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Biomedical subjects

M López

Publications and source records attributed to M López.

At least 73 records · Page 4Linked to original sources
PubMed2001

Analysis of phenolic constituents of biological interest in red wines by high-performance liquid chromatography.

We describe a reversed-phase HPLC method that uses gradient elution and diode array detection to determine four biologically active phenolic constituents of red wines: gallic acid, trans-resveratrol, quercetin and rutin. The method permits direct injection without sample pre-treatment. ODS Hypersil served as the stationary phase; the gradient was formed by acetic acid, methanol, and water. Each analysis required an equilibration period of 10 min and a run time of 50 min for completion. Previously, total phenols were analysed according to the Folin-Ciocalteu method, using gallic acid as the standard, and the results are given as gallic acid equivalent.

Calibration↗
PubMed2001

Prepro-orexin mRNA levels in the rat hypothalamus, and orexin receptors mRNA levels in the rat hypothalamus and adrenal gland are not influenced by the thyroid status.

Orexins are two recently discovered neuropeptides that play an important role in the regulation of food intake and in the regulation of the sleep-wake cycle. In this work we examined the effects of thyroid hormones on prepro-OX expression in the rat hypothalamus, and OXRs expression in the rat hypothalamus and adrenal gland. Hypo- and hyperthyroidism were induced in adult male rats, and the levels of hypothalamic prepro-OX and OXRs mRNA, and adrenal OXRs mRNA were determined using semiquantitative reverse transcription-polymerase chain reaction and/or in situ hybridization. Our results indicate that thyroid status affects neither prepro-OX in the hypothalamus nor hypothalamic and adrenal gland OXRs expression.

Adrenal Glands↗
PubMed2001

Results of salvage surgery for local or regional recurrence after larynx preservation with induction chemotherapy and radiotherapy.

BACKGROUND: After treatment of locally advanced laryngeal carcinomas with induction chemotherapy and radiotherapy, some patients suffer a local or regional failure of the tumor, and salvage surgery is required. The aim of this study was to review the results of such salvage surgery in this group of patients. METHODS: A retrospective study of a cohort of 110 patients diagnosed between 1989 and 1996 with a locally advanced laryngeal carcinoma (T3-T4) treated with induction chemotherapy and radiotherapy was performed. The results of salvage surgery in patients with a local and/or regional failure of the treatment were analyzed. RESULTS: Forty-two patients presented a local and/or a regional recurrence of the tumor: 26 patients in the larynx, eight in the neck, and a further eight in both in the larynx and the neck. Salvage surgery was carried out in 28 patients (67%), consisting of total laryngectomies with neck dissections (24 cases), endoscopic resection of the tumor (one case), and radical neck dissections (three cases). Five-year adjusted survival for the 42 patients was 38%. Five-year survival for the 28 patients treated with salvage surgery was 57%. Five patients had postoperative complications: four had pharyngo-cutaneous fistulas and one had wound infection. CONCLUSIONS: After a local and/or regional recurrence, 67% of patients with advanced laryngeal carcinoma treated with induction chemotherapy and radiotherapy were candidates to salvage surgery. Five-year adjusted survival for this group of patients was 57%.

Cohort Studies↗
PubMed2001

Improvement of the synthesis of diphenylmethyl 7beta-(o-hydroxy)benzylideneamino-3-hydroxymethyl-3-cephem-4-carboxylate.

The title product (I) is synthesized currently from 7-aminocephalosporanic acid, and diphenyldiazomethane (DDM) is used as a protective reagent of the acid function for further reactions. When DDM was prepared from benzophenone hydrazone by reaction with chloramine T, it was resulted impure by p-toluenesulfonamide, formed as side product, which cannot be removed during the final purification step carried out according to the literature procedure. Two simple methods are proposed here to obtain I with the suitable degree of purity necessary for a drug.

Benzene↗
PubMed2001

PCR-PRINS-FISH analysis of structurally abnormal sex chromosomes in eight patients with Turner phenotype.

According to cytogenetic analysis, about 50% of Turner individuals are 45,X. The remaining cases have a structurally abnormal X chromosome or are mosaics with a second cell line containing a normal or abnormal sex chromosome. In these mosaics, approximately 20% have a sex marker chromosome whose identity cannot usually be determined by classical cytogenetic methods, requiring the use of molecular techniques. Polymerase chain reaction (PCR), primed in situ labeling (PRINS), and fluorescence in situ hybridization (FISH) analyses were performed in 8 patients with Turner syndrome and 45,X mosaic karyotypes to determine the origin and structure of the marker chromosome in the second cell line. Our data showed that markers were Y-derived in 2 patients and X-derived in the remaining 6 patients. We were also able to determine the breakpoints in the two Y chromosomes. The use of cytogenetic and molecular techniques allowed us to establish unequivocally the origin, X or Y, of the marker chromosomes in the 8 patients with Turner phenotype. This study illustrates the power of resolution and utility of combined cytogenetic and molecular approaches in some clinical cases.

Adolescent↗
PubMed2001

Regulation of serine-type exoproteinases by endogenous inhibitors present in exoantigens of the mycelial form of Paracoccidioides brasiliensis.

We have partially characterized some biochemical properties of exoproteinases secreted into culture medium by the mycelial form of Paracoccidioides brasiliensis, a dimorphic fungus that causes human disease in Latin America. Proteinase activity was analyzed in solid- and liquid-phase systems using zymography and Azocoll, respectively. Minimal or no gelatinase activity was observed by zymography in the crude filtrates among proteins with a relative mobility greater than 200 kDa. When the crude filtrate was fractionated by isoelectric focusing or ion exchange chromatography, we observed striking activation of gelatinases, both those of high apparent molecular mass and alkaline isoelectric points (pI), as well as those of lower molecular mass and acidic pI. The apparent high molecular mass gelatinases, pI 10, showed optimal activity at pH 7.0. They were totally inhibited by phenylmethylsulfonylfluoride and partially inhibited by incubation with previously neutralized fractions of pI 5.4 and 6.1. The latter inhibition could be reversed by exposure to 10% isopropanol. These results provide evidence of regulatory mechanisms controlling proteinase activity in secreted proteins. The principal mechanism appears to be the formation of reversible complexes with endogenous inhibitors.

Antigens, Fungal↗
PubMed2001

Cellular localization of orexin receptors in human pituitary.

Orexins-A and -B are hypothalamic peptides derived from a precursor called prepro-orexin and relationated with the stimulation of food intake. They act on G protein receptors named orexin receptor 1 (OX(1)R) and orexin receptor 2 (OX(2)R), respectively. In the present study, we used RT-PCR and immunohistochemical techniques to detect the presence of OX(1)R and OX(2)R in human pituitary. A band of the expected size for both OX(1)R and OX(2)R was shown in human pituitary by RT-PCR. The cellular localization of OX(1)R and OX(2)R was carried out using histological techniques. By consecutive sections we demonstrated that OX(1)R was present in acidophil, diffusely distributed cells, which represent the half of the total adenohypophysis cell population. As was expected, these cells were shown to coexpress GH. OX(2)R was found in the pars intermedia and in clusters of basophil cells of the anterior pituitary, which coexpress ACTH. These results were confirmed by double immunofluorescence techniques. We also found focal positivity in axon terminals of neurohypophysis, more intense for OX(2)R than for OX(1)R. In conclusion, these results demonstrated for the first time that OX(1)R and OX(2)R were expressed by somatotrope and corticotrope cells, respectively.

Aged↗
PubMed2001

In situ hybridization of somatolactin transcripts in the pituitary glands from acclimatized carp (Cyprinus carpio).

We isolated and cloned a carp somatolactin SL DNA fragment, of which 78% of the nucleotides were identical to the corresponding salmon SL sequence. The results obtained upon Northern blot hybridization of carp pituitary RNA allowed the identification of two transcripts as described for other fish. When the content of SL transcripts in pituitary sections from summer- and winter-acclimatized carp was quantified by in situ hybridization assays, we found no significant differences between the two seasons. In salmonids, plasma SL reaches higher levels in summer than in winter in synchrony with the water temperature cycle; in the eurythermal carp, however, the complex adaptive responses imposed by seasonal environmental changes do not seem to include the regulation of the somatolactin detected with the probe used at the transcriptional level in pituitary glands.

Acclimatization↗
PubMed2001

Cellular localization of orexin receptors in human pituitary.

Orexins-A and -B are hypothalamic peptides derived from a precursor called prepro-orexin and relationated with the stimulation of food intake. They act on G protein receptors named orexin receptor 1 (OX(1)R) and orexin receptor 2 (OX(2)R), respectively. In the present study, we used RT-PCR and immunohistochemical techniques to detect the presence of OX(1)R and OX(2)R in human pituitary. A band of the expected size for both OX(1)R and OX(2)R was shown in human pituitary by RT-PCR. The cellular localization of OX(1)R and OX(2)R was carried out using histological techniques. By consecutive sections we demonstrated that OX(1)R was present in acidophil, diffusely distributed cells, which represent the half of the total adenohypophysis cell population. As was expected, these cells were shown to coexpress GH. OX(2)R was found in the pars intermedia and in clusters of basophil cells of the anterior pituitary, which coexpress ACTH. These results were confirmed by double immunofluorescence techniques. We also found focal positivity in axon terminals of neurohypophysis, more intense for OX(2)R than for OX(1)R. In conclusion, these results demonstrated for the first time that OX(1)R and OX(2)R were expressed by somatotrope and corticotrope cells, respectively.

Aged↗
PubMed2000

Repeated surgical embryo recovery and embryo production in rabbits.

The purpose of this study was to assess the embryo production after repeated surgical recovery of embryos in Gigante de España does. A total of 195 ovulatory treatments and embryo recoveries were performed from 1995 to 1999. Ovulation was induced by an intramuscular injection of 20 microg GnRH immediately after mating. Each doe was induced to ovulate up to four consecutive times at intervals of at least 50 days. Embryos were surgically collected from oviducts 68-69 h post-coitus. An average of 8.6 corpora lutea and 6.4 recovered embryos (90% of them classified as viable) were recorded from the 195 treatments. The process seemed to be less efficient in the fourth treatment, with a drop of more than two recovered and viable morulae with reference to the third (P<0.05 for both parameters) or the second recovery (P<0.1 and P<0.05, respectively). More than 20 recovered embryos and 18 viable embryos per donor doe were recorded considering the three first ovulatory treatments performed in 33% of the does (30/90). Results indicate that the methodology used in the present study could be an efficient way to maximize in vivo embryo production from rabbits.

Animals↗
PubMed2000

Comparison of the instrumental reinforcer devaluation effect in two strains of rats (Wistar and Lister).

In this experiment, the effect of the reinforcer devaluation upon instrumental performance was analysed in two strains of rats (Wistar and Lister): Food deprived rats were trained to press a lever for sucrose pellets in a single session. Immediately after the fulfilment of this session, half of the Wistar and Lister rats received an injection of lithium chloride (LiCl), while the remaining animals were injected with a sodium chloride (NaCl) solution. A subsequent extinction test showed that the subjects who had received immediate LiCl did not press the lever as often as those injected with NaCl. No differences in response suppression were found between the two strains of rats. These results also show that a single devaluation experience is sufficient for an impact on instrumental performance.

Journal Article↗
PubMed2000

The action of Lonomia achelous caterpillars venom on human factor V.

The bleeding syndrome produced by contact with the Lonomia achelous caterpillars is characterized by a decrease of fibrinogen, factor XIII, plasminogen, and factor V with normal platelets. In this study, we report the effect of crude hemolymph and some semipurified chromatographic fractions on human factor V. Incubation of factor V with crude hemolymph resulted in an increase in procoagulant activity, followed by a subsequent decline in factor V activity. Identical results were obtained with fraction I, whereas with fraction II there was only a decrease in activity reaching its minimum at 30 minutes. fraction III did not modify the activity of factor V. All concentrations of fraction I tested produced an initial rise and subsequent fall in activity. However, at lower relative concentrations of fraction I, more sustained increases in activity were observed. The activator and inactivator activities present in fraction I show differences in temperature and pH stability, susceptibility to different inhibitors, and in SDS/PAGE pattern. The factor V activator is a thermostable protein, with maximum activity at acid pH and is inhibited by o-phenantroline, EDTA, and EGTA, while the factor V inactivator is thermolabile, presents maximum activity at basic pH, precipitates at pH 5.0, and is completely inhibited by iodoacetic acid and TLCK. It is partially blocked by diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, and p-chloromercuribenzoic acid. These results suggest that the activator should be a metallo-proteinase, while the inactivator is a serine or cysteine proteinase with a serine, histidine, or cysteine residue in the active site.

Anticoagulants↗
PubMed2000

Leptin regulation of prepro-orexin and orexin receptor mRNA levels in the hypothalamus.

The aim of this study was to determine the effects of leptin treatment on prepro-orexin and orexin receptor expression in the rat hypothalamus. Adult male rats, food-deprived for 48 and 72 h, were treated one time with vehicle or leptin (10 microg, icv). Prepro-orexin mRNA content was measured by semiquantitative RT-PCR, Northern blot, and in situ hybridization; orexin receptor 1 and 2 mRNA content was quantified by Northern blot and/or semiquantitative RT-PCR. Our results indicate that leptin inhibits a fasting-induced increase in prepro-orexin mRNA and orexin receptor 1 mRNA levels in the rat hypothalamus, while orexin receptor 2 mRNA levels were unchanged in all situations evaluated. These data provide direct evidence for an additional mechanism of adaptation of the hypothalamus to food deprivation and for a new effect of leptin in the regulation of food intake.

Animals↗
PubMed2000

Synthesis and biological evaluation of 3,4-diaryloxazolones: A new class of orally active cyclooxygenase-2 inhibitors.

A series of 3,4-diaryloxazolones were prepared and evaluated for their ability to inhibit cyclooxygenase-2 (COX-2). Extensive structure-activity relationship work was carried out within this series, and a number of potent and selective COX-2 inhibitors were identified. The replacement of the methyl sulfone group on the 4-phenyl ring by a sulfonamide moiety resulted in compounds with superior in vivo antiinflammatory properties. In the sulfonamide series, the introduction of a methyl group at the 5-position of the oxazolone ring gave rise to very COX-2-selective compounds but with decreased in vivo activity. Selected 3,4-diaryloxazolones exhibited excellent activities in experimental models of arthritis and hyperalgesia. The in vivo activity of these compounds was confirmed with the evaluation of their antipyretic effectiveness and their ability to inhibit migration of proinflammatory cells. As expected from their COX-2 selectivity, most of the active compounds lacked gastrointestinal toxicity in vivo in rats after a 4-day treatment of 100 mg/kg/day. Within this novel series, sulfonamides 9-11 have been selected for further preclinical evaluation.

Animals↗
PubMed2000

Endoscopic laser surgery in the treatment of radiation failure of early laryngeal carcinoma.

BACKGROUND: This study was undertaken to analyze the local control rates and survival of patients undergoing endoscopic laser surgery for squamous cell carcinoma of the larynx as a salvage procedure for recurrent tumors after radiotherapy. METHODS: Twenty-four patients underwent endoscopic laser surgery for radiation failure at our hospital from 1988 to 1995. All patients were men, between 45 and 88 years old. The localization of the recurrence was in the supraglottis in 3 cases and in the glottis in 21 cases. The postirradiation recurrences were staged rT1 in 18 patients and rT2 in 6 patients. RESULTS: Local recurrence after laser surgery developed in six patients (25%) who had a total laryngectomy. Voice preservation was thus achieved in 75% of patients. No patient died because of the laryngeal cancer, and the observed 5-year survival rate was 76%. CONCLUSIONS: These results indicate that in selected cases, endoscopic laser surgery may be used successfully for treating radiation failure of early laryngeal carcinomas.

Actuarial Analysis↗