Domingo González-Lamuño

Does melatonin induce apoptosis in MCF-7 human breast cancer cells in vitro?

Melatonin inhibits proliferation of the estrogen‐responsive MCF‐7 human breast cancer cells. The objective of this work was to assess whether melatonin not only regulates MCF‐7 cell proliferation but also induces apoptosis. In this experiment we used 1,25‐dihydroxycholecalciferol (D3) as a positive control because it inhibits MCF‐7 cell proliferation and induces apoptosis. MCF‐7 cells were cultured with either 1 nM melatonin, 100 nM D3 or its diluent to determine their effects on cell proliferation, cell viability, cell‐cycle phase distribution, population of apoptotic cells, and expression of p53, p21WAF1, bcl‐2, bcl‐XL and bax proteins. After 24 or 48 hr of incubation, both melatonin and D3‐treatment significantly decreased the number of viable cells in relation to the controls, although no differences in cell viability were observed between the treatments. The incidence of apoptosis, measured as the population of cells falling in the sub‐G1 region of the DNA histogram, or by the TUNEL reaction, was similar in melatonin‐treated and control cells whereas, as expected, apoptosis was higher among cells treated with D3 than in controls. The expression of p53 and p21WAF1 proteins significantly increased after 24 or 48 hr of incubation with either melatonin or D3. No significant changes in bcl‐2, bcl‐XL and bax mRNAs were detected after treatment with melatonin whereas in D3‐treated cells, a significant drop in bcl‐XL was observed. These data support the hypothesis that melatonin reduces MCF‐7 cell proliferation by modulating cell‐cycle length through the control of the p53–p21 pathway, but without clearly inducing apoptosis.

A new pentaplex system to study short tandem repeat markers of forensic interest on X chromosome

A new method has been optimised to amplify five X chromosome short tandem repeat (STR) markers of interest in forensic medicine: human phosphoribosyl transferase (HPRTB), DXS101, androgen receptor (ARA), DXS7423 and DXS8377. Markers were conveniently amplified in a single PCR reaction with fluorochrome-labelled primers, which allowed the analysis of fragment sizes after injection into a capillary electrophoresis system. The most common alleles of each locus were sequenced and used in a control ladder to type unknown samples.

Henoch-Schönlein purpura in northern Spain: clinical spectrum of the disease in 417 patients from a single center.

AbstractThe severity of clinical features and the outcomes in previous series of patients reported with Henoch-Schönlein purpura (HSP) vary greatly, probably due to selection bias. To establish the actual clinical spectrum of HSP in all age groups using an unselected and wide series of patients diagnosed at a single center, we performed a retrospective review of 417 patients classified as having HSP according to the criteria proposed by Michel et al. Of 417 patients, 240 were male and 177 female, with a median age at the time of disease diagnosis of 7.5 years (interquartile range [IQR], 5.3–20.1 yr). Three-quarters of the patients were children or young people aged 20 years or younger (n = 315), and one-quarter were adults (n = 102). The most frequent precipitating events were a previous infection (38%), usually an upper respiratory tract infection, and/or drug intake (18.5%) shortly before the onset of the vasculitis. At disease onset the most common manifestations were skin lesions (55.9%), nephropathy (24%), gastrointestinal involvement (13.7%), joint symptoms (9.1%), and fever (6.2%). Cutaneous involvement occurring in all patients, mainly purpuric skin lesion, was the most common manifestation when the vasculitis was fully established, followed by gastrointestinal (64.5%), joint (63.1%), and renal involvement (41.2%). The main laboratory findings were leukocytosis (36.7%), anemia (8.9%), and increased serum IgA levels (31.7%). The most frequent therapies used were corticosteroids (35%), nonsteroidal antiinflammatory drugs (14%), and cytotoxic agents (5%). After a median follow-up of 12 months (IQR, 2–38 mo), complete recovery was observed in most cases (n = 346; 83.2%), while persistent, usually mild, nephropathy was observed in only 32 (7.7%) cases. Relapses were observed in almost a third of patients (n = 133; 31.9%).In conclusion, although HSP is a typical vasculitis affecting children and young people, it is not uncommon in adults. The prognosis is favorable in most cases, depending largely on renal involvement.

A clinical and experimental overview of sirenomelia: insight into the mechanisms of congenital limb malformations

Sirenomelia, also known as sirenomelia sequence, is a severe malformation of the lower body characterized by fusion of the legs and a variable combination of visceral abnormalities. The causes of this malformation remain unknown, although the discovery that it can have a genetic basis in mice represents an important step towards the understanding of its pathogenesis. Sirenomelia occurs in mice lacking Cyp26a1, an enzyme that degrades retinoic acid (RA), and in mice that develop with reduced bone morphogenetic protein (Bmp) signaling in the caudal embryonic region. The phenotypes of these mutant mice suggest that sirenomelia in humans is associated with an excess of RA signaling and a deficit in Bmp signaling in the caudal body. Clinical studies of sirenomelia have given rise to two main pathogenic hypotheses. The first hypothesis, based on the aberrant abdominal and umbilical vascular pattern of affected individuals, postulates a primary vascular defect that leaves the caudal part of the embryo hypoperfused. The second hypothesis, based on the overall malformation of the caudal body, postulates a primary defect in the generation of the mesoderm. This review gathers experimental and clinical information on sirenomelia together with the necessary background to understand how deviations from normal development of the caudal part of the embryo might lead to this multisystemic malformation.

MCT1 genetic polymorphism influence in high intensity circuit training: a pilot study.

Monocarboxylate Transporter 1 (MCT1) mediates the transport of the main fraction of lactate across the sarcolemma. A common polymorphic MCT1 variant has been identified, but its role in high intensity exercise performance has not been defined. We investigated the influence of MCT1 A1470T polymorphism (rs1049434) on lactate accumulation after high intensity circuit training. Ten men aged 20-26 performed three controlled circuit training (CWT) sessions at 60%, 70%, and 80% of the 15 repetition maximum (15RM), in non-consecutive days. CWT included three sets of a circuit of eight exercises, obtaining lactate measurements immediately after each set had been completed. Two independent variables were analysed: MTC1 genotypes according to the presence or absence of the A1470T polymorphism, and the intensity of circuit training. Genotype distributions were in Hardy-Weinberg equilibrium, being 30% wild-type, 50% heterozygotes, and 20% mutated homozygotes. Mean lactate concentration at 80% of 15RM were significantly higher than the mean lactate values at the other intensities (p<0.01). Significant differences between genetic groups were found in the lactate accumulation slope at 80% of 15RM (p=0.02) and in the maximal lactate concentration reached by all subjects in the study (L(max)) (p=0.03). The carriers of the A1470T polymorphism in the MTC1 gene seem to exhibit a worse lactate transport capability into the less active muscle cells for oxidation.

Birth weight and blood lipid levels in Spanish adolescents: Influence of selected APOE, APOC3 and PPARgamma2 gene polymorphisms. The AVENA Study

BackgroundThere is increasing evidence indicating that genes involved in certain metabolic processes of cardiovascular diseases may be of particular influence in people with low body weight at birth. We examined whether the apolipoprotein (APO) E, APOC3 and the peroxisome proliferator-activated receptor-γ-2 (PPARγ2) polymorphisms influence the association between low birth weight and blood lipid levels in healthy adolescents aged 13–18.5 years.MethodsA cross-sectional study of 502 Spanish adolescents born at term was conducted. Total (TC) and high density lipoprotein cholesterol (HDLc), triglycerides (TG), apolipoprotein (apo) A and B, and lipoprotein(a) [Lp(a)] were measured. Low density lipoprotein cholesterol (LDLc), TC-HDLc, TC/HDLc and apoB/apoA were calculated.ResultsLow birth weight was associated with higher levels of TC, LDLc, apoB, Lp(a), TC-HDLc, TC/HDLc and apoB/apoA in males with the APOE ε3ε4 genotype, whereas in females, it was associated with lower HDLc and higher TG levels. In males with the APOC3 S1/S2 genotype, low birth weight was associated with lower apoA and higher Lp(a), yet this association was not observed in females. There were no associations between low birth weight and blood lipids in any of the PPARγ2 genotypes.ConclusionThe results indicate that low birth weight has a deleterious influence on lipid profile particularly in adolescents with the APOE ε3/ε4 genotype. These findings suggest that intrauterine environment interact with the genetic background affecting the lipid profile in later life.

Variables Affecting BMI Evolution at 2 and 5 Years after Vertical Banded Gastroplasty

Background: Vertical banded gastroplasty (VBG) has been found to result in significant reduction in body mass index (BMI) during the first postoperative year.We investigated the impact of some intrinsic and extrinsic factors on long-term BMI evolution in morbidly obese patients who underwent VBG, with the aim of establishing a long-term weight-loss prognosis. Methods: 67 consecutive morbidly obese patients who underwent VBG were followed for 2 years; of these, 34 were followed 3 more years, for a total follow-up of 5 years. BMI was monitored and correlated with demographic (preoperative BMI, obese relatives, age and gender) and lifestyle variables (physical activity, habitual dietary transgression and occupational status). Results: Global BMI fell from 47.5 at the time of the intervention to 32.1 when patients were examined 12 months after surgery. From the second year, an upward trend was observed, and at 5 years, mean BMI was above 35, considered in the high-risk range. Modifiable variables affecting lifestyle have shown significantly favorable effects on BMI evolution. Among intrinsic variables, BMI before surgery and obese parents also affect long-term evolution. Conclusion: Different variables should be considered in order to establish a long-term weight-loss prognosis for each patient, thus making it possible to act more specifically on modifiable variables.

Influence of the MCT1-T1470A polymorphism (rs1049434) on blood lactate accumulation during different circuit weight trainings in men and women

OBJECTIVES To analyze the effect of the MCT1 T1470A polymorphism (rs1049434) on venous blood lactate levels in men and women, during three different circuit weight training protocols. DESIGN Cross-sectional laboratory study. METHODS 14 women and 15 men, all caucasian and moderately active, performed three circuit training sessions (Weight Machine Protocol, Free Weight Protocol and Combined Protocol) at 70% of the 15 repetition maximum and 70% of the heart rate reserve, in non-consecutive days. The sessions included three sets of a circuit of eight exercises. Venous lactate measurements were obtained after each set and during the recoveries between sets (i.e. in min 3, 5, 7 and 9). One-way analysis of covariance and one-way analysis of covariance with repeated measures were used to determine differences among genotypes (AA, TA and TT) in lactate levels. RESULTS In men, the AA group had higher lactate values than the TT group in all the measures (p ≤ 0.03) except for the average lactate during the Weight Machine Protocol, in which a borderline significant difference was found (p=0.07). We did not observe differences across genotypes in females. CONCLUSIONS Our data suggest an influence of the MCT1 polymorphism on lactate transport across sarcolemma in males. Future studies on lactate transport and metabolism should take into account the gender-specific results.

Apolipoprotein C-III and E polymorphisms and cardiovascular syndrome, hyperlipidemia, and insulin resistance in renal transplantation.

Hyperlipidemia and insulin resistance frequently develop after renal transplantation, contributing to cardiovascular disease. Individual differences in response based upon genetic variations in proteins regulating lipidic and glucose tolerance metabolism could be expected. In the general population, the S2 allelic variant of the apoprotein (apo) C‐III gene has been associated with hypertriglyceridemia and an insulin resistant state, whereas the E4 allele of the apo E has been associated with hypercholesterolemia and atherosclerosis. Its influence in renal transplant patients remains to be seen. In order to assess the impact of apo E and C‐III major polymorphisms on atherosclerotic vascular disease, lipid profile and impaired glucose tolerance in renal transplant patients, we studied 110 consecutively examined patients undergoing kidney transplantation (age range 24–73 years). Atherosclerotic complications were detected in 25% of patients, with age, male sex and hypercholesterolemia being significant atherosclerotic risk factors. Among the male patients with E4 allele, the odds ratio for coronary disease and global atherosclerosis were 10.2 (95% CI) and 6.4 (95% CI), respectively. There were no significant differences in the frequency of any of the polymorphisms among patients with dyslipidemia and impaired glucose tolerance. As the number of patients in our sample was small, larger studies are needed to verify these issues. While in the studied population C‐III polymorphism appears to have little association with the prevalence of atherosclerotic complications, E4 allele should be considered as a genetic marker of coronary artery disease and global atherosclerosis in renal transplant patients.

Effect of the Ala12 allele in the PPARγ-2 gene on the relationship between birth weight and body composition in adolescents: The AVENA study

The intent of this study was to assess whether the effect of birth weight on later body composition is modified by Pro12Pro, Pro12Ala, and Ala12Ala genotypes of the peroxisome proliferator–activated receptor γ-2 (PPARγ-2) gene. The PPARγ-2 gene polymorphism was genotyped in 273 adolescents aged 13–18.5 y, born at term and whose birth weight was known. They were selected from a cross-sectional multicenter study conducted in five Spanish cities in 2000–2002. Body mass index (BMI) was calculated from weight and height measurements, and body composition and fat distribution were estimated from skinfold thickness. A total of 229 subjects (111 males and 118 females) carried the Pro12Pro genotype and 44 (22 males and 22 females) the Pro12Ala and Ala12Ala PPARγ-2 genotypes. In the Pro12Pro group, birth weight Z score was positively associated with both fat-free mass (FFM) (p < 0.05) and fat mass (FM) (p < 0.05), but these relationships disappeared after controlling for age, gestational age, socioeconomic status (SES), physical activity, Tanner stage, sex, and BMI. In the Ala12 group, birth weight Z score was positively associated with FFM (p < 0.01), and this relationship remained significant after controlling for confounding variables (p < 0.05). Small body weight at birth may program lower FFM in adolescents carrying the Ala12 allele in the PPARγ-2 gene.