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Biomedical subjects

R Pujol

Publications and source records attributed to R Pujol.

At least 73 records · Page 4Linked to original sources

X-ray-induced synaptonemal complex damage during meiotic prophase in female fetuses of Rattus norvegicus.

The different types of damage (synaptic anomalies, chromosome reorganizations and nucleolar fragmentation) observed in oocytes from female rat fetuses irradiated at 14, 16 and 18 days of gestation (d.g.), respectively, indicates the existence of a special sensitivity at the different stages of prophase to X-rays. At the highest dose (5 Gy), we observed a decrease in some of the synaptonemal complex indicators of chromosomal damage, probably reflecting a selection against cells with the highest degree of chromosome anomalies.

Animals↗

[Risk factors of arteriopathy of the lower extremities: lipid and not lipid factors].

BACKGROUND: The role of lipoproteins as markers of peripheral arterial disease (PAD) is not well defined. METHODS: We measured both lipid and non-lipid risk factors in 51 male patients with angiographically proven PAD and in 56 control subjects. The independent association of risk factors with PAD was evaluated by means of a multiple logistic regression analysis. RESULTS: The levels of cholesterol bound to high density lipoprotein (HDLc) and to its subfraction HDL2 were lower and triglycerides were higher in patients than in control subjects (1.0 +/- 0.3 vs 1.2 +/- 0.3, p < 0.003; 0.4 +/- 0.2 vs 0.5 +/- 0.3, p < 0.03; and 1.8 +/- 1.2 vs 1.3 +/- 0.7, p < 0.02, respectively). Total cholesterol and LDLc levels were similar in both groups. In the multiple logistic regression analysis that was done with lipid parameters, a statistically significant association of triglycerides (OR = 1.73; CI95% = 1.06-2.80) and HDLc (OR = 0.15; CI95% = 0.05-0.50) with PAD was observed, while HDL subfractions and apolipoproteins were not significantly associated. In the multiple logistic regression analysis that was done with non-lipid parameters, hypertension (OR = 5.35; CI95% = 1.86-15.4) and smoking (packs-year) (OR = 1.04; CI95% = 1.10-1.06) were the only significantly associated with PAD. When lipid and non-lipid parameters were included in the regression analysis, a statistically significant association between hypertension, smoking and HDLc with PAD was observed. CONCLUSIONS: Among lipid risk factors, a low HDLc and high triglycerides, and among non-lipid risk factors hypertension and smoking, are significantly and independently associated with lower limb arteriopathy.

Adult↗

Cloning of the genes encoding two murine and human cochlear unconventional type I myosins.

Several lines of evidence indicate a crucial role for unconventional myosins in the function of the sensory hair cells of the inner ear. We report here the characterization of the cDNAs encoding two unconventional type I myosins from a mouse cochlear cDNA library. The first cDNA encodes a putative protein named Myo1c, which is likely to be the murine orthologue of the bullfrog myosin I beta and which may be involved in the gating of the mechanotransduction channel of the sensory hair cells. This myosin belongs to the group of short-tailed myosins I, with its tail ending shortly after a polybasic, TH-1-like domain. The second cDNA encodes a novel type I myosin Myo1f which displays three regions: a head domain with the conserved ATP- and actin-binding sites, a neck domain with a single IQ motif, and a tail domain with the tripartite structure initially described in protozoan myosins I. The tail of Myo1f includes (1) a TH-1 region rich in basic residues, which may interact with anionic membrane phospholipids; (2) a TH-2 proline-rich region, expected to contain an ATP-insensitive actin-binding site; and (3) a SH-3 domain found in a variety of cytoskeletal and signaling proteins. Northern blot analysis indicated that the genes encoding Myo1c and Myo1f display a widespread tissue expression in the adult mouse. Myo1c and Myo1f were mapped by in situ hybridization to the chromosomal regions 11D-11E and 17B-17C, respectively. The human orthologuous genes MYO1C and MYO1F were also characterized, and mapped to the human chromosomal regions 17p18 and 19p13.2-19p13.3, respectively.

Amino Acid Sequence↗

Protein kinase C may be involved in synaptic repair of auditory neuron dendrites after AMPA injury in the cochlea.

A suitable model of sudden deafness occurring after acoustic trauma or ischemia, is obtained in guinea pigs by an acute intracochlear perfusion of 200 microM alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA), a glutamate analog. By overloading the AMPA/kainate receptors, located post-synaptically to inner hair cells (IHCs), it induces a massive swelling of primary auditory neuron dendrites, which disconnects the IHCs. This synaptic uncoupling and the resulting hearing loss are followed by a progressive regrowth of dendrites, which make new synapses with IHCs, leading to a functional recovery of auditory responses that is completed after 5 days. Knowing the role of protein kinase C in neuroplastic events, we studied the expression of its isoforms alpha,beta(I,II) and gamma, respectively pre- and post-synaptic, in auditory neurons at various times after AMPA administration. In untreated cochleas, we observed an expression of PKC alpha,beta(I,II) and gamma in cell bodies of primary auditory neurons. After the intracochlear administration of AMPA, both isozymes were transiently overexpressed, with a peak at 3-6 h, followed by a decrease after about 24 h. At this point in time immuno-electron microscopy revealed some regrowing dendrites immunoreactive for PKCgamma. Five days after AMPA, when the auditory responses were restored, PKCgamma levels were still elevated in ganglion cell bodies.

Animals↗

Development of the organ of Corti in horseshoe bats: scanning and transmission electron microscopy.

The late prenatal and early postnatal development of the organ of Corti were studied in the horseshoe bat (Rhinolophus rouxi) by using scanning and transmission electron microscopy. Arrangements and dimensions of stereocilia bundles, together with their contacts with the tectorial membrane, were found to be adult-like shortly before birth, and thus before the biological onset of hearing (3-5 days after birth). During the first postnatal week, there were baso-apical gradients in disappearing kinocilia on inner hair cells (IHC), microvillis of supporting cells, and marginal pillars. The lower basal cochlear turn was mature with respect to these regressing structures at 3 days after birth, the apical turn at 10 days after birth. At birth, cytodifferentiation was found to be completed, and the tunnel of Corti and innermost spaces of Nuel had opened. The ultrastructure of IHCs was not markedly different from that at later ages. In outer hair cells (OHC), the adult-like regular arrangement of a single layer of subsurface cisternae and pillars was seen as soon as protrusions of supporting cells had withdrawn from the lateral wall of OHCs (basal turn at birth and throughout the cochlea 2 days after birth). Numerous efferent endings contacted the somata of IHCs up to the second postnatal week. Since the medial olivocochlear system is absent in horseshoe bats, the adult-like innervation pattern of OHCs was established at the biological onset of hearing. During the first 2 postnatal weeks, the cytoskeleton of pillar and Deiters cells, and the specialized Deiters cups developed. The organ of Corti appeared adult-like at 14 days, apart from the persistence of a reduced tympanic cover layer attached to the basilar membrane. Morphological data support physiological findings that the first broadly tuned auditory responses arise from the basal turn. The distinct low to high frequency gradient in development of sensitivity during the first 2 postnatal weeks of the horseshoe bat was not, however, matched by morphological gradients, and it would appear that the development of the cytoskeleton of supporting cells contributed to the establishment of tuning in the auditory fovea. Adult-like morphology of the organ of Corti coincided with the emergence of sharply tuned responses from the auditory fovea, but there was no clear-cut correlate for the shift in tuned foveal frequency representation that occurred during the following 3 weeks.

Animals↗

Cutaneous involvement in sarcoidosis. Relationship to systemic disease.

Sarcoidosis is an antigen-mediated disease defined by granuloma formation in different organs. It involves mainly the mediastinal and peripheral lymph nodes, lungs, eyes, skin, liver, and spleen. Cutaneous lesions of sarcoidosis may be specific, showing histologically noncaseating granulomas, or nonspecific, most typically erythema nodosum. Frequently, both types of skin lesions are the means of presentation of the disease and may contribute to the diagnosis. A workup for systemic sarcoidosis should be undertaken in every patient with sarcoid cutaneous granulomas. Some types of cutaneous lesions have prognostic significance. Lupus pernio and plaques are associated with more severe systemic involvement and more chronic course, while erythema nodosum is the hallmark of acute and benign disease.

Acute Disease↗

Identification of preferentially expressed cochlear genes by systematic sequencing of a rat cochlea cDNA library.

107 expressed sequence tags (ESTs) from a rat cochlea cDNA library were identified by systematic sequencing coupled to database selection and RT-PCR analysis of novel sequences. This approach led us to select a clone, pCO8, showing no significant homology with any database sequence, that corresponds to a mRNA whose expression is restricted to the cochlea, except for traces detected in brain. Additional clones with novel sequences enriched in the cochlea were also found. ESTs bearing significant homologies with database sequences (63 out of 107) were classified according to the putatively encoded protein. They include tissue-specific genes not previously described in the cochlea as well as known genes from other species. We performed in situ hybridization in cochlear tissues to localize the pCO8 mRNA and that of clone pCO6 which is 100% homologous to the delayed rectifier potassium channel drk1. We found that both mRNAs were exclusively expressed in the cellular body of the primary auditory neurons from the spiral ganglion of the cochlea. These results indicate that this approach is an efficient way to identify novel genes that could be of importance in cochlear function.

Animals↗

Resistance to hyperosmotic stress in boar spermatozoa: the role of the ionic pumps and the relationship with cryosurvival.

The resistance to changes in the osmolarity of boar sperm was used to measure the resistance of boar sperm cells against freezing/thawing. Semen was incubated for 5 min in different solutions ranging from about 600 mOsm to about mOsm, and at 4 degrees C, 16 degrees C or 37 degrees C (undisturbed media). This undisturbed media was constituted by NaCl, glycerol or glucose. This semen was then placed in an isoosmotic solution (disrupted solutions). Incubation in undisturbed media did not alter the percentages of viability or altered acrosomes, except when the initial incubation has been at 37 degrees C and with osmolarities above 1000 mOsm. Viability and altered acrosome statistics were strongly modified in disrupted media. These effects are dependent upon the initial osmolarity of the media, but not upon the temperature. Pre-incubation with ouabain or amiloride did not affect spermatozoa incubated at 16 degrees C in a 2211 mOsm, NaCl medium. However, in sperm incubated in this 2211 mOsm medium and then rapidly placed in an isoosmotic solution, ouabain induced a decrease in viability and an increase in altered acrosomes. Amiloride did not affect the response of cells to the disrupted medium. Some significant correlations were observed among the percentages of altered acrosomes after hyperosmotic stress and some quality parameters of the fresh boar semen, especially the motion parameters. Although the resistance to hyperosmotic stress could be a valuable parameter in assessing fresh quality analysis, its usefulness in frozen-thawed semen is compromised, since other factors beside osmotic changes are involved in the resistance of boar semen to freezing-thawing. The NA+/K+, ouabain-dependent ATP-ase activity seems to be related to the mechanisms of resistance to hyperosmotic stress in boar sperm.

Acrosome↗

Implication of NMDA type glutamate receptors in neural regeneration and neoformation of synapses after excitotoxic injury in the guinea pig cochlea.

In the adult mammalian cochlea, the ability of nerve fibres to regenerate has been observed following disruption of the organ of Corti by various means, or transsection of the cochlear nerve in the internal auditory meatus. Based upon the implication of glutamate as a neurotransmitter at synapses between sensory hair cells and terminal dendrites of the auditory nerve in the mammalian cochlea, we have developed, in a previous study, an in vivo model of neural regeneration and formation of synapses after the destruction of the afferent nerve endings by local application of the glutamate agonist alpha-amino-3-hydroxy-5-methyl-isoxazol-propionic acid (AMPA). In situ hybridization experiments performed during the re-innervation process revealed an overexpression of mRNA coding for NR1 subunit of N-methyl-D-aspartate (NMDA) receptors in the spiral ganglion neurons, suggesting that these receptors are implicated in neural regenerative processes. The present study has been designed to study the functional implication of NMDA receptors in the regrowth and synaptic repair of auditory dendrites in the guinea pig cochlea, by blocking the NMDA receptors during the period of normal functional recovery. In a first set of experiments, we recorded compound action potential after acute perilymphatic perfusion of cumulative doses (0.03-10mM) of DL 2-amino-5-phosphonovalerate (D-AP5), a NMDA antagonist, to determine the efficiency of the drug. In a second set of experiments, the auditory dendrites were destroyed by local application of the glutamate agonist AMPA. The blockage of NMDA by the antagonist D-AP5 applied with an osmotic micropump delayed the functional recovery and the regrowth of auditory dendrites. The findings of our study support the hypothesis that, in addition to acting as a fast transmitter, glutamate has a neurotrophic role via the activation of NMDA receptors.

2-Amino-5-phosphonovalerate↗

Clinical skills assessment with standardized patients.

Previous projects (Combell I & II) to assess clinical skills were conducted in medical schools in Catalonia, in order to introduce a model of such an assessment using standardized patients (SP). The aim of this study (Combell III) was to measure selected characteristics of our model. Seventy-three medical students in the final year at the Bellvitge teaching unit of the University of Barcelona participated in a clinical skills assessment (CSA) project that used 10 SP cases. The mean group scores for the four components of clinical skills for each day of testing were studied, and ratings for each student in the 10 sequential encounters were checked. The study also compared the clinical skills scores with their academic grades. The total case mean score (mean score of history-taking, physical examination and patient notes scores) was 51.9%, and the mean score for communication skills was 63.6%. The clinical skills scores over the 8 testing days showed no day-to-day differences. The study did not find differences among the sequential encounters for each student (training effect). There was a lack of correlation between clinical skills scores and academic grades. The project demonstrated the feasibility of the method for assessing clinical skills, confirmed its reliability, and showed that there is no correlation between scores with this method and academic examinations that mainly reflect knowledge.

Clinical Competence↗

Synaptic repair mechanisms responsible for functional recovery in various cochlear pathologies.

In some cochlear pathologies, temporary hearing loss can be followed by complete or partial functional recovery. Our previous findings suggest the involvement of an excitotoxic (glutamate-related) disruption of inner hair cell (IHC)-auditory nerve synapses, followed by synaptic regeneration. It is essential to understand the molecular mechanisms responsible for this synaptic repair if new therapeutic strategies are to be developed. In guinea pig cochleas, acute synaptic excitotoxic damage (mimicking what occurs with acoustic trauma or local ischemia) is achieved by locally applying AMPA, a glutamate agonist. This results in a total disruption of all IHC-auditory dendrite synapses, together with a disappearance of cochlear potentials. Within the next 5 days, however, a recovery of both the normal pattern of IHC innervation and the physiological responses is observed. The fact that the blockage of the NMDA receptors during functional recovery delayed the regrowth of neurites and the restoration of hearing suggests that glutamate plays a neurotrophic role via activation of NMDA receptors. Experiments are in progress to investigate, among other factors, the role of other glutamate receptor subunits. A reversible in vivo antisense strategy is being developed to overcome the lack of specificity of some antagonists. First results bode well for future pharmacological therapies in cochlear pathologies where glutamatergic synapses are likely to be involved; i.e., noise trauma, ischemia-related sudden deafness, and neural presbycusis.

Animals↗

[An analysis of the impact of the COMBELL project on clinical medical competence. Competencia clínica Bellvitge].

OBJECTIVE: The most frequent tests for evaluate medical education in our country, do not analyze clinical competence. The aims of this project was to introduce a method for the assessment of clinical skills in two different groups of students using the simulation methodology with standardized patients and to compare with classical methods of assessment. METHODS: Seventy three last year medical students were evaluated using 10 standardized patients encounters in April 1995 (COMBELL III project) and the other group of last year medical students was assessed in March 1996 using the same group of standardized patients (COMBELL IV project). RESULTS: The global score for the COMBELL III, was 50.6% +/- 4.8 lower to COMBELL IV, 55.6% +/- 6 (p = 0.0001), remarking the improving of the item of physical examination (39.6% +/- 7.7 versus 48.1 +/- 9.6 p < 0.0001). Personal intercommunication also improved, 63.1% +/- 11.4 versus 70.4% +/- 12.2 (p = 0.0008). We did not find differences between academic grades and did not find correlations between clinical competence assessment and academic grades in COMBELL III but we found correlations in COMBELL IV. CONCLUSIONS: Our results show that when this new assessment method was introduces in our medical school the clinical competence levels improved.

Clinical Competence↗

Effect of X-rays on germ cells in female fetuses of Rattus norvegicus irradiated at three different times of gestation.

We analyzed the effect of X-rays on the number of recoverable germ cells in female rat fetuses irradiated at three different times of gestation: 14 days (oogonia), 16 days (leptotene) and 18 days (zygotene). Our results show that irradiation significantly decreases the number of germ cells, and increases the incidence of synaptonemal complex fragmentation. The different sensitivity of the germ cells to irradiation could depend on the meiotic stage at the time of irradiation.

Animals↗

Fate of fumonisins B1 and B2 in steeped corn kernels.

Corn kernels naturally contaminated with fumonisins B1 and B2 were steeped by dipping the whole kernels in either water or a 0.3% solution of sodium bisulphite (pH = 5.7) for 6, 12, 24, 36 and 48 h. The toxin content of both the solid and the liquid were analysed for each time period. A different rate of interchange between the solid matrix and the water solution was found for FB1 and FB2 and some differences were found in the fate of both fumonisins between the two steeping systems studied. In addition, a preliminary study of the distribution of FB1 and FB2 between the germ and the remaining part of the kernel was carried out.

Carboxylic Acids↗

Are the pulmonary function tests and the markers of activity helpful to establish the prognosis of sarcoidosis?

The prognosis of sarcoidosis is difficult to establish. It depends mainly on the persistence of activity over time and the degree of functional impairment of the involved organs. The aim of this study was to analyze whether the pulmonary function tests and the more commonly used markers of activity, such as serum angiotensin-converting enzyme (SACE) and gallium-67 scan, are helpful to evaluate the prognosis of sarcoidosis, besides the clinical data. Over a 14-year period (1974-1987), 209 patients were diagnosed as having sarcoidosis at the Bellvitge Hospital, a 1,000-bed teaching hospital in Barcelona, Spain. Clinical, radiological, pulmonary function tests, and activity markers (SACE and gallium-67 scan) data at diagnosis were collected and classified as variables, and a definition of disease activity was established. One hundred sixteen patients were on follow-up, had all the variables available and were included in the statistical analysis. A Cox proportional-hazard regression model identified the following variables as independently influencing the persistence of activity over time: absence of erythema nodosum [risk ratio (RR) = 2.78; 95% confidence interval (CI): 1.48-5.18], age > or = 40 years (RR = 1.67; 95% CI: 1.008-1.04), SACE level > or = mean + 2 SD U/ml (RR = 1.45; CI: 0.99-1.07), hyperglobulinemia (RR = 2.47; CI: 0.98-6.24), forced vital capacity < 80% predicted (RR = 2.17; CI: 0.97-4.85), and male sex (RR = 1.8; CI: 0.95-3.45). We conclude that the pulmonary function tests and the SACE level but not the gallium scan are helpful to identify the factors predicting persistence of activity in sarcoidosis. Therefore, we recommend to add these tests to the initial clinical evaluation of patients with sarcoidosis in order to establish the prognosis and improve the therapeutic approach.

Adult↗

Periarticular ankle sarcoidosis: a variant of Löfgren's syndrome.

OBJECTIVE: To focus more attention on the syndrome characterized by periarticular ankle inflammation associated with bilateral hilar adenopathy--some authors regard it as a variant of the Löfgren syndrome, while other suggest it is a distinct clinical entity-- we present a series of 33 cases of periarticular ankle sarcoidosis. METHODS: Sarcoidosis was diagnosed in 330 patients at Bellvitge Hospital over a 20 year period. The medical charts of patients who presented with periarticular ankle inflammation (swelling of both ankles with acute inflammatory signs and preserved articular motion) were reviewed. RESULTS: 33 patients (10%) were identified. Periarticular ankle inflammation began during the spring in more than one-half (54.5%). The mean age was 33 years and patients were predominantly women (66.6%). Most cases (78.8%) were stage I on chest radiograph. All patients had thoracic gallium scans showing increased hilar/mediastinal uptake. Increased characteristic parotid, lacrimal, and/or submandibular uptake was found in 11 of 13 patients who had head scans. Four of 6 whose scans included the legs had bilateral ankle uptake. Erythema nodosum was present concomitantly in 36.3% of patients. No granulomas were found in 4 biopsies of periarticular ankle tissue. All 24 patients who were followed had inactive disease one year after diagnosis. CONCLUSION: The association of periarticular ankle inflammation with bilateral hilar adenopathy is an acute form of sarcoidosis that follows a benign course to total remission. It should be regarded as a variant of the Löfgren syndrome.

Adult↗