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Biomedical subjects

A Nieto

Publications and source records attributed to A Nieto.

At least 37 records · Page 2Linked to original sources
PubMed2003

Steroid hormone profile of canine inflammatory mammary carcinoma: a preliminary study.

Inflammatory mammary carcinoma (IMC) is the most aggressive spontaneous type of mammary malignant tumor both in women and dogs. Latest studies in dogs indicate that different endocrine mechanisms seem to be involved in inflammatory carcinomas (IMCs). The aim of the present study was to characterize the steroid hormone profile of inflammatory carcinoma, and to compare it with mammary dysplasias, benign tumors and other malignant tumors. Eighty-six mammary samples (10 normal mammary tissue, 21 dysplasias, 26 benign, 22 malignant, and 7 IMC) from 30 female dogs were used. Hormone levels of progesterone (P4), 17beta-estradiol (E2), androstenedione (A4), dehydroepiandrosterone (DHEA), and estrone sulphate (E1SO4) in tissue homogenates were measured by enzyme immunoassays (EIAs) techniques, previously validated for this species. IMC displayed the following steroid profile: P4: 13.80+/-0.56 microg/g; E2: 675.19+/-33.00 ng/g; A4: 631.73+/-70.73 microg/g; DHEA: 702.22+/-89.93 microg/g, and E1SO4: 2.84+/-0.32 mg/g. All of these hormones were significantly higher (P<0.001) compared with the hormone steroid profile determined for malignant, benign, dysplasias, and normal mammary tissue. The most relevant finding was the increased levels, two or three times, of both DHEA and E1SO4 in IMC respect to other groups (P<0.001). These results, together with the highest immunohistochemical expression of P450scc found in IMC, suggest the hypothesis that an autocrine mechanism could be especially involved in the development of canine inflammatory carcinoma.

Androstenedione↗
PubMed2003

Gender differences in rising trends in cutaneous malignant melanoma in Spain, 1975-98.

BACKGROUND: Cutaneous malignant melanoma (CMM) morbidity and mortality rates have increased over the past several decades all around the world, but some developed countries have experienced recent declines in mortality from CMM among the young and middle-aged and even a reversal in the long-term increase in deaths attributable to CMM. OBJECTIVES: To describe CMM mortality trends, by age and gender, in Spain between 1975 and 1998. METHODS: Crude, age-adjusted, truncated, cumulative, age-specific and potential years of life lost rates of CMM mortality were calculated by gender. Age and period Poisson regression models were fitted to gender- and age-specific CMM mortality rates. RESULTS: In males there were marked increases in all CMM mortality rates between 1975 and 1994, followed by their decrease during 1995-98. In contrast, rates rose in females between 1975 and 1998. There were annual increases of 13% in the risk of dying among males and 33% among females. Relative risks of CMM mortality increased with age in both genders: (i) 21.75 (95% confidence interval, CI 17.32-27.30) in males, and (ii) 28.27 (95% CI 24.99-31.97) in females in the > or = 70-year-old group, when compared with those < 35 years. CONCLUSIONS: CMM mortality rates continue to rise in females while males have experienced a recent reversal of this trend in Spain. Emphasizing the danger of overexposure to ultraviolet radiation may be especially important in females.

Adolescent↗
PubMed2003

A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease.

Several recombinant clones expressing antigens from Echinococcus granulosus were isolated previously from a parasite cDNA library using cystic hydatid disease (CHD) patients' sera or rabbit hyperimmune antiserum against a lipoproteic fraction from bovine cyst fluid. Six of these antigens were expressed in Escherichia coli and the purified recombinant proteins were tested in enzyme-linked immunosorbent assay (ELISA) for specific IgG with a panel of sera from patients with surgically confirmed (n = 58) or immunologically diagnosed (n = 71) CHD. Sera from clinically normal individuals (n = 203) and sera from individuals with other helminthic infections (n = 65) were assayed for the assessment of specificity. A cut-off value was determined by receiver-operating-characteristic plots for each antigen. A recombinant antigen B subunit (AgB8/2) presented the highest sensitivity (93.1%), considering the group of sera from patients with CHD surgically confirmed, and specificity (99.5%) and is proposed as the basis for an immunodiagnostic test. The other recombinant antigens tested presented sensitivities between 58.6% and 89.7%, and three of them were considered of complementary value. In subclass-specific ELISA, different IgG isotypes showed dominance in the response for each of the recombinant antigens. There was a clear predominance of IgG4 response for all antigens tested, indicating that this would be the subclass of choice to be assessed for these recombinant proteins.

Animals↗
PubMed2003

BRCA1 expression in canine mammary dysplasias and tumours: relationship with prognostic variables.

BRCA1 is a nuclear phosphoprotein that participates in the regulation of the cell cycle. The role of the BRCA1 gene in canine mammary tissue and mammary tumours has not been studied. The present study examined immunohistochemically the expression and intracellular distribution of BRCA1 protein in two normal, seven dysplastic and 44 neoplastic canine mammary glands and its relationship with clinical and pathological variables and other prognostic parameters. Strong nuclear immunolabelling of BRCA1 protein was observed in the epithelial cells of the normal mammary glands and mammary dysplasias. The majority of benign tumours, and more especially of malignant tumours, showed a significant reduction in the nuclear expression of BRCA1 protein and an increase in cytoplasmic expression. Loss of BRCA1 expression was associated with high proliferation marker Ki-67 and ER-alpha negative tumours. The reduction and aberrant distribution of BRCA1 in canine mammary tumours were significantly associated with malignant characteristics. The results may indicate that BRCA1 has a role in the malignant behaviour of these tumours.

Adenocarcinoma↗
PubMed2003

Oral and oropharyngeal cancer in Spain: influence of dietary patterns.

Cancer of the oral cavity and oropharynx is a major public health problem in Spain. Tobacco and alcohol have been identified as the two major risk factors for oral cancer in most western populations. Other risk factors include diets low in fruits and vegetables, but the impact of dietary habits on the risk of these cancers has never been assessed in Spain. The objective of this study was to elucidate the role of dietary habits in oral and oropharyngeal cancer, a multicentric case-control study was conducted in three areas of Spain (Barcelona, Granada and Sevilla) between 1996 and 1999. Cases were 375 patients (71 women), with incident, histologically confirmed cancer of the oral cavity or oropharynx, and controls were 375 subjects (71 women) admitted to hospitals for conditions unrelated to smoking or alcohol drinking. Data were analysed using multivariate logistic regression procedures. After allowance for education, tobacco and alcohol use, a significant inverse association with the risk of oral and oropharyngeal cancer was found for total consumption of total green vegetables (OR 0.54, 95% CI = 0.34-0.87) and total fruit (OR 0.52, 95% CI = 0.34-0.79) with significant trends in risk. We found that the protective effect of each of these food items was consistently larger among current smokers and among heavy alcohol drinkers, following a multiplicative effect model. In conclusion, this study provides further support to the beneficial effect of high intake of vegetables and fruits on the risk of developing cancers of the oral cavity and oropharynx in Spain, particularly among current smokers and heavy alcohol drinkers.

Adult↗
PubMed2003

NGF and TGF-beta mRNA expression during pregnancy in a rat corneal wound healing model.

BACKGROUND: Growth factors seem to play a major role in corneal wound healing and TGF-beta seems to be associated with abnormal healing after corneal surgical procedures. Few studies have analysed the role of NGF and TGF-beta on corneal wound healing during pregnancy. The aim of the present study was to create an animal model to evaluate the expression of NGF and TGF-betas during corneal wound healing in two groups: control and pregnant rats. METHODS: Corneal mRNA for NGF and the three isoforms of TGF-beta were analysed by RT-PCR, in a time-course experiment on different days after epithelial wounding (2, 7, 14 days) in pregnant and control groups RESULTS: The results show high corneal mRNA expression for NGF and TGF-beta1 without any variation throughout the healing process or pregnancy evolution. However, we detected a different expression of corneal mRNAs for TGF-beta2 and TGF-beta3 in the control group. This data was not detected in the pregnant group. DISCUSSION: Our results suggested that pregnancy could have a relevant role on TGF-beta2 and TGF-beta3 mRNA expression during the corneal wound healing process. Additional research should be performed to corroborate these findings.

Animals↗
PubMed2003

Immunohistochemical expression of p53, fibroblast growth factor-b, and transforming growth factor-alpha in feline vaccine-associated sarcomas.

Fifty feline sarcomas associated with vaccine-site injection were evaluated to determine the immunohistochemical expression of p53 protein, basic fibroblast growth factor (FGF-b), and transforming growth factor-alpha (TGF-alpha). Forty-one tumors (82%) were fibrosarcomas (FS), eight (16%) were malignant fibrous histiocytomas (MFH), and one (2%) was a chondrosarcoma (CS). Overexpression of p53 protein was observed in the nuclei of tumor cells in 28 (56%) sarcomas; FGF-b expression was found in the cytoplasm of tumor cells in 40 (80%) feline sarcomas, but the staining was more intense in the spindle-shaped cells of FS than in polygonal or round cells of MFH. The single CS faintly expressed FGF-b. The majority of feline vaccine-associated sarcomas (43 of 50, 86%) expressed moderate or intense staining for TGF-alpha in the cytoplasm of tumor cells. Heterogeneous immunolabeling for p53, FGF-b, and TGF-alpha was present in neoplastic, multinucleated giant cells. Intense expression of FGF-b was statistically associated with younger cats (P < 0.01) and with tumors with nodular growth patterns (P = 0.02). In addition, sarcomas negative for p53 protein expressed FGF-b more frequently than did p53-positive tumors (P = 0.04). The frequency of FGF-b immunostaining was significantly higher in sarcomas with intense expression of TGF-alpha (P = 0.05). Immunohistochemical detection of p53 protein, FGF-b, and TGF-alpha suggests that these growth-regulating proteins may play different roles in the development of sarcomas associated with vaccine sites.

Animals↗
PubMed2002

Gastric toxicity of racemic ketoprofen and its enantiomers in rat: oxygen radical generation and COX-expression.

OBJECTIVE AND DESIGN: The gastric toxicity of racemic-ketoprofen and its enantiomers (S(+)- and R(-)-ketoprofen), oxygen free radical generation and neutrophil infiltration in response to damage were evaluated in rats. Changes in prostaglandin synthesis, cyclooxygenase expression and glutathione metabolism were also studied. MATERIALS AND METHODS: Studies were performed in Wistar rats. Drugs were given by oral administration: racemic-ketoprofen (100, 50 and 25 mg/kg body weight); S(+) and R(-)-ketoprofen (50, 25 and 12.5 mg/kg body weight). Determinations were made of gastric mucosal injury, lipid peroxidation, xanthine oxidase, myeloperoxidase and superoxide dismutase activities, glutathione levels, glutathione peroxidase and glutathione reductase activities, gastric prostaglandin synthesis (PGE2 levels) and COX-expression. RESULTS: Racemic-ketoprofen dose-dependently exhibited the highest toxicity. In contrast, S(+)-ketoprofen at half the dose of the racemic compound proved to be less ulcerogenic. R(-)-ketoprofen was also less ulcerogenic, but when administered as the racemate exacerbated gastric ulceration caused by S(+)-ketoprofen. Drug administration produced significant increases in lipid peroxidation levels and xanthine-oxidase and a decrease in superoxide dismutase activity. Nevertheless the racemate induced the highest disturbances in oxidative metabolism. No changes in myeloperoxidase values and glutathione metabolism were found. Cyclooxygenase-1 immunoreactivity was observed and did not differ from that in control rats. Cyclooxygenase-2 could also be expressed after treatments. CONCLUSIONS: R(-)-ketoprofen and S(+)-ketoprofen have a comparable gastric toxicity and they both have a better gastric toxicity profile as compared to the racemate. In addition to inhibition of prostaglandin synthesis, damage resulted in an increase of cyclooxygenase-2 protein expression. Oxygen radicals, including superoxide anions, could also be implicated.

Animals↗
PubMed2002

Assessment of profilin as an allergen for latex-sensitized patients.

BACKGROUND: The presence of the actin-binding protein, profilin, has been demonstrated in natural latex extracts; but the clinical significance of this molecule as an allergen for latex-allergic patients is not clear. We studied the allergenic relevance of isolated latex natural and recombinant profilin, by in vivo and in vitro techniques, in two populations of spina bifida children (SB) and adults allergic to latex (AL). METHODS: Profilin is present in small amounts in latex extracts obtained from low ammoniated (LA) natural latex. Its purification by affinity chromatography resulted difficult due to Heb v 1 unspecific binding. Therefore a method was developed to obtain natural profilin from natural latex, combining affinity chromatography (PLP, poly-L-proline Sepharose column) and previous ammonium sulfate fractionation. Alternatively, latex c-serum containing a low amount of Hev b 1 and a relatively higher profilin content could be used. Recombinant latex profilin isoform (rHev b 8) was cloned by PCR amplification. The entire coding region of Hev b 8 was subcloned into the expression vector pKN172 and a non-fusion form of Hev b 8 was expressed in Escherichia coli BL21 (DE3). Purified recombinant protein was obtained after a single passage through PLP-Sepharose column. RESULTS: Natural and recombinant purified Hev b 8 were tested cutaneously by intradermoreaction (ID) in 17 SB and 14 AL patients. They were positive in 15 SB and 14 AL patients. No wheals were produced when tested in nonatopic control patients. Only 42% of sera from latex-allergic patients revealed specific IgE titers of class 1 or higher by enzyme immunoassay and only 39% of them exhibited IgE binding by SDS-PAGE immunoblotting with any natural or recombinant Hev b 8 forms. CONCLUSION: It seems that profilin is a relevant allergen for both groups of patients from a frequency point of view, but with scarce presence in natural latex extracts and raw sources, with a subsequent low IgE induction capacity.

Adult↗
PubMed2002

Metastatic lymphangiosarcoma in a horse.

A lymphangiosarcoma with metastases was found in a horse that presented with respiratory distress and edema in the ventral thorax and abdomen. The necropsy revealed diffuse edema in the subcutaneous connective tissue. Mediastinal, mesenteric, iliac, and renal lymph nodes were enlarged and white with soft, yellowish necrotic areas. Histologic examination revealed numerous channels and disorganized vessels lined by large polyhedral, polymorphic cells. Tumor metastases were observed in the spleen, lungs, and kidneys. Immunohistochemical evaluation of the tumor cells demonstrated positive staining for factor VIII, vimentin, and keratin. Laminin was scarce, and collagen IV staining was negative, consistent with a discontinuous or absent basement membrane.

Animals↗
PubMed2001

Detection of estrogen alpha and progesterone receptors and cell proliferation in the uterus during early pregnancy in the goat.

The objectives of this study were to investigate in the goat uterus the expression of estrogen-alpha (ER alpha) and progesterone receptors (PR) and their relationship to proliferation indices (Ki-67) during peri-implantation on Days 22 to 30 post coitum (pc). Immunohistochemical methods were used to quantify ER alpha and PR for luminal and deep regions of the endometrium and of the myometrium. On Day 22 pc cell proliferation was only observed in the luminal epithelium. On Day 24 pc, high cell proliferation indices were seen in luminal epithelium and proliferation began in the luminal stroma and glands. There was a positive correlation between Ki-67 and total ER alpha score in the luminal epithelium (r = 0.53, P < 0.01). Levels of PR scores were highly correlated with Ki-67 indices in luminal epithelium (r = 0.74, P < 0.01) and stroma (r = 0.70, P < 0.01). No Ki-67 expression was observed in deep glands, stroma or myometrium on any of the days studied. Results indicate that patterns of ER alpha and PR expression differ markedly, and that there was a high correlation between PR expression and cell proliferation in the caprine uterus during the peri-implantation period.

Animals↗
PubMed2001

Comparative evaluation of the CD4+CD8+ and CD4+CD8- lymphocytes in the immune response to porcine rubulavirus.

The porcine immune system is unique in the expression of CD4+CD8+ (double-positive, DP) lymphocytes. These cells have been associated with immunological memory due to their gradual increase with age, the expression of memory phenotype and their ability to respond to recall viral antigen. This work analyzes the biological function of CD4+CD8- and CD4+CD8+ lymphocytes in the immune response to porcine rubulavirus (PRv). CD4+CD8- cells isolated from pigs 3 weeks after infection with porcine rubulavirus proliferated in response to homologous virus and generated lymphoblasts which were predominantly of the CD4+CD8+ phenotype, whereas stimulation with mitogen induced proliferation but did not switch the phenotype. CD4+CD8- lymphocytes isolated after 10 weeks of infection proliferated in response to phytohemagglutinin (PHA) but did not proliferate in response to homologous virus and did not change their phenotype, whereas CD4+CD8+ lymphocytes proliferated in response to PHA and to viral antigen. The cytokine profile of both lymphocyte populations showed the presence of IL-2 and IL-10 transcripts, quantitation demonstrated that CD4+CD8+ cells expressed mainly IL-10, whereas CD4+CD8- lymphocytes expressed primarily IL-2. Our results show that CD4+CD8- lymphocytes in the early phase of porcine rubulavirus infection can be converted to double-positive cells expressing IL-10 in an antigen-dependent manner, and that CD4+CD8- T-cells late in infection do not acquire CD8.

Animals↗
PubMed2001

Rheumatoid arthritis in southern Spain: toward elucidation of a unifying role of the HLA class II region in disease predisposition.

OBJECTIVE: To evaluate the contributions of HLA-DQ and -DR polymorphisms to susceptibility to rheumatoid arthritis (RA) in a population in southern Spain, and to compare the value of the shared epitope (SE) and RA protection (RAP) models in accounting for the HLA class II region's contribution to RA predisposition. METHODS: One hundred sixty RA patients and 153 healthy controls were typed for HLA-DRB1 and -DQB1 using high-resolution DNA techniques. Distributions of predisposing DRB1 alleles in patients and control subjects according to the SE model were compared with distributions of predisposing DQ and protective DERAA-positive DRBI alleles according to the RAP model. RESULTS: DQ3 (DQBI*03 and *04 combined with DQA1*03) and DQ5 (DQB1*0501/DQA1*0101) alleles predisposed individuals to RA independently of SE-positive DRB1 alleles. DQ3/3-homozygous individuals had the strongest risk of developing RA. DQ3 molecules predisposed to RA more than did DQ5 molecules. The weaker predisposition mediated by DQ5 included the DRB1*1001-carrying haplotype; no DRB1*1001-homozygous patients were observed. DRBI*0401 played a unique role in the contribution of DQ3-DR4 haplotypes to RA, in spite of its low frequency in southern Spain. CONCLUSION: The low prevalences of RA and of mild disease observed in Spain, and in southern Europe in general, can be explained in great part by the low frequency of DQ3-DR4 haplotypes, especially those carrying DRB1*0401. However, the overall distribution of HLA-DQ and -DR alleles in RA patients compared with control subjects is similar to that in other European and North American populations. A model involving both DQ and DR can best account for the contribution of HLA to RA.

Alleles↗
PubMed2001

O-glycosylation in Echinococcus granulosus: identification and characterization of the carcinoma-associated Tn antigen.

In the present work we demonstrate that the cancer-associated O-glycosylated Tn antigen (GalNAc-O-Ser/Thr) is expressed by the cestode Echinococcus granulosus. This antigen was detected in both larval and adult worm extracts, with the highest specific activity observed in the adult excretion/secretion preparation. Histochemical analysis showed that Tn is preferentially expressed in the parenchyma in both parasite stages and the external part of tegument in adult worms. A similar pattern was observed for sialyl-Tn, a related O-linked antigen. Tn glycoproteins from protoscoleces were resolved by SDS-PAGE in two main components of 43 and 49 kDa. After purification, this material was reactive with lectins which bind GlcNAc/sialic acid, GalNAc, and T antigen. In a preliminary evaluation, high levels of Tn antigen were detected in serum samples from patients with hydatid cyst, suggesting that the measure of Tn in serum could be a biomarker of this disease, although extensive work is necessary in order to determine the clinical usefulness of this assay. The results reported here, the first evidence of O-glycosylation pathways in E. granulosus and the presence of Tn antigen in cestodes, suggest that the evaluation of O-glycosylated antigens might give new insights in the host-parasite relationship.

Adult↗
PubMed2001

Distribution of HERV-LTR elements in the 5'-flanking region of HLA-DQB1 and association with autoimmunity.

We established the detailed polymorphism of the 5'-flanking region and the first exon of the human leukocyte antigen (HLA)-DQB1 alleles. One hundred and forty-five Spanish rheumatoid arthritis (RA) patients and 200 healthy voluntary blood donors from southern Spain along with 42 B-cell lines were analyzed for the presence of the retrovirus-derived long terminal repeats (LTRs) LTR3, LTR5, and LTR13. LTR3 positivity was always associated with certain DQB1 alleles, i.e., *0302, *0402, *0601, *0202, and *0305. Sequencing analysis of the 5'-flanking region of DQB1*0301, *0303 and *0502 alleles in homozygous B-cell lines showed the absence of LTR3 and a massive deletion of 5635 base pairs. The undetected deletion in the flanking region of some DQB1 alleles and a lack of stratification for HLA typing explain previously reported associations of the LTR3 element with RA and type I diabetes (IDDM). LTR5 showed identical distribution to LTR3, consistent with a previously suggested LTR3-LTR5 tandem arrangement. LTR13 positivity was associated with DQB1*0302, *0303, and *0402 alleles. Distributions of the LTR elements in all B-cell lines, RA patients, and controls could be explained entirely by linkage disequilibrium with DQB1 alleles, independently of the haplotypes carrying them. LTR elements are known to regulate gene expression. Therefore, a possible involvement of LTR13 in the association of DQB1*0302, *0303, and *0402 with IDDM requires further investigation. The sequencing results of the DQB1 first exon demonstrated that DQB1*0601 was generated by a recombination event between a DR53 and a non-DR53 haplotype. Our results shed new light on the phylogeny of the HLA region and the possible contribution of DQB1 to susceptibility to autoimmunity.

Alleles↗
PubMed2001

Identification of circulating antigens, including an immunoglobulin binding protein, from Toxoplasma gondii tissue cyst and tachyzoites in murine toxoplasmosis.

Here we describe the identification of Toxoplasma gondii circulating antigens in sera of BALB/c mice experimentally infected with either the virulent RH strain, or the cystogenic WTD1 strain or with an isolate from a human patient. The circulating antigens were identified by immunoblot in tachyzoite (RH strain) and in tissue cyst (ME-49 strain) crude antigens, using antibodies produced by immunisation of BALB/c mice with homologous sera from infected animals. The most relevant tachyzoite antigen identified are in the following four clusters of 109-94, 67-57, 35-31 and 28-21 kDa. Tissue cyst-specific circulating antigens, like the 18 kDa one, were detected in sera from mice infected with the cystogenic strains. These immune sera, after depletion of tachyzoite specific antibodies, recognised three tissue cysts antigens with Mr of 120, 79 and 48 kDa, and a cluster of antigens in the range of 68-53 kDa. We produced monoclonal antibodies by fusion of myeloma cells with lymphocytes from the mouse immunised with circulating antigens from the RH strain. One of the clones (3A11/H12) obtained, secretes IgG(1) and recognises a peptide epitope from a tachyzoite 67 kDa protein. This parasite protein also binds irrelevant mouse IgG(1) as well as immunoglobulins from other species. The reactivity with non-specific antibodies was inhibited by preincubation with 2% normal mouse and goat serum, while the reaction with the monoclonal antibody 3A11/H12 was not. Furthermore, a biotinylated F(ab')(2) of an irrelevant mouse IgG(1) did not show any reactivity while the F(ab')(2) of the monoclonal antibody 3A11/H12 reacts specifically with the 67 kDa antigen suggesting that this circulating antigen is a putative Fc binding protein.

Animals↗