DNA-Methylation Signatures of Tobacco Smoking in a High Cardiovascular Risk Population: Modulation by the Mediterranean Diet.

Biomarkers based on DNA methylation are relevant in the field of environmental health for precision health. Although tobacco smoking is one of the factors with a strong and consistent impact on DNA methylation, there are very few studies analyzing its methylation signature in southern European populations and none examining its modulation by the Mediterranean diet at the epigenome-wide level. We examined blood methylation smoking signatures on the EPIC 850 K array in this population (n = 414 high cardiovascular risk subjects). Epigenome-wide methylation studies (EWASs) were performed analyzing differential methylation CpG sites by smoking status (never, former, and current smokers) and the modulation by adherence to a Mediterranean diet score was explored. Gene-set enrichment analysis was performed for biological and functional interpretation. The predictive value of the top differentially methylated CpGs was analyzed using receiver operative curves. We characterized the DNA methylation signature of smoking in this Mediterranean population by identifying 46 differentially methylated CpGs at the EWAS level in the whole population. The strongest association was observed at the cg21566642 (p = 2.2 × 10-32) in the 2q37.1 region. We also detected other CpGs that have been consistently reported in prior research and discovered some novel differentially methylated CpG sites in subgroup analyses. In addition, we found distinct methylation profiles based on the adherence to the Mediterranean diet. Particularly, we obtained a significant interaction between smoking and diet modulating the cg5575921 methylation in the AHRR gene. In conclusion, we have characterized biomarkers of the methylation signature of tobacco smoking in this population, and suggest that the Mediterranean diet can increase methylation of certain hypomethylated sites.

Simple sugar intake and cancer incidence, cancer mortality and all-cause mortality: A cohort study from the PREDIMED trial.

OBJECTIVE: To examine associations between intake of simple sugars and cancer incidence, cancer mortality, and total mortality in a prospective cohort study based on the PREDIMED trial conducted from 2003 to 2010. METHODS: Participants were older individuals at high cardiovascular risk. Exposures were total sugar, glucose and fructose from solid or liquid sources, and fructose from fruit and 100% fruit juice. Cancer incidence was the primary outcome; cancer mortality and all-cause mortality were secondary outcomes. Multivariable-adjusted, time-dependent Cox proportional hazard models were used. RESULTS: Of 7447 individuals enrolled, 7056 (94.7%) were included (57.6% women, aged 67.0 ± 6.2 years). 534 incident cancers with 152 cancer deaths and 409 all-cause deaths were recorded after a median follow-up of 6 years. Intake of simple sugars in solid form was unrelated to outcomes. Higher cancer incidence was found per 5 g/day increase in intake of liquid sugars, with multivariable-adjusted HR of 1.08 (95% CI, 1.03-1.13) for total liquid sugar, 1.19 (95% CI, 1.07-1.31) for liquid glucose, 1.14 (95% CI, 1.05-1.23) for liquid fructose, and 1.39 (95% CI, 1.10-1.74) for fructose from fruit juice. Cancer and all-cause mortality increased to a similar extent with intake of all sugars in liquid form. In categorical models, cancer risk was dose-related for all liquid sugars. CONCLUSIONS: Simple sugar intake in drinks and fruit juice was associated with an increased risk of overall cancer incidence and mortality and all-cause mortality. This suggests that sugary beverages are a modifiable risk factor for cancer and all-cause mortality.

Dietary vitamin D intake and colorectal cancer risk: a longitudinal approach within the PREDIMED study.

PURPOSE: We evaluated whether the intake of dietary vitamin D is associated with the incidence of both colorectal cancer (CRC) and colon cancer in the framework of the PREDIMED cohort of older adults at high cardiovascular risk. METHODS: We analyzed data from 7216 men and women (55-80 years) without CRC at baseline from the PREvención con DIeta MEDiterránea study. Baseline consumption of vitamin D was assessed using a validated 137-item food frequency questionnaire. Cox proportional hazards ratios (HRs) of CRC and colon cancer incidence were estimated for quartiles and per 1-SD of baseline vitamin D intake. RESULTS: During a median follow-up of 6 years, we documented 97 incident CRC cases after the exclusion of subjects with no baseline dietary data and/or outliers of energy intake. A non-significant HRs and 95% confidence intervals (CIs) of CRC for the comparison of extreme quartiles (4th vs 1st) of vitamin D intake were observed [0.55 (0.30-1.00), P for trend = 0.072], whereas it was significant for colon cancer incidence alone [0.44 (0.22-0.90), P for trend = 0.032]. However, this association became significant in CRC and colon cancer incidence, after excluding 391 subjects consuming baseline vitamin D and/or calcium medication or prescribed supplements [0.52 (0.28-0.96) and 0.41 (0.12-0.85), respectively]. CONCLUSION: A higher dietary intake of vitamin D was significantly associated with a reduced CRC risk in individuals at high cardiovascular risk.

Association between the 2018 WCRF/AICR and the Low-Risk Lifestyle Scores with Colorectal Cancer Risk in the Predimed Study.

Limited longitudinal studies have been conducted to evaluate colorectal cancer (CRC) incidence based on the updated 2018 World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) recommendations or other global lifestyle indices, and none in aged populations at high cardiovascular risk. We aimed to assess the association between CRC incidence and adherence to two emerging lifestyles indices (2018 WCRF/AICR score and another low-risk lifestyle (LRL) score comprising smoking status, alcohol consumption, physical activity, diet, and body mass index) in the Spanish PREvencion con DIeta MEDiterranea (PREDIMED) cohort. We studied 7216 elderly men and women at high cardiovascular risk. The 2018 WCRF/AICR and LRL scores were calculated. Multivariable Cox proportional regression models were fitted to estimate the HRs (hazard ratios) and 95% confidence intervals (CIs) for incident CRC events. During a median interquartile range (IQR) follow-up of 6.0 (4.4-7.3) years, 97 CRC events were considered. A significant linear association was observed between each 1-point increment in the WCRF/AICR score (score range from 0 to 7) and CRC risk (HR (95% CI) = 0.79 (0.63-0.99)). Similarly, each 1-point increment in the LRL score (score range from 0 to 5) was associated with a 22% reduction in CRC risk (0.78 (0.64-0.96)). Adhering to emergent lifestyle scores might substantially reduce CRC incidence in elderly individuals. Further longitudinal studies, which take different lifestyle indexes into account, are warranted in the future.

Genome-Wide Association Study for Serum Omega-3 and Omega-6 Polyunsaturated Fatty Acids: Exploratory Analysis of the Sex-Specific Effects and Dietary Modulation in Mediterranean Subjects with Metabolic Syndrome.

Many early studies presented beneficial effects of polyunsaturated fatty acids (PUFA) on cardiovascular risk factors and disease. However, results from recent meta-analyses indicate that this effect would be very low or nil. One of the factors that may contribute to the inconsistency of the results is that, in most studies, genetic factors have not been taken into consideration. It is known that fatty acid desaturase (FADS) gene cluster in chromosome 11 is a very important determinant of plasma PUFA, and that the prevalence of the single nucleotide polymorphisms (SNPs) varies greatly between populations and may constitute a bias in meta-analyses. Previous genome-wide association studies (GWAS) have been carried out in other populations and none of them have investigated sex and Mediterranean dietary pattern interactions at the genome-wide level. Our aims were to undertake a GWAS to discover the genes most associated with serum PUFA concentrations (omega-3, omega-6, and some fatty acids) in a scarcely studied Mediterranean population with metabolic syndrome, and to explore sex and adherence to Mediterranean diet (MedDiet) interactions at the genome-wide level. Serum PUFA were determined by NMR spectroscopy. We found strong robust associations between various SNPs in the FADS cluster and omega-3 concentrations (top-ranked in the adjusted model: FADS1-rs174547, p = 3.34 × 10-14; FADS1-rs174550, p = 5.35 × 10-14; FADS2-rs1535, p = 5.85 × 10-14; FADS1-rs174546, p = 6.72 × 10-14; FADS2-rs174546, p = 9.75 × 10-14; FADS2- rs174576, p = 1.17 × 10-13; FADS2-rs174577, p = 1.12 × 10-12, among others). We also detected a genome-wide significant association with other genes in chromosome 11: MYRF (myelin regulatory factor)-rs174535, p = 1.49 × 10-12; TMEM258 (transmembrane protein 258)-rs102275, p = 2.43 × 10-12; FEN1 (flap structure-specific endonuclease 1)-rs174538, p = 1.96 × 10-11). Similar genome-wide statistically significant results were found for docosahexaenoic fatty acid (DHA). However, no such associations were detected for omega-6 PUFAs or linoleic acid (LA). For total PUFA, we observed a consistent gene*sex interaction with the DNTTIP2 (deoxynucleotidyl transferase terminal interacting protein 2)-rs3747965 p = 1.36 × 10-8. For adherence to MedDiet, we obtained a relevant interaction with the ME1 (malic enzyme 1) gene (a gene strongly regulated by fat) in determining serum omega-3. The top-ranked SNP for this interaction was ME1-rs3798890 (p = 2.15 × 10-7). In the regional-wide association study, specifically focused on the FADS1/FASD2/FADS3 and ELOVL (fatty acid elongase) 2/ELOVL 5 regions, we detected several statistically significant associations at p < 0.05. In conclusion, our results confirm a robust role of the FADS cluster on serum PUFA in this population, but the associations vary depending on the PUFA. Moreover, the detection of some sex and diet interactions underlines the need for these associations/interactions to be studied in all specific populations so as to better understand the complex metabolism of PUFA.

Candidate Gene and Genome-Wide Association Studies for Circulating Leptin Levels Reveal Population and Sex-Specific Associations in High Cardiovascular Risk Mediterranean Subjects.

Leptin is a hormone crucial in the regulation of food intake and body-weight maintenance. However, the genes and gene variants that influence its plasma levels are still not well known. Results of studies investigating polymorphisms in candidate genes have been inconsistent, and, in addition, very few genome-wide association studies (GWAS) have been undertaken. Our aim was to investigate the genes and gene variants most associated with plasma leptin concentrations in a high-cardiovascular-risk Mediterranean population. We measured plasma leptin in 1011 men and women, and analyzed the genetic factors associated using three approaches: (1) Analyzing the single nucleotide polymorphisms (SNPs) reported in a GWAS meta-analysis in other populations (including an SNP in/near each of these LEP, SLC32A1, GCKR, CCNL, COBLL1, and FTO genes); (2) Investigating additional SNPs in/near those genes, also including the RLEP gene; and (3) Undertaking a GWAS to discover new genes. We did not find any statistically significant associations between the previously published SNPs and plasma leptin (Ln) in the whole population adjusting for sex and age. However, on undertaking an extensive screening of other gene variants in those genes to capture a more complete set of SNPs, we found more associations. Outstanding among the findings was the heterogeneity per sex. We detected several statistically significant interaction terms with sex for these SNPs in the candidate genes. The gene most associated with plasma leptin levels was the FTO gene in men (specifically the rs1075440 SNP) and the LEPR in women (specifically the rs12145690 SNP). In the GWAS on the whole population, we found several new associations at the p < 1 × 10-5 level, among them with the rs245908-CHN2 SNP (p = 1.6 × 10-6). We also detected a SNP*sex interaction at the GWAS significance level (p < 5 × 10-8), involving the SLIT3 gene, a gene regulated by estrogens. In conclusion, our study shows that the SNPs selected as relevant for plasma leptin levels in other populations, are not good markers for this Mediterranean population, so supporting those studies claiming a bias when generalizing GWAS results to different populations. These population-specific differences may include not only genetic characteristics, but also age, health status, and the influence of other environmental variables. In addition, we have detected several sex-specific effects. These results suggest that genomic analyses, involving leptin, should be estimated by sex and consider population-specificity for more precise estimations.

A Guide to Applying the Sex-Gender Perspective to Nutritional Genomics.

Precision nutrition aims to make dietary recommendations of a more personalized nature possible, to optimize the prevention or delay of a disease and to improve health. Therefore, the characteristics (including sex) of an individual have to be taken into account as well as a series of omics markers. The results of nutritional genomics studies are crucial to generate the evidence needed so that precision nutrition can be applied. Although sex is one of the fundamental variables for making recommendations, at present, the nutritional genomics studies undertaken have not analyzed, systematically and with a gender perspective, the heterogeneity/homogeneity in gene-diet interactions on the different phenotypes studied, thus there is little information available on this issue and needs to be improved. Here we argue for the need to incorporate the gender perspective in nutritional genomics studies, present the general context, analyze the differences between sex and gender, as well as the limitations to measuring them and to detecting specific sex-gene or sex-phenotype associations, both at the specific gene level or in genome-wide-association studies. We analyzed the main sex-specific gene-diet interactions published to date and their main limitations and present guidelines with recommendations to be followed when undertaking new nutritional genomics studies incorporating the gender perspective.

Mastectomía ahorradora de piel y complejo areola-pezón con reconstrucción inmediata: una técnica en auge en el tratamiento del cáncer de mama / Nipple-sparing mastectomy with immediate reconstruction: An increasing technique for breast cancer treatment

Introducción: La mastectomía ahorradora de piel y pezón (MAP-CAP) es una técnica quirúrgica cada vez más empleada en el tratamiento del cáncer de mama. Nuestro objetivo es describir la técnica quirúrgica y exponer nuestros resultados. Métodos: Se analizaron retrospectivamente 46 MAP-CAP con reconstrucción inmediata con implante definitivo, realizadas en 35 pacientes con cáncer de mama. Resultados: Se realizaron 46 MAP-CAP entre 2010 y 2015. En 9 casos se realizó una mastectomía contralateral reductora de riesgo por mutaciones en los genes BRCA o acúmulo de riesgo familiar. La edad media fue de 48 años. El tipo histológico más frecuente fue el carcinoma ductal infiltrante (58%). El 26% de las neoplasias fueron carcinoma ductal in situ. Se evidenció multicentricidad en el 44,7% de los casos. Se administró neoadyuvancia en el 41% de los casos y radioterapia adyuvante (RT) en 24 casos (63%). En 4 casos aparecieron complicaciones postoperatorias (8,7%). En 13 casos se produjo una contractura capsular, habiéndose administrado RT en todos ellos. En 7 casos (15%) se requirió una segunda intervención quirúrgica para recambio de la prótesis inicial por contractura o alteraciones estéticas significativas. Existió un único caso de recidiva regional en axila (2,2%), con una mediana de seguimiento de 44 meses. Conclusiones: La MAP-CAP con prótesis definitiva es una técnica oncológicamente segura, con una tasa de recidivas similar a la mastectomía clásica. La administración de RT eleva la tasa de contractura capsular, implicando una segunda intervención en el 15% de los casos, por lo que debemos restringir su administración a los casos estrictamente necesarios

Introduction: Nipple-sparing mastectomy (NSM) is a surgical procedure that is increasingly used for the treatment of breast cancer. Our aim was to describe the technique performed at our institution and to present our results. Methods: We retrospectively analysed data from 46 NSM with immediate breast reconstruction with definitive implants performed in 35 patients with breast cancer. Results: We performed 46 NSMs between 2010 and 2015. Nine of these patients received a contralateral prophylactic NSM because of BRCA mutations or high familial cancer risk. The median age was 48 years. The most frequent histologic type was ductal invasive carcinoma (58%). A total of 26% of the neoplasms were ductal carcinoma in situ. Neoadjuvant treatment was administered to 41% of the patients and adjuvant radiotherapy was administered in 24 (63%). Early complications appeared in 4 patients (8.7%). Capsular contracture occurred in 13 patients. All of them had received radiotherapy. In 7 of the 46 patients (15%), a second surgery to change the initial prosthesis was required because of high-grade capsular contracture or to improve cosmetic results. There was only one case of regional recurrence (2.2%) in the axilla, with a median of follow-up of 44 months. Conclusions: NSM is an oncologically safe technique for breast cancer, with a similar recurrence rate to classic mastectomy. Radiotherapy increases the rate of capsular contracture, requiring a second intervention in 15% of patients. Consequently, its use should be restricted to cases in which it is strictly necessary

Dairy product consumption and risk of colorectal cancer in an older mediterranean population at high cardiovascular risk.

Prospective studies have reported an inverse association between the consumption of total dairy products and milk and the risk of colorectal cancer (CRC). Nonetheless, there is little and inconsistent evidence regarding subtypes of dairy product and CRC risk. We assessed the associations between the consumption of total dairy products, their different subtypes and CRC risk in older Mediterranean individuals at high cardiovascular risk. We analyzed data from 7,216 men and women (55-80 years) without CRC at baseline from the PREvención con DIeta MEDiterránea study. Individuals were recruited between 2003 and 2009 and followed up until December 2012. At baseline and yearly thereafter, consumption of total and specific dairy products was assessed using a validated 137-item food-frequency questionnaire. Cox proportional hazards ratios (HRs) of CRC incidence were estimated for tertiles of mean consumption of dairy products during the follow-up. During a median [interquartile range] follow-up of 6.0 [4.4-7.3] years, we documented 101 incident CRC cases. In the multivariable-adjusted models, HRs and 95% confidence intervals (CIs) of CRC for the comparison of extreme tertiles of total dairy product and low-fat milk consumption were 0.55 (95% CI: 0.31-0.99; p-trend = 0.037) and 0.54 (95% CI: 0.32-0.92; p-trend = 0.022), respectively. No significant associations with other dairy products (whole-fat and low-fat dairy products; total, low-fat and whole-fat yogurt; cheese; total, low-fat and whole-fat milk; concentrated full-fat dairy products, sugar-enriched dairy products and fermented dairy products) were found. A high consumption of total dairy products and low-fat milk was significantly associated with a reduced CRC risk.

Effects of the Ser326Cys Polymorphism in the DNA Repair OGG1 Gene on Cancer, Cardiovascular, and All-Cause Mortality in the PREDIMED Study: Modulation by Diet.

BACKGROUND: Oxidatively induced DNA damage, an important factor in cancer etiology, is repaired by oxyguanine glycosylase 1 (OGG1). The lower repair capacity genotype (homozygote Cys326Cys) in the OGG1-rs1052133 (Ser326Cys) polymorphism has been associated with cancer risk. However, no information is available in relation to cancer mortality, other causes of death, and modulation by diet. OBJECTIVE: Our aim was to evaluate the association of the OGG1-rs1052133 with total, cancer, and cardiovascular disease (CVD) mortality and to analyze its modulation by the Mediterranean diet, focusing especially on total vegetable intake as one of the main characteristics of this diet. DESIGN: Secondary analysis in the PREDIMED (Prevención con Dieta Mediterránea) trial is a randomized, controlled trial conducted in Spain from 2003 to 2010. PARTICIPANTS/SETTING: Study participants (n=7,170) were at high risk for CVD and were aged 55 to 80 years. INTERVENTION: Participants were randomly allocated to two groups with a Mediterranean diet intervention or a control diet. Vegetable intake was measured at baseline. MAIN OUTCOME MEASURES: Main outcomes were all-cause, cancer, and CVD mortality after a median follow-up of 4.8 years. STATISTICAL ANALYSES: Multivariable-adjusted Cox regression models were fitted. RESULTS: Three hundred eighteen deaths were detected (cancer, n=127; CVD, n=81; and other, n=110). Cys326Cys individuals (prevalence 4.2%) presented higher total mortality rates than Ser326-carriers (P=0.009). The multivariable-adjusted hazard ratio for Cys326Cys vs Ser326-carriers was 1.69 (95% CI 1.09 to 2.62; P=0.018). This association was greater for CVD mortality (P=0.001). No relationship was detected for cancer mortality in the whole population (hazard ratio 1.07; 95% CI 0.47 to 2.45; P=0.867), but a significant age interaction (P=0.048) was observed, as Cys326Cys was associated with cancer mortality in participants <66.5 years (P=0.029). Recessive effects limited our ability to investigate Cys326Cys×diet interactions for cancer mortality. No statistically significant interactions for total or CVD mortality were found for the Mediterranean diet intervention. However, significant protective interactions for CVD mortality were found for vegetable intake (hazard ratio interaction per standard deviation 0.42; 95% CI 0.18 to 0.98; P=0.046). CONCLUSIONS: In this population, the Cys326Cys-OGG1 genotype was associated with all-cause mortality, mainly CVD instead of cancer mortality. Additional studies are needed to provide further evidence on its dietary modulation.

[Radiosurgery treatment of acoustic neuromas. Our experience]. / Tratamiento del neurinoma del acústico con radiocirugía. Nuestra experiencia.

OBJECTIVE: To evaluate the results and side effects obtained using radiosurgery to treat acoustic neuromas. MATERIAL AND METHOD: Between 1999 and 2004 we treated 30 patients with unilateral acoustic neuromas with a mean age of 54 years old (31-76), a mean follow-up of 34 months (12-54), and tumour size between 4 and 38 millimetres (mean, 16.5). All of them were treated with radiosurgery (24 Gamma Knife and 6 LINAC) with doses of 1200-1300 cGy. RESULTS: Tumour growth control rate (smaller or same size) was 93 % (28/30). In 11 patients there was a post-treatment hearing loss (36.6 %). Trigeminal (3/30) and facial (2/30) neuropathy was transient. CONCLUSIONS: Although microsurgery is the treatment of choice for acoustic neuromas, we consider radiosurgical treatment a valid alternative for selected patients (the elderly, co-morbid conditions, small size, contralateral hearing loss...).

Tratamiento del neurinoma del acústico con radiocirugía. Nuestra experiencia / Radiosurgery treatment of acoustic neuromas. Our experience

Objetivo: Valorar los resultados obtenidos, así como los efectos secundarios derivados del tratamiento con radiocirugía del neurinoma del acústico. Material y método: Entre los años 1999 y 2004, hemos tratado a 30 pacientes afectos de neurinoma del acústico (NA) unilateral, con una media de edad de 54 (31-76) años, un tiempo medio de seguimiento de 34 (12-54) meses y un tamaño que osciló en 4-38 (media, 16,5) mm. Se practicó radiocirugía a todos ellos (24 Gamma Knife y 6 LINAC) a dosis de 1.200-1.300 cGy. Resultados: La tasa de control del crecimiento tumoral fue del 93 % (28/30). Un 36,6 % (11/30) presentó empeoramiento de su nivel de audición postratamiento. La afección del V par (3/30), así como la del VII (2/30), fue transitoria. Conclusiones: Aunque la microcirugía sigue siendo el tratamiento de elección para los NA, consideramos la radiocirugía como una alternativa válida en pacientes seleccionados (edad avanzada, comorbilidad asociada, pequeño tamaño, hipoacusia contralateral, entre otros)

Objective: To evaluate the results and side effects obtained using radiosurgery to treat acoustic neuromas. Material and method: Between 1999 and 2004 we treated 30 patients with unilateral acoustic neuromas with a mean age of 54 years old (31-76), a mean follow-up of 34 months (12-54), and tumour size between 4 and 38 millimetres (mean, 16.5). All of them were treated with radiosurgery (24 Gamma Knife and 6 LINAC) with doses of 1200-1300 cGy. Results: Tumour growth control rate (smaller or same size) was 93 % (28/30). In 11 patients there was a post-treatment hearing loss (36.6 %). Trigeminal (3/30) and facial (2/30) neuropathy was transient. Conclusions: Although microsurgery is the treatment of choice for acoustic neuromas, we consider radiosurgical treatment a valid alternative for selected patients (the elderly, co-morbid conditions, small size, contralateral hearing loss...)