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The detection of IgG aquaporin-4 antibodies in the serum of patients with Neuromyelitis optica (NMO) has dramatically improved the diagnosis of this disease and its distinction from multiple sclerosis. Recently, a group of patients have been described who have an NMO spectrum disorder (NMOsd) and who are seronegative for AQP4 antibodies but positive for IgG aquaporin-1 (AQP1) or myelin oligodendrocyte glycoprotein (MOG) antibodies. The purpose of this study was to determine whether AQP1 and MOG could be considered new biomarkers of this disease, and if point mutations in the gDNA of AQP4, AQP1 and MOG genes could be associated with the etiology of NMOsd. We evaluated the diagnostic capability of ELISA and cell-based assays (CBA), and analyzed their reliability, specificity, and sensitivity in detecting antibodies against these three proteins. The results showed that both assays can recognize these antigen proteins under appropriate conditions, but only anti-AQP4 antibodies, and not AQP1 or MOG, appears to be a clear biomarker for NMOsd. CBA is the best method for detecting these antibodies, and serum levels of AQP4 antibodies do not correlate with the progression of this disease. So far, the sequencing analysis has not revealed a genetic basis for the etiology of NMOsd, but a more extensive analysis is required before definitive conclusions can be drawn.

CAPN3 mutations cause a limb girdle muscular dystrophy. Functional characterization of novel mutations facilitates diagnosis of future cases. We have identified a novel (c.1992 + 2T>G) CAPN3 mutation that disrupts the donor splice site of intron 17 splicing out exon 17, with mRNA levels severely reduced or undetectable. The mutation induces a strong change in the 3D structure of the mRNA which supports no-go mRNA decay as the probable mechanism for RNA degradation. The mutation was identified in two unrelated Roma individuals showing a common ancestral origin and founder effect. This is the first Roma CAPN3 mutation to be reported., This project has been founded by ISCIII and FEDER “a way to achieve Europe”; Grant number PI16/00612(MC‐S) and PI16/01843 (CP). MC‐S was supported by ISCIII (JR15/00042) and Junta de Andalucia‐Consejeria de Salud (B‐0005‐2017).

SummaryRecently, we reported that the polymorphism 1132T>C (Gene-Bank: AF519768.1) of the NOS3 gene was associated with susceptibility to metabolic syndrome (MS) in hypertensive patients. This suggests that other genes such as CAV1, whose product (CAV1) regulates eNOS activity, could also be related to this phenotype.In this work we investigated the following:i) whether CAV1 is a quantitative trait locus of clustering of atherothrombotic traits associated with MS; ii) whether CVA1 is associated with hypertension or MS in hypertensive patients; and iii) whether genetic interaction between NOS3 and CAV1 is involved in the susceptibility or protection to hypertension associated with MS.To carry out the study, we genotyped 285 randomly selected individuals and 175 hypertensive patients, all of them ≤ 60 years old, with two polymorphisms of the CAV1 gene: the 22285 C>T and the 22375–22375 del AC (GeneBank AF125348), and the 1132T>C polymorphism of the NOS3 gene. To perform sample genotyping, we used pyrosequencing and FRET techniques.The 22285 C-22375–22375 del (Cd) haplotype of CAV1 gene was associated with low levels of blood pressure in the general population. Moreover, it was a genetic protection factor against MS in hypertensive patients. In addition, we found no evidence of gene-gene interaction between NOS3 and CAV1 genes with regard to that phenotype.

We analyzed two polymorphisms (–9C>G and IVS1+905A>G) within the BMP15 gene in women from Spain with polycystic ovary syndrome (PCOS). In this study, the BMP15 gene does not seem to be associated with PCOS. Nonetheless, we observed in both markers a genetic association with anovulation or infertility in these patients.

Germline mutations in different genes are associated with premature ovarian failure (POF, OMIM 311360), defined as premature menopause with amenorrhea occurring before the age of 40 years along with increased gonadotropin concentrations [follicle-stimulating hormone (FSH) >40 IU/L]. A new candidate gene, bone morphogenetic protein 15 ( BMP15 ) has been investigated in POF. In a family affected by hypergonadotropic ovarian failure, a mutation in the pro-region of the BMP15 gene (Y235C) was found in 2 affected sisters (1), and 3 linked markers within the BMP15 gene (−673C>T, −9C>G and IVS1 + 905A>G) are associated with high follicle production in women undergoing recombinant FSH stimulation (2). Several heterozygous variations affecting the pro-region and mature peptide of the BMP15 gene have been identified in women with POF (3)(4)(5), but A180T was the only variant found in all reported studies, occurring with relatively high frequency in POF women (8 of 502, 1.6%) but not at all in control groups. To clarify the role of the A180T allele in early menopause and ovarian failure, we used a pyrosequencing protocol (Biotage) to evaluate the A180T …

Objective Age at natural menopause (ANM) can be considered a complex parameter that depends on the interaction of multiple factors. In the present study, the role of interaction between genetic variants within estrogen synthesis and signalling pathways in the ANM in Spanish women is studied. Material and methods Nine single nucleotide polymorphisms (SNPs) located at different candidate genes related to the estrogen signalling pathway were analysed in 1980 Spanish postmenopausal women. Results Independently, none of the nine markers were significantly associated with early ANM. Only heterozygosis at the NRIP rs2229741 locus could be associated with early menopause; however, this marker does not maintain statistical significance. In contrast, linear regression analysis suggests several epistatic interactions including these markers in relation to ANM, especially between ESR2, NRIP1 and BMP15. The genetic variant that appears most in these interactions is that of the BMP15 rs3897937. It was observed that AA-TC combined genotype for NRIP-BMP15 (rs3897937), respectively, appears to be associated with a lower ANM than other possible combinations of these SNP (46.1±5.9 versus 50.4±3.3; P = 0.002). In the multilocus analysis, the multigenic interaction formed by ESR2 (AA), BMP15 rs3897937 (TC) and NRIP1 (AA) has the lower ANM (45.37±6.8 versus 48.69±5; P = 0.038). Conclusions The results suggest that epistatic interactions of estrogen-related alleles may contribute to variance in ANM in Spanish women. Moreover, BMP15 and NRIP1 also appear as attractive candidate genes for premature menopause but require further investigation to confirm them.

Papillary thyroid cancer (PTC) accounts for 80% of all thyroid malignancies, and genetic alterations associated to its etiology remain largely unknown. Chromosomal band 11q13 seems to be one of the most frequently amplified regions in human cancer, providing several candidate genes that need detailed characterization. The aim of our study was to investigate the existence of allelic imbalance at EMSY, CAPN5, and PAK1, as candidate genes within 11q13.5-q14 region using a single nucleotide polymorphism-based analysis. We selected a panel of 9 polymorphisms that were analyzed in 41 thyroid carcinoma samples, their contralateral non-pathological tissue and 178 controls from the general population. We did not detect allelic imbalance at these loci in our series. However, we observed a difference in the EMSY-haplotype distribution among PTC patients when compared to controls (odds ratio=2.00; p=0.02). We conclude that 11q13.5-q14 is not imbalanced in PTC, but there is evidence suggesting that EMSY might be of relevance in PTC etiology.

Cardiovascular disease is the leading cause of morbidity and mortality in the industrialized world. Familial aggregation of cardiovascular risk factors is a frequent finding, but genetic factors affecting its presentation are still poorly understood. The calpain 10 gene (CAPN10) has been associated with type 2 diabetes (T2DM), a complex metabolic disorder with increased risk of cardiovascular disease. Moreover, the CAPN10 gene has been associated with the presence of metabolic syndrome (MS) in T2DM and in polycystic ovary syndrome (PCOS). In this work, we have analysed whether the polymorphisms UCSNP44, -43, -19 and -63 are related to several cardiovascular risk factors in the context of MS. Molecular analysis of CAPN10 gene was performed in 899 individuals randomly chosen from a cross-sectional population-based epidemiological survey. We have found that CAPN10 gene in our population is mainly associated with two indicators of the presence of insulin resistance: glucose levels two hours after a 75-g oral glucose tolerance test (OGTT) and HOMA values, although cholesterol levels and blood pressure values are also influenced by CAPN10 variants. In addition, the 1221/1121 haplogenotype is under-represented in individuals that fulfil the International Diabetes Federation (IDF) diagnostic criteria for MS. Our results suggest that CAPN10 gene is associated with insulin resistance phenotypes in the Spanish population.

Sexual dimorphism in blood pressure (BP) regulation has been observed both in humans and experimental animals, and estrogens have been shown to contribute to this epidemiological observation. A key enzyme in determining estrogen levels is aromatase cytochrome P450. The aim of this study was to evaluate the role of the gene encoding aromatase, CYP19A1 , as an independent risk factor for hypertension and its relationship with systolic and diastolic BP measures. We genotyped 2 polymorphisms within the CYP19A1 gene, IVS4 rs11575899 and 3′UTR rs10046, in 3448 individuals. In quantitative analysis, we observed significant associations between the 2 polymorphisms and BP values in women, being these associations dependent on BMI and independent of menopause status. The case–control analysis revealed that the most prominent associations were found for nonobese women in diastolic hypertension (DHT): the IVS4_22 and 3 ′ UTR_11 are risk genotypes (OR=1.61, P =0.027 and OR=1.59, P =0.012, respectively), whereas IVS4_11 and 3 ′ UTR_22 genotypes have a protective effect against DHT (OR=0.63, P =0.009, and OR=0.61, P =0.020, respectively). Haplotype analysis confirmed the above associations: among nonobese women the haplotype 21 is overrepresented in hypertensive women (OR=1.33, P =0.004, for DHT and OR=1.25, P =0.026, for systolic hypertension, SHT) and, conversely, the haplotype 12 protects against hypertension (OR=0.78, P =0.015 for DHT and OR=0.82, P =0.04 for SHT). Our study has shown that the CYP19A1 gene may be involved in the genetic regulation of BP in women. This effect is dependent on BMI and independent of menopause status, suggesting that this action is mainly driven by aromatase activity in fat tissue.

The application of pharmacogenetics and pharmacogenomics to assisted reproductive techniques will help clinicians to improve the efficacy of hormone treatments that are being routinely applied during assisted reproductive technique protocols. Genetic markers involving controlled ovarian hyperstimulation pharmacogenetics are being isolated within follicle-stimulating hormone and estrogen receptor signaling pathways using the candidate gene approach. Furthermore, the information obtained during controlled ovarian hyperstimulation pharmacogenetics studies could be applied to other estrogen-related diseases, such as osteoporosis, breast cancer, essential hypertension and many other diseases related to estrogen production or its mechanism of action. The theory that estrogen-related diseases may share some risk factors with controlled ovarian hyperstimulation efficacy, and side effects linked to genetic markers, is discussed.

Objective Controlled ovarian stimulation (COS) using recombinant follicle-stimulating hormone (rFSH) is the main treatment in assisted reproduction. We performed a pharmacogenetic analysis of bone morphogenetic protein 15 (BMP15) gene using single nucleotide polymorphisms (SNPs) in COS. We also investigated the role of the BMP15 gene in ovarian hyperstimulation syndrome (OHSS). Methods We analysed different intragenic SNPs located within the BMP15 gene in 307 women treated with rFSH, evaluating its involvement in COS outcome. Results First, we analysed two polymorphisms, by applying different tests for genetic association, and we found a minimum P-value in patients producing ≥12 follicles in COS (high responders) in both polymorphisms of the BMP15 gene. Using bi-directional DNA sequencing, we identified two additional single nucleotide DNA variants. Second, we conducted association studies with all polymorphisms together, and noticed that none of them seemed to fully explain the association of the BMP15 gene with over-response to rFSH. However, N103S missense mutation is predicted to disrupt the secondary structure of human BMP15 protein and is weakly associated with OHSS. This coding mutation of the BMP15 gene may partially explain the results obtained during our research. Using Thesias software, we reconstructed haplotypes with the four intragenic variants and calculated their frequencies in normal and over-responders to rFSH. The haplotype TGGA was over-represented in high responders when compared with the rest of patients. Moreover, this association was higher in patients with OHSS, with a significant global haplotypic effect of the BMP15 gene. Conclusion Our results suggest a direct relationship between increased follicle production during COS and BMP15 alleles in response to rFSH in humans. The use of BMP15 markers to prevent OHSS is also a possibility that requires thorough evaluation.

The calpain family is a group of cysteine proteases unique in their dependency on calcium to attain functionally active forms. Calpains are involved in a wide range of cellular calcium-regulated functions, including signal transduction, cell proliferation and differentiation, and apoptosis. Moreover, altered calpain activity has been observed in several human diseases. Specific calpain inhibitors hold promise for the treatment of neuromuscular and neurodegenerative diseases in which calpains have been shown to be upregulated (e.g. Parkinson's disease and Duchenne muscular dystrophy). Conversely, calpain activators could be a useful approach for those diseases where reduced calpain activity has been observed, such as type 2 diabetes or metabolic syndrome.

Controlled ovarian hyperstimulation (COH) is a routine treatment employed in most assisted reproductive techniques (ARTs). The existence of genetic factors involved in COH has been suspected. The main challenge for clinicians involved in ART is COH cycle cancellation, which usually occurs due to two opposing situations. On the one hand, there is the presence of a poor response during COH treatment, and on the other there is the presentation of a side effect related to gonadotropin hypersensitivity (ovarian hyperstimulation syndrome [OHSS]). Evidence for an association between single nucleotide polymorphisms and COH outcome has been obtained during the last decade. The genetic dissection of both extreme phenotypes of COH will be the main objective of this review. The development of predictive panels useful for the clinical management of COH is currently underway, and will improve the clinical management of patients undergoing ART.

Objective To examine whether polymorphisms within the ESR1 , FSHR , ESR2 , CYP19A1 , and NRIP1 genes are susceptibility factors for human male idiopathic infertility and to test the joint effects of these genes on male reproductive function. Design Genetic association study of male infertility with polymorphisms, using both single-gene and multilocus approaches. Setting Private and public fertility units and a private center for biomedical research. Patient(s) One hundred four Spanish men with azoospermia or severe oligozoospermia and 95 unselected race-matched healthy controls from the same geographic region. Intervention(s) Peripheral blood extraction, DNA purification, and ESR1 g.938T>C, FSHR Ser680Asn, ESR2 *39A>G, CYP19A1 *19C>T, and NRIP1 Gly75Gly polymorphism analyses. Main Outcome Measure(s) Single-gene statistical analyses and multilocus statistical analyses with Sumstat, Permutation and Model-free analysis, and Estimating Haplotypes software. Result(s) We observed an excess of homozygous infertile men for the ESR1 g.938T>C marker. Multilocus analyses detected genetic interaction between the five candidate gene markers that are influential over male infertility. In addition, we detected a five-loci protector genetic pattern with a frequency of 9.4% in controls but absent in infertile men. Conclusion(s) Our results support a relevant role for the estrogenic pathway, notably the ESR1 gene, in human male reproductive function and advocate a complex trait model for male infertility.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are fatal neurodegenerative syndromes that belong to the same clinicopathological spectrum.1,2 Frontotemporal lobar degeneration is a primary dementia characterized by early behavioral, language, and extrapyramidal changes, while symptoms of ALS are the result of the degeneration of motor neurons. Both syndromes may occur within the same family or even the same patient. Previously, linkage analyses revealed a 3.7-Mb region on 9p21 associated with familial ALS/FTLD,3–10 and genome-wide association studies suggested a major risk factor in the same locus for sporadic ALS and FTLD.11–15 Recently, 2 research groups independently explained this locus by a pathological noncoding hexanucleotide (G4C2)30–1600 repeat expansion in the chromosome 9 open reading frame 72 (C9orf72) gene of unknown function.16,17 Based on the allele frequencies in cases vs controls, the first studies suggested that expansions with more than 30 repeats should be considered pathological, while alleles with less than 20 repeats are wild type.16 However, a reliable cutoff for the pathological alleles remains to be established by additional studies (eg, segregation, neuropathological, or functional studies). Furthermore, the contribution of intermediate-size alleles (20–29 repeats) to disease pathology has not yet been evaluated. The expansion is the most frequent cause of ALS and FTLD identified to date. In the Finnish population, 46% of patients with familial ALS, 21% of patients with sporadic ALS, and 29% of patients with sporadic FTLD have the expansion.16 DeJesus-Hernandez et al17 reported the expansion in 24% of patients with familial ALS, 4% of patients with sporadic ALS, and 12% of patients with familial FTLD. In the Flanders-Belgian cohort, the mutation was observed in 47% of patients with familial ALS, 5% of patients with sporadic ALS, and 16% of patients with familial FTLD.18 The pathological mechanism associated with the expansion is currently unknown except that the expansion leads to a 50% reduction of C9orf72 messenger RNA expression,17,18 and the brain pathology in mutation carriers is associated with possibly toxic nuclear RNA foci, as well as TAR DNA-binding protein 43 (TDP-43) and p62 inclusions.17,19 The clinical phenotype appears to be highly heterogeneous in reported mutation carriers.20 Following the discovery of this novel mutation, many questions are yet to be addressed. What is the expansion frequency in other ALS/FTLD cohorts? Are there any clinical features that can discriminate between patients with and without the C9orf72 expansion? What is a reliable cutoff for the pathological repeat number? What is the role of alleles with intermediate repeat sizes or variability in the region flanking the G4C2 repeat? Could the expansion account for other neurodegenerative diseases, such as Alzheimer disease (AD) or Parkinson disease (PD)? These questions were investigated by the current study. The expansion frequency was estimated in a comprehensive case-control sample set consisting of 2224 individuals (patients with FTLD, ALS, AD, and PD and healthy controls). To our knowledge, this is the first case-control study using a 2-step genotyping strategy that allowed for the analysis of genotype information for the alleles with less than 50 repeats. Clinical data were analyzed to understand the phenotype spectrum observed in mutation carriers.

The present research is aimed at assessing the role of 3 estrogen receptor alpha (ESR1) gene variants in late onset Alzheimer's disease (AD) susceptibility. One thousand one hundred thirteen unrelated late onset sporadic AD patients, 1109 healthy controls and 121 neurologically healthy elderly controls were used to carry out case-control genetic association studies with ESR1 rs3844508, rs2234693, and ESR1 noncoding deletion 1 (ESR1-NCD1) polymorphisms. Thirty-five healthy male samples were used for molecular analyses. The rs2234693 polymorphism is associated with AD in our population (odds ratio [OR], 1.29; p = 0.008). The rs3844508 marker confers protection against AD in males (OR, 0.57; p = 0.001) and the deletion ESR1-NCD1 is a risk factor for AD in women (OR, 1.67; p < 0.001). Molecular analyses on ESR1-NCD1 indicate that this deletion confers a higher response to estradiol activity on ESR1 receptor and it is also associated with differential expression of ESR1 isoforms. Our results support the involvement of ESR1 gene in AD and point to the existence of sexual dimorphism for ESR1 markers. In addition, carriers of ESR1-NCD1 deletion could overrespond to estradiol action.

Context Obesity is a multifactorial disorder, that is, a disease determined by the combined effect of genes and environment. In this context, polygenic approaches are needed. Objective To investigate the possibility of the existence of a crosstalk between the CALPAIN 10 homologue CALPAIN 5 and nuclear receptors of the peroxisome proliferator-activated receptors family. Design Cross-sectional, genetic association study and gene-gene interaction analysis. Subjects The study sample comprise 1953 individuals, 725 obese (defined as body mass index ≥ 30) and 1228 non obese subjects. Results In the monogenic analysis, only the peroxisome proliferator-activated receptor delta (PPARD) gene was associated with obesity (OR = 1.43 [1.04–1.97], p = 0.027). In addition, we have found a significant interaction between CAPN5 and PPARD genes (p = 0.038) that reduces the risk for obesity in a 55%. Conclusion Our results suggest that CAPN5 and PPARD gene products may also interact in vivo.