Validación de la Age Universal I-E Scale en sujetos mexicanos / Validation of the Age Universal I-E Scale in Mexican Subjects

Se evalúan las propiedades psicométricas de la Escala I-E-12 en tres muestras de sujetos mexicanos: religiosos (N = 78), no religiosos (N = 148) y una muestra mixta (N = 226). Mediante análisis factorial exploratorio se obtuvo una estructura compuesta por tres factores, siendo la estructura de grupos religiosos la que explicó el 63.5 % de la varianza con una consistencia interna alfa de entre 0.70 y 0.88 para la escala total y sus diferentes factores. La escala evaluada en sujetos mexicanos muestra resultados consistentes en buena medida con lo reportado por Simkin y Etchezahar (2013) en la exploración de la I-E 12, en el contexto argentino.

The work evaluates the psychometric properties of the scale I-E-12 in three Mexican samples: religious (N = 78), non religious (N = 148) and composite sample (N = 226). After the exploratory factor analysis applied to each of the samples, there was a structure composed of three factors being the structure of religious groups which he explained the 63.5 % of the variance with an alpha internal consistency between 0.7 and 0.88 for the full scale and its various factors. The scale assessed in Mexican subjects shows consistent results in good measure to those reported by Simkin and Etchezahar (2013), in the exploration of the I-E-12 in the argentine context.

Frequent HLA class I alterations in human prostate cancer: molecular mechanisms and clinical relevance.

Reduced expression of HLA class I is an important immune escape mechanism from cytotoxic T cells described in various types of malignancy. It often correlates with poor prognosis and resistance to therapy. However, current knowledge about the frequency, underlying molecular mechanisms, and prognostic value of HLA class I and II alterations in prostate cancer (PC) is limited. Immunohistochemical analysis demonstrated that 88 % of the 42 studied cryopreserved prostate tumors have at least one type of HLA alteration as compared to adjacent normal prostate epithelium or benign hyperplasia. Total loss of HLA-I expression found in 50 % of tumors showed an association with increased incidence of tumor relapse, perineural invasion, and high D'Amico risk. The remaining HLA-I-positive tumors demonstrated locus and allelic losses detected in 26 and 12 % of samples, respectively. Loss of heterozygosity at chromosome 6 was detected in 32 % of the studied tumors. Molecular analysis revealed a reduced expression of B2M, TAP2, tapasin and NLRC5 mRNA in microdissected HLA-I-negative tumors. Analysis of twelve previously unreported cell lines derived from neoplastic and normal epithelium of cancerous prostate revealed different types of HLA-I aberration, ranging from locus and/or allelic downregulation to a total absence of HLA-I expression. The high incidence of HLA-I loss observed in PC, caused by both regulatory and structural defects, is associated with more aggressive disease development and may pose a real threat to patient health by increasing cancer progression and resistance to T-cell-based immunotherapy.

Immune escape of cancer cells with beta2-microglobulin loss over the course of metastatic melanoma.

Cancer cells escape T-cell-mediated destruction by losing human leukocyte antigen (HLA) class I expression via various mechanisms, including loss of beta2-microglobulin (ß2m). Our study illustrates the immune escape of HLA class I-negative tumor cells and chronological sequence of appearance of tumor ß2m gene mutation in successive lesions obtained from a patient with metastatic melanoma. We observed a gradual decrease in HLA expression in consecutive lesions with few HLA-negative nodules in the primary tumor and the emergence of a totally negative lesion at later stages of the disease. We detected loss of ß2m in ß2m-negative nests of the primary tumor caused by a combination of two alterations: (i) a mutation (G to T substitution) in codon 67 in exon 2 of ß2m gene, producing a stop codon and (ii) loss of the second gene copy by loss of heterozygosity (LOH) in chromosome 15. The same ß2m mutation was found in a homogeneously ß2m-negative metastasis 10 months later and in a cell line established from a biopsy of a postvaccination lymph node. Microsatellite analysis revealed the presence of LOH in chromosomes 6 and 15 in tumor samples, showing an accumulation of chromosomal loss at specific short tandem repeats in successive metastases during disease progression. HLA loss correlated with decreased tumor CD8+ T-cell infiltration. Early incidence of ß2m defects can cause an immune selection and expansion of highly aggressive melanoma clones with irreversible genetic defects causing total loss of HLA class I expression and should be taken into consideration as a therapeutic target in the development of cancer immunotherapy protocols.

Immune infiltrates are prognostic factors in localized gastrointestinal stromal tumors.

Cancer immunosurveillance relies on effector/memory tumor-infiltrating CD8(+) T cells with a T-helper cell 1 (TH1) profile. Evidence for a natural killer (NK) cell-based control of human malignancies is still largely missing. The KIT tyrosine kinase inhibitor imatinib mesylate markedly prolongs the survival of patients with gastrointestinal stromal tumors (GIST) by direct effects on tumor cells as well as by indirect immunostimulatory effects on T and NK cells. Here, we investigated the prognostic value of tumor-infiltrating lymphocytes (TIL) expressing CD3, Foxp3, or NKp46 (NCR1) in a cohort of patients with localized GIST. We found that CD3(+) TIL were highly activated in GIST and were especially enriched in areas of the tumor that conserve class I MHC expression despite imatinib mesylate treatment. High densities of CD3(+) TIL predicted progression-free survival (PFS) in multivariate analyses. Moreover, GIST were infiltrated by a homogeneous subset of cytokine-secreting CD56(bright) (NCAM1) NK cells that accumulated in tumor foci after imatinib mesylate treatment. The density of the NK infiltrate independently predicted PFS and added prognostic information to the Miettinen score, as well as to the KIT mutational status. NK and T lymphocytes preferentially distributed to distinct areas of tumor sections and probably contributed independently to GIST immunosurveillance. These findings encourage the prospective validation of immune biomarkers for optimal risk stratification of patients with GIST.

Analysis of HLA-ABC locus-specific transcription in normal tissues.

We developed a novel human leukocyte antigen HLA-ABC locus-specific quantitative real-time polymerase chain reaction (PCR) to determine the locus-specific gene expression of HLA-ABC in peripheral blood leukocytes (PBLs, n = 53), colon mucosa (n = 15), and larynx mucosa (n = 15). Laser-assisted tissue microdissection allowed us to study the selected cells without interference from surrounding stroma. We report evidence on the specificity of the technique, describing the HLA-ABC locus-specific gene expression patterns found in the PBLs and two solid tissues studied. PBLs showed a higher gene expression of HLA-B than of HLA-A or HLA-C (p = 4.7 × 10(-10) and p = 1.6 × 10(-6), respectively). In solid tissue, HLA-A and HLA-B gene expressions were similar and HLA-C expression lower. In particular, in larynx mucosa, significant differences were found between HLA-A and HLA-C expressions and between HLA-B and HLA-C expressions (p = 6.5 × 10(-4) and p = 8.1 × 10(-4), respectively). The same differences were observed in colon mucosa, but significance was not reached (p = 0.08 and p = 0.06, respectively). Differences in locus-specific regulation may be related to the control of cytotoxic responses of NK and CD8 positive T cells. Gene expression of HLA-ABC specific locus showed no intra-individual variability, but there was a high inter-individual variability. This may result from differences in the expression of common regulatory factors that control HLA-ABC constitutive expression.

Estimación de la masa grasa por DXA y el modelo de cuatro compartimentos en púberes mexicanos de 9 a 14 años / Estimation of body fat by DXA and the four compartment model in Mexican youth

Se evaluó la precisión y exactitud en la estimación de la grasa corporal (%) por absorciometría dual de rayos X (DXA Lunar-DPX-MD) comparado con el modelo de cuatro compartimentos (4C) en 32 púberes (F=16) de 9 a 14 años. El sesgo entre la DXA y el modelo de 4C fue de -3.5% de grasa (r=0.25; p=0.171) con un intervalo de confianza de -1.9 a -5.1 (p=0.050). Los límites de concordancia al 95% fueron de +5% a -12% de grasa. El coeficiente de correlación de concordancia fue de pc=0.85. La prueba de exactitud por análisis de regresión mostró que el intercepto y la pendiente de las estimaciones de grasa corporal por DXA fueron diferentes al modelo de 4C (p>0.05). La precisión evaluada con el valor de R2 mostró que la DXA explicó el 83% de la varianza de la grasa corporal por el modelo de 4C con un error de 4.1%. El error total como medida de exactitud fue de 5.6%. La exactitud grupal evaluada por análisis de varianza no mostró interacción entre el método (DXA-4C) y el análisis por separado del sexo, el estado puberal y la presencia de sobrepeso. No obstante, hubo efecto del método (p=0.043) en presencia de sobrepeso (p<0.001). En conclusión, los resultados muestran que el uso de la DXA comparado con el modelo de 4C no es equivalente en púberes mexicanos. Sin embargo, estos datos no limitan el uso de la DXA en estudios de composición corporal y su relación con anormalidades metabólicas.

The objective of this study was to validate the estimation of body fat (%BF) by DXA (Dual-Energy X-Ray AbsorciomDPX-MD) against the four compartment model (4C) of body composition in 32 Mexican pubertal girls and boys (aged 9 - 14y; F=16). The mean of the difference between DXA and 4C model was -3.5 %BF (p=0.171). The limits of agreement (95% ± 2 SD) were +5% to -12%BF. The precision of estimated limits of y the confidence intervals were -1.9% to -5.1%BF (P=0.050). The concordance correlation coefficient was pc= 0.85. The test of accuracy for coincidence of slop intercepts between DXA and the 4C model showed no coincidence (p< 0.05). The precision by R2 explained 83% of the variance (SEE, 4.1 %). The individual accuracy assess by the total error was 5.6%. The group mean accuracy by two way analysis of variance of body fat did not show interaction between method (DXA-4C model) and separate analysis of gender and overweight. However, there was an effect of method (p=0.043) in the presence of overweight (p<0.001). In conclusion, the estimation of percent of body fat by DXA was not precise and accurate in a group of Mexican children. However, results do not limit the utility of DXA for the measurements of body composition and its relation with health outcomes, especially in follow up studies.

Regulation of clusterin expression in human cancer via DNA methylation.

BACKGROUND/AIMS: Clusterin has attracted much recent attention because of its association with tumorigenesis and the progression of human carcinomas. The present study was designed to examine the role of clusterin methylation as an indicator of clusterin expression in tumor cell lines and breast tissue samples. METHODS: For this purpose, we used methylation-sensitive restriction analysis followed by PCR. RESULTS: None of the non-tumoral breast samples showed expression of clusterin by immunohistochemistry, and a methylated state was found in the promoter region of the gene. However, a demethylated state was found in 5 of 6 analyzed carcinoma cell lines. Four of 5 demethylated cell lines presented moderate to strong expression of clusterin, while no expression was detected in the unmethylated cell line. The inverse correlation found in most cell lines between clusterin expression and promoter methylation was also found in most human tumors analyzed (p < 0.001). Thus, a methylated state was present in 14 carcinomas, 12 of them with a null expression of clusterin, while a demethylated state was detected in 7 breast tumor samples, with 5 of them presenting strong expression. CONCLUSIONS: We conclude that clusterin expression is under epigenetic control via methylation of its promoter.

HLA and melanoma: multiple alterations in HLA class I and II expression in human melanoma cell lines from ESTDAB cell bank.

Altered HLA class I and class II cell surface expression has been reported in many types of malignancy and represents one of the major mechanism by which tumour cells escape from T lymphocytes. In this report, we review the results obtained from the study of constitutive and IFN-gamma-induced expression of HLA class I and II molecules in 91 human melanoma cell lines from the European Searchable Tumour Cell Line Database, and compare them with published data on HLA expression in other types of cancer. Various types of alterations in HLA class I cell surface expression were found in a high percentage (67%) of the studied cell lines. These alterations range from total to selective HLA class I loss and are associated with beta2-microglobulin gene mutations, transcriptional downregulation of HLA class I genes and antigen processing machinery components, or with the loss of heterozygosity in chromosome 6. The most frequently observed phenotype is selective downregulation of HLA-B locus, reversible after treatment with IFN-gamma. The expression of constitutive- or IFN-gamma induced-surface expression of at least one HLA class II locus is positive in 71.5% of the analysed cell lines. Four different HLA class II expression phenotypes were defined, and a positive correlation between the expression of class I and II molecules is discussed. More detailed information on the HLA expression patterns and others immunological characteristics of these melanoma cell lines can be found on the following website http://www.ebi.ac.uk/ipd/estdab .

Characterization of HLA class I altered phenotypes in a panel of human melanoma cell lines.

BACKGROUND: Altered HLA class I cell surface expression is one of the major mechanisms by which tumor cells escape from T lymphocytes. Immunohistochemistry-defined phenotypes of lost HLA class I expression have been described in human solid tumors, nut less information is available on melanoma cell lines. OBJECTIVES: To describe the frequency and distribution of different types of HLA class I antigen alterations in 91 melanoma cell lines from the European Searchable Tumour Cell and Databank (ESTDAB). METHODS: The HLA class I expression was assessed by flow cytometry and HLA genotyping. RESULTS: We found various types of HLA class I cell surface alterations in about 67% of the melanoma cell lines. These alterations range from total to selective HLA class I loss due to loss of heterozygosity (LOH), haplotype loss, beta2-microglobulin gene mutation, and/or total or selective down-regulation of HLA class I molecules. The most frequently observed phenotype is down-regulation of HLA-B locus that was reversible after treatment with IFN -gamma. CONCLUSIONS: In general, HLA class I alterations in the majority of the cells analyzed were of regulatory nature and could be restored by IFN-gamma. Analysis of the frequency of distinct HLA class I altered phenotypes in these melanoma cell lines revealed specific differences compared to other types of tumors.

Role of altered expression of HLA class I molecules in cancer progression.

HLA class I antigens play a key role in immune recognition of transformed and virally infected cells via binding to the peptides of "non-self" or aberrantly expressed proteins and subsequent presentation of the newly formed "HLA-I-peptide" complex to T lymphocytes. Consequently, a chain of immune reactions is initiated leading to tumor cell elimination by cytotoxic T cells. Altered tumor expression of HLA class I is frequently observed in various types of malignancies. It represents one of the main mechanisms used by cancer cells to evade immunosurveillance. Because of immune selection, HLA class I-negative variants escape and lead to tumor growth and metastatic colonization. Loss or downregulation of HLA class I antigens on tumor cell surface is a factor that limits clinical outcome of peptide-based cancer vaccines aimed to increasing specific anti-tumor activity of cytotoxic T lymphocytes. Thus, gaining more knowledge regarding frequency of HLA class I defect, its tissue specificity, and underlying molecular mechanisms may help designing appropriate therapeutic strategies in cancer treatment. Here, we describe various types of HLA class I alterations found in different malignancies and molecular mechanisms that underlie these defects. We also discuss a correlation between HLA class I defects cancer progression in melanoma patients with poor clinical response to autologous vaccination.

Distinct mechanisms of loss of IFN-gamma mediated HLA class I inducibility in two melanoma cell lines.

BACKGROUND: The inability of cancer cells to present antigen on the cell surface via MHC class I molecules is one of the mechanisms by which tumor cells evade anti-tumor immunity. Alterations of Jak-STAT components of interferon (IFN)-mediated signaling can contribute to the mechanism of cell resistance to IFN, leading to lack of MHC class I inducibility. Hence, the identification of IFN-gamma-resistant tumors may have prognostic and/or therapeutic relevance. In the present study, we investigated a mechanism of MHC class I inducibility in response to IFN-gamma treatment in human melanoma cell lines. METHODS: Basal and IFN-induced expression of HLA class I antigens was analyzed by means of indirect immunofluorescence flow cytometry, Western Blot, RT-PCR, and quantitative real-time RT-PCR (TaqMan(R) Gene Expression Assays). In demethylation studies cells were cultured with 5-aza-2'-deoxycytidine. Electrophoretic Mobility Shift Assay (EMSA) was used to assay whether IRF-1 promoter binding activity is induced in IFN-gamma-treated cells. RESULTS: Altered IFN-gamma mediated HLA-class I induction was observed in two melanoma cells lines (ESTDAB-004 and ESTDAB-159) out of 57 studied, while treatment of these two cell lines with IFN-alpha led to normal induction of HLA class I antigen expression. Examination of STAT-1 in ESTDAB-004 after IFN-gamma treatment demonstrated that the STAT-1 protein was expressed but not phosphorylated. Interestingly, IFN-alpha treatment induced normal STAT-1 phosphorylation and HLA class I expression. In contrast, the absence of response to IFN-gamma in ESTDAB-159 was found to be associated with alterations in downstream components of the IFN-gamma signaling pathway. CONCLUSION: We observed two distinct mechanisms of loss of IFN-gamma inducibility of HLA class I antigens in two melanoma cell lines. Our findings suggest that loss of HLA class I induction in ESTDAB-004 cells results from a defect in the earliest steps of the IFN-gamma signaling pathway due to absence of STAT-1 tyrosine-phosphorylation, while absence of IFN-gamma-mediated HLA class I expression in ESTDAB-159 cells is due to epigenetic blocking of IFN-regulatory factor 1 (IRF-1) transactivation.

Identification of different tumor escape mechanisms in several metastases from a melanoma patient undergoing immunotherapy.

The cytotoxic activity of T cells selects the outgrowth of tumor cells that escape from immune surveillance by different strategies. The different mechanisms that interfere with immune recognition and limit vaccination efficiency are still poorly understood. We analysed six cell lines established from different metastases of melanoma patient UKRV-Mel-20 for specific characteristics known to have an impact on the tumor-T cell interaction: (1) alterations in the HLA class I phenotype, (2) expression of Fas/CD95, and (3) expression of specific cytokines and chemokines. One of the cell lines, UKRV-Mel-20f, exhibited an HLA class I haplotype loss and just this cell line was also characterised by the expression of Fas/CD95 and of relatively high levels of proinflammatory chemokines suggesting that the cytotoxic activity of tumor-infiltrating T cells might have selected the outgrowth of this tumor cell variant. All other cell lines analysed showed no alterations in HLA class I expression, but, in contrast to UKRV-Mel-20f, expressed much lower levels of Fas/CD95 and of proinflammatory chemokines and some of them produced high levels of immunosuppressive TGF-beta1. These results suggest that in patient UKRV-Mel-20, tumor cells interfere with T cell recognition by different strategies which might partially explain why this patient did not have a clinical response to an autologous tumor cell vaccine.

[Total content and dialyzable iron and zinc in foods from northern and Southern Mexico]. / Contenido total y disponibilidad in vitro de hierro y zinc en alimentos de mayor consumo en Sonora y Oaxaca, México.

Developing countries diets are based on a variety of plant foods that often are the main suppliers of important amounts of iron (Fe) and zinc (Zn). The objectives of this study were 1) to measure the total and dialyzable amounts of Fe and Zn in foods from Northern Mexico (Sonora) and from Southern Mexico (Oaxaca) and 2) to evaluate the effect of meat content of diets on the dialyzable amount of Fe and Zn. Methods to calculate the total dialyzable amount of Fe and Zn, were those of the AOAC and of Shen et al. Total Fe in e northern Mexican foods went from 0.78 +/- 0.0 to 11.59 +/- to 0.03 mg/ 100g (dry weight, DW); in southern Mexican foods the same micronutrient amounts were 0.86 +/- 0.18 to 8.8 +/- 0.57 mg/100 g (BS). Total Zn values were 0.91 +/- 0.00 to 13.58 +/- 0.05 mg/100 g (DW) in Sonora, and 0.64 +/- 0.18 to 20.80 +/- 0.33 mg/100 g (DW) in Oaxaca. In northern Mexico, foods dialyzable Fe had values from 0.1 +/- 0.04% to 10.6 +/- 0.36% and for Zn from 4.0 +/- 0.21% to 55.32 +/- 0.14%. Meanwhile, the range of values of dialyzable Fe for foods from Oaxaca were from 0.22 +/- 0.06% to 9.40 +/- 0.14% for and from 2.41 +/- 0.26% to 54.27 +/- 1.49% for dialyzable Zn. The average value for dialyzable Fe was higher in the foods that contained meat or meat products (p= 0.001).

Contenido total y disponibilidad in vitro de hierro y zinc en alimentos de mayor consumo en Sonora y Oaxaca México / Total content and dialyzable iron and zinc in foods from Northern and Southern Mexico

La dieta en países en vías de desarrollo está constituida por cereales y legumbres que si bien aportan cantidades importantes de hierro (Fe) y zinc (Zn), resultan ser minerales de baja disponibilidad. Los objetivos del presente trabajo fueron: 1) cuantificar el contenido total y dializable de Fe y Zn en alimentos del norte (Sonora) y del sur (Oaxaca) de México; 2) evaluar el efecto de la presencia de carne sobre los valores de Fe y Zn dializables. La cuantificación de Fe y Zn total y dializable se hizo por el método de la AOAC y de Shen et al, respectivamente. Los resultados obtenidos para Fe total en alimentos de Sonora presentaron un rango de 0.78±0.0 a 11.59±0.03 mg/100g (base seca, BS); y en los de Oaxaca de 0.86±0.18 a 8.8±0.57 mg/100g (BS). Para Zn total los valores variaron de 0.91±0.00 a 13.58±0.05 mg/100g (BS) en Sonora y de 0.64±0.18 a 20.80±0.33 mg/100 g (BS) en Oaxaca. En los alimentos de Sonora se tuvieron valores de 0.1±0.04 por ciento a 10.6±0.36 por ciento para Fe dializable y de 4.0±0.21 por ciento a 55.32±0.14 por ciento para Zn dializable. En los de Oaxaca el rango fue de 0.22±0.06 por ciento a 9.40±0.14 por ciento para Fe dializable y de 2.41±0.26 por ciento a 54.27±1.49 por ciento para Zn dializable. Se obtuvo un promedio mayor en el contenido de Fe dializable en los alimentos que contenían carne, respecto a los que no la contenían y entre el frijol pinto y las tortillas de maíz de Sonora respecto al frijol negro y tortillas de maíz rurales de Oaxaca. El contenido de Zn dializable no mostró diferencias significativas entre los alimentos de las dos regiones

High frequency of homozygosity of the HLA region in melanoma cell lines reveals a pattern compatible with extensive loss of heterozygosity.

Malignant transformation of cells is frequently associated with abnormalities in human leukocyte antigen (HLA) expression. MHC class I loss or down-regulation in cancer cells is a major immune escape route used by a large variety of human tumours to evade antitumour immune responses mediated by cytotoxic T lymphocytes. The goal of our study was to explore HLA genotyping and phenotyping in a variety of melanoma tumour cell lines. A total of 91 melanoma cell lines were characterised for HLA class I and II genotype. In addition, 61 out of the 91 cell lines were also analysed for HLA class I and II cell surface molecule expression by flow cytometry. Unexpectedly, we found that 19.7% of the melanoma cell lines were homozygous for HLA class I genotypes, sometimes associated with HLA class II homozygosity (8.79%) and sometimes not (10.98%). The frequency of homozygosity was significantly higher compared with the control groups (1.6%). To identify the reasons underlying the high frequency of HLA homozygosity we searched for genomic deletions using eight pairs of highly polymorphic microsatellite markers covering the entire extended HLA complex on the short arm of chromosome 6. Our results were compatible with hemizygous deletions and suggest that loss of heterozygosity on chromosome arm 6p is a common feature in melanoma cell lines. In fact, although autologous normal DNA from the patients was not available and could not be tested, the retention in some cases of heterozygosity for a number of microsatellite markers would indicate a hemizygous deletion. In the rest of the cases, markers at 6p and 6q showed a single allele pattern indicating the probable loss of part or the whole of chromosome 6. These results led us to conclude that loss of heterozygosity in chromosome 6 is nonrandom and is possibly an immunologically relevant event in human malignant melanoma. Other well-established altered HLA class I phenotypes were also detected by flow cytometry that correspond to HLA class I total loss and HLA-ABC and/or specific HLA-B locus down-regulation.

Analysis of HLA-E expression in human tumors.

Downregulation of MHC class Ia molecule expression is a widespread mechanism used by tumor cells to escape antitumor T-cell-mediated immune responses. However, it is not known why NK cells cannot lyse these MHC class-Ia-deficient tumor targets. Tumors must select additional routes of escape from NK cells. An attractive hypothesis is that the aberrant expression of nonclassical HLA class Ia molecules in tumors provides the required inhibitory signal to NK cells, rendering tumor cells resistant to NK lysis. To analyze the possible role of HLA-E molecules in providing tumor cells with an NK escape mechanism, we studied the cell surface expression of this HLA class Ib molecule in a variety of tumor cell lines with well-defined HLA class Ia alterations. Tests were done with the monoclonal antibody 3D12 recognizing cell surface HLA-E molecules. Our results indicate that HLA-E was mainly detected in leukemia-derived cell lines. In addition, HLA-E was detected in tumor cell lines of different origin. This expression was related with the availability of free beta(2)-microglobulin (beta(2)m) in the cytoplasm of tumor cells. Expression was detected in tumor cell lines showing an imbalance in heavy chain/beta(2)m expression, particularly in tumor cell lines with alterations in the expression of heavy-chain genes. Several lines of evidence favor these conclusions: (1) In the FM55 and NW145 melanoma tumor systems, the reduction in HLA class Ia expression paralleled the increased cell surface detection of HLA-E. (2) A cervical tumor (808) and a melanoma cell line (R22.2) expressing a single HLA-A1 allele also expressed HLA-E. (3) The addition of human beta(2)m to tumor cell lines that expressed the HLA-E(G) allele increased HLA-E cell surface expression. (4) There was no HLA-E cell surface expression in tumor cell lines with total loss of HLA class Ia expression, including cell lines with low transcription of HLA class I heavy chains or with beta(2)m mutations. Our findings suggest that the biological consequences of these cumulative genetic and molecular changes in tumor cells lead to the appearance of HLA-E in a limited number of tumor cell lines with peculiar phenotypic and genotypic characteristics, namely: HLA-class Ia downregulation, free beta(2)m and HLA-E(G) genotype. The aberrant HLA-E expression might be of particular biological relevance in those HLA tumor phenotypes that express a single HLA-A allele when NK inhibition is markedly reduced due to the downregulation of HLA-B and -C alleles.

Complete loss of HLA class I antigen expression on melanoma cells: a result of successive mutational events.

Alterations in the surface expression of HLA class I molecules have been described as a strategy of tumors to evade recognition by cytotoxic T cells. We detected complete loss of HLA class I antigen presentation for 2 tumor cell lines from 1 melanoma patient, the first originated from a regional lymph node lesion diagnosed simultaneously with the primary tumor and the second established 8 months later from a metastatic pleural effusion sample. Antigen presentation was not inducible with IFN-gamma but could be restored after transfection of tumor cells with b2m cDNA, indicating a defect in b2m expression. Analysis of the nature of this defect revealed that it originated from at least 2 mutational events affecting both copies of the b2m gene: a microdeletion of 498 bp in one b2m gene, including its entire exon 1, and a macrodeletion involving the entire copy of the second b2m gene. Microsatellite analysis pointed to the macrodeletion by demonstrating LOH for several specific markers on the long arm (q) of chromosome 15. Structural imbalance of 15q was verified by FISH. FISH studies also indicated the coexistence of a structurally abnormal variant of chromosome 15q with 2 apparently entire chromosomes 15q harboring the homozygous b2m microdeletion. Block of b2m expression in tumor cells builds a barrier to immunotherapy of cancer patients, and its early incidence should be of major consideration in the development and design of immunotherapeutic strategies.

[Ascorbic acid consumption and serum levels in smokers and non-smokers adult men in Hermosillo, Sonora, México]. / Consumo de ácido ascórbico y niveles séricos en hombres adultos fumadores y no fumadores de la CD. de Hermosillo, Sonora, México.

Ascorbic acid is one of the important antioxidant nutrients that can aid in the prevention of oxidative cellular damage. Adequate dietary intake is essential as humans can not synthesize this vitamin. It has been reported that smokers require higher dietary intakes to maintain their serum levels. The objective of this study was to determine serum levels of ascorbic acid in young male smokers and non smokers in the city of Hermosillo, Sonora, Mexico. In addition, their dietary intake of ascorbic acid was determined by a 24 h dietary recall. The dietary intake of ascorbic acid in 12 smokers was 64 +/- 11 mg/d and in 13 non smokers it was 70 +/- 12 mg/d. The smokers in this study did not meet the dietary recommendation of 100 mg/d. Serum ascorbic acid values in smokers and non smokers were 24.2 +/- 6.9 mumol/L and 30.9 +/- 3.7 mumol/L respectively. No significant difference was found among the 2 groups. Although the average serum ascorbic acid values fell within the range considered normal, 50% of the smokers had individual values that were below 23 mumol/L, indicating that these subjects have hipovitaminosis. A positive correlation between intake and serum levels was obtained for smokers (r = 0.71; p = 0.03). The results of this study suggest smokers may be at increased risk for chronic diseases due to their low intake and low serum levels of ascorbic acid.

Contenido y absorción del calcio proveniente de la dieta del noroeste de México: una retrospectiva bibliográfica / Content and absorption of calcium in diets of northwestern México: a look at the literature

El calcio es un mineral indispensable para el crecimiento y desarrollo del esqueleto y de los dientes en el hombre. Durante la edad adulta la actividad ósea se limita al mantenimiento de la masa esquelética para lo cual se requiere cubrir adecuadamente la ingestión de calcio. El objetivo de este escrito es realizar una retrospectiva de la información bibliográfica sobre contenido y biodisponibilidad de calcio proveniente de la dieta del Noroeste de México. Los trabajos dietarios mostraron que las cantidades de calcio (1164 mg) en las dietas del Noroeste de México cubren la recomendación de la RDA para adultos mayores de 25 años y que sus principales aportadores son las tortillas de maíz, las tortillas de harina de trigo y los frijoles. Los valores de disponibilidad de calcio de las dietas conteniendo esos alimentos aumentaron con tan solo agregar pequeñas cantidades de productos aportadores de calcio de origen animal. Esa diferencia en la disponibilidad del calcio se observó aún cuando uno de los estudios in vivo se realizó con un número mínimo de mujeres, por lo que podemos concluir que existe la necesidad de evaluar el efecto del calcio aportado por las dietas del Noroeste de México directamente en la población, considerando, además, otros factores que intervienen en el mantenimiento del esqueleto (ejercicio, edad, sexo, etc)

AMEU aspiración manual endouterina / AMEU endouterine manual aspiration

La estadística que se realizó en el Instituto Maternologico Percy Boland desde 28 de septiembre de 1999 al 3 de febrero del 2000, con un total de 178 pacientes arrojo las siguientes estadísticas detalladas en la investigación