the system. Furthermore, the molecular weight of the polymer forming the nanoparticles did not change. Finally, it was observed that the destabilizing effects caused by the adsorption of LZM onto the nanocapsules can be prevented by previous adsorption of the cationic poly(amino acid) poly-L-lysine. Using this approach the adsorption of LZM was hindered and its consequences avoided.

Adsorption↗

PubMed1997

Chronic local tissue reactions, long-term immunogenicity and immunologic priming of mice and guinea pigs to tetanus toxoid encapsulated in biodegradable polymer microspheres composed of poly lactide-co-glycolide polymers.

Immunogenicity of tetanus toxoid (TT) encapsulated in biodegradable polymer microspheres composed of poly lactide (PLA) or poly lactide-co-glycolide (PLGA) polymers was evaluated in mice and guinea pigs for 1 year. Microsphere formulations made from polymers differing in molecular weight and composition elicited significantly higher IgG antibody levels than soluble TT in mice. The antibody levels elicited by microsphere formulations in mice and guinea pigs were similar to those elicited by a single injection of AlPO4 adsorbed TT. Immunogenicity was not consistently better with a particular polymer composition, molecular weight or microsphere size. However, animals primed with TT-containing microspheres showed significantly higher anamnestic response to a low dose booster 1 year after priming than those primed with AlPO4 adsorbed TT. Microspheres made from low molecular weight PLGA polymer showed a minimal local tissue reaction 1 year after injection. In contrast, aluminum adjuvant formed local granulomas which persisted for 1 year after injection. Microsphere formulations used in this study released a small fraction of antigenic TT during in vitro release studies due to denaturation of TT during encapsulation and hydration of microspheres. Nevertheless, strong priming of immune responses were seen. It remains to be demonstrated whether stabilization of TT would lead to more immunogenic microsphere formulations.

Albumins↗

PubMed1997

Duodenal leishmaniasis diagnosed by biopsy in two HIV-positive patients.

We describe two cases of duodenal leishmaniasis in patients with human immunodeficiency virus (HIV) infection, diagnosed by light and electron microscopy. The patients presented nonspecific signs and symptoms, blood cultures were sterile, and serological tests for Leishmania spp. were negative. Endoscopy showed normal-appearing mucosa in one patient and possible peptic duodenitis in the other patient. In these patients, the parasite was only detected in a duodenal biopsy specimen. In view of the unusual location of the parasite and the fact that the diagnostic and dissemination of the disease was established by means of conventional biopsy, this is not a routine procedure for the diagnosis of leishmaniasis because the classic procedures require the demonstration of antibodies and visualization in bone marrow, lymph nodes, liver and/or spleen aspirates. We decided to report these two cases to call attention to the possible finding of Leishmania amastigotes in biopsies from intestinal mucosa in HIV infected patients.

Adult↗

PubMed1997

Chitosan and chitosan/ethylene oxide-propylene oxide block copolymer nanoparticles as novel carriers for proteins and vaccines.

PURPOSE: The aim of this study was to investigate the interaction between the components of novel chitosan (CS) and CS/ethylene oxide-propylene oxide block copolymer (PEO-PPO) nanoparticles and to evaluate their potential for the association and controlled release of proteins and vaccines. METHODS: The presence of PEO-PPO on the surface of the nanoparticles and its interaction with the CS was identified by X-ray photoelectron spectroscopy (XPS). The mechanism of protein association was elucidated using several proteins, bovine serum albumin (BSA), and tetanus and diphtheria toxoids, and varying the formulation conditions (different pH values and concentrations of PEO-PPO), and the stage of protein incorporation into the nanoparticles formation medium. RESULTS: BSA and tetanus and diphtheria toxoids were highly associated with CS nanoparticles partly due to electrostatic interactions between the carboxyl groups of the protein and the amine groups of CS. PEO-PPO also interacted electrostatically with CS, thus competing with the proteins for association with CS nanoparticles. A visible amount of PEO-PPO was projected towards the outer phase of the nanoparticles. Proteins were released from the nanoparticles at an almost constant rate, the intensity of which was closely related to the protein loading. Furthermore, the tetanus vaccine was released in the active form for at least 15 days. CONCLUSIONS: CS and CS/PEO-PPO nanoparticles prepared by a very mild ionic crosslinking technique are novel and suitable systems for the entrapment and controlled release of proteins and vaccines.

Animals↗

PubMed1997

Pressure modulates the reactivity of isolated rabbit epicardial arteries.

1. The magnitude of responses to prostaglandin F2 alpha (PGF2 alpha) and adenosine (ADO) in pressurized rabbit epicardial coronary arteries (367 +/- 44 microns o.d.) with intact endothelium was assessed when they developed spontaneous tone. 2. The arteries were cannulated and changes in arterial diameter registered with an automated video perfusion system. The vessels, in the stabilization period at 60 mmHg, were divided into two groups, one exposed to a longitudinal stretch of +20% of unstretched initial length (Lo), and the other to +35% Lo, which developed spontaneous tone. In both cases, diameter-pressure curves were obtained by changing the intravascular pressure from 30 to 120 mmHg in aleatory steps of 30 mmHg (dp/dt = 15 mmHg s-1). 3. Diameter reduction of the arteries with stretch of +35% Lo in response to 1 microM PGF2 alpha was greater than that with stretch of +20% Lo. Likewise, increase of the arteries that had +35% Lo in response to 1 microM ADO was greater than with +20% Lo. 4. Intravascular flow (40 microliters min-1) increased the tone level of arteries. The addition of PGF2 alpha enhanced this tone which was similar to that obtained with a stretch of +35% Lo and no flow, whereas the effect of ADO was increased. 5. These data indicate that the vasomotor responses of PGF2 alpha and ADO are modulated by the degree of longitudinal stretch in epicardial arteries.

Adenosine↗

PubMed1997

Glyoxylate oxidoreductase activity and glyoxylate-like molecules in the motor, somatosensory, cyngulate and olfactory areas of the rat frontoparietal cortex. Histoenzymological and immunohistochemical study.

In the present paper the presence of glyoxylate-oxidoreductase (GOR, GLYO-DH) activity and glyoxylate-like molecules has been investigated in several areas of the rat frontoparietal cortex-motor, somatosensory, cyngulate and olfactory areas-, using the histoenzymological and immunocytochemical methods. Antibodies against glyoxylate-like molecules have been obtained using a glyoxylate-BSA conjugate as immunogen. GOR activity as well as immunostaining for glyoxylate-like molecules were observed in the periphery of the perikaryon and neuronal processes of scattered neurons in the above mentioned cortical areas. At the electron microscopic level positive immunoreaction was found associated to synaptic vesicles in axon terminals, and also within glial cells. These findings reveal both the presence of glyoxylate-like molecules and glyoxylate metabolism in neurons of the motor, somatosensory, cyngulate and olfactory areas of the rat frontoparietal cortex. The presence of such molecules associated to synaptic vesicles within the axon terminals suggest the possible involvement of these molecules in some type of neurotransmission.

Alcohol Oxidoreductases↗

PubMed1997

Simultaneous detection of antigens and specific DNA sequences of human papillomavirus in uterine cervical biopsy specimens. Description of a double-labelling technique.

We studied 70 uterine cervical biopsy specimens with a histological diagnosis compatible with human papillomavirus (HPV) infection. We carried out immunohistochemical and in situ hybridization techniques and selected 22 specimens that had given a positive result with both techniques. We then used a double-labelling technique (a combination of immunohisto-chemistry and in situ hybridization) to detect simultaneously viral antigens and specific gene sequences of HPV. With this technique we found three different cell types in the tissue: (1) cells with black nuclei, positive by immunohisto-chemistry and in situ hybridization; (2) cells with red nuclei, negative by immunohistochemistry and positive by in situ hybridization; and (3) cells with light blue nuclei, negative by both techniques. In this study we describe this technique; and we believe that the simultaneous detection of viral antigens and specific gene sequences of HPV may be very useful in the study of the virus-cell interaction.

Antigens, Viral, Tumor↗

PubMed1996

Hepatic Morphological alterations induced by zidovudine (ZDV) in an experimental model.

Zidovudine (AZT) inhibits HIV replication. Many studies have demonstrated its toxic myopathic effect in both HIV-positive patients treated with the drug and experimental animal models. So far hepatic lesions induced by AZT have not been reported. In our study, an experimental rat model was used in which the rats were administered AZT (1 mg/ml) in drinking water; histological and ultrastructural alterations were observed in the liver of treated animals and compared with the findings in control animals. The histological alterations detected were turbid swelling, vacuolar degeneration and microvacuolar fatty degeneration of panlobular distribution; these lesions were progressively greater as the duration of treatment increased. The ultrastructural alterations detected involved the mitochondria (similar to those described in cardiac muscle), smooth and rough endoplasmic reticulum (SER and RER), and the accumulation of fat and glycogen in the hepatocytes of treated animals. The histopathological and ultrastructural findings in our experimental model suggest hepatotoxicity induced by AZT or its catabolites in treated, as compared to control animals.

Animals↗

PubMed1996

Comparative in vitro evaluation of several colloidal systems, nanoparticles, nanocapsules, and nanoemulsions, as ocular drug carriers.

Three different colloidal carriers, namely, nanoparticles and nanocapsules made of poly-epsilon-caprolactone and submicron emulsions, were designed, and their capacity for increasing the comeal penetration of drugs was investigated. The three systems differed in their inner structure and composition, but they had a similar size (200-250 nm) and a negative superficial charge (-16 to -42 mV). Indomethacin, which was used as a model drug, was dispersed at a molecular level within the colloidal systems, no chemical interaction between the polymer and the drug being detected. Release of the encapsulated indomethacin occurred very rapidly upon high dilution in a buffered medium and was independent of the composition of the system. The in vitro comeal penetration of the encapsulated indomethacin was more than 3-fold that of the commercial eye drops. This increased penetration was similar for the three formulations investigated, which therefore excludes the influence of the inner structure or chemical composition of the colloidal systems on the comeal penetration of indomethacin. Thus, it could be stated that the main factor responsible for the favorable comeal transport of indomethacin is the colloidal nature of these carriers rather than their inner structure or composition.

Animals↗

PubMed1996

Pulsed controlled-released system for potential use in vaccine delivery.

Biodegradable polymeric devices intended to provide a viable route for single-dose vaccination were developed using controlled-release technology. One of the main challenges in the development of these devices was to overcome several water-mediated inactivation processes that occur in conventional polymeric systems. Our strategy was focused on the prevention of antigen exposure to environmental conditions. For this purpose a microencapsulation process was designed and optimized to provide an inert and insulated environment for the bioactive material inside controlled-release systems. Tetanus toxoid (TT) was used as a model antigen. The systems consist of core-wall microcapsule structures in which the antigenic protein is entrapped into oil-based cores of TT surrounded by outer polymer shells made of poly(D,L-lactide-co-glycolide), thus potentially protecting the bioactive material against deleterious conditions. Furthermore, using these microcapsules, pulses of immunochemically detected TT were programmed to release at two different times (3 and 7 weeks), as corroborated by in vitro release studies. The engineering of these specific antigen release properties was possible by careful selection of the copolymer composition and molecular weight. The final formulations were characterized with respect to morphology, structure, size distribution, and amount of immunochemically detected TT encapsulated. The new systems offer the potential to control the manner and timing of delivery. Over 92% of the TT released over a 63 day period from these microcapsules was immunochemically detected.

Capsules↗

PubMed1996

Polyester nanocapsules as new topical ocular delivery systems for cyclosporin A.

PURPOSE: Nanocapsules composed of an oily core (Migliol 840) (MG) surrounded by a poly-epsilon-caprolactone (PECL) coat were evaluated as potential vehicles for the topical ocular administration of cyclosporin A (CyA). METHODS: A 2(3) experimental factorial design was applied to optimize the coating of the oily nanodroplets by a solvent displacement technique and to encapsulate a high dose of CyA. The variables investigated were: volume of oil (MG), amount of polymer (PECL), and volume of the organic solvent (acetone) used to dissolve the polymer. RESULTS: Nanocapsules had a mean size in the range of 210-270 nm, a negative zeta potential (between -55 and -60 mV) and a maximum loading capacity of 50% (CyA/PECL ratio). These highly loaded nanocapsules displayed a thick spongeous polymer coating around the oily nanodroplets. The corneal levels of CyA were up to 5 times higher for the encapsulated CyA than for the oily solution of CyA. In addition, these levels remained significantly higher than those of the control group (oily solution) for up to 3 days. Furthermore, the area-under-the-curve (AUC) values were significantly increased for the encapsulated CyA (319.98) with respect to the oily control (74.34). CONCLUSIONS: The CyA-loaded nanocapsules are shown to be interesting vehicles for the improvement of the ocular penetration of CyA.

Administration, Topical↗

PubMed1996

Effect of ageing and hypertension on endothelial modulation of ouabain-induced contraction and sodium pump activity in the rat aorta.

OBJECTIVE: To investigate the endothelial modulation of the rat thoracic aorta sodium-potassium ATPase activity and its possible alteration by ageing and hypertension. METHODS: Male spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) aged 5 weeks, 3, 6, 12 and 18 months were anaesthetized. Their aortae were dissected and divided into cylindrical segments and vasomotor responses obtained in segments with or without endothelium were recorded on a polygraph. RESULTS: Endothelium removal increased ouabain responses in young (aged 3 and 6 months) WKY rat aortic segments, but reduced those obtained in old (aged 18 months) WKY rat segments and in segments from SHR aged 3, 6 and 12 months. An enhancement of contractions in response to ouabain with age was observed in both strains, but it occurred at earlier ages in vessels from SHR. At ages 3 and 6 months, responses to ouabain were higher in intact vessels from SHR than they were in those from WKY, but were lower in endothelium-denuded vessels. In aortic segments from 6-month-old WKY rats and SHR, ouabain reduced or abolished potassium-induced relaxations in segments with or without endothelium, respectively. In vessels from SHR and WKY rats aged 18 months, ouabain abolished potassium-induced relaxations both in intact and in endothelium-denuded vessels. CONCLUSIONS: Endothelium of WKY rat aorta might release a factor in response to ouabain that modulates its contraction negatively. Endothelium of SHR and old WKY rat aorta releases a contracting factor that modulates ouabain contractions positively. Thus, the inhibitory effect of endothelium on contractions induced by ouabain might be lost as a consequence both of hypertension and of age, being replaced by an endothelium-dependent contracting factor that facilitates ouabain responses.

Aging↗

PubMed1996

Endothelium stimulates the vascular smooth muscle cell Na/K pump of normotensive and hypertensive rats by a protein kinase C pathway.

OBJECTIVE: To investigate the mechanisms involved in the endothelial stimulation of the vascular smooth muscle cell Na/K pump and their possible alteration by hypertension. METHODS: The Na/K pump activity of vascular smooth muscle cells (VSMC) from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) was studied using the radioactive analogue of K+ 86Rb+. Conditioned medium of bovine endothelial aortic cells was used to investigate the endothelial modulation of VSMC Na/K pump activity. RESULTS: Conditioned medium enhanced VSMC Na/K pump activity (ouabain-sensitive 86Rb+ uptake), this effect being higher in SHR cells. This stimulatory effect was neither modified in Na(+)-loaded cells from both rat strains nor inhibited by the Na/H exchange blocker amiloride. Permeable analogues of cyclic adenosine and guanosine monophosphates did not modify the baseline VSMC Na/K pump activity of WKY rats and SHR, and subsequently the guanylate cyclase inhibitor methylene blue did not alter the conditioned medium-induced stimulation of the pump. However, the Ca(2+)-channel inhibitor nifedipine reduced the Na/K pump stimulation by conditioned medium, this decrease being higher in WKY rat than in SHR VSMC. Moreover, treatment with phorbol 12,13-dibutyrate for 24 h or with the protein kinase C inhibitor staurosporine for 15 min reduced the conditioned medium-induced Na/K pump activation in both VSMC cultures. CONCLUSIONS: Na/K pump stimulation by conditioned medium of endothelial cells is mediated mainly via activation of protein kinase C in VSMC from either WKY or SHR VSMC. However, SHR VSMC show some alterations in their intracellular signalling pathways.

Animals↗

PubMed1996

Noradrenergic transmission in the tail artery of hypertensive rats transgenic for the mouse renin gene Ren-2.

1. The aim of the present study was to analyse the noradrenergic transmission in the tail artery of hypertensive rats transgenic for the mouse renin gene Ren-2 (TGR) in comparison with its control, the Sprague-Dawley (SD) rat. 2. Electrical field stimulation (EFS) of vascular segments produced frequency-dependent vasoconstrictions that were significantly greater in TGR arteries. 3. These contractions were abolished by tetrodotoxin (0.1 microM). Phentolamine (50 nM) and prazosin (1 - 10 nM) produced an inhibition of these responses that was significantly greater in SD arteries, whereas that produced by yohimbine (0.5-1 microM) was higher in TGR arteries. In both strains, propranolol (1 microM) potentiated the responses to EFS, and this increase was observed at lower frequencies in TGR arteries. 4. The EFS-evoked [3H]-noradrenaline (NA) release was significantly greater in TGR than in SD rats. However, NA (10 nM-10 microM) reduced and yohimbine and phentolamine (10 nM-10 microM) increased the tritium outflow to a similar degree in both strains. 5. Exogenous NA also induced greater vasoconstriction in TGR arteries. 6. These results suggest the existence in TGR tail artery of an increase in: (a) NA-release and alpha 2-adrenoceptor-mediated contractions, which could contribute to the elevated blood pressure in these rats; and (b) beta-adrenoceptor-mediated vasodilatations, which may be a mechanism to counteract high blood pressure.

Animals↗

PubMed1996

Improved ocular bioavailability of indomethacin by novel ocular drug carriers.

The ability of different drug carriers to improve the ocular bioavailability of drugs was investigated in the rabbit eye. The assayed drug carriers were suspensions of nanoparticles, nanocapsules and microparticles made of poly-epsilon-caprolactone (PECL) and a submicron emulsion. Results indicated that the three submicron systems, nanoparticles, nanocapsules and emulsion, increased more than 3-fold the indomethacin concentration in the cornea, aqueous humour and iris-ciliary body at 0.5 and 1 h post-instillation. Furthermore, an increased indomethacin ocular bioavailability of 300% was observed after instillation of the submicron systems in comparison with the value obtained for a commercial solution. In contrast, the microparticles hardly increased the ocular bioavailability of indomethacin. The mechanism of interaction of the colloidal carriers with the corneal epithelium was investigated by confocal laser scanning microscopy. Confocal images indicated that submicron particles penetrate into the corneal epithelium cells by an endocytic mechanism. The similar behaviour of the three colloidal carriers suggests that any of their specific ingredients (PCEL, lecithin and oil) acts as a penetration enhancer or an endocytotic stimulator. On the other hand, the favourable ocular penetration of indomethacin when encapsulated in the colloidal carriers, but not in the microparticles, led us to assume that the colloidal nature of these carriers is the main factor responsible for the increased ocular bioavailability of indomethacin. PECL nanoparticles and nanocapsules as well as submicron emulsions are shown to be novel corneal drug carriers, thus representing a useful approach for increasing the ocular bioavailability of drugs.

Analysis of Variance↗

PubMed1996

Labelling pattern obtained by non-isotopic in situ hybridization as a prognostic factor in HPV-associated lesions.

In the study of infection of the lower female genital tract caused by human papillomavirus (HPV), one of the main concerns is the search for prognostic factors to predict the evolution of premalignant low- and high-grade squamous intraepithelial lesions. This study has evaluated the prognostic usefulness of the patterns of positive reaction obtained by non-isotopic in situ hybridization (NISH), referred to as diffuse, punctate, or mixed 'labelling patterns'. The study examined 141 vulvar and uterine cervical biopsy specimens that were positive for HPV by a NISH screening technique and that had the following histological diagnoses: low-grade squamous intraepithelial lesion (LSIL; n = 87); high-grade squamous intraepithelial lesion (HSIL, n = 40); and squamous cell carcinoma (SCC n = 14). Typing of all the specimens was carried out by NISH with DNA probes specific for HPV types 6/11 (low risk), 16/18 (high risk), and 31/33/51 (intermediate risk), and the labelling pattern observed in each specimen was recorded. Statistical analysis of the results showed that there was a significant difference in the distribution of labelling patterns, both by lesion diagnosis (P < or = 0.004) and by infecting viral type (P < or = 10(-6)). Lesions with a punctate or mixed pattern are considered more likely to undergo malignant evolution and consequently have a worse prognosis than lesions with a diffuse pattern.

Carcinoma, Squamous Cell↗

PubMed1996

Strategies for stabilising tetanus toxoid towards the development of a single-dose tetanus vaccine.

The stability of tetanus toxoid (TT) has been investigated for the purpose of enhancing its immunogenicity when encapsulated in TT-polylactide-co-glycolide (PLGA) microspheres. In this paper we describe our simulations of various potential inactivating events during microsphere processing and release of the antigen. These include: i) processing aqueous TT solutions in the presence of organic solvents, ii) exposing lyophilized TT to moisture, and iii) incubating vaccine with the degrading PLGA. At 37 degrees C, TT began to aggregate in solution a few hours after the addition of either methylene chloride or ethyl acetate to form a single oil-water interface. Similarly, exposure of the lyophilized vaccine to elevated humidity caused the antigen to lose solubility. Previous analysis of moisture-induced aggregates has revealed that formaldehyde, which is stored in labile linkages in the vaccine following its detoxification, is the precursor to the reactive species in the principal aggregation pathway [1]. Methods to inhibit this mechanism, such as blocking nucleophilic amino groups of TT with succinic anhydride, were verified. Succinylation of TT resulted in the incorporation of 10-fold greater antigenically active vaccine in PLGA microspheres relative to the unmodified TT following processing by double-emulsion/solvent evaporation with ethyl acetate, strongly suggesting the formaldehyde-mediated aggregation pathway also occurs during the deleterious conditions of microsphere processing. Incubation of solutions of TT in the presence of excess blank (unloaded) poly (D,L) lactide (mol. wt. 2000) microspheres led to a dramatic reduction in pH (approximately 3.2 units after one day at 45 degrees C) and simultaneous precipitation of TT. Stabilisation in the presence of the degrading polymer is likely to be the final obstacle before controlled-release preparations can be formulated to release antigenically active TT over extended time periods. Hence, mechanistic analyses as described here may be crucial for the development of effective single-dose vaccines.

Delayed-Action Preparations↗