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Biomedical subjects

M Rodríguez

Publications and source records attributed to M Rodríguez.

At least 73 records · Page 4Linked to original sources

The use of ischaemic vessels as prostheses or tissue engineering scaffolds after cryopreservation.

OBJECTIVE: to evaluate the condition of organ donor arteries subjected to prolonged cold-ischaemia followed by cryopreservation, for their possible use as vascular grafts. MATERIALS AND METHODS: fresh specimens of human iliac artery from organ donors were used as controls. These arteries were divided into two portions, one of which was cryopreserved in an automated freezer. A further group of arteries was immersed in Wisconsin solution and kept for 4 days at 4 degrees C (cold-ischaemia). After this period, the arteries were also cut into two, and one of these portions was cryopreserved. All the cryopreserved arterial segments were stored for a month and then subjected to automated gradual thawing. The thawed specimens were evaluated by light microscopy, scanning and transmission electron microscopy, immunohistochemical analysis (MMPs, elastin, CD31, von Willebrand factor) and the in situ detection of fragmented DNA (TUNEL method). RESULTS: the most marked changes induced by cryopreservation were partial vessel deendothelialisation and morphological changes in cells of the intima that were in the process of detachment. No significant changes were observed in the medial layer, other than discrete elastic fibre fragmentation. Following cold-ischaemia, the endothelium was the most affected layer, with large denuded areas and exposure of the fibroelastic layer. Increased MMP-2 expression was also noted after cold-ischaemia. When subjected to both cold-ischaemia and cryopreservation, a large proportion of endothelial cells showed positivity for the TUNEL technique, however, no significant difference was observed between the ischaemic and the ischaemic/cryopreserved specimens. CONCLUSIONS: prolonged cold-ischaemia causes some additional damage to the arterial wall compared to cryopreservation alone. However, the structural component of the ischaemic vessel remains in a condition that is suitable for subsequent cryopreservation and use as a vessel substitute or a scaffold for tissue engineering.

Adult↗

Determination of triazines in water samples by high-performance liquid chromatography with diode-array detection.

Triazines are widely used herbicides that can be detected in the environment at trace level. A preconcentration step is necessary to determinate them before analysis. In this study, carbonaceous and polymeric adsorbents are compared with C18 for the solid-phase extraction of simazine, atrazine, and propazine in water samples in order to quantitate their levels by high-performance liquid chromatography using photodiode-array detection.

Chromatography, High Pressure Liquid↗

Role of individual and contextual effects in injury mortality: new evidence from small area analysis.

OBJECTIVE: To analyse the role of individual and contextual variables in injury mortality inequalities from a small area analysis perspective, looking at the data for the city of Barcelona (Spain) for 1992-98. SETTING: Barcelona (Spain). METHODS: All injury deaths in residents older than 19, which occurred in the period 1992-98 were included (n=4393). Age and sex specific mortality rates were calculated for each educational level and each cause of death (traffic injuries, falls, drug overdose, suicide, other injuries). The contextual variables included were the proportion of men unemployed, and the proportion of men in jail, in each neighbourhood. Multilevel Poisson regression models were fitted using data grouped by age, educational level, and neighbourhood for each sex. RESULTS: Death rates were higher in males, at the extremes of the age distribution (under 44 and over 74 years), and for lower educational levels. The results of the Poisson multilevel models indicate that inequalities by educational level follow a gradient, with higher risks for the population with no schooling, after having adjusted for the contextual variables of the neighbourhood. Such inequalities were more important in the youngest age group (20-34 years), as relative risk of 5.41 (95% confidence interval (CI) 3.9 to 7.4) for all injury causes in males and 4.38 (95% CI 2.3 to 8.4) in females. The highest relative risks were found for drug overdose. There was a contextual neighbourhood effect (the higher the deprivation, the higher the mortality) after having taken into account individual variables. CONCLUSION: The findings underscore the need to implement injury prevention strategies not only at the individual level taking into account socioeconomic position, but also at the neighbourhood level.

Adult↗

Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures.

The subcommissural organ (SCO), an ependymal (glial) circumventricular organ, releases glycoproteins into the cerebrospinal fluid; however, the regulation of its secretory activity is largely unknown. To identify neuroactive substances that may regulate SCO activity, we investigated immunocytochemically identified bovine SCO cells by means of calcium imaging. This analysis was focused on: (1) serotonin (5HT) and substance P (SP), immunocytochemically shown to be present in axons innervating the bovine SCO; and (2) ATP, known to activate glial cells. 5HT had no effect on the intracellular calcium concentration ([Ca(2+)](i)), and its precise role remains to be clarified. SP elicited rises in [Ca(2+)](i) in approx. 30% and ATP in even 85% of the analyzed SCO cells. These effects were dose-dependent, involved NK(3) and P2Y(2) receptors linked to G protein and phospholipase C (PLC) activation, and could not be mimicked by forskolin or 8-bromo-cAMP. In 50% of the SP-sensitive cells, the increases in [Ca(2+)](i) comprised calcium release from thapsigargin-sensitive intracellular stores and an influx of extracellular calcium via protein kinase C (PKC)-induced opening of L-type voltage-gated calcium channels (VGCCs). In the remaining SP-sensitive cells, the increase in [Ca(2+)](i) was caused exclusively by influx of extracellular calcium via VGCCs of the L-type. In all ATP-sensitive cells the increase in [Ca(2+)](i) involved calcium release from thapsigargin-sensitive intracellular stores and a PKC-mediated influx of extracellular calcium via L-type VGCCs. Our data suggest that SP and ATP are involved in regulation of the activity of SCO cells.

Adenosine Triphosphate↗

The deep mesencephalic nucleus as an output center of basal ganglia: morphological and electrophysiological similarities with the substantia nigra.

The deep mesencephalic nucleus (DMN) is a large midbrain reticular region between the superior colliculus, the substantia nigra compacta, the periaqueductal gray, and the medial geniculate body. Although some data suggest that it is involved in nociception and visceral control, its functions remain unclear. In the present study, by using morphological (combination of anterograde and retrograde tracers with immunocytochemistry and in situ hibrydization) and electrophysiological (firing activity and transynaptic response to striatal stimulation) methods, we show that a subpopulation of DMN cells shares many morphological and electrophysiological characteristics with those of the substantia nigra reticulata (SNR). These similarities include the following: 1) firing rate, firing pattern, and conduction velocity; 2) expression of GAD65, GAD67, and PV; 3) excitatory and inhibitory inputs from the striatum; and 4) projections to the ventral thalamus, superior colliculus, and pedunculopontine tegmental nucleus. Some differences were also found. In comparison with SN, DMN cells and striatal afferents are more sparsely distributed and they show conspicuous contralateral projections to the thalamus and superior colliculus. This suggests that, similarly to the SNR, the DMN acts as an output center of basal ganglia and probably facilitates the inter-hemispheric regulation of these centers.

Action Potentials↗

Plasma ionized calcium and parathyroid hormone concentrations in horses after endurance rides.

OBJECTIVE: To evaluate changes in plasma ionized calcium (Ca2+) and parathyroid hormone (PTH) concentrations in horses competing in endurance rides. DESIGN: Longitudinal clinical study. ANIMALS: 28 horses. PROCEDURE: Venous blood samples were obtained from horses before and after racing 80 km. Plasma pH and concentrations of Ca2+, PTH, inorganic phosphorus, albumin, lactate, and magnesium were measured. RESULTS: Overall, a significant decrease in mean (+/- SD) plasma Ca2+ concentration (from 6.44 +/- 0.42 to 5.64 +/- 0.42 mg/dl) and a significant increase in plasma PTH concentration (from 49.9 +/- 30.1 to 148.1 +/- 183.0 pg/ml) were found after exercise. Exercise also resulted in significant increases in plasma inorganic phosphorus, albumin, and lactate concentrations. No changes in plasma magnesium concentration or pH were detected after exercise. Plasma PTH concentration was not increased after exercise in 8 horses; in these horses, plasma PTH concentration decreased from 58.2 +/- 26.3 to 27.4 +/- 22.4 pg/ml, although plasma Ca2+ concentration was also decreased. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma Ca2+ concentration was decreased after racing for 80 km, compared with values obtained before racing. In most horses, an increase in plasma PTH concentration that was commensurate with the decrease in plasma Ca2+ was detected; however, some horses had decreased plasma PTH concentrations.

Animals↗

Synthesis, structure, and redox and catalytic properties of a new family of ruthenium complexes containing the tridentate bpea ligand.

We have prepared a new family of ruthenium complexes containing the bpea ligand (where bpea stands for N,N-bis(2-pyridyl)ethylamine), with general formula [Ru(bpea)(bpy)(X)](n+) (2, X = Cl(-); 3, X = H(2)O; 4, X = OH(-)), and the trisaqua complex [Ru(bpea)(H2O)(3)](2+), 6. The complexes have been characterized through elemental analyses, UV-vis and (1)H NMR spectroscopy, and electrochemical studies. For complex 3, the X-ray diffraction structure has also been solved. The compound belongs to the monoclinic P2(1)/m space group, with Z = 2, a = 7.9298(6) A, b = 18.0226(19) A, c = 10.6911(8) A, and beta = 107.549(8) degrees. The Ru metal center has a distorted octahedral geometry, with the O atom of the aquo ligand placed in a trans position with regard to the aliphatic N atom of the bpea ligand so that the molecule possesses a symmetry plane. NMR spectra show that the complex maintains its structure in aqueous solution, and that the corresponding chloro complex also has a similar structural arrangement. The pH dependence of the redox potential for the complex [Ru(bpea)(bpy)(H2O)](PF(6))(2) is reported, as well as the ability of the corresponding oxo complex to catalyze the oxidation of benzylic alcohol to benzaldehyde in both chemical and electrochemical manners.

Journal Article↗

Gradual thawing improves the preservation of cryopreserved arteries.

This study was designed to test a slow, controlled, automated process for the thawing of cryopreserved arteries, whereby specimen warming is synchronized with the warming of its environment. Segments of minipig iliac artery, 4-5 cm in length, were subjected to controlled, automated cryopreservation in a biological freezer at a cooling rate of 1 degrees C/min to -120 degrees C, followed by storage in liquid nitrogen at -196 degrees C for 30 days. Following storage, the arterial segments were subjected to rapid (warming rate of approximately 100 degrees C/min) or gradual (1 degrees C/min) thawing. Thawed specimens were processed for light microscopy and for scanning and transmission electron microscopy, Cell death was determined by the TUNEL method. Metalloproteinase (MMP) expression was estimated by immunohistochemical analysis. Most of the cryopreserved vessels subjected to rapid thawing showed spontaneous fractures, mainly microfractures, whereas these were absent in slowly thawed specimens. In rapidly thawed vessels, the proportion of damaged cells was double that observed in those thawed more gradually. Increased intensity and extent of MMP-2 expression was shown by rapidly thawed specimens. The slow-thawing protocol tested avoids the formation of spontaneous fractures and microfractures and the accumulation of fluid within the arterial wall tissue. This results in improved tissue preservation.

Animals↗

Dopamine cell degeneration induced by intraventricular administration of 6-hydroxydopamine in the rat: similarities with cell loss in parkinson's disease.

In an attempt to find a convenient rat model to study cell vulnerability in Parkinson's disease, we have investigated the cell-loss profile in different midbrain dopaminergic nuclei and subnuclei of rats injected with 6-hydroxydopamine (6-OHDA) in the third ventricle. Following administration of different doses (5-1000 microgram) of 6-OHDA, motor behavior was evaluated and tyrosine hydroxylase-immunostained neurons were counted in the A8 group and different subdivisions of A9 and A10 groups. Animals developed hypokinesia, repetitive chewing movements, and catalepsia. Signs of cell degeneration were evident from the first day after injection, reaching the definitive pattern at the end of the first week. There was a similar degeneration in both brain sides, the A9 group showing the highest degree of cell-loss, followed by A8 and A10 groups. In the A9 group, the degeneration mostly affected those subgroups located in its ventral, lateral, and posterior regions. In the A10 group the degeneration mainly affected the parabrachial pigmented nucleus, the paranigral nucleus and the ventral tegmental area. This topographic pattern of degeneration is very similar to that previously described in Parkinson's disease, suggesting that this model may be a useful tool in the study of the cell vulnerability mechanisms in this neurodegenerative disorder. In addition, our results also showed that small dopaminergic neurons are more resistant to degeneration than the large ones. In some DA subgroups, the cells that contained calbindin but not calretinin were less vulnerable to the neurotoxic effect of 6-OHDA.

Animals↗

Structural and phylogenetic analysis of the gamma-actin encoding gene from the penicillin-producing fungus Penicillium chrysogenum.

The nucleotide sequence of a 2994-bp genomic fragment, including the gamma-actin encoding gene from Penicillium chrysogenum, has been determined, showing an open reading frame (ORF) of 1756 bp interrupted by five introns with fungal consensus splice-site junctions. The 5' untranslated region contains a consensus TATA box, five CAAT motifs, and two large pyrimidine stretches. The predicted protein (375 amino acids) revealed high identity to gamma-actins from fungi (>90%), and gene phylogenies support the grouping of P. chrysogenum actin close to those from the majority of the filamentous fungi. The actA gene is present as a single copy in the genome of P. chrysogenum, and its expression is constitutive during penicillin fermentation, showing a single 1.4-kb transcript.

5' Untranslated Regions↗

Effect of the thawing process on cryopreserved arteries.

This study was designed to explore the changes that occur in cryopreserved pig arteries following different thawing procedures, before and after being placed in an in vitro flow circuit. Segments of minipig iliac artery were cryopreserved in complete minimal essential medium plus 10% dimethylsulphoxide and stored in liquid nitrogen at -196 degrees C for 30 days. Three study groups were established according to whether the arterial specimens were fresh (control, n = 20), cryopreserved and rapidly thawed (RT) at 37 degrees C (n = 22) or cryopreserved and subjected to controlled, automated slow thawing (ST) (n = 22). Half of the specimens of each group were subsequently placed in the flow circuit for 72 hr. Evaluation was made of morphological and ultrastructural changes. Cell damage was established using the TUNEL method. All cryopreserved specimens showed endothelial denudation that was most extensive in those subjected to rapid thawing. Slowly thawed specimens showed improved cell viability and organization of the vessel wall, compared to those thawed rapidly. Under conditions of flow, the damage induced by the freezing/thawing process was enhanced. These findings suggest that (a) slow thawing of cryopreserved arteries results in improved preservation of the structure and viability of vessels, and (b) the damage induced by freezing/thawing is enhanced when vessels are subjected to flow in an in vitro circuit.

Animals↗

Chlamydia pneumoniae DNA in the arterial wall of patients with peripheral vascular disease.

BACKGROUND: Chlamydia pneumoniae is a human respiratory pathogen that has recently been related to the genesis of symptomatic atherosclerosis. C. pneumoniae has been studied more widely in relation to coronary atherosclerosis than to peripheral arterial occlusive disease (PAOD). The present study aimed to retrospectively analyze the presence of C. pneumoniae DNA in patients with PAOD. MATERIALS AND METHODS: A seminested PCR method was applied on 85 samples from 71 patients with PAOD secondary to surgical treatment. The control group comprised 50 patients with chronic superficial venous insufficiency who required varicose resection surgery. RESULTS: The number of patients, number of samples studied and percentage of patients found to be positive in the PCR study were 17, 18 and 59%, respectively, for arteries of the lower extremities; 15, 16 and 60% for aneurysm of the abdominal aorta; 22, 23 and 73% for carotid stenosis and 17, 18 and 65% for aortic stenosis. C. pneumoniae DNA was found in six external pudendal arteries (12%) of the control group, significantly lower than the incidence in the patient group (p < 0.0001). CONCLUSION: A causal relationship between chronic C. pneumoniae infection and PAOD cannot be ruled out. On the contrary, the high incidence of C. pneumoniae DNA detected in our patients suggests that C. pneumoniae infection may play some role in the pathogenesis of peripheral vascular disease.

Aged↗

Delayed P300 during Sternberg and color discrimination tasks in poor readers.

The P300 ERP component was studied in poor and normal readers, using Sternberg and color discrimination (Spaceships) tasks. During the first one, subjects must decide if a probe item belongs or not to a set of digits previously presented. In the second one, the participants must shoot violet spaceships with one key and other than violet spaceships with another key. There were no significant differences between groups with respect to reaction times, but a larger proportion of errors was observed in poor readers. Longer P300 latencies were recorded for poor readers than controls in both tasks. P300 amplitudes showed topographical differences between the two groups: Poor readers' P300 is larger frontally during the Sternberg task, but smaller at posterior sites during the Spaceships task. These results suggest that poor readers may have deficiencies during the early processing stage, such as visual stimulus evaluation. Poor readers also appear to have deficits for classifying and memorizing visual stimuli.

Child↗

[Microbiologic profile in symptomatic pregnant women's genital secretions in Gran Buenos Aires, Argentina].

BACKGROUND: Establish the prevalence of microorganisms associated with genital colonization in symptomatic pregnant women. In order to review the evolution of frequent pathogens ecology and adjust the laboratory design, in a population attended at the public health Hospital, in the Great Buenos Aires. METHODS: Vaginal and endocervical samples, were explored for specific detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Streptococcus agalactie, Trichomonas vaginalis, Candida spp., Mycoplasma hominis, Ureaplasma urealyticum and bacterial vaginosis (VB).Direct methods, culture, inmunodetection and PCR, were employed. In 1999; 198 women, with genital discharge, were studied. Age in the group range from 16 to 42 years old (Median 27 years old). RESULTS: In 51 cases (25.7%) none of the above microorganisms or bacterial vaginosis were detected. In 30 cases (15.1%) bacterial vaginosis was diagnosed. Frequency of detection was: Ureaplasma urealyticum, 49,5%; Candida spp., 34,3%; Mycoplasma hominis, 14.1%; Streptococcus agalactie, 4.5%; Trichomonas vaginalis, 3.5%; Chlamydia trachomatis, 2.5%. No detection of Neisseria gonorrhoeae was demonstrated. CONCLUSIONS: There is a relevant frequency of bacterial vaginosis. On the other hand, lower prevalence of the Trichomonas vaginalis and Chlamydia trachomatis and also the absence of Neisseria gonorrhoeae was demonstrated. Culture for Streptococcus agalactie (at birth) and detection of Chlamydia trachomatis. must be extended to all pregnant women. Study of species and drug sensitivity of Candida spp., and detection of Ureaplasma urealyticum, Mycoplasma hominis and Neisseria gonorrhoeae, have to be explored under specific clinical requirement.

Adolescent↗