Radio-induced apoptosis of peripheral blood CD8 T lymphocytes is a novel prognostic factor for survival in cervical carcinoma patients.

BACKGROUND AND PURPOSE: A close relationship exists between immune response and tumor behavior. This study aimed to explore the associations between radiation-induced apoptosis (RIA) in peripheral blood lymphocytes (PBL) and clinical pathological variables. Furthermore, it assessed the role of RIA as a prognostic factor for survival in cervical carcinoma patients. PATIENTS AND METHODS: Between February 1998 and October 2003, 58 consecutive patients with nonmetastatic, localized stage I-II cervical carcinoma who had been treated with radiotherapy (RT) ± chemotherapy were included in this study. Follow-up ended in January 2013. PBL subpopulations were isolated and irradiated with 0, 1, 2 and 8 Gy then incubated for 24, 48 and 72 h. Apoptosis was measured by flow cytometry and the ß value, a parameter defining RIA of lymphocytes, was calculated. RESULTS: Mean follow-up duration was 111.92 ± 40.31 months. Patients with lower CD8 T lymphocyte ß values were at a higher risk of local relapse: Exp(B) = 5.137, confidence interval (CI) 95 % = 1.044-25.268, p = 0.044. Similar results were observed for regional relapse: Exp(B) = 8.008, CI 95 % = 1.702-37.679, p = 0.008 and disease relapse: Exp(B) = 6.766, CI 95 % = 1.889-24.238, p = 0.003. In multivariate analysis, only the CD8 T lymphocyte ß values were found to be of prognostic significance for local disease-free survival (LDFS, p = 0.049), regional disease-free survival (RDFS, p = 0.002), metastasis-free survival (MFS, p = 0.042), disease-free survival (DFS, p = 0.001) and cause-specific survival (CSS p = 0.028). CONCLUSION: For the first time, RIA in CD8 T lymphocytes was demonstrated to be a predictive factor for survival in cervical carcinoma patients.

Plasminogen activator inhibitor-1 (PAI-1) expression in relation to hypoxia and oncoproteins in clinical cervical tumors.

PURPOSE: Explore the role of plasminogen activator inhibitor-1 (PAI-1) in cervical cancer and its relationship to hypoxia and the expression of p53, Ku70/80, and cyclin D1. MATERIAL AND METHODS: The expression of PAI-1, cyclin D1, and p53, together with tumor oxygenation, were determined in 43 consecutive patients suffering from localized cervical carcinoma. Oncoprotein expression was determined by immunohistochemistry. Tumor oxygenation was measured using a polarographic probe system, "pO2 histography." RESULTS: PAI expression was considered negative in 32.6% and overexpressed in 18.6% of cases. Cyclin D1 showed a median expression of 5.0 (range 0-70). We observed a positive association between PAI expression and altered p53 (p = 0.049) and cyclin D1 (p = 0.020). An inverse association was detected between PAI and Ku70/80 expression (p = 0.042). Cyclin D1 staining increased according to tumor volume (r = 0.314, p = 0.009). We did not observe a significant association between PAI and hypoxia or other clinicopathological parameters. CONCLUSION: The present results show that PAI-1 overexpression is associated with nonhomologous end-joining DNA repair down-regulation (low Ku70/80 expression) and with increased p53 and cyclin D1 expression, and they suggest that PAI-1 plays a role in the tumor behavior in cervical carcinoma.

Human biomonitoring of persistent organic pollutants in elderly people from the Canary Islands (Spain): A temporal trend analysis from the PREDIMED and PREDIMED-Plus cohorts.

The population of the Canary Islands has been exposed to high levels of persistent organic pollutants (POPs). Biomonitoring studies are essential to know the temporal trend in residue levels, particularly of substances banned decades ago. The purpose of this study was to analyze the distribution of plasma concentrations of 59 POPs in 175 participants from the PREDIMED-Plus trial (2014-2016), and to compare them with the distribution of these POPs in 343 participants in the PREDIMED trial (2006-2009). All participants had metabolic syndrome. No difference in the distribution of age, gender or BMI was observed between trials. POPs were determined by gas chromatography-mass spectrometry. Density plots -POP Geoffrey Rose curves- were used to represent the full population distribution of each compound. Three out of 59 POPs were detected and quantified in ≥95% of the samples (p,p'-DDE, median = 694.7 ng/g lipid; HCB, median = 57.0 ng/g lipid; and ß-HCH, median = 75.7 ng/g lipid). PCB congeners 138, 153 and 180 were detected in 64.6, 40.0 and 88.0% of the samples. Females showed highest concentrations of organochlorine pesticides and those subjects who lost ˃ 5 kg showed significant higher plasma concentrations of POPs. In a range of 6-14 years, plasma concentrations of POPs decreased 3.3-21.6 fold, being notable the decrease of 28.7-fold observed for HCB among women. Despite this sharp decline, levels of POPs are still higher than those reported in other regions, since one third of the subjects included in the present report had high concentration of more than three pollutants. Continuous biomonitoring studies are required to know the evolution of the levels of residues and to evaluate the effectiveness of environmental policies.

The effect of in vivo growth hormone treatment on blood gene expression in adults with growth hormone deficiency reveals potential biomarkers to monitor growth hormone therapy.

OBJECTIVE: Growth hormone (GH) replacement therapy is presently utilized in the treatment of adult GH deficiency (AGHD). Adult responses to GH treatment are highly variable and, apart from measurement of IGF-I, few tools are currently available for monitoring GH treatment progress. As GH receptors are expressed in certain blood cell types, changes in gene expression in peripheral blood can reflect perturbations induced as a result of GH therapy. DESIGN/PATIENTS: We have carried out a pilot study to identify GH-responsive genes in blood, and have assessed the utility of GH-responsive genes in monitoring GH therapy in AGHD. Blood was collected from ten women diagnosed with AGHD syndrome both before and 4 weeks after initiation of GH substitutive therapy. RNA was extracted from peripheral blood mononuclear cells (PBMCs) and changes in response to GH were detected using microarray-based gene analysis. RESULTS: All patients responded to GH replacement therapy, with serum levels of IGF-I increasing by an average of 307% (P = 0.0003) while IGFBP-3 increased by an average of 182% (P = 0.0002). Serum levels of triglycerides, LDL-C, HDL-C, APOA1 or APOB did not change after 1 month of GH treatment. By contrast, we detected an increase in Lp(a) serum levels (P = 0.0149). Using a stringent selection cutoff of P

Biomonitoring of 45 inorganic elements measured in plasma from Spanish subjects: A cross-sectional study in Andalusian population.

Heavy metals and other toxic elements are frequently detected in humans. Rare earth elements (REE) have arisen as a novel group of substances considered as emerging pollutants due to its dependence for high tech industry. We designed a study aimed to conduct the biomonitoring a total of 45 inorganic elements in the population of Andalusia (Spain). A total of 419 participants were recruited and their plasma samples analyzed. Concentration of elements, including elements in the ATSDR's priority pollutant list and REE were measured by ICP-MS in the blood plasma of participants. Arsenic, copper, lead, selenium, antimony, strontium, and bismuth were detected in ˃98% of subjects. Median values of arsenic, mercury and lead were 1.49, 1.46, and 5.86 ng/mL, respectively. These concentrations did not exceed reference values published by international agencies. We observed a positive correlation between age and plasma concentrations of arsenic, mercury, antimony and strontium. Sum of elements was lower in the group of subjects younger than 45 years old (P = 0.002). Positive correlations were observed between body mass index (BMI) and plasma concentrations of barium, cerium, osmium, tin, and ytterbium. 7 out of 26 REEs showed a percentage of detection ≥ 90%. Bismuth, yttrium, and cerium were quantified at the highest concentrations (median value = 7.7, 0.19, and 0.16 ng/mL, respectively). We found that plasma levels of 6 REEs were higher among males, and a positive correlation between REEs and age was detected. The present results suggest a potential interaction with the human physiology that deserves additional research. Given the high persistence of these elements in the environment, and the significant technological dependence on them, future studies are needed to elucidate the potential sources of exposure and possible adverse effects on health, especially in the most vulnerable populations.

Ethanol levels in legally autopsied subjects (2016-2017): Update of data and epidemiological implications in relation to violent deaths in Canary Islands (Spain).

The aim of this study was to analyze the prevalence of ethanol in individuals (277) subjected to a mandatory medico-legal autopsy in the Institute of Legal Medicine of Las Palmas (Canary Islands, Spain) during 2016-2017, comparing the results with data published in 2015. Blood and/or vitreous humor samples were analyzed by gas chromatography. 31.8% of the individuals were positive to ethanol. We observed a decrease in the prevalence of ethanol among males (p = 0.002). While the prevalence of ethanol was reduced, the concentration among those positive subjects has increased. 11.9% of the series died in a traffic accident and the percentage of positive to ethanol decreased in relation to 2015 (64.3, 25.0, and 35.3%). The number of suicides remains constant, although the age of the suiciders has been significantly reduced (p = 0.022). The results of this study indicate that ethanol is still heavily involved in non-natural deaths.

The impact of red and processed meat consumption on cancer and other health outcomes: Epidemiological evidences.

Meat is one of the staples of the human diet, which provides high-quality nutrients, but that also constitutes a relevant source of cholesterol and saturated fatty acids. Epidemiologic studies have linked consumption of red or processed meat with obesity, type 2 diabetes, cardiovascular diseases, and cancers. Most epidemiological studies suggest that a high intake of meat, especially processed meat, is associated with increased colorectal cancer risk. Potential reasons for the association between high meat intake and colorectal cancer risk include some chemicals naturally contained in meat, or generated by the processing and cooking. From the literature it can be concluded that there is enough epidemiological evidence linking processed meat intake and colorectal cancer risk, but there is limited evidence regarding unprocessed red meat intake and the disease. On the contrary, there is only limited evidence linking meat intake with other diseases such as cardiovascular disease, type 2 diabetes or other cancers. Nevertheless, the literature suggest that dietary intervention may be a promising approach for prevention of cancers of the colon, esophagus, liver, stomach and bladder, and type 2 diabetes and cardiovascular disease which still need to be confirmed by further well designed prospective studies and experimental research.

Organochlorine Pesticides Exposure and Bladder Cancer: Evaluation from a Gene-Environment Perspective in a Hospital-Based Case-Control Study in the Canary Islands (Spain).

The incidence of bladder cancer has increased significantly since the 1950s. Pesticide exposure has been linked with increasing bladder cancer incidence, although the evidence is inconclusive. However, most epidemiological studies did not evaluate the potential role played by the organochlorine pesticides, the most widely used pesticides in Western countries from the 1940s to the 1970s. Organochlorine pesticides were banned in the late 1970s because of their persistence in the environment and their carcinogenic and mutagenic effects. Organochlorine pesticides were employed in huge amounts in the Spanish archipelago of the Canary Islands; the authors, therefore, evaluated the role played by organochlorine pesticides exposure on bladder cancer. Serum levels of the most prevalent organochlorine pesticides used in the agriculture of these Islands (dichlorodiphenyltrichloroethane [p,p'-DDT], and its metabolites dichlorodiphenyldichloroethylene [p,p'-DDE] and dichlorodiphenyldichloroethane [p,p'-DDD], hexachlorobenzene, hexachlorocyclohexane isomers, aldrin, dieldrin, endrin, heptachlor, cis-chlordane, trans-chlordane, α- and ß-endosulfan, endosulfan sulfate, methoxychlor, and mirex) were measured in 140 bladder cancer cases and 206 controls. GST-M1 and GST-T1 gene polymorphisms were genotyped by polymerase chain reaction (PCR)-based methods. These results showed that serum levels of organochlorine pesticides did not increase bladder cancer risk. On the contrary, total burden of hexachlorocyclohexanes was found to be negatively associated to bladder cancer (odds ratio [OR] = 0.929, 95% confidence interval [CI]: 0.865-0.997; P = .041). This effect disappeared when the distribution of the gluthathione S-transferase polymorphisms was introduced in the statistical model. These results indicate that organochlorine pesticides are not a risk factor for bladder cancer. However, these findings provide additional evidence of gene-environment interactions for organochlorine pesticides and bladder cancer and reinforce the relevance of genes encoding xenobiotic-metabolizing enzymes in bladder cancer.

Association between single-nucleotide polymorphisms in DNA double-strand break repair genes and prostate cancer aggressiveness in the Spanish population.

BACKGROUND: Novel predictors of prognosis and treatment response for prostate cancer (PCa) are required to better individualize treatment. Single-nucleotide polymorphisms (SNPs) in four genes directly (XRCC5 (X-ray repair complementing defective repair in Chinese hamster cells 5) and XRCC6 (X-ray repair complementing defective repair in Chinese hamster cells 6)) or indirectly (PARP1 and major vault protein (MVP)) involved in non-homologous end joining were examined in 494 Spanish PCa patients. METHODS: A total of 22 SNPs were genotyped in a Biotrove OpenArray NT Cycler. Clinical tumor stage, diagnostic PSA serum levels and Gleason score at diagnosis were obtained for all participants. Genotypic and allelic frequencies were determined using the web-based environment SNPator. RESULTS: (XRCC6) rs2267437 appeared as a risk factor for developing more aggressive PCa tumors. Those patients carrying the GG genotype were at higher risk of developing bigger tumors (odds ratio (OR)=2.04, 95% confidence interval (CI) 1.26-3.29, P=0.004), present higher diagnostic PSA levels (OR=2.12, 95% CI 1.19-3.78, P=0.011), higher Gleason score (OR=1.65, 95% CI 1.01-2.68, P=0.044) and D'Amico higher risk tumors (OR=2.38, 95% CI 1.24-4.58, P=0.009) than those patients carrying the CC/CG genotypes. Those patients carrying the (MVP) rs3815824 TT genotype were at higher risk of presenting higher diagnostic PSA levels (OR=4.74, 95% CI 1.40-16.07, P=0.013) than those patients carrying the CC genotype. When both SNPs were analyzed in combination, those patients carrying the risk genotypes were at higher risk of developing D'Amico higher risk tumors (OR=3.33, 95% CI 1.56-7.17, P=0.002). CONCLUSIONS: We believe that for the first time, genetic variants at XRCC6 and MVP genes are associated with risk of more aggressive disease, and would be taken into account when assessing the malignancy of PCa.

New biological markers in the decision of treatment of head and neck cancer patients.

Head and neck squamous cell carcinoma is the sixth most common cancer type worldwide. Also the 5-year survival rate of less than 50 % seems to be lower than other cancer types. There are some reasons behind this high mortality rate; one of them is the lack of knowledge about the biology and genomic instability behind the carcinogenic processes. These biological features could condition the failure of frontline treatment, in which case rescue treatment should be used, representing an overtreatment for the patients. For years many biological factors have been tested as prognostic and predictive factors in relation to treatment with a modest success. To find appropriate tests which could be used in the context of the individualized treatment decision, we have reviewed new biological markers, not only in tumor tissue, but also in normal tissue from head and neck carcinoma patients.

Molecular biomarkers in the decision of treatment of cervical carcinoma patients.

Cervical carcinoma is the third most common cancer in women worldwide. The programs developed for early detection have made that most patients are diagnosed in early stages. Treatment for those patients consists of conservative techniques as surgery or radical radiotherapy; however, the decision between those two therapies is still controversial. Even though in many cases this decision varies according to classical associated risk factors (i.e. tumor stage or age), in the clinical practice, a significant number of patients treated by surgery also receive post-surgery radiotherapy, with the consequent over-treatment and toxic effects. Since response to treatments is conditioned by individual factors, the use of new biological markers as novel predictive factors for both tumor and normal tissues could help clinicians to choose the best treatment schedule for each patient individually. Based on the experience of our institution, we have reviewed the new biological markers in cervical carcinoma patients treated by radiotherapy.

Polychlorobiphenyls and organochlorine pesticides in conventional and organic brands of milk: occurrence and dietary intake in the population of the Canary Islands (Spain).

The population of the Spanish archipelago of the Canary Islands has been studied in depth regarding its levels of contamination by organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs). Foodstuffs of animal origin, such as milk, are prominent contributors to the body burden of these contaminants. As this population presents one of the highest milk-intake in Spain and Europe, we evaluated the level of OCs and PCBs in 26 commercially available brands of milk (16 conventional and 10 organic brands) present in the market of these Islands, in order to estimate the relevance of milk as a source of these chemicals for the Canary population. Our findings showed that hexachlorobenzene, trans-chlordane, and PCB153 were present in almost all the samples with independence of the type of milk. For both types of milks, the concentration of OCs was very low, showing organic milks lower levels than conventional ones. As a consequence, the estimated daily intake for OCs was lower than the tolerable daily intake (TDI) established by International Agencies. The levels of PCBs in milk were also found to be very low, but, in this case, the situation was the opposite: there were higher levels of PCBs in organic than in conventional brands of milk. Unexpectedly, levels of dioxin-like PCBs (DL-PCBs) reached values higher than 25 pg WHO-TEQ g(-1) fat in percentile 75 for both types of milk indicating the existence of a number of brands of milk highly contaminated by these toxicants. Consequently, the population who consume the most contaminated milk brands could have estimated daily intakes well above the recommended TDI (2 pg WHO-TEQ kg(-1)b.w.d(-1)) established by European Union Authorities. These results are of concern if we consider the well known adverse health effects exerted by dioxin-like compounds.

Gene polymorphisms in TYMS, MTHFR, p53 and MDR1 as risk factors for breast cancer: a case-control study.

Breast cancer (BC) is a complex disease influenced by environmental and genetic factors. The disease has important genetic and environmental components, most of them are still unknown. An important role of gene polymorphisms related to the risk of developing BC has been reported. However, the results have been controversial. We investigated the association of TSER, MTHFR C677T, p53 codon 72 and MDR1 C3435T gene polymorphisms with breast carcinoma in women from Canary Islands (Spain). Blood samples collected from 135 patients with BC and 304 healthy controls all of them Caucasian, were analyzed through polymerase chain reaction-restriction fragment length polymorphism. Subsequently, a structured questionnaire including patient history and risk factors in relation to BC development was filled out. Allelic frequencies of these genetic variations were: TSER, (2) 0.55 and (3) 0.45 in cases, 0.49 and 0.51 respectively in controls (P=0.240); MTHFR C677T, (C) 0.63 and (T) 0.37 in cases, 0.60 and 0.40 respectively in controls (P=0.568); p53 Arg72Pro, (Arg) 0.74 and (Pro) 0.26 in cases and controls (P=0.910); MDR1 C3435T, (C) 0.52 and (T) 0.48 in cases, 0.55 and 0.45 respectively in controls (P=0.523). We did not observe any gene polymorphism as a risk factor to develop BC. A statistical association was observed between p53 codon 72 polymorphism and family history of breast cancer in both groups, as well as between MDR1 C3435T and smoking habits in cases (P<0.05). Gene polymorphisms vary by regions. The present study contributes to the characterization of the genetic pattern of the Canary population.

Role of pituitary hormones on 17alpha-ethinylestradiol-induced cholestasis in rat.

Estrogens cause intrahepatic cholestasis in susceptible women during pregnancy, after administration of oral contraceptives, or during postmenopausal hormone replacement therapy. 17alpha-Ethinylestradiol (EE) is a synthetic estrogen widely used to cause experimental cholestasis in rodents with the aim of examining molecular mechanisms involved in this disease. EE actions on the liver are thought to be mediated by estrogen receptor alpha (ERalpha) and pituitary hormones. We tested this hypothesis by analyzing metabolic changes induced by EE in livers from hypophysectomized (HYPOX) and hypothyroid rats. Microarray studies revealed that the number of genes regulated by EE was increased almost 4-fold in HYPOX rat livers compared with intact males. Little overlap was apparent between the effects of EE in intact and HYPOX rats, demonstrating that pituitary hormones play a critical role in the hepatic effects of EE. Consistently, hypophysectomy protects the liver against induction by EE of serum bilirubin and alkaline phosphatase, two markers of cholestasis and hepatotoxicity and modulates the effects of EE on several genes involved in bile acid homeostasis (e.g., FXR, SHP, BSEP, and Cyp8b1). Finally, we demonstrate a novel mechanism of action of EE through binding and negative regulation of glucocorticoid receptor-mediated transcription. In summary, pituitary- and ERalpha-independent mechanisms contribute to development of EE-induced changes in liver transcriptome. Such mechanisms may be relevant when this model of EE-induced cholestasis is evaluated. The observation that the pharmacological effects of estrogen in liver differ in the absence or presence of the pituitary could be clinically relevant, because different drugs that block actions of pituitary hormones are now available.

Contaminantes orgánicos persistentes en plasma de tortugas bobas (Caretta caretta) varadas en las Islas Canarias / Plasma leves of persistent organic pollutans in loggerhead sea turtles (caretta caretta) stranded in the Canary Islands

En el presente estudio se ha evaluado el grado de contaminación por contaminantes orgánicos persistentes (COPs) en 193 ejemplares de tortuga boba (Caretta caretta) varadas en las Islas Canarias entre 2007-2011. La cuantificación en plasma de los niveles de pesticidas organoclorados (POCs), bifenilos policlorados (PCBs) e hidrocarburos aromáticos policíclicos (PAHs) se realizó mediante GC-MS. Todas las muestras analizadas presentaron niveles cuantificables de alguno de los COPs incluidos en el estudio. El grupo de COPs que presentó mayores niveles fue el de los PAHs (alcanzando la carga total de PAHs 6,45 ng/ml), siendo el fenantreno el hidrocarburo más frecuentemente detectado y a concentraciones más altas, lo que indica el origen petrogénico de estos contaminantes. La contaminación por PCBs alcanzó niveles menores (3,84 ng/ml), predominando el grupo de los hexaclorobifenilos (PCB-153 y PCB- 138 principalmente). Los niveles de contaminación por POCs fueron también bajos alcanzando valores de 1,67 ng/ml, siendo el principal metabolito del DDT, el p,p´-DDE el compuesto más frecuentemente detectado (89,6%) y a más altas concentraciones (0,68 ng/ml). Fue evidente una asociación inversa entre el tamaño de las tortugas y la carga de PCBs y PAHs. Asimismo existieron niveles más altos de contaminación por COPs en los años 2009 y 2010. Este trabajo evalúa por vez primera la presencia de PAHs en sangre de tortugas varadas y nuestros resultados parecen indicar que esta metodología y esta especie animal pueden ser muy útiles para monitorizar la presencia de contaminación por derivados del petróleo en el medio acuático (AU)

This study assessed the degree of contamination by persistent organic pollutants (POPs) present in 193 specimens of loggerhead turtle (Caretta caretta) stranded in the Canary Islands between 2007-2011. Quantification of plasma levels of organochlorine pesticides (POCs), polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) were performed by GC-MS. All samples tested showed measurable levels of several of the POPs included in the study. PAHs was the group that showed the highest levels (total burden of PAHs = 6.45 ng/ml), being phenanthrene the compound most frequently detected and at higher concentrations, indicating the petrogenic origin of these contaminants. PCBs contamination reached lower levels (3.84 ng/ml), dominating the group of hexachlorobiphenyls (PCB-153 and PCB-138 in particular). The pollution levels were also low in the subgroup of POCs, reaching values of 1.67 ng/ml, and being the main metabolite of DDT, p, p'-DDE the compound most frequently detected (89.6%) and at highest concentrations (0.68 ng/ml). There was a clear inverse association between the size of the turtles and the burden of PCBs and PAHs. There were also higher levels of POPs in 2009 and 2010 than in the other years. This study evaluates for the first time the presence of PAHs in stranded turtles blood and our results suggest that this methodology and this animal species can be very useful for monitoring the presence of petroleum derivatives contamination in the aquatic environment (AU)