Biomedical subjects
Publications and source records attributed to M Izquierdo.
The ameloblastomas are benign odontogenic tumors locally aggressive that between 15-20% are located in the upper maxillary. The maxillary ameloblastomas have worse forecast than their mandibular homologous upon presenting greater index of postsurgery relapses and a greater percentage of malignancy. Their proximity to nasal cavity, orbit and skull base suppose a risk added by the probability of extension to these structures.
BACKGROUND: Limitations of the isolated perfused rat heart model for heart preservation studies include short study time due to the lack of stability of the preparation. We aimed to develop a new experimental model based on heterotopic heart transplantation in the rat to achieve simultaneous (31)P magnetic resonance spectroscopy (MRS) and functional study of the transplanted heart during early and late blood reperfusion. METHODS: Twenty-five Lewis rats underwent heterotopic abdominal isograft heart transplantation and were randomized in two groups. Hearts were harvested after cardioplegic arrest induced with Centre de Résonance Magnétique Biologique et Médicale (CRMBM) solution and then stored at 4 degrees C for a total ischemic time of 3 hours. Graft contractility measurement and simultaneous (31)P MRS were performed after 1 hour and 24 hours of blood reperfusion, respectively, in groups I (n = 12) and II (n =13). RESULTS: Contractility improved during reperfusion. The mean rate pressure product plus or minus standard error of mean increased from 11,373 +/- 1,377 mm Hg/min in group I to 24,363 +/- 3,860 mm Hg/min in group II (P = 0.003), while mean dP/dtmax increased from 1,642 +/- 173 mm Hg/sec to 2,571 +/- 333 mm Hg/sec, respectively (p = 0.03). Simultaneously, both the phosphocreatine/adenosine triphosphate (ATP) and inorganic phosphate/ATP ratios decreased from group I to group II (p = 0.025 and p = 0.015, respectively), suggesting regeneration of the intracellular pool of ATP in group II. CONCLUSIONS: Simultaneous functional and metabolic studies of the transplanted heart are feasible in rats. Improvement in contractility during late reperfusion is contemporary with significant changes in energetic metabolism. Our model should be useful for the further improvement of heart preservation, which may result in significant clinical progress.
The use of DNA-based methodologies in identification of hake species belonging to the Merluccius genus was shown to be successful. A short fragment of the left hypervariable domain of the mitochondrial control region was amplified, sequenced, and digested from 11 hake species. The hake-specific PCR product, due to its limited size, was obtained in a variety of tissue samples with different levels of DNA concentration and degradation, including sterilized food products. On the basis of this phylogenetically informative 156-bp sequence were selected four restriction enzymes (ApoI, DdeI, DraIII, and MboII) that allow the hake species discrimination. Species identification by phylogenetic analysis of sequences or by PCR-RFLP methodologies is useful in a variety of scenarios including authentication of thermally processed food, detection of food components, and species determination of individuals whose morphological characters are removed.
Identification of flatfish species using a DNA-based methodology was studied. The polymerase chain reaction was employed to obtain a 464 bp amplicon from mitochondrial cytochrome b gene. The sequences from this fragment belonging to 24 species were analyzed using a genetic distance method, and polymorphic sites were determined. The fragment was found to be highly polymorphic (231 sites), and this permitted the differentiation of most of the species. Phylogenetic tree construction was employed to allow the identification of flatfish species. As a result, each species was grouped in a well-differentiated clade, except for two pairs: Limanda ferruginea and L. limanda, and Solea impar and S. lascaris, which could not be differentiated. On the basis of the sequences obtained, restriction enzymes were selected to provide specific restriction profiles, which allow the differentiation of 21 species of flatfish in a faster and less expensive manner than sequencing. This polymerase chain reaction-restriction fragment length polymorphism methodology (PCR-RFLP) was tested using commercial samples.
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Duct endoscopy is a recent technique used for a direct view of the breast ductal system. The aim of this study is to determine any morphological changes in breast tissue attributable to low-pressure irrigation with saline solution that the technique requires. A total of 26 breast biopsies from patients who underwent ductal endoscopy before surgery were compared with 26 breast specimens from the retroareolar region. Breast specimens from duct endoscopy showed more frequent epithelial detachment (73%), epithelial loss (35%), periductal clefts (77%), stromal disaggregation (46%) and displacement of epithelial cells into the stroma (27%) than the control group in which epithelial detachment was seen in 4% of patients, periductal clefts in 15%, and stromal disaggregation in 15%. Epithelial loss and epithelial displacement where not seen in the control group. Although low-pressure fluid perfusion used for duct endoscopy induced morphological changes in breast tissue, these can easily be distinguished from malignancy, and are most likely to occur as the result of duct rupture.
PURPOSE: To examine maximal strength, power and muscle cross-sectional area, maximal and submaximal cycling endurance characteristics, and serum hormone concentrations of testosterone (T), free testosterone (FT), and cortisol (C) in middle-aged and elderly men. METHODS: Maximal knee extension force (isometric; MIF(KE)), power-load curves during concentric actions with loads ranging from 15% to 70% of 1 RM half-squat (1RM(HS)), muscle cross-sectional area of quadriceps femoris (CSA(QF)), workload, heart rate and lactate accumulation during incremental cycling, and serum hormone concentrations were measured in 26 middle-aged (M42 yr) and 21 elderly men (M65 yr). RESULTS: The 1RM(HS) (14%), MIF(KE) (24%) and CSA(QF) (13%) were lower in M65 than in M42 (P < 0.05-0.01). Power during submaximal actions was lower (P < 0.05-0.001) in M65 than in M42, but the differences disappeared when expressed relative to CSA(QF). Serum FT was in M42 higher (P < 0.05) than in M65. Maximal workload, maximal heart rate and peak blood lactate during cycling in M65 were 31%, 11%, and 20% lower than in M42 (P < 0.01). During submaximal cycling blood lactate rose more rapidly with increasing workload in M65 than in M42 (P < 0.05-0.01), but the differences disappeared when expressed relative to CSA(QF). Significant correlations existed between individual values of serum FT:C ratio, C and T, and those of muscle strength and maximal workload. CONCLUSION: Declines in maximal strength, muscle mass, and endurance performance seem to take place with increasing age, although muscle power and demand for aerobic energy per unit of muscle tissue during submaximal loads remain similar. The balance between anabolic and catabolic hormones in aging people over the years may be associated with age-related decreased strength and declines in maximal cycling workload.
Effects of 16-wk strength training on maximal strength and power performance of the arm and leg muscles and serum concentrations [testosterone (T), free testosterone (FT), and cortisol] were examined in 11 middle-aged (M46; 46 +/- 2 yr) and 11 older men (M64; 64 +/- 2 yr). During the 16-wk training, the relative increases in maximal strength and muscle power output of the arm and leg muscles were significant in both groups (P < 0.05-0.001), with no significant differences between the two groups. The absolute increases were higher (P < 0.01-0.05) in M46 than in M64 mainly during the last 8 wk of training. No significant changes were observed for serum T and FT concentrations. Analysis of covariance showed that, during the 16-wk training period, serum FT concentrations tended to decrease in M64 and increase in M46 (P < 0.05). However, significant correlations between the mean level of individual serum T and FT concentrations and the individual changes in maximal strength were observed in a combined group during the 16-wk training (r = 0.49 and 0.5, respectively; P < 0.05). These data indicate that a prolonged total strength-training program would lead to large gains in maximal strength and power load characteristics of the upper and lower extremity muscles, but the pattern of maximal and power development seemed to differ between the upper and lower extremities in both groups, possibly limited in magnitude because of neuromuscular and/or age-related endocrine impairments.
Retroviral vectors have long been used in a wide variety of gene transfer applications but have certain drawbacks, such as small cargo size, limited tropism, and low titers. HSV expression vectors overcome these disadvantages, but, because they persist in target cells as nonreplicative episomes, they are not retained in all the progeny of dividing cells. Chimeric HSV/AAV products that can mediate transgene integration in human mitotic cells have been constructed, but, to date, genetic modification of dividing cells in animal models using HSV products has not been possible. Here, we report the construction of hybrid HSV/retroviral vectors that exhibit up to 50-fold higher transgene integration efficiency compared to vectors containing only HSV-1 components. Efficient integration of a retroviral transgene cassette encoding pac in human cells required expression of the Moloney murine leukemia virus gag-pol genes, but in murine cells, could also be mediated by endogenous activities, albeit at a lower level. Gene delivery was equally efficient in BHK21, a cell line resistant to retroviral infection, and transgene retention and expression were observed to be stable for least one month in Hs683 human glioma cells. These vectors have wide applications for the genetic modification of many cell types.
We have combined the picornavirus foot-and-mouth disease virus (FMDV) 2A sequence and the internal ribosome entry sites (IRESes) from encephalomyocarditis virus (ECMV) and avian reticuloendotheliosis virus type A (REV-A) to construct tricistronic and tetracistronic vectors. All the polycistronic constructs show high titers and expression of the genes inserted. Clones have been obtained in which cells simultaneously express the three or four genes carried by the polycistronic vectors.
Many of the strategies developed in the last few years to treat cancer by gene therapy are based on putative killer-suicide genes whose products convert a prodrug into a toxic compound. When the therapy is applied to humans, a vector carrying the killer gene is first inoculated into the tumor of the patient, who 1 week later receives the corresponding prodrug that will selectively kill the cells able to process it to its toxic derivative. A strategy that obviates the need for a prodrug to destroy the cancer cells would be preferable because the patient would only need one treatment instead of two consecutive ones. In the following study, we describe the construction of retroviral vectors in which a reporter or a toxin gene (either the Pseudomonas exotoxin or the Ricinus communis toxin, ricin) is placed under the control of the thyroid hormone (T3) regulatable promoter of the rat myelin basic protein (MBPp). We demonstrate that the expression of these genes under the control of MBPp is regulated by T3 in vitro and in vivo. In vitro, the MBPp is switched off when T3 is removed from the serum of the culture medium, allowing the production of retroviruses carrying the toxic gene. In vivo, the toxin gene bearing retroviruses is capable of eradicating experimentally induced brain tumors in Wistar rats. The gene therapy strategy described here does not require the use of a prodrug to destroy the neoplastic cells.
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BACKGROUND: Adrenoleukodystrophy (ALD) is characterised by wide phenotypic variation, and there is no marker to predict the onset of cerebral demyelination. The indications for therapeutic approaches depend largely on the onset of cerebral demyelination. OBJECTIVE: To evaluate the brain spectroscopic pattern in normal-appearing white matter (NAWM) in patients with various phenotypes of ALD to determine if these abnormalities could be of useful prognostic value. MATERIALS AND METHODS: Spectroscopic imaging acquisition mode (MRSI, 16 x 16 voxels) was performed in 20 patients with ALD, including 7 neurologically asymptomatic patients without detectable demyelination on MRI, 3 patients with early signs of cerebral demyelination, 8 patients with adrenomyeloneuropathy (AMN) and 2 patients with cerebral ALD who had previously undergone bone marrow transplantation. Controls were 22 healthy subjects. In all patients, four voxels entirely located in the juxtaventricular NAWM were studied. The ratios NAA/Cho, NAA/ CPC and Cho/CPC for the four ROIs were measured in the patient population and compared with control values. Results. In spite of a large distribution of ratios, the statistical tests did not show any significant difference between the ratios within NAWM in the patient population compared with control values. Means of ratios in the left posterior (LP) voxel compared normal subjects were (a) in neurologically asymptomatic ALD patients (n = 7) 2.27 +/- 0.63 for NAA/CPC, 2.21 +/- 0.75 for NAA/ Cho, 1.06 +/- 0.28 for Cho/CPC, (b) in patients with early signs of demyelination (n = 3) 3.43 +/- 0.85 for NAA/ CPC, 2.47 +/- 0.32 for NAA/Cho, 1.37 +/- 0.16 for Cho/CPC and (c) in AMN patients (n = 8) 1.47 +/- 0.53 for NAA/CPC, 2.17 +/- 1.58 for NAA/ Cho, 0.83 +/- 0.32 for Cho/CPC. Conclusions. The study did not show significant differences in metabolite ratios between patients and controls. The large distribution of results precludes the possibility of detecting small variations. Part of this distribution can be due to the CSI method. Longitudinal spectroscopic studies, preferentially using monovoxel spectroscopy, are clearly needed.
Toxic oil syndrome (TOS) was an epidemic which broke out in Spain in 1981, caused by the ingestion of rapeseed oil denatured with 2% aniline and sold illegally as edible oil. More than 20,000 people were affected and mortality rate was 8.4%. Genetic susceptibility appears to be involved in the pathology of this disease. Several reports have described association between the chronic stage of the disease and DR-DQ antigens (DR3, DR4, DR2 and DQ8). In the present work, we have reassessed the HLA class II antigens in a well-designed case-control study. Triplets of subjects (n=265) composed by chronic patients (n=117), non-affected family members (n=71) and non-related controls (n=77) were studied. Also, HLA class II antigens were analyzed in patients who had died from TOS (n= 34) and in TOS control patients who died from other non-TOS related causes (n=13). Regarding surviving patients no significant association was found between HLA and disease. In contrast, an increase in phenotypic frequency of DR2 antigen, was found in patients who had died from TOS (73.5%) compared with the whole study group: TOS-affected alive patients (25.6%, corrected P<0.001), non-affected family members (28.5%, corrected P<0.001), non-related controls (23.9%, corrected P<0.001) and dead controls (38.4%, P=0.03).
Delta9-Tetrahydrocannabinol, the main active component of marijuana, induces apoptosis of transformed neural cells in culture. Here, we show that intratumoral administration of Delta9-tetrahydrocannabinol and the synthetic cannabinoid agonist WIN-55,212-2 induced a considerable regression of malignant gliomas in Wistar rats and in mice deficient in recombination activating gene 2. Cannabinoid treatment did not produce any substantial neurotoxic effect in the conditions used. Experiments with two subclones of C6 glioma cells in culture showed that cannabinoids signal apoptosis by a pathway involving cannabinoid receptors, sustained ceramide accumulation and Raf1/extracellular signal-regulated kinase activation. These results may provide the basis for a new therapeutic approach for the treatment of malignant gliomas.
PURPOSE: This investigation examined the effects of creatine (Cr) supplementation on intermittent high-intensity exercise activities specific to competitive soccer. METHODS: On two occasions 7 d apart, 17 highly trained male soccer players performed a counter-movement jump test (CMJT), a repeated sprint test (RST) consisting of six maximal 15-m runs with a 30-s recovery, an intermittent endurance test (IET) consisting of forty 15-s bouts of high-intensity running interspersed by 10-s bouts of low-intensity running, and a recovery CMJT consisting of three jumps. After the initial testing session, players were evenly and randomly included in a CREATINE (5 g of Cr, four times per day for 6 d) or a PLACEBO group (same dosage of maltodextrins) using a double-blind research design. RESULTS: The CREATINE group's average 5-m and 15-m times during the RST were consistently faster after the intervention (0.95 +/- 0.03 vs 0.97 +/- 0.02 s, P < 0.05 and 2.29 +/- 0.08 vs 2.32 +/- 0.07 s, P = 0.07, respectively). Neither group showed significant changes in the CMJT or the IET. The CREATINE group's recovery CMJT performance relative to the resting CMJT remained unchanged postsupplementation, whereas it tended to decrease in the PLACEBO group. CONCLUSION: In conclusion, acute Cr supplementation favorably affected repeated sprint performance and limited the decay in jumping ability after the IET in highly trained soccer players. Intermittent endurance performance was not affected by Cr.
The follow-up of HBV markers in selected high infection risk populations, in patients from the hemodialysis and peritoneal dialysis services was used to assess the effectiveness of a special vaccination program. Viral infection markers were studied in prevalence cross sections of the whole population of patients, and also by recording the reports of clinical cases of hepatitis B occurred during that period in those groups of patients. The prevention program consisted of the vaccination of all patients negative to the viral markers and the indication of vaccination for the new cases during the period of the kidney disease, just before the start of the treatment at the hemodialysis unit; besides all the persons susceptible to infection that had already been included in the program, regardless of the stage of the disease. The results show the benefit of the vaccination in these patients, but it is more effective in the period before the treatment with dialysis where there is a lower possibility of being exposed to the virus and the immune system is still competent. Once the program was established, after a follow up o 6 years, there have been no reports of new cases of hepatitis B and the incidence of the disease has been declining.