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Biomedical subjects

E Carrillo

Publications and source records attributed to E Carrillo.

At least 19 recordsLinked to original sources
PubMed2006

Immunogenicity of the P-8 amastigote antigen in the experimental model of canine visceral leishmaniasis.

The P-8 proteoglycolipid complex (P-8 PGLC), an amastigote antigen of Leishmania pifanoi, has been demonstrated to induce protection in mouse models, as well as to induce Tc1/Th1-like cellular responses in American cutaneous leishmaniasis patients. Because the immunization with P-8 PGLC in the murine model does not appear to be genetically restricted, we have studied the reactivity of the P-8 PGLC in Leishmania infantum infected dogs. In this study, it is shown that PBMC from experimentally infected dogs (asymptomatic, oligosymptomatic) significantly proliferated in response to soluble leishmanial antigen (SLA) or the P-8 PGLC. Further, quantification of the gene expression induced by the stimulation with P-8 in asymptomatically infected dogs showed an up-regulation of IFN-gamma and TNF-alpha, which were three to 4-fold higher than that induced by soluble Leishmania antigen (SLA). While no measurable induction of IL-10 was observed, low levels of IL-4 mRNA were observed in response to both P-8 and SLA antigens. Thus, our studies establish that P-8 is recognized by infected canines and elicits a potentially curative/protective Th1-like immune response. The identification of Leishmania antigens that elicit appropriate immune responses across different host species (humans, canine) and disease manifestations (cutaneous or visceral) could be an advantage in generating a general vaccine for leishmaniasis.

Animals↗
PubMed2006

Induction of drug resistance in embryonal rhabdomyosarcoma treated with conventional chemotherapy is associated with HLA class I increase.

Effectiveness of conventional cytotoxic treatment of rhabdomyosarcoma (RMS) may be limited by the development of multidrug resistance (MDR) mediated by mdr1 gene. This gene codes for P-glycoprotein (P-gp) which has been related to a immunoregulatory function. Modulation of HLA expression by P-gp has been described in different types of tumor cells including RMS. However, very little is known about biological implications of the P-gp expression in RMS patients treated with conventional chemotherapy. In order to study the problem, we used embryonal RMS tissue samples from treated patients. Our results indicated that positive RMS samples to mdr1 show higher HLA class I expression than those which were negative to mdr1 PCR, what indicates a significant correlation between the expression of both molecules. In addition, we developed two resistant RMS cell lines (A-204-1 and 2) using similar concentrations of actinomycin D as are plasma levels in clinical situation. Both resistant cell lines showed mdr1 expression and an increase of HLA class I expression which was dose-dependent. These results demonstrated that conventional chemotherapy of embryonal RMS is able to induce resistance which can modulate HLA class I expression and suggest that immunological studies of these tumors may be necessary to the design new specific therapeutic strategies.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2005

Modulation of Ki-67 expression and morphological changes induced by gef gene in MCF-7 human breast cancer cells.

New therapeutic strategies are required to overcome the limitations of conventional breast cancer treatment. Suicide gene therapy offers a potential approach to this type of tumour, since systems based on the use of prodrugs may present some drawbacks related to toxicity, drug release and bioavailability. The gef gene has cell-killing functions in Escherichia coli and does not depend on the use of a prodrug for its action, making it an attractive target for suicide gene therapy. We created a gef-overexpressing human breast cancer cell line (MCF-7TG) by transfecting the gef gene under the control of a pMAMneo promotor. Dexamethasone-induction of gef gene expression in MCF-7TG cells produced a significant decrease in Ki-67 expression, which is a known proliferation marker. In addition, annexin-V-FITC and propidium iodide assays showed the presence of apoptotic cell death, which was confirmed by scanning electron microscopy. The most significant finding was the presence of "craters" in the cell membrane, as previously described in other apoptotic breast cancer cells. These results demonstrate the ability of the gef gene to down regulate Ki-67 expression and induce apoptosis in a breast cancer cell line, suggesting its potential application as a new gene therapy strategy for this type of tumor.

Apoptosis↗
PubMed2004

Regulation of muscle Cav1.1 channels by long-term depolarization involves proteolysis of the alpha1s subunit.

The effects of long-term depolarization on frog skeletal muscle Cav1.1 channels were assessed. Voltage-clamp and Western-blot experiments revealed that long-term depolarization brings about a drastic reduction in the amplitude of currents flowing through Cav1.1 channels and in the levels of the alpha1s subunit, the main subunit of muscle L-type channels. The decline of both phenomena was prevented by the action of the protease inhibitors E64 (50 microM) and leupeptin (50 microM). In contrast, long-term depolarization had no effect on beta1, the auxiliary subunit of alpha1s. The levels of mRNAs coding the alpha1s and the beta1 subunits were measured by RNase protection assays. Neither the content of the alpha1s nor the beta1 subunit mRNAs were affected by long-term depolarization, indicating that the synthesis of Cav1.1 channels remained unaffected. Taken together, our experiments suggest that the reduction in the amplitude of membrane currents and in the alpha1s subunit levels is caused by increased degradation of this subunit by a Ca2+-dependent protease.

Animals↗
PubMed2003

Inhibition of growth and induction of apoptosis in human breast cancer by transfection of gef gene.

The gef gene has cell-killing functions in Escherichia coli. To evaluate the feasibility of using this gene as a new strategy for cancer therapy, we transfected it in MCF-7 cells derived from breast cancer (MCF-7TG). The gef gene was cloned in a pMAMneo vector under the control of a mouse mammary tumour virus promoter, inducible by dexamethasone (Dex), and was transfected with liposomes. After selection and induction, expression of the gef gene was confirmed by reverse transcription-polymerase chain reactions (RT-PCR) and Western blot. Cell viability was determined with a haemocytometre and the sulphorodamine B colorimetric assay, and the cell cycle was studied by propidium iodide (PI) staining. Annexin V-FITC and PI assays were used to evaluate apoptosis, which was confirmed by electron microscopy. In comparison with MCF-7 parental cells and MCF-7 cells transfected with an empty vector, MCF-7TG cells induced with Dex showed a significant decrease in proliferation rate, which was associated with evidence of apoptosis. Morphological findings confirmed apoptosis and showed a typical pattern of mitochondrial dilation. Furthermore, the cell cycle was characterised by premature progression from G(1) to S phase and G(2) delay. Our results show that the gef gene was able to decrease proliferation in a breast cancer cell line, and induce apoptosis. These findings suggest that the gef gene is a potential candidate for tumour therapy.

Animals↗
PubMed2003

Contractile regulatory proteins tropomyosin and troponin-T as indicators of the modulatory role of retinoic acid.

Retinoic acid (RA), the active metabolite of vitamin A, plays a significant role in regulating cardiac form and function throughout the life of the organism. Both cardiac morphogenesis and myocardial differentiation are affected by alterations in RA homeostasis. In order to test the effect of all-trans RA and 13-cis RA on cardiomyocyte differentiation, we studied the level and the subcellular compartmentalization of alpha-tropomyosin and troponin-T proteins in cultures of chick embryo cardiomyocytes obtained from Hamburger and Hamilton's (HH) stage 22, 32 and 40 embryos. The retinoids increased the levels of alpha-tropomyosin and troponin-T in the cytoplasmic and cytoskeletal fractions of cells at all three stages of development. The greatest increases in alpha-tropomyosin occurred in the cytoplasmic fraction in HH22 cells cultured for 24 h with all-trans RA or 13-cis RA, whereas the greatest increases in troponin-T were found in the cytoplasmic fraction of HH32 cells exposed to retinoids for 24 h. In cultures treated for 48 h with retinoids, the levels of alpha-tropomyosin and troponin-T showed significant increases in the cytoplasmic compartment of cells treated in HH32-with respect to the control values. These findings are further evidence that RA plays a modulating role in the formation and reorganization of sarcomeric proteins during the process of cardiomyocyte maturation.

Animals↗
PubMed2003

Modulation of HLA class I expression in multidrug-resistant human rhabdomyosarcoma cells.

An abnormal HLA expression has been detected in some tumors including rhabdomyosarcoma (RMS). Classical cytotoxic treatment of these tumors, the most common childhood soft tissue malignancy, may induce multidrug resistance (MDR) associated with the expression of a 170-kDa membrane-associated glycoprotein (P-glycoprotein). In order to analyse the connection between modulation of HLA expression and the development of the MDR phenotype mediated by P-glycoprotein in RMS, we used three resistant RMS cell lines; two of these resistant cell lines (TE.32.7.DAC and RD-DAC) were established by in vitro exposure to actinomycin D, a drug of choice in the treatment of RMS; the resistant RMS- GR cell line was established from an embryonal RMS tumor after polychemotherapy. Our results showed that all the resistant cell lines showed a significant increase in the expression of HLA class I surface antigens in comparison to drug-sensitive cells. Blockade of P-glycoprotein with verapamil led to a decrease in HLA class I expression in RMS resistant cell lines. However, no modulation of HLA class II expression was observed in any of the three analyzed cell lines. These findings support the hypothesis that the development of resistance mediated by mdr 1/P-glycoprotein, directly influences the expression of HLA class I in RMS cells, inducing to upregulation. This effect may be relevant to the application in RMS of immunotherapy against tumor-associated antigens presented by HLA class I molecules.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

Gender differences in cardiovascular and electrodermal responses to public speaking task: the role of anxiety and mood states.

Gender moderates psychophysiological responses to stress. In addition to the hormonal background, different psychological states related to social stressors, such as anxiety and mood, could affect this response. The purpose of this study was to examine the existence of gender differences in the cardiovascular and electrodermal responses to a speech task and their relationship with anxiety and the mood variations experienced. For this, non-specific skin conductance responses (NSRs), heart rate (HR), and finger pulse volume (FPV) were measured at rest, and during preparation, task and recovery periods of an academic career speech in undergraduate men (n=15) and women (n=23), with assessment of changes in the state version of the State-Trait Anxiety Inventory (STAI-S) and in the Profile of Mood States (POMS) questionnaires. Men and women did not differ in trait anxiety, hostility/aggressiveness, or in the appraisal of the task, which were evaluated with the trait version of the State-Trait Anxiety Inventory (STAI-T), the Buss and Durkee Hostility Inventory (BDHI), and a self-report elaborated by ourselves, respectively. Women had higher FPV in all periods except during the task, and were more reactive to the stressor in state anxiety, and in the amplitude of NSRs. No gender differences for HR and for the frequency of NSRs were found. Anxiety and mood states were differently related to cardiovascular and electrodermal measurements in men and women. Further studies should consider the hormonal variations in addition to the psychological dimensions, in order to offer a more integrative perspective of the complex responses to stress.

Adult↗
PubMed2001

Induction of sexual activity of male creole goats in subtropical northern Mexico using long days and melatonin.

The aim of this study was to determine whether the sexual activity of local male Creole goats in subtropical Mexico can be induced during the non-breeding season by a long-day treatment followed by insertion of two melatonin implants. The experiment was carried out in the Laguna region in the State of Coahuila, Mexico (26 degrees N). Fourteen male goats were allocated to two balanced groups (n = 7 each) according to body and testicular weights. Males were kept together in two separate groups and fed lucerne hay for ad libitum intake and 300 g of commercial concentrate and had free access to water and mineral blocks. The control group remained in open sheds under natural photoperiod and ambient temperature conditions. The experimental group was placed in a light-proof building and exposed to 2.5 mo of long days (16 h of light/d) from November 1 to January 15. On January 16, each male received two s.c. melatonin implants and was exposed to natural photoperiodic changes in an open shed. In the control group, testicular weight exhibited seasonal variations; the highest value occurred on May 30 (146 +/- 10 g). Treated males reached maximum testicular weight earlier (March 15; 147 +/- 11 g), and sperm quality from January to March was higher than that observed in the control group (P < 0.05). Treatment caused an increase in LH pulse frequency (2.0 +/- 0.5 vs 0.3 +/- 0.2 pulse/8 h in February, 4.6 +/- 1.1 vs 0.1 +/- 0.1 pulse/8 h in March; experimental vs control group, respectively). In the control group, plasma testosterone remained low until mid-June and increased thereafter to remain elevated until the end of the study. In the experimental group, elevated plasma testosterone was observed from February to April and from July to November. Treating male goats in subtropical latitudes with artificial long days and melatonin can induce an intense sexual activity during the natural nonbreeding season.

Animals↗
PubMed2001

Sex ratio and breeding of white-lipped peccaries Tayassu pecari (Artiodactyla: Tayassuidae) in a Costa Rican rain forest.

White-lipped peccaries are non-seasonal breeders in South America, but little is known about their reproduction in Central America. There are few studies about the sex ratio of this species in the field. We studied the reproduction and sex ratio of white-lipped peccaries during 200 hours of field observation of four radiomarked and two unmarked herds, from July 1996 to April 1997, in Corcovado National Park, Costa Rica. Sex ratio data of three additional, radiomarked herds observed in 1998 were also included. We recorded numbers of mountings, presence of newborns and numbers of nursing interactions. The peccaries showed a distinct reproductive seasonality, with one mating peak in February and another in July. The greatest number of newborns and the peak in nursing activity were observed during July and August, when fruit availability for the peccaries was high. The adult sex ratio was significantly female biased (1.4:1-1.8:1), also in contrast with South American populations.

Animals↗
PubMed2000

Association of bovine DRB3 alleles with immune response to FMDV peptides and protection against viral challenge.

We have analysed the influence of bovine MHC (BoLA) polymorphism on the immune response and degree of protection induced by peptide vaccines against foot-and-mouth disease (FMD) in cattle. The peptides used for animal immunisation were: A (VP1(138-156)), AT (peptide A linked to VP1(21-40)) and ACT (peptide A, linked to VP1(196-209) and VP1(21-40)). Sixteen different DRB3 types were found among the 46 cattle analysed by PCR-RFLP typing. No absolute correlation was observed, for any type, with the serum neutralising titres (SNT) values and the protection induced. However, among the most common haplotypes present, associations were observed between expression of different types with the levels of SNT and/or protection induced by peptides A and ACT. Thus, types DRB3.2*1, 3 and 7 were associated with increased levels of protection. In contrast, types DRB3.2*12 and 18 were associated non-protection, and DRB3.2*12 was also associated with low SNT titres. Overall, the results indicate that the polymorphism in BoLA class II molecules affects both the immune response and protection induced by potential FMD peptide vaccines.

Alleles↗
PubMed2000

Protective immunity against foot-and-mouth disease virus induced by a recombinant vaccinia virus.

We report the construction of a recombinant vaccinia virus expressing the precursor for the four structural proteins of FMD virus (FMDV) (P1) strain C3Arg85 using a procedure for isolation of recombinant vaccinia viruses based solely on plaque formation. Adult mice vaccinated with this recombinant vaccinia virus elicited high titers of neutralizing antibodies against both the homologous FMDV and vaccinia virus, measured by neutralization assays. Liquid phase blocking sandwich enzyme-linked immunosorbent assays (ELISAs) using whole virus as antigen showed high total antibody titers against homologous FMDV, similar to those induced by the conventional inactivated vaccine. When ELISAs were carried out with heterologous strains A79 or O1Caseros as antigens, sera from animals vaccinated with the recombinant virus cross-reacted. Mice boosted once with the recombinant vaccinia virus were protected against challenge with infectious homologous virus. These results indicate that recombinant vaccinia viruses are efficient immunogens against FMDV when used as a live vaccine in a mouse model.

Animals↗
PubMed2000

Development of chick cardiomyocytes: modulation of intermediate filaments by basic fibroblast and platelet-derived growth factors.

Recent studies suggest that peptide growth factors play a functional role in cardiac muscle. To test whether embryonic cardiac muscle is a target for regulation by basic fibroblast growth factor and platelet-derived growth factor, we analyzed the effects of these peptides on the expression of the intermediate filaments desmin and vimentin at the subcellular level during development. Sodium dodecyl sulfate-gel electrophoresis, immunoblotting and fluorescence-activated cell sorting analysis were used to study the effect of basic fibroblast growth factor and platelet-derived growth factor on cultures of chick cardiomyocytes during development. Cytoplasmic and cytoskeletal concentrations of desmin and vimentin were dependent on the stage of embryonic development and on the type of growth factor added to the culture. The most significant finding was the increase in desmin expression in the cytoplasmic and cytoskeletal compartments after treatment with basic fibroblast growth factor (10 ng/ml) of chick heart cells at Hamburger and Hamilton stage 19. In more mature stages, basic fibroblast growth factor did not modify the levels of desmin expression. However, this factor led to a progressive deceleration in the rate of increase in vimentin expression. Platelet-derived growth factor increased vimentin expression in all stages studied, the greatest increases appearing in early stages of heart development. Our findings support the hypothesis that basic fibroblast growth factor plays a role in cardiomyocyte differentiation during the early stages of development, whereas platelet-derived growth factor has a dedifferentiating effect.

Animals↗
PubMed1998

[The health costs of alcoholism].

OBJECTIVE: To evaluate the economic effect in terms of health costs of alcohol abuse in Spain. DESIGN: The most up-to-date available, secondary sources of information were used. The period for the costs calculation was a natural year, with the prices adjusted to pesetas in 1996. The focus was based on the prevalence of cases per period. All the information referred to the whole of Spain. SETTING: Health costs were broken down into out-patient attendance, hospital emergencies, admissions, treatment at special centres and other health expenditure. MEASUREMENTS AND MAIN RESULTS: The total health cost attributable to alcoholism was 177,084 million pesetas, broken down as follows: a) cost of hospital admissions: 93,644 million pesetas (52.88%); b) cost of out-patient attendance: 34,600 million (19.53%); c) cost of treatment at special centres: 18,029 million (10.18%); d) cost of hospital emergencies: 10,481 million (5.91%); and e) other health expenditure: 20,330 million pesetas (11.48%). CONCLUSIONS: The figures arrived at do not cover the total cost of alcohol abuse, since a conservative approach was adopted. They stand as a minimum of the expenditure in our country. Some are direct costs on health authorities, whereas others can be managed as a cost-opportunity problem.

Alcoholism↗
PubMed1998

Optimization of the tyrosinase mRNA probe to detect circulating melanocytes with reverse transcription and polymerase chain reaction.

It was recently suggested that reverse transcription polymerase chain reaction (RT-PCR)-based detection of tyrosinase messenger RNA (mRNA) in peripheral blood is useful in the early detection of circulating tumor cells, since tyrosinase is thought to be a melanocyte-specific marker. However, the sensitivity of detection of these cells in circulation is controversial, some authors reporting 0% effectiveness, others obtaining 100% efficacy. We developed a modification of a technique to process blood samples to detect tyrosinase mRNA, and tested the method with 50 samples from as many patients with histologically confirmed malignant melanoma in different stages. Whole blood was processed by discarding the plasma and extracting RNA from density gradient-isolated peripheral blood lymphocytes. The RNA samples were tested with a sensitive nested primer RT-PCR assay. Sensitivity was tested using RNA extracted from SK-mel-1 human melanoma cells diluted serially with peripheral blood obtained from healthy control subjects. A lymph node from a patient with confirmed disseminated melanoma served as the positive control. Our technique was able to detect tyrosinase mRNA in samples from the 37 patients with progressive metastatic melanoma. The test detected tyrosinase mRNA from both the melanoma cell line and the positive lymph node. Our method to extract RNA from whole blood improves the specificity and sensitivity of tyrosinase mRNA detection by RT-PCR. The test should be of use in determining the prognosis of patients with melanoma, and in deciding when to initiate early treatment in patients with malignant melanoma.

Humans↗
PubMed1997

RUG-III and resource allocation: comparing the relationship of direct care time with patient characteristics in five countries.

BACKGROUND: resource use by different types of patients is of increasing interest to health care services all over the world. Case-mix systems that group together individuals with similar patterns of resource use have been developed to address these questions. Resource Utilization Groups version III (RUG-III) was developed in the USA to address the issue in the care of elderly people and has been validated in a number of countries. METHOD: this paper synthesizes the results of RUG-III validation studies performed in the USA, Japan, Spain, Sweden and England and Wales, showing the consistency of the system in spite of different skill-mix and total time spent with patients. Data from the validation studies of five countries were compared. Percentage of time given by trained nurses and mean nursing time per patient was compared overall and between selected RUG-III groups. RESULTS: mean time per patient ranged from 84.4 min per day in Japan, to 155.6 min in England and Wales. Trained nurse time ranged from 7.5% of total time in the USA to 53.2% of total time in England and Wales. The inter-group relationship was very similar in all countries. The RUG-III system appears robust in a wide variety of settings and countries. Future research should address the relationship between skill-mix and total time spent with patients with respect to outcome and quality of care.

Activities of Daily Living↗