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This study was designed to analyse possible associations between DNA polymorphisms in the 5-hydroxytryptamine (5-HT; serotonin) 5-HT2A receptor and the 5-HT transporter (5-HTT) genes, and myocardial infarction (MI). 5-HT has been shown to be involved in cardiovascular pathophysiology. In addition to platelet aggregation and vascular contraction, 5-HT induces hyperplasia of artery smooth muscle cells. Recently, a 5-HT transporter gene polymorphism has been associated with MI. To determine the influence of genetic variation at the 5-HT2A receptor (T102C polymorphism) and the 5-HTT (insertion/deletion polymorphism) on the risk of developing early MI, we genotyped 210 MI patients of 60 years old) to analyse the effects of these polymorphisms on the age at which the first MI episode occurred. The 5-HT2A receptor polymorphism was not associated with MI in our population. In addition, since the 5-HT2A receptor gene and genotype frequencies did not differ between patients with early and late onset of MI, this polymorphism does not appear to have an effect on age at the first MI episode. Gene and genotype frequencies for the 5-HTT promoter did not differ between patients 60 years old compared with patients

Plasmacytoma of the calvarium is very rare, accounting forthe 0.7 % of all plasmacytomas [1–5]. The most commonlocations are the thoracic and lumbar spine (30–40 % ofcases). Involvement of the hip, femurs, and ribs is lessfrequent [6].Plasmacytoma of the calvarium can be indistinguish-able from other lytic lesions, especially metastatic dis-ease. Meningioma, eosinophilic granuloma, anddermoid cysts can have similar appearances, althoughusually in a different clinical context [10, 11]. Histo-pathological examination and laboratory tests are nec-essary to make a definitive diagnosis of plasmacytoma(Fig. 1).The typical features on imaging are a lytic lesion on X-rayexamination(Fig.1 question) [1, 6]. On computedtomography (CT), plasmacytoma presents as a lyticpunched-out lesion with interspersed residual thickened bonytrabeculae. Cortical thinning and focal destruction are typical(Fig. 2 question) [2]. On magnetic resonance imaging (MRI),bone plasmacytoma of the calvarium presents as an intra-medullary lesion of low to intermediate intensity on T1-weighted images and high intensity on T2-weighted images.Contrast enhancement is usually mild [4–9].In the presented case, the calvarium plasmacytomapresents a “mini-brain” appearance, due to the presence oflow-intensity hypertrophic residual trabeculae interspersedin a relatively homogeneous mass, mimicking the brainsulci.Major et al. first described the “mini-brain” appearanceof plasmacytoma in a series of ten patients with spinal

Background and aims Recent genomic surveys have identified IL12B as susceptibility locus for psoriasis (Ps). Our aim was to replicate the association between IL12B SNPs and Ps. In addition, we sequenced the IL12B gene in several patients to identify new variants that could explain the disease-risk. Results A total of 304 Ps-patients and 422 healthy controls (all Caucasian Spanish) were genotyped for three IL12B polymorphisms. SNP rs6887695 (GG genotype) was significantly associated with Ps (p = 0.002; OR = 1.60, 95% CI = 1.19–2.16). This genotype was also more frequent among patients with severe psoriasis (p = 0.03). Sequencing of 30 patients with the risk genotype identified several IL12B reported SNPs. Allele and genotype frequencies for two putative functional variants (rs3213120 and rs3213119) did not differ between patients and controls. Conclusions Our study confirmed rs6887695 as a risk factor for Ps. No other IL12B variants that could explain this association were found in our patients.

In the last two decades, food scientists have advanced the development of new technologies for preservation of fresh and processed fruits and vegetables. The scientific achievements in this area are driven by increased consumption of these products and increasing consumer demand for fresh, safe, healthy, and natural products. This chapter describes the use of natural antimicrobials as an alternative to synthetic disinfectants to assure safety, preserve quality, and extend the shelf life of fruits and vegetables.

Es una Carta al editor: 4 páginas, 2 tablas, 22 referencias bibliográficas. This is a pre-copyedited, author-produced PDF of an article accepted for publication in Brain following peer review. The version of record Brain 138(Pt 12):e400. (2015) is available online at: http://dx.doi.org/10.1093/brain/awv175, This study was supported in part by grants from Instituto de Salud Carlos III (PI12/01311) and MINECO (SAF2014-59469-R).

This chapter describes the use of natural antimicrobials as an alternative to synthetic disinfectants in order to assure safety, preserve quality, and extend the shelf life of fruits and vegetables. It also discusses secondary responses induced by the application of natural agents (antioxidant and flavoring effects).

Background Recent studies have shown that exogenous administration of vascular endothelial growth factor (VEGF) is protective against cyclosporine A (CsA) renal toxicity. No data are available, however, on the possible role of endogenous VEGF. Our objective was to examine whether endogenous VEGF has a significant role in the renal response against CsA toxicity. Methods In vivo, we used high-dose (50-150 mg/kg/day) CsA +/- specific goat anti-mouse VEGF blocking monoclonal antibody (alpha-VEGF) in mice. In vitro, we exposed mouse tubular cells (MCT) to CsA +/- alpha-VEGF. Results alpha-VEGF markedly enhanced CsA renal toxicity, inducing severe tubular damage and increased blood urea nitrogen. In animals treated with CsA + alpha-VEGF, damage progressed to generalized tubular injury (histology) and apoptosis (terminal deoxynucleotide transferase-mediated dUTP nick-end labeling) with associated anemia and reticulocytosis (18 days of treatment). CsA + alpha-VEGF treatments strikingly increased tubular VEGF and Bcl-xL proteins. In vitro, autocrine production of VEGF by MCT was identified by Western blot. Of specific interest, CsA toxicity in MCT increased significantly in the presence of alpha-VEGF. Conclusions Endogenous VEGF has a relevant role in the renal tubular defense against CsA toxicity. Blockade of the VEGF effect by alpha-VEGF results in clear-cut intensification of the tubular injury and appearance of regenerative anemia in the CsA + alpha-VEGF-treated animals. The occurrence of both in vivo and in vitro effects of VEGF blockade provides evidence of a direct protective effect of VEGF on the tubular cell.

Local inflammatory processes associated with amyloid plaques would contribute to the progression of late-onset Alzheimer disease (LOAD). Tumor necrosis factors alpha (TNF(alpha)) and beta (LT(alpha)) are inflammatory cytokines involved in the local immune response occurring in the central nervous system of LOAD patients. Genetic variation at these genes could contribute to the risk of developing AD or influence the age at the onset of the disease. We genotyped 315 LOAD patients and 400 healthy controls for DNA-polymorphisms in the genes encoding TNF(alpha) (-308 G/A, -238G/A) and LT(alpha) (Asn26Thr). Carriers of -308A showed a mean age at onset 3 years younger than noncarriers of this allele (P = 0.019). Our data suggest an effect of the TNF(alpha)-308 polymorphism on the age at onset of late AD. This represents additional evidence of the importance of genetic variation at the proinflammatory components in the origin and progression of this common neurodegenerative disease.

In chronic hepatitis C (CHC), the interlobular bile ducts, particularly the small bile ducts, show a variety of abnormalities collectively known as hepatitisassociated bile duct damage. This type of bile duct lesion is found in up to 90% of the liver biopsies performed in these patients (1, 2). Usually, these lesions are different from that observed in primary biliary cirrhosis (PBC). However, cases of CHC associated with true PBC, with (3– 6) and without (7–9) positive anti-mitochondrial antibodies (AMA), have been reported. In the former cases, interferon (IFN) therapy was ineffective (3) or even triggered cholestatic damage (4 – 6). In this paper, we report the case of a female patient with CHC and PBC-like duct lesions, negative AMA and positive anti-nuclear antibodies (ANA), fulfilling the criteria for autoimmune cholangitis (AC).

OBJECTIVE To investigate the association between apolipoprotein E (ApoE) genotype and schizophrenia. METHOD We genotyped 106 schizophrenic out-patients [Diagnostic Statistic Manual IV (DSM-IV) criteria] and 250 healthy volunteers (hospital staff and blood donors) from Asturias (Northern Spain). The ApoE genotypes (epsilon2, epsilon3, epsilon4-alleles) were determined after polymerase chain reaction (PCR) amplification, followed by digestion with the restriction enzyme Cfol and electrophoresis on a 4% agarose gel. RESULTS No significant differences in ApoE-allele frequencies between patients and controls was found, although an increased 64-frequency was recorded in patients compared with controls [9.0% vs. 6.2%, P = 0.124; odds ratio (OR) = 1.49; 95% confidence interval (CI) = 0.82-2.70]. ApoE-genotype frequencies did not differ between both groups. The mean age of onset for schizophrenic patients that carried the epsilon4-allele was not significantly different from that of patients without this allele. CONCLUSION Variation in the ApoE gene was not associated with the development of schizophrenia in our population. ApoE-genotypes did not modify the age of onset of the disease.

To the Editor: MicroRNAs (miRNAs)1 regulate cardiac growth and conduction and play an important role in cardiac diseases (1). Several miRNAs are differentially produced in cardiac hypertrophic tissue, compared with normal tissue, and may contribute to the development of cardiomyocyte hypertrophy (2, 3). Hypertrophic cardiomyopathy (HCM) is frequently familial and caused by mutations in sarcomeric genes. To our knowledge, no study has reported the miRNA production profile in HCM tissues with sarcomeric gene mutations. To better define the molecular changes in HCM, we defined the production of well-characterized miRNAs in left ventricular (LV) heart tissue from 5 patients who underwent a cardiac transplantation and a control heart tissue (human LV tissue, Ambion/Applied Biosystems), and we compared their production profiles. Two of the patients were familial HCM patients who were carriers of missense mutations in the MYH7 2 (myosin, heavy chain 7, cardiac muscle, beta) gene (Val822Met and Arg453Cys). Three patients were cases of sporadic LV hypertrophy secondary to heart valve disease. The study was approved by the Ethical Committee of Hospital Universitario Central Asturias (HUCA), and all of the patients provided written informed consent. We isolated total RNA with TRIzol (Invitrogen) and …

Edible films can be used as carriers for antimicrobial compounds to assure food safety andquality; in addition, pathogenesis of food bacteria is related to a cell to cell communicationmechanism calledquorum sensing(QS). Oregano essential oil (OEO) has proved to beuseful as food antimicrobial; however, its foodapplications can be compromised by thevolatile character of its active constituents.Therefore, formulation of edible films containingOEO can be an alternative to improve its food usages. QS inhibitory activity of OEO andpectin-OEO films was evaluated usingChromobacterium violaceumas bacterial model.Additionally, antibacterial activity was tested againstEscherichia coliO157:H7,SalmonellaCholeraesuis,Staphylococcus aureus,andListeria monocytogenes. OEO was effective toinhibit bacterial growth at MIC of 0.24 mg/mL for all tested bacteria and MBC of 0.24,0.24, 0.48, and 0.24 mg/mL againstE. coliO157:H7,S.Choleraesuis,S. aureus, andL.monocytogenes, respectively. Pectin-films incorporated with 36.1 and 25.9 mg/mL of OEOshowed inhibition diameters of 16.3 and 15.2 mm forE. coliO157:H7; 18.1 and 24.2 mm forS.Choleraesuis; 20.8 and 20.3 mm forS. aureus; 21.3 and 19.3 mm forL. monocytogenes,respectively. Pectin-OEO film (15.7 mg/mL) was effective againstE. coliO157:H7 (9.3 mm),S. aureus(9.7 mm), andL. monocytogenes(9.2 mm), but not forS.Choleraesuis. Allconcentrations of OEO (0.0156, 0.0312, 0.0625 and 0.125 mg/mL) and pectin-OEO films(15.7, 25.9 and 36.1 mg/mL) showed a significant anti-QS activity expressed as inhibition ofviolacein production byC. violaceum. Additionally, the application of pectin-OEO films waseffective reducing total coliforms, yeast, and molds of shrimp and cucumber slices storedat 4◦C during 15 d.These results demonstrated the potential of pectin films enriched withOEO as food related microorganisms and QS inhibitors Fil: Alvarez, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingeniería. Departamento de Ingeniería Química. Grupo de Investigación en Ingeniería En Alimentos; Argentina Fil: Ortega Ramirez, Luis Alberto. Centro de Investigación en Alimentación y Desarrollo. Hermosillo; México Fil: Gutierrez Pacheco, Maria Melissa. Centro de Investigación en Alimentación y Desarrollo. Hermosillo; México Fil: Bernal Mercado, Thalia. Centro de Investigación en Alimentación y Desarrollo. Hermosillo; México Fil: Rodriguez Garcia, Isela. Centro de Investigación en Alimentación y Desarrollo. Hermosillo; México Fil: Ponce, Alejandra Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingeniería. Departamento de Ingeniería Química. Grupo de Investigación en Ingeniería En Alimentos; Argentina Fil: Moreira, Maria del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingeniería. Departamento de Ingeniería Química. Grupo de Investigación en Ingeniería En Alimentos; Argentina Fil: Roura, Sara Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingeniería. Departamento de Ingeniería Química. Grupo de Investigación en Ingeniería En Alimentos; Argentina Fil: Ayala Zavala, Jesús Fernando. Centro de Investigación en Alimentación y Desarrollo. Hermosillo; México

The deregulation of several microRNAs (miRNAs) has been associated with neurodegenerative processes, including Parkinson's disease (PD). Our aim was to characterize the level of miRNAs in the substantia nigra (SN) of PD patients and healthy donors. This is an important issue to characterize new putative markers and therapeutic targets for PD. RNA was extracted from the SN of postmortem PD (n=8) and healthy (n=4) subjects, and the level of 733 human miRNAs was assayed with TaqMan low-density arrays (TLDAs). Overall, there was a miRNA downregulation in the SN of patients. The mean level of 11 miRNAs was significantly different (p

Nitric oxide (NO) influences susceptibility to infection and hemodynamic failure (HF) in sepsis. NOS3 and NOS2 SNPs might modify plasma nitrite/nitrate (NOx) levels, sepsis development, hemodynamics and survival.90 severely septic and 91 non-infected ICU patients were prospectively studied. NOS3 (E298D), NOS3 (-786 T/C), NOS3 (27 bp-VNTR), and NOS2A (exon 22) SNPs and plasma NOx levels were assessed.21 patients (11.6%) died, 7 with sepsis. TT homozygotes and T allele carriers of NOS3 (E298D) and AG carriers of the NOS2A (exon 22) SNPs were more frequent among septic compared to non-infected ICU patients (p 0.05). Plasma NOx was higher in septic, especially in septic with hemodynamic failure (HF) or fatal outcome (p 0.006). Plasma NOx was higher in carriers of the T allele of the NOS3 (E298D) SNP (p = 0.006). Sepsis independently associated with HF, increased NOx, peripheral neutrophils, and fibrinogen levels, decreased prothrombin and the presence of the NOS3 (E298D) and NOS2A (exon 22) SNPs. A low APACHE II score was the only variable associated with sepsis survival. NOx was independently associated with sepsis, HF, decreased neutrophils and higher APACHE.NOS3 (E298D) and NOS2A (exon 22) SNPs, individually and in combination, and plasma NOx, associated with sepsis development. NOx associated with HF and fatal outcome.

Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) are the most common genetic determinants of familial and sporadic Parkinson's disease (PD). Most of the mutational screenings analyzed the exon-coding sequence. Our aim was to determine whether LRRK2 3' untranslated region (UTR) variants were associated with the risk of developing PD in a large cohort of patients (n=743) and controls (n=523) from Spain. We identified a total of 12 3'UTR variants (two new). Single-nucleotide polymorphism (SNP) rs66737902 C allele was overrepresented in patients (P=0.005; odds ratio=1.47). This SNP was in linkage disequilibrium with the p.R1441G mutation, but the association remained significant among mutation-negative cases. We found a significant lower level of the LRRK2 transcript in the Substantia nigra (SN) of PD postmortem donors (n=9) who were rs66737902 C carriers (P=0.01). This SNP was predicted to affect a binding site for miR-138-2-3p. We showed that this microRNA was expressed in all the SN samples. In conclusion, we found a significant association between SNP rs66737902 and the risk of developing PD. This effect on PD risk could be explained by differences in LRRK2 transcript levels between the two alleles.

Gitelman's syndrome (GS) is a rare recessive disorder caused by mutations in the renal salt-handling genes SLC12A3 and CLCNKB. Our aim was to develop a next-generation sequencing (NGS) procedure for these genes based on two-tubes multiplex amplification of DNA pools and semiconductor sequencing with the Ion Torrent Personal Genome Machine (PGM). We created one pool with DNA from 20 GS patients previously Sanger sequenced for the coding exons of SLC12A3. A total of 13 mutations present in 11 of these patients were used as control variants to validate the NGS procedure. The full coding sequence of SLC12A3, CLCNKB and CLCNKA was amplified in only two Ampliseq tubes and processed and sequenced with the PGM. Large SLC12A3 and CLCNKB deletions were ascertained through multiplex ligation-dependent probe amplification in some patients. With the exception of the SLC12A3 exon 9, all the amplicons were successfully read and 12 of the 13 control variants were detected. The analysis of CLCNKB showed four putative mutations in the GS pool that were further assigned to specific patients. Two patients were heterozygous compounds for a single-nucleotide mutation and a large deletion at SLC12A3 or CLCNKB. We reported a NGS procedure that would facilitate the rapid and cost-effective large-scale screening of the three renal salt-handling genes. In addition to characterize the mutational spectrum of GS patients, the described procedure would facilitate the rapid and cost-effective screening of these genes at a population scale.

Summary: Esophageal varices are a frequent complication among patients with liver cirrhosis. Nitric oxide and other vasoactive molecules regulate the vascular tone in both the liver microcirculation and the systemic and splanchnic circulation. Several genes that encode proteins involved in the maintenance of vascular tone, such as the endothelial-constitutive nitric oxide synthase (ecNOS), the angiotensinogen (AGT), the angiotensin-converting enzyme (ACE), and the angiotensin II receptor type 1 (AT1R) are polymorphic, and these polymorphisms have been associated with several cardiovascular diseases. Our aim was to define a possible role for DNA polymorphisms at these genes in the risk of developing esophageal varices among patients with alcoholic cirrhosis. We analyzed 145 male patients with liver cirrhosis. Patients and 200 healthy controls were genotyped by polymerase chain reaction for the ACE-I/D, the AGT-M235T, the AT1R-A1166C, and the ecNOS-4/5 (intron 4) polymorphisms. Ninety-five patients had varices and 50 did not show this complication. Carriers of the ACE-I allele (ID +II genotypes) were at a significantly higher frequency among patients with varices (p = 0.013). Patients without varices had a higher frequency of the ecNOS-4 allele compared with patients with varices (p = 0.026). ACE-I carriers +ecNOS-55 were at a significantly higher frequency (p = 0.0012; odds ratio = 3.19; 95% CI = 1.55–6.55) among patients with varices (51 of 95, 54%) compared with patients without (18 of 50, 36%). Allele and genotype frequencies for the AGT and AT1R polymorphisms did not differ between the two groups. The genotypes associated with an increased risk for varices have been linked to higher plasma levels of nitric oxide and reduced levels of ACE. These genotypes could have a vasodilatory effect in the systemic and splanchnic circulation, thus favoring the development of portocollaterals.

In order to determine the role of two polymorphisms in the factor VII gene (R353Q and intron 7 hypervariable region) in the susceptibility to develop early myocardial infarction, a total of 175 patients with acute myocardial infarction aged 50 years or less (mean age 41±7 years) and 200 controls (average age 42±6) without cardiovascular disease were genotyped for these polymorphisms. Gene and genotype frequencies did not differ between patients and controls. Although the 353-QQ genotype was at a higher frequency among controls (4%) compared to patients (1%), the difference did not reach statistical significance. Carriers of the H7-allele (intron 7 polymorphism) were at a slightly higher frequency among patients (51 vs. 43%; P =0.19; OR=1.36; 95% CI=1.09–1.70). Our data suggest a lack of association between both polymorphisms in the factor VII gene and early myocardial infarction in our population.

Autocrine expression of VEGF has been detected in endothelial cells under hypoxia or oxidative stress. However, the functional significance of this VEGF autocrine expression remains undefined. To analyze the role of autocrine VEGF in the endothelial response against injury, cultured bovine aorta endothelial cells (BAEC) were challenged with potentially cytotoxic substances with different chemical structure and pharmacologic properties, namely cytochalasin D (CyD), hydrogen peroxide (H2O2) and cyclosporine A (CsA). Our results revealed that: i. In particular conditions, exposure to potentially cytotoxic agents as CyD, H2O2 or CsA results in significant BAEC cytoprotection rather than injury. ii. The response to the 3 agents is shifted to a cell damaging pattern in the presence of a specific anti VEGF monoclonal antibody (mAb). iii. CyD and H2O2 markedly stimulate the autocrine expression of VEGF mRNA and VEGF protein. In conclusion, the present study reveals a protective mechanism of endothelial cells against injury involving autocrine VEGF production. Moreover, the occurrence of a significant increase in VEGF expression accompanying this defensive mechanism is further disclosed.

The N-terminal region of parathyroid hormone (PTH) and PTH-related protein (PTHrP) interacts with a common PTH/PTHrP receptor in osteoblasts. These cells synthesize PTHrP, but its role in bone turnover is unclear. Intermittent treatment with N-terminal PTHrP or PTH stimulates bone growth in vivo, possibly by increasing local bone factors. In addition, C-terminal PTHrP (107-139), which does not bind to the PTH/PTHrP receptor, appears to affect bone resorption in vivo and in vitro, although its effect on bone formation in vivo remains controversial. Bone angiogenesis is an often overlooked but critical event in the process of bone remodeling. Recently, PTH (1-34) has been shown to induce gene expression of vascular endothelial growth factor (VEGF), a potent angiogenic factor, by osteoblastic cells. However, no data are available on the effect of PTHrP (107-139) on VEGF expression in these cells. Using semiquantitative reverse transcription followed by PCR, we found that PTHrP (107-139), between 10 nM and 1 pM, increased VEGF mRNA in human osteoblastic (hOB) cells from trabecular bone. This effect of this agonist, at 10 nM, was maximal (fivefold for VEGF(165), and twofold for VEGF(121), compared to control) within 1 to 4 h. This effect was similar to that induced by PTHrP (1-34) in these cells, as well as in human osteosarcoma MG-63 cells, using Northern blot analysis. Moreover, the effect of both peptides, added together at 100 pM, was not higher than that observed with each peptide alone in hOB cells. The effects of PTHrP (107-139) and that of PTHrP (1-34) were abolished by actinomycin D in hOB cells. In these cells, the protein kinase C inhibitor staurosporine, but not the protein kinase A inhibitor H89, inhibited the increase in VEGF mRNA induced by 10 nM PTHrP (107-139). PTHrP (107-139), at 10 nM, also stimulated cytosolic VEGF immunostaining in hOB cells, and VEGF secretion into the medium conditioned by hOB or MG-63 cells for 24 h, which was (ng/mg protein): 10 +/- 1 or 5 +/- 3 (control), respectively, and 21 +/- 1 or 11 +/- 2 (PTHrP [107-139]-stimulated), respectively. Furthermore, medium conditioned by these cells for 24 h in the presence of 10 nM PTHrP (107-139), with or without 10 nM PTHrP (1-34), increased about 30% bovine aortic endothelial cell (BAEC) growth at 48 h. This effect was inhibited by adding a specific anti-VEGF antibody to the BAEC incubation medium. These findings demonstrate that the C-terminal domain of PTHrP induces expression and secretion of VEGF, a main angiogenic factor, in hOB cells and MG-63 cells. This relationship between PTHrP and VEGF has potential implications for both bone vascularization and bone formation, and neoangiogenesis in PTHrP-producing tumors.

Mitochondrial dysfunction has been implicated in Parkinson's disease (PD). The nuclear encoded transcription factors A, B1 and B2 are essential for mitochondrial DNA replication. Sequence variants at the genes encoding TFAM, TFB1M and TFB2M could contribute to the risk of developing PD. Here, we searched for TFB1M and TFB2M nucleotide variants in a cohort of PD-patients (n=300) and healthy controls (n=200) from Spain. Single strand conformation analysis and direct sequencing were used to determine the variation at all the coding exons of the two genes. In addition to previously reported polymorphisms, we found several rare variants in patients and controls. Allele frequencies for all the nucleotide changes did not differ between patients and controls. Our work suggests that DNA variants in TFB1M and TFB2M did not contribute to the risk for PD in our population.

OBJECTIVE Familial multiple endocrine neoplasia type 1 (MEN1) is an hereditary dominant trait characterized by tumours of the parathyroids, anterior pituitary and endocrine pancreatic glands, among others. The MEN1 gene has recently been cloned, and MEN1-mutations have been identified in several families as well as in a number of sporadic cases. The aim of this study was to search for mutations in a large MEN1-family in order to define the clinical heterogeneity among mutation-carriers. We also analysed DNA from several tumour tissues in order to test the 'two hit' model for inactivation of the MEN1 gene. PATIENTS AND METHODS We searched for mutations in the MEN1-gene in members of a large MEN1-family. A total of 11 affected and 4 healthy at risk individuals were analysed. DNA was obtained and exons 1 to 10 of the MEN1-gene were PCR-amplified and subjected to automated-direct sequencing. In addition, we isolated DNA from parathyroid tumours of two family members, and compared this DNA with that of the normal tissue counterpart to define if the normal copy of the MEN1-gene was deleted. RESULTS G to A change at nucleotide 7640 (exon 10) that would convert Trp to Stop at codon 471 was found. This mutation was identified in eleven affected individuals, as well as in four healthy (asymptomatic) family members. These patients showed a wide spectrum of clinical symptoms and ages of presentation. Comparison of normal and tumour DNAs showed the loss of the normal (non-mutated) copy of MEN1 gene in the tumour tissue. CONCLUSION The different ages of disease presentation and the heterogeneity of symptoms among carriers of the Trp471Stop mutation, which would lead to the synthesis of a truncated non-functional protein, suggest that clinical heterogeneity is a characteristic of MEN1 independent of the type of mutation. Finally, the lack of amplification of the normal MEN1-allele on DNA from parathyroid tumours of two family members indicates that MEN1 is a tumour suppressor gene, the second hit that inactivates the normal copy in mutation carriers being a deletion.

Only a small percentage of long-term alcoholics develop liver cirrhosis. Genetic and non-genetic factors have been implicated in the risk of developing this disease. Among the genetic factors, case-control studies suggest an association with some polymorphisms at the alcohol dehydrogenase and cytochrome P450IIE1 genes. N-Acetyltransferase-2 metabolizes multiple compounds, transforming some of them to organ-toxic compounds and others into non-toxic molecules. Slow- and rapid-acetylator individuals exist in most human populations, and the mutations responsible for the slow-acetylator genotype have been determined. Slow acetylators, who should be at higher risk of developing breast cancer, and fast acetylators, who have an increased risk of developing colon cancer, can be characterized by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) genotyping. GSTM1 is another detoxifying enzyme for which functional interindividual differences, on the basis of gene polymorphisms, have been described.We conducted a case-control study in which 120 alcoholic cirrhotics, 30 long-term non-cirrhotic alcoholics, and 200 healthy controls were genotyped for polymorphisms (RFLPs) at the ADH2, P450IIE1, and NAT2 genes. PCR, followed by restriction enzyme digestion, was performed. The homozygous deletion of the GSTM1 gene was also PCR-analyzed. Genotype frequencies were statistically compared.Frequencies for the ADH2, P450IIE1, and GSTM1 polymorphisms did not differ between patients and controls. Individuals homozygous for the NAT2*5 allele, which is the most frequent slow-acetylator (SA) allele and shows the lowest acetylator activity, were at a significantly decreased frequency among the cirrhotic patients compared with controls (9% versus 16%; P = 0.042). The frequency of this SA genotype was significantly increased (40%) in long-term alcoholics who did not develop cirrhosis (P = 0.0041 compared with controls; P= 0.000017 compared with cirrhotics).According to our results, NAT2 activity may be a factor that determines the risk of developing alcoholic liver cirrhosis, and slow acetylators would be protected.

Objective: to examine the association between coronary artery disease and polymorphisms at the angiotensin-converting enzyme (ACE) and angiotensin II type 1 receptor (AT1R) genes. Methods : A total of 181 patients younger than 50 years and 240 controls from the same homogeneous Caucasian population (Asturias, Northern Spain) were genotyped (using polymerase chain reaction) for the ACE insertion/deletion (ACE-I/D) and the AT1R A/C transversion (AT1R-A/C) (3′-untranslated region) polymorphisms. Results : The DD-genotype was at a non-significant higher frequency among patients (50%) than in controls (41%). No difference between the two groups was found for the AT1R-genotypes. Distribution of ACE-genotypes according to AT1R-genotypes showed a significant association between ACE-DD and AT1R-CC and early coronary disease. Among the CC patients 58% were DD, compared to 21% among the controls ( p =0.02; OR=5.32, 95%CI=1.45, 19.51). We determined the distribution of these genotypes among the hypertensive and non-hypertensive patients. Frequencies of ACE- or AT1R-genotypes did not differ between the two groups. However, we found an interaction between the DD- and CC-genotypes in the group of normotensives. Among the CC patients, 13% of the hypertensives and 75% of the normotensives were DD ( p =0.014). Conclusions: Our results indicate a synergistic contribution of ACE and AT1R polymorphisms to the risk of coronary artery disease. This gene–gene interaction could have clinical implications. Approximately 2% of individuals in our population are CC+DD, and the genotyping of both polymorphisms could identify those with a high relative risk for coronary artery disease.

We analysed a group of Spanish intravenous drug users and controls to determine the role of mutations at the chemokine receptor-5/HIV-1 cofactor (CCR5), previously implicated in resistance to HIV-1 infection, and CXCR4 genes in susceptibility to HIV-1 infection. The complete coding sequence of both genes was amplified by the polymerase chain reaction from genomic DNA of 50 seropositive slow progressors and 10 long-term non-progressors, and analysed by the single-strand conformation polymorphism technique in a search for mutations. No mutation in CXCR4 was found, and delta ccr5 was the only mutation identified at the CCR5 gene. We genotyped (delta ccr5 allele) 150 HIV-1+ intravenous drug users and 250 healthy controls from the same population (Asturias, Northern Spain). Patients were divided into rapid progressors, presenting an event indicating progression to the acquired immunodeficiency syndrome (AIDS) in the 2 years after infection (100 patients), and slow progressors, remaining asymptomatic for 2-10 years (50 patients). The frequencies of the delta ccr5 allele were 0.105 and 0.040 in controls and HIV-1+ patients, respectively. Eighteen per cent of the controls (45/250) and 8% (12/150) of the patients carried the delta ccr-5 allele (P=0.013). The frequency of delta ccr5 carriers among rapid and slow disease progressors was 3 and 15%, respectively. A highly significant difference was found between rapid progressors and controls (P=0.0014). No patient (0/150) was delta ccr5 homozygous compared with 1% among controls. Thus, the delta ccr5 allele (the only CCR5 mutation found in our HIV-1 patients) was rare among seropositive intravenous drug users, suggesting that the absence of this mutation confers an advantage to the virus when infecting cells in vivo. In addition, patients carrying the delta ccr5 allele tend to show a slow progression towards HIV-1-related disease, remaining asymptomatic for longer periods of time.

The aim of this study is to investigate the association between three polymorphisms of the interleukin-1 (IL-1) gene complex and schizophrenia. We genotyped 228 outpatients with schizophrenia (DSM-IV criteria) and 419 unrelated healthy controls. The following polymorphisms were analyzed: IL-1α −889 C/T, IL-1β +3953 C/T, and IL-1RA (86 bp)n. No significant differences in genotype or in allelic distribution of the Il-1α, IL-1β, and IL-1RA polymorphisms were found. Estimated haplotype frequencies were similar in both groups. Our data do not suggest that genetically determined changes in the IL-1 gene complex confer increased susceptibility for schizophrenia. © 2005 Wiley-Liss, Inc.

Matrix-metalloproteases (MMPs) and their tissue-inhibitors (TIMPs), modulated by different single nucleotide polymorphisms (SNPs), are critical in sepsis development. Ninety ICU severely septic and 91 ICU uninfected patients were prospectively studied. MMP-1 (−1607 1G/2G), MMP-3 (−1612 5A/6A), MMP-8 (−799 C/T), MMP-9 (−1562 C/T) and MMP-13 (−77A/G) SNPs were genotyped. Plasma MMPs (-1, -2, -3, -8, -9, -10, -13) and TIMPs (-1,-2,-4) were measured. AA homozygotes and A allele carriers of MMP-13 (−77 A/G) and 1G2G carriers of the MMP-1 (−1607 1G/2G) SNPs frequencies were different between septic and uninfected patients (p < 0.05), as well as plasma MMP-3, -8, -9 -10 and TIMP-2 levels (p < 0.04). No differences in MMPs levels among MMP-13 or MMP-1 SNPs genotypes carriers were observed. The area under the ROC curve for MMP-8 in the diagnosis of sepsis was 0.87 (95% CI 0.82–0.92) and that of CRP was 0.98 (0.94–0.998), whereas the area of MMP-9 in the detection of non-septic state was 0.73 (0.65–0.80), p < 0.0001 for all curves. Sepsis associated with increased MMP-8 and decreased MMP-9 levels in multivariate analysis (p < 0.0002). We report for the first time an association between MMP-13 and MMP-1 SNPs and sepsis. An independent association of MMP-8 and MMP-9 levels with sepsis was also observed.

Mutations in different genes encoding sarcomeric proteins are responsible for 50-60% of familial cases of hypertrophic cardiomyopathy (HCM); however, the genetic alterations causing the disease in one-third of patients are currently unknown. Here we describe a case with familial HCM of unknown cause. Whole-exome sequencing reveals a variant in the gene encoding the sarcomeric protein filamin C (p.A1539T) that segregates with the disease in this family. Sequencing of 92 HCM cases identifies seven additional variants segregating with the disease in eight families. Patients with FLNC mutations show marked sarcomeric abnormalities in cardiac muscle, and functional analysis reveals that expression of these FLNC variants resulted in the formation of large filamin C aggregates. Clinical studies indicate that FLNC-mutated patients have higher incidence of sudden cardiac death. On the basis of these findings, we conclude that mutations in the gene encoding the sarcomeric protein filamin C cause a new form of familial HMC.

This work studies apple juice concentration by reverse osmosis (RO) using polyamide tubular membranes at different operating conditions. Permeate flux has been predicted by using the solution-diffusion model combined with the film model. Good agreement was found between the experimental values and the ones calculated by using both models. Pressure was found to be the most important variable controlling the process. Physico-chemical analyses were made in order to evaluate the quality of concentrated apple juice.

Autosomal recessive mutations in the parkin gene are the predominant cause of familial, early-onset parkinsonism; missense mutations involving one or a few nucleotides, exonic deletions and duplications have been described. Here we report a family with two affected brothers. Direct sequencing of parkin did not detect mutations, but semi-quantitative analysis identified a novel exonic rearrangement of exons 2-4. Both patients were homozygous for unique genomic triplications of the parkin gene.

Mutations in the alpha-synuclein ( SNCA ) gene cause autosomal dominant Parkinson's disease (PD). Common SNCA polymorphisms have been associated with the risk of developing PD. Abnormal expression and post-translational modification of SNCA has been found in PD-brains. In addition to a full length transcript ( SNCA -140) there are three short isoforms ( SNCA -98, -112, and -126) that could be prone to aggregation. The association between SNCA polymorphisms and PD could be explained through an increased expression of these alternative transcripts. Our aim was to measure the different SNCA transcripts in the substantia nigra (SN), cerebellum (CB), and occipital cortex (OC) from PD-patients ( n = 9) and healthy subjects ( n = 6). In addition, we determined whether two SNCA polymorphisms (SNPs rs356165 and rs11931074) were related to differences in transcript isoform expression. PD brain tissues showed higher levels of the three short transcripts in the SN, but only SNCA -112 and SNCA -98 were significantly increased in the CB of patients vs. controls ( p = 0.02, p = 0.03). The genotyping of a large cohort of PD-patients and controls showed that haplotype rs356165-A + rs11931074-G had a protective effect (OR = 0.71; CI = 0.59–0.83), while the G-T haplotype increased the risk for PD (OR = 1.44; CI = 1.06–1.96). We did not find significant differences for the SNCA levels between the haplotypes. In conclusion, we found statistically significant higher levels of the SNCA -112 and SNCA -98 transcripts in the CB of PD brains, and a trend toward higher levels of the short transcript isoforms in the SN of PD brains.

DNA variants at the genes encoding cardiac channels have been associated with inherited arrhythmias and the QT interval in the general population. Next generation sequencing technologies would be of special interest to uncover the genetic variation at these genes. The amplification and sequencing of DNA pools (instead of single individuals) would facilitate the rapid and cost-effective screening of large amounts of individuals. However, this pooling strategy could result in a signal of the rare variants below the detection capacity. To validate this approach, a pool of 20 individuals with known rare unique variants in five genes was amplified in only two tubes and sequenced using the non optical semi-conductor (Ion Torrent PGM, Life Technologies) technology. We show that this could be an effective strategy for the screening of large cohorts. Among others, this would facilitate the discovery of new sequence variants linked to cardiac arrhythmia in the general population.

In recent years, endoscopic ultrasonography (EUS)-guided techniques have been developed as alternatives to surgical, radiologic, or conventional endoscopic approaches for the treatment or palliation of several digestive diseases. The use of EUS guidance allows the therapeutic area to be targeting more precisely, with a possible clinical benefit and less morbidity. Nevertheless, the risks persist and must be taken into consideration. This review gives an overview of the complications observed with the most established procedures of therapeutic EUS. The PubMed and Embase databases were used to search English language articles on interventional EUS. The studies considered for inclusion were those reporting on complications of EUS-guided celiac plexus block (EUS-CPB), EUS-guided celiac plexus neurolysis (EUS-CPN), drainage of fluid pancreatic and pelvic collections, and EUS-guided biliary and pancreatic drainage (EUS-BD and EUS-PD). Variations in methodology and design in most studies made a thorough statistical analysis difficult. Instead, a frequency analysis of complications and a critical discussion were performed. Although EUS-guided celiac plexus injection causes mainly mild and transient complications, growing experience shows that EUS-CPN is not as benign a procedure as previously thought. Most of the major complications have been observed in patients with chronic pancreatitis. The findings show that EUS-guided drainage of fluid collections is a safe procedure. Complications occur more often after the drainage of pancreatic abscesses and necrosis. Although the heterogeneity of studies dealing with pancreatobiliary drainage makes the evaluation of risks after these procedures difficult, complications after EUS-BD and EUS-PD are relatively frequent and can be severe. The technical complexity and the lack of specifically designed devices may account for their complication rates. Clinicians can consider EUS-guided celiac injection and EUS-guided drainage of fluid collections to be safe alternatives to surgical and radiologic interventions. Well-designed prospective trials are needed to assess the risks of EUS-BD and EUS-PD accurately before they are broadly advocated after a failed endoscopic retrograde cholangiopancreatography (ERCP).

The CYP3A5*3 and CYP3A4*1B alleles have been related with tacrolimus (Tac) dose requirements. The rare CYP3A4*22 variant has also been associated with a significantly lower Tac dose. We genotyped the three single-nucleotide polymorphisms in 206 kidney-transplanted patients who received Tac as the primary immunosuppressor. CYP3A5*1 and CYP3A4*1B allele carriers received a significantly higher Tac dose (P0.01) compared with wild-type homozygotes. We did not find significant differences between the CYP3A4*22 genotypes, either nominally or according to the CYP3A5 genotype (expressers vs. nonexpressers). Sequencing of CYP3A4 coding exons in a total of 15 patients revealed only one nonreported missense change (p.P227T) in one patient. We concluded that CYP3A5*3 and CYP3A4*1B were the main determinants of the Tac dose-adjusted blood concentration in our cohort of renal-transplanted patients.

4 páginas, 1 figura, a tabla. Los autores pertenecen a The dementia genetic Spanish consortium (DEGESCO)., A non-synonymous genetic rare variant, rs75932628-T (p.R47H), in the TREM2 gene has recently been reported to be a strong genetic risk factor for Alzheimer's disease (AD). Also, rare recessive mutations have been associated with frontotemporal dementia (FTD). We aimed to investigate the role of p.R47H variant in AD and FTD through a multi-center study comprising 3,172 AD and 682 FTD patients and 2,169 healthy controls from Spain. We found that 0.6% of AD cases carried this variant compared to 0.1% of controls (odds ratio [OR]=4.12, 95% confidence interval [CI]: 1.21-14.00, P=0.014). A meta-analysis comprising 32,598 subjects from four previous studies demonstrated the large effect of the p.R47H variant in AD risk (OR=4.11, 95% CI: 2.99-5.68, P=5.27x10-18). We did not find an association between p.R47H and age of onset of AD or family history of dementia. Finally, none of the FTD patients harbored this genetic variant. These data strongly support the important role of p.R47H in AD risk and suggest that this rare genetic variant is not related to FTD., This study was supported by grants from Instituto de Salud Carlos III (PI12/01311 and 12/00013), grants from the Ministry of Science (SAF2010-15558, SAF2009-10434), Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED, Spain), Consolider (CSD2010-00045), and the Department of Health of the Government of Navarra (refs. 13085 and 3/2008). CR held during the period 2009-2013 a “Torres Quevedo” fellowship from the Spanish Ministry of Science and Technology, co-financed by the European Social Fund. Fundació ACE researchers are indebted to Trinitat Port-Carbó and her family who are supporting Fundació ACE scientific programs.

Interleukin-8 is a chemokine with a potent neutrophil chemoattractant activity. In humans, two different cDNAs encoding human IL8 receptors designated IL8RA and IL8RB have been cloned. IL8RA binds IL8, while IL8RB binds IL8 as well as other alpha-chemokines. Both human IL8Rs are encoded by two genes physically linked on chromosome 2. The IL8RA and IL8RB genes have open reading frames (ORF) lacking introns. By direct sequencing of the polymerase chain reaction products, we sequenced the IL8R genes of cell lines from four non-human primates: chimpanzee, gorilla, orangutan, and macaca. The IL8RB encodes an ORF in the four non-human primates, showing 95% - 99% similarity to the human IL8RB sequence. The IL8RA homologue in gorilla and chimpanzee consisted of two ORF 98% - 99% identical to the human sequence. The macaca and orangutan IL8RA homologues are pseudogenes: a 2 base pair insertion generated a sequence with several stop codons. In addition, we describe the physical linkage of these genes in the four non-human primates and discuss the evolutionary implications of these findings.

The polymorphism of HLA-B27 alleles is located in the peptide-anchoring motif. In recent years, fundamental insights have been made into the molecular aspects of HLA-B27-restricted presentation. Subtle differences in peptide binding fine specificity are especially interesting for closely related HLA-B27 alleles that have differential association with ankylosing spondylitis. Bacterial infection has been suggested to play a role in the pathogenesis of HLA-B27-associated disease. Remarkable progress has been made in identifying peptides derived from bacteria that can be presented by HLA-B27. Despite the mechanisms proposed to explain B27-associated diseases, there are no clear correlations between peptide sequence, differential binding to B27 subtypes, and recognition by peptide-specific T cell receptors. Furthermore, new transgenic models have now been developed that we hope will allow a clearer view of the function of B27 and the mechanisms involved in the pathogenesis of spondyloarthropathies.

Obesity is a growing problem in developed countries, and surgery is the most effective treatment in terms of weight loss and improving medical comorbidity in a high proportion of obese patients. Despite the advances in surgical techniques, some patients still develop acute and late postoperative complications, and an endoscopic evaluation is often required for diagnosis. Moreover, the high morbidity related to surgical reintervention, the important enhancement of endoscopic procedures and technological innovations introduced in endoscopic equipment have made the endoscopic approach a minimally-invasive alternative to surgery, and, in many cases, a suitable first-line treatment of bariatric surgery complications. There is now evidence in the literature supporting endoscopic management for some of these complications, such as gastrointestinal bleeding, stomal and marginal ulcers, stomal stenosis, leaks and fistulas or pancreatobiliary disorders. However, endoscopic treatment in this setting is not standardized, and there is no consensus on its optimal timing. In this article, we aim to analyze the secondary complications of the most expanded techniques of bariatric surgery with special emphasis on those where more solid evidence exists in favor of the endoscopic treatment. Based on a thorough review of the literature, we evaluated the performance and safety of different endoscopic options for every type of complication, highlighting the most recent innovations and including comparative data with surgical alternatives whenever feasible.

The hereditary spastic paraplegias (HSPs), a group of neurodegenerative movement disorders, are among the genetically most heterogeneous clinical conditions. Still, the more than 50 forms known so far apparently explain less than 80% of cases. The present study identified two large HSP families, which seemed to show an autosomal recessive and an X-linked inheritance pattern. A set of genetic analyses including exome sequencing revealed plausible mutations only when assuming incomplete/sex-dependent penetrance of adjacent alterations in the autosomal dominant HSP gene ATL1 (c.1243C>T and c.1244G>A, respectively). By screening of additional HSP patients for the presence of these alterations, we identified three more cases and obtained additional evidence for reduced penetrance. Bisulfate sequencing and haplotype analysis indicated that c.1243C and c.1244G constitute a mutational hotspot. Our findings suggest that misinterpretation of inheritance patterns and, consequently, misselection of candidate genes to be screened in gene-focused approaches contribute to the apparently missing heritability in HSP and, potentially, in other genetically heterogeneous disorders.

The present study is aimed to develop a process for production of functionalized scaffolds impregnated with natural compounds extracted from Patagonian Usnea lichen. A setup for an integrated supercritical CO2 extraction of natural compounds with posterior impregnation on solid matrices (polycaprolactone, PCL) is developed and presented here. In order to establish optimized operating conditions, supercritical extraction of Usnea as well as sorption kinetics and resulting material properties have been studied separately first. Usnea extracts isolated by supercritical carbon dioxide at 30 MPa and 40 °C have shown strong antibacterial activity with values of the minimum inhibitory concentration (MIC) ranging from less than 1.25 μg/mL to 320 μg/mL against Listeria innocua and Methicillin-resistant Staphylococcus (MRS) strains. Useful scaffolds of PCL for tissue engineering containing a porous structure with pore diameters between 150 and 340 μm can be obtained when PCL is exposed to carbon dioxide at 35 °C and 15 MPa. The degree of crystallinity of functionalized PCL was shown to be influenced by the incorporated antibacterial agent. The presented results showed that the impregnated PCL samples are promising bactericidal compounds against L. innocua. Screening of antibacterial activity of functionalized PCL against a Methicillin-resistant Staphylococcus aureus (MRSA) strain showed a higher activity when a low bacterial inoculum level (2 × 104 Colony-forming Units/mL [CFU/mL]) was assayed. Fil: Fanovich, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigación en Ciencia y Tecnología de Materiales (i); Argentina; Universidad Nacional de Mar del Plata. Facultad de Ingenieria; Argentina; Fil: Ivanovic, J.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigación en Ciencia y Tecnología de Materiales (i); Argentina; University of Belgrade, Faculty of Technology and Metallurgy; Serbia; Fil: Misic, D.. University of Belgrade, Faculty of Veterinary Medicine; Seribia; Fil: Alvarez, María Victoria. Universidad Nacional de Mar del Plata. Facultad de Ingenieria; Argentina; Fil: Jaeger, P.. Hamburg University of Technology, Inst Thermal Process Engineering; Alemania; Fil: Zizovic, I.. University of Belgrade, Faculty of Technology and Metallurgy; Serbia; Fil: Eggers, Rudolf. Hamburg University of Technology, Inst Thermal Process Engineering; Alemania

Antimicrobial properties of chitosan (CH) coatings and CH enriched with bioactive compounds (BC) and essential oils (EO) were determined by in vitro and in vivo assays on minimally processed broccoli. The efficiency of CH plus BC/EO in improving the safety of broccoli was tested against the native microflora. Also, its effects on the survival of Escherichia coli and Listeria monocytogenes inoculated in broccoli were evaluated. In vitro assays performed in tea tree, rosemary, pollen and propolis demonstrated significant inhibitory effects on E. coli and L. monocytogenes counts while pomegranate and resveratrol presented reduced activity. In vivo application of these BC on broccoli exerted a bacteriostatic effect on mesophilic and psychrotrophic populations except for rosemary. The application of CH alone or enriched with BC/EO resulted in a significant reduction in mesophilic and psychrotrophic counts. Between days 5 and 7, significant reductions (2.5 log) were observed in samples treated with CH þ BC. The enrichment with BC improved the antimicrobial action of CH. The application of these coatings did not introduce deleterious effects on the sensory attributes of broccoli. CH coatings enriched with BC/EO were a good alternative for controlling not only the microorganisms present in broccoli, but also the survival of E. coli and L. monocytogenes. Fil: Alvarez, María Victoria. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ponce, Alejandra Graciela. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Moreira, Maria del Rosario. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The pathological hallmark of Parkinson’s disease (PD) is the loss of dopaminergic neurons and the presence of Lewy bodies and Lewy neurites in the substantia nigra pars compacta, corpus striatum and brain cortex [1]. PD is a complex disease caused by the interaction of genetic/ inherited and environmental/acquired risk factors [2]. MicroRNAs (miRNAs) are small RNAs that control gene expression by binding to the 30 UTRs of mRNAs [3]. Postmortem analysis of brain tissues and in vitro studies have identified several miRNAs implicated in PD. MiR-133b was shown to be downregulated in PD brains and to promote the survival of dopaminergic neurons [4]. MiR-433 was related with PD through targeting the FGF20, which in turn regulates the expression of a-synuclein [5]. MiR-7 targets a-synuclein and could regulate oxidative stress and cell death, while miR-184 and let-7 regulate dopaminergic neurons survival and activity [6, 7]. Recently, a miRNA profiling of PD brains identified early downregulation of miR-34b/c, modulating mitochondrial function in areas with pathological affectation [8]. Plasma (circulating) miRNAs have been proposed as biomarkers for several diseases and aging [9, 10]. Our aim was to characterize the plasma miRNA profile in PD patients and healthy controls, to determine its usefulness as a biomarker for PD. The study was approved by the Ethical Committee of Hospital Universitario Central de Asturias (HUCA) in accordance with the ethical standards of the Declaration of Helsinki and all the participants signed an informed consent. The study cohort consisted of sexand age-matched healthy controls (n = 25; mean age 67.6; 52 % males) and patients (n = 31; mean age 63.9; 55 % males) who fulfilled the PD-clinical diagnosis criteria [11]. None of the patients were receiving drugs for PD-treatment or had a diagnosis of cardiovascular or tumor disease. Full details of the experimental procedure are available as supplementary material. Briefly, blood was collected in tubes with EDTA, centrifuged and plasma was aliquoted (350 ll). A total of 2 pg of a synthetic Arabidopsis thaliana miRNA (Ath-miR-159a; 5 ll of a 0.4 pg/ll dilution) was immediately added and each aliquot stored at -80 C until use. Ath-miR-159a was used as control of the extraction process (supplementary material). Total plasma RNA was extracted (TRIzol LS Reagent, Ambion) and resuspended in 25 ll of RNAse-free water. Five ll of each sample were retrotranscribed (RT) with the Megaplex RT primers Human pool A and TaqMan microRNA Reverse transcription kit (Applied Biosystems). Three ll of the RT product were pre-amplified with the Megaplex Preamp primers Human pool A and TaqMan Universal Master Mix no AmpErase UNG (Applied Biosystems). All the preamplifications were assayed with a custom Ath-miR-159a Taqman assay in an ABI 7500 Real-Time PCR (Applied Electronic supplementary material The online version of this article (doi:10.1007/s00415-013-6900-8) contains supplementary material, which is available to authorized users.

Fil: Alvarez, Maria Victoria. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingenieria. Departamento de Ingenieria Quimica. Grupo de Investigacion en Ingenieria en Alimentos; Argentina

Background Tacrolimus (Tac) is mainly metabolized by cytochrome P450 3A isoenzymes. In a cohort of heart transplant recipients, we investigated the effect of CYP3A5, CYP3A4, and ABCB1/MDR1 polymorphisms on Tac dose requirements and the risk of developing new-onset diabetes after transplantation (NODAT). Methods A total of 65 heart transplant recipients were genotyped for 3 single nucleotide polymorphisms (SNPs) in the CYP3A5 (SNP rs776746), CYP3A4 (SNP rs2740574), and ABCB1 (SNP rs104564). The mean Tac dose values were compared between the genotypes. Results CYP3A5*3 homozygotes (nonexpressers; n = 55, 85%) received significantly higher Tac dose compared with CYP3A5*1 carriers (expressers). No different NODAT frequencies were found between the genotypes. Conclusions The CYP3A5 polymorphism was the main determinant of Tac dose requirements among heart transplant recipients. This common functional polymorphism had no influence on the risk of developing NODAT.

The MYH9 gene encodes a protein that is expressed in the kidney glomerular podocytes. MYH9 single nucleotide polymorphisms (SNPs) have been linked to the risk for chronic kidney disease (CKD) and end stage renal disease. Our aim was to determine whether MYH9 SNPs were associated with renal disease in Spanish Caucasians. The RENASTUR cohort consisted of 592 Spanish Caucasians, aged 55-85 years. They were genotyped for SNPs rs3752462 and rs4821480, which tagged haplotype E. The main values between individuals with a glomerular filtration rate (eGFR)

PRKN mutations have been linked to Parkinson’s disease (PD). Most of the mutational screenings have focused on the coding exons. The 3′ untranslated region (UTR) could also harbor functionally relevant nucleotide changes. We performed a mutational screening of PRKN in a cohort of early-onset PD patients (n = 235) from Spain. We found 16 mutations (five new): 16 patients (7 %) carried two mutations and only one mutation was found in 28 (12 %). Patients with two mutations had significantly lower mean age (30 ± 9 years) compared to patients with one (40 ± 7) or no mutation (42 ± 7). We found a total of 15 nucleotide variants (three new) in the 3′ UTR region. The frequency of carriers of the rare rs62637702 G allele (*94A/G) was significantly lower among the patients compared to healthy controls (n = 418) (0.03 vs. 0.004; p < 0.001), suggesting a protective role for this allele. In order to investigate the basal effect of this variant, we performed luciferase assays. No different basal activity was observed between the two alleles. In conclusion, the rs62637702 polymorphism was associated with PD. This could be a surrogate marker for disease risk, in linkage disequilibrium with other non-identified functional variant.