HW Equilibrium Research Articles

Association of R1939W and P1987R variants of Otoferlin (OTOF) gene with severe to profound nonsyndromic sensorineural hearing loss in Pakistani subjects.

To find possible association of R1939W and P1987R variants of OTOF gene with severe to profound NSSHL in cochlear implant subjects. It was a case control study, conducted from June 2021 to February 2022, comprising 50 cases of severe to profound NSSHL who had received cochlear implant from ENT Department, CMH Rawalpindi and 50 age-matched healthy controls from PEMH Rawalpindi. Blood samples were collected from all the subjects, followed by DNA extraction and allele-specific polymerase chain reaction, performed at Multi-disciplinary Laboratory of Department of Biochemistry and Molecular Biology, Army Medical College Rawalpindi. Statistical analysis was done using 'SPSS' and 'XLSTAT', followed by genetic analysis using 'SNPstat'. Mean age of the cases was 5.96 ± 4.62 years (N=50), comprising 58% males and 42% females. All had bilateral and prelingual HL. Parental consanguinity was 72%, whereas 62% cases had a positive family history of deafness. Alleles of R1939W and P1987R were not associated with NSSHL, as shown by their p values of 0.56 and 0.89 respectively. For R1939W ORs were 0.71 (dominant model) and 0.80 (overdominant model), indicating negative association with NSSHL. Regarding P1987R OR was 0.96 (log-additive model). Genotypes of both variants were not in HW Equilibrium (p <0.0001), whereas their alleles showed high LD (D'=0.92). High percentage of parental consanguinity was observed among cochlear implant candidates. The OTOF variants R1939W and P1987R were found to have protective roles against NSSHL in study population.

Population genetic structure of turbot (Scophthalmus maximus L., 1758) in the Black Sea.

Turbot, Scophthalmus maximus, is a commercially important demersal flatfish species distributed throughout the Black Sea. Several studies performed locally with a limited number of specimens using both mitochondrial DNA (mtDNA) and microsatellite markers evidenced notable genetic variation among populations. However, comprehensive population genetic studies are required to help management of the species in the Black Sea. In the present study eight microsatellite loci were used to resolve the population structure of 414 turbot samples collected from 12 sites across the Black Sea. Moreover, two mtDNA genes, COI and Cyt-b, were used for taxonomic identification. Microsatellite markers of Smax-04 and B12-I GT14 were excluded from analysis due to scoring issues. Data analysis was performed with the remaining six loci. Loci were highly polymorphic (average of 17.8 alleles per locus), indicating high genetic variability. Locus 3/20CA17, with high null allele frequency (>30%), significantly deviated from HW equilibrium. Pairwise comparison of the FST index showed significant differences between most of the surveyed sampling sites (P < 0.01). Cluster analysis evidenced the presence of three genetic groups among sampling sites. Significant genetic differentiation between Northern (Sea of Azov and Crimea) and Southern (Turkish Black Sea Coast) Black Sea sampling sites were detected. The Mantel test supported an isolation by distance model of population structure. These findings are vital for long-term sustainable management of the species and development of conservation programs. Moreover, generated mtDNA sequences would be useful for the establishment of a database for S. maximus.

MALDI–TOF-MS-based high throughput genotyping of mutations associated with body measurement traits in cattle

Exploration of genes in relation to body measurement traits through large-scaled mutation identification is highly conductive for the genomics-assisted breeding of superior productivity cattle. In this investigation, 31 objective mutations were genotyped synchronously in 384 yellow cattle of 8 breeds through the application of optimized MALDI-TOF-MS and multiplex PCR techniques. High genotyping rate was obtained as well as greatly decreased cost which was below one thirtieth of the routine analysis. Results from genotyping revealed 23 mutations as valid mutations in the studied cattle population with gene heterozygosity and effective allele number varying from 0.0052 to 0.4998 and 1.0052 to 1.9991, respectively. Among the 23 effective mutations, 12 was classified as moderate polymorphism (0.25 < PIC < 0.5) while the other 11 belonged to low polymorphism (PIC < 0.25), 7 mutations did not obey the HW equilibrium (p < 0.05) and linkage mainly appeared between mutations of UCP2 and PTHR1 genes. Furthermore, 8 body measurement traits in the 384 cattle were recorded to validate their association with tag mutations, and significant correlations were found in 12 mutations of 9 genes including PTHR1, CDK6, IHH, HHIP, GHRL, COL1A1, INS, GDF5 and UCP2, of which, PTHR1 was proved to be the most potential contributor to bone modeling in cattle. Results highlight the potential application value of 12 novel mutations in enhancing cattle production traits as well as the high genotyping rate achieved by MALDI-TOF-MS coupled with multiplex PCR technique.

Türkiye’de yetiştirilen dört keçi populasyonunda Sinir Büyüme Faktörü (NGF) geninde SacII ve MboII polimorfizmlerinin belirlenmesi

Nerve Growth Factor Gene (NGF) has important functions in the maintenance of viability and proliferation of ovarian cells. Recent studies on caprine NGF gene revealed two SNPs (determined by SacII/C291A and MboII/A705G polymorphisms) related with body length and litter size in goats. This study aimed to determine SacII and MboII polymorphisms of NGF in native Turkish goat populations including Turkish Hair (HAI), Kabakulak (KBK), Honamlı (HNM) and Norduz (NRD) by using PCR-RFLP. Amplificated PCR fragments of 808 were digested with SacII, while amplificated PCR fragments of 319 bp were digested with MboII restriction enzyme in order to detect different genotypes of NGF gene. The “A” allele frequency ranged from 0.500 (HAI, HNM and KBK) to 0.950 (NRD), while the “C” allele frequency ranged from 0.025 (NRD) to 0.500 (HAI, HNM and KBK) in NGF/SacII polymorphism. The “A” allele frequency ranged from 0.213 (HAI) to 1.000 (NRD), while the “G” allele frequency ranged from 0.000 (NRD) to 0.787 (HAI) in NGF/MboII polymorphism. Deviation from HW equilibrium was significant in HNM goat population (P &amp;lt; 0.05). In this study, polymorphisms of caprine NGF gene in native Turkish goat populations were revealed for the first time. The results obtained from this study showed that NGF/SacII polymorphisms could be used for body length in NRD population while NGF/MboII polymorphisms could be used for litter size in HAI, HNM and KBK populations in MAS studies.

Allele and Genotype Frequencies of the Κappa-Casein (CSN3) Locus in Macedonian Holstein-Friesian Cattle

Abstract The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P&lt;0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.

Abstract 23: BDNF val 66 met Genotype is Associated With Greater Brain Atrophy After Stroke

Background: Genetic factors may be useful to understand differences in outcomes post-stroke. We studied an imaging measure of brain atrophy in relation to two genotypes that may be associated with differences in stroke recovery, the val 66 met genotype for brain-derived neurotrophic factor (BDNF) and the ApoE e4 genotype. Each genotype has been associated with brain atrophy in non-stroke populations. The current study hypothesized that each genotype is associated with brain atrophy after stroke. Methods: The ICARE Study examined behavioral outcomes after stroke in relation to motor therapies. Of 361 ICARE enrollees, genetic and neuroimaging data were available in 127. The volume of the ventricles and the brain were measured, and brain atrophy expressed as the Ventricle-Brain Ratio (VBR). For MRI scans, VBR was first extracted from the scans using ALVIN then manually refined. For CT scans, VBR was extracted manually. All VBR measurements were verified by a neuroimaging expert. VBR was examined in relation to the two genotypes of interest, BDNF val 66 met and ApoE e4, controlling for the 3 covariates employed in ICARE: motor deficits, enrollment site, and time post-stroke at study entry. Results: There were 61 MRI and 66 CT scans. These were acquired 5±11 days post-stroke, which did not vary according to either genotype. Median ventricle volume=26.1 cc; brain volume=1,163 cc; and VBR=0.024. The BDNF val 66 met genotype was present in 23 subjects; ApoE e4, in 41; both were in HW equilibrium. Presence of the BDNF val 66 met genotype was associated with greater atrophy: median VBR increased 1.97-fold when this genotype was present vs. absent (p=0.01), controlling for above covariates; and remained significant (p=0.04) when also controlling for age. This BDNF gene-imaging correlate did not extend to gene-behavioral findings. The ApoE e4 genotype was not related to VBR (p=0.88). Conclusions: Median degree of brain atrophy is 97% greater in patients with stroke when the BDNF val 66 met genotype is present, at least for the mild-moderately impaired subjects enrolled in ICARE. Understanding the biology of inter-subject differences in brain anatomy after stroke can provide insights into patient heterogeneity and inform efforts to individualize stroke recovery therapies.

Genetic Variations at 15 Forensically Relevant Microsatellite Loci (STRs) in the Three Major Ethno‐Linguistic Population Groups in Nigeria

This study is to identify genetic markers that will show distinct genetic variability among the three major ethno‐linguistic population groups in Nigeria that can help establish ethnic origin of forensic evidential materials. Samples from 315 unrelated individuals representing the three populations; ‘Hausa’, ‘Ibo’ and ‘Yoruba’ were analyzed using 15 Microsatellite loci (STRs) from Applied Biosystems. Analyses of inter‐ and intra‐population diversity were carried out using average heterozygosity, allele range (base pair), most frequent allele per a locus, Fixation index (Fst), Forensic and Paternity parameters. Multi‐locus genotype frequencies were utilized for testing conformity with Hardy‐Weinberg equilibrium (HW). 7 loci was found to conform to Hardy Weinberg equilibrium after X2 analysis. However, Bonferoni corrections show all loci to be in HW equilibrium, equally, exact test showed all loci to be in conformity with HW. Forensic and paternity parameters across the loci showed adequate discriminatory power for forensic identification of individuals as well as sufficient exclusionary power for parentage analysis. Average heterozygosity Fixation Index and Nei's standard genetic distance gave the least distance between the Ibo and Yoruba ethic groups. Allele frequency distribution comparisons at all loci analyzed show substantially no significant difference at 5% level of confidence between the Nigerian population and the FBI African American population allele frequencies. Ethnically blinded test from twenty samples among the three ethnic groups show a high degree of certainty in ethnic origin inferences from unknown samples. The study show Genetic variations between the three populations with marked phylogenetic relationships.Support or Funding InformationUniversity of Lagos, Lagos Nigeria

POLYMORPHISM STEAROYL-COA DESATURASE (SCD) GENE AND ASSOCIATON WITH CHARACTERISTICS MEAT IN BALI CATTLE

Stearoyl-CoA desaturase (SCD) is an enzyme produced by SCD gene which is responsible for a conversion of saturated fatty acids (SFA) to mono-unsaturated fatty acid (MUFA) in adipose tissue. This enzyme affects the fats in intramuscular so having influence on marbling. The purpose of this study was to obtain the polymorphisms of the SCD gene and their associations with meat quality traits in Bali cattle. The number of samples used were 48 heads of cattle consisted of 24 bulls and 24 cows from BPTU-HMT Bali cattle in the province of Bali. The SCD gene has been amplified using forward primer 5’-ACC CCT TGG TGT GTG GTT GTT CTT C-3 ‘and reverses primer 5'-CCT GAC GAT ACT ATG TTT CTA CTT C-3'. The polymorphisms of the SCD gene were identified by direct sequencing method. Meat quality traits such as thick of longissimus dorsi (TLD), thick of back fat (TBF), thick of fat rump (TFR), thick of rump (TR), marbling score (MS), and the percentage of intramuscular fat (PIMF) were analyzed using the Veterinary Ultrasound Scanner. To determine Hardy-Weinberg equilibrium status, both allele and genotype frequencies were analyzed using GENEPOP program (V3.2). Association of the SCD gene SNP and meat quality traits was analyzed by GLM. This result showed that there were 5 monomorphic SNPs (c.10153A&gt;G, c.10318C&gt;A, c.10329C&gt;T, g.10394G&gt;A, g.10486A&gt;C) and 3 polymorphic SNPs (g.10360G&gt;A, g.10428C&gt;T, g. 10487G&gt;A) were in HW equilibrium. Association analysis showed that g.10428C&gt;T SNP significantly affected marbling score (MS) and percentage of intramuscular fat (PIMF) (P&lt;0.05). Based on these results, g.10428C&gt;T SNP of the SCD gene may be used as a candidate marker to select meat quality traits in Bali cattle.

P022 HLA-DRB1 and HLA-DQA1/B1 alleles in association with leprosy compared to household contacts

Aim The aim of this study was to investigate the association between HLA-DRB1 and DQA1 / DQB1 genetic polymorphisms in leprosy patients and their domiciliary contacts from Southern Brazil. Methods This study was conducted with 183 leprosy patients and 238 healthy controls that had prolonged contact with the patient but are not blood relatives. The polymorphisms DRB1 and DQA1 / DQB1 genes were analyzed by PCR-SSOR, using the Luminex® technology. The DQA1 / DQB1 and HLA-DRB1 (High Resolution) genotypes were used LABType® SSO Kit. Statistical analysis was performed using the Chi-square with the Yates’s or Fisher’s exact test, and Bonferroni correction ( Pc ) was also applied. This study was approved by the Ethics Committees. Results The sample evaluated was in HW Equilibrium. The allelic group frequencies showed that DQA1 ∗ 03, DQB1 ∗ 03 and DRB1 ∗ 04 were significantly higher in the control group than in patients leprosy per se ( DQA1 ∗ 03 = 17.2% vs. 11.8%, OR = 0.6, 95% CI 0.4–0.95, P = 0.026, Pc = 0.16), ( DQB1 ∗ 03 = 33.2% vs. 26.5%, OR = 0.7, 95% CI 0.5–0.98, P = 0.043, Pc = 0.25) and ( DRB1 ∗ 04 = 14.9% vs. 8.2%, OR = 0.5, 95% CI 0.3–0.8, P = 0,003, Pc = 0.046), respectively. A protective association was also observed between the multibacillary (MB) clinical form and the DRB1 ∗ 04 and DQA1 ∗ 03 alleles, while DQA1 ∗ 1 was associated with susceptibility (47.3% vs. 39.5%, OR = 1.4, 95% IC 1.0–1.8, P = 0.038, Pc = 0.232). The DRB1 ∗ 14- DQA1 ∗ 01- DQB1 ∗ 06 haplotype was associated with susceptibility to leprosy per se (1.1% vs. 0%, OR = 11.8, 95% IC 1.95 – undefined, P = 0.03, Pc = 0.39) and to the MB clinical form (1% vs. 0%, OR = 11.3, 95% IC = 1.4 – undefined, P = 0.043, Pc = 0.56). In addition, DRB1 ∗ 15:02 and DQB1 ∗ 06:01 alleles were associated with susceptibility to leprosy per se and to the MB form. On the other hand, DQA1 ∗ 03:01 (1.4% vs. 5.0%, OR = 0.3, 95% IC = 0.9–0.7, P = 0.004, Pc = 0.03) and DQB1 ∗ 03:02 (4.4% vs. 10.5%, OR = 0.4, 95% IC = 0.2–0.7, P = 0.001, Pc = 0.009) suggests a protective association to leprosy per se and the MB form compared to controls. Conclusion This study confirmed that polymorphisms in DQA1/DQB1 and DRB1 genes may influence the development/protection of leprosy and also have some influence in clinical forms of the disease.

Forensic-paternity effectiveness and genetics population analysis of six non-CODIS mini-STR loci (D1S1656, D2S441, D6S1043, D10S1248, D12S391, D22S1045) and SE33 in Mestizo and Amerindian populations from Mexico

Background: STRs are powerful tools intensively used in forensic and kinship studies.Aim: In order to assess the effectiveness of non-CODIS genetic markers in forensic and paternity tests, the genetic composition of six mini short tandem repeats—mini-STRs—(D1S1656, D2S441, D6S1043, D10S1248, D12S391, D22S1045) and the microsatellite SE33 in Mestizo and Amerindian populations from Mexico were studied.Subjects and methods: Using multiplex polymerase chain reactions and capillary electrophoresis, this study genotyped all loci from 870 chromosomes and evaluated the statistical genetic parameters.Results: All mini-STRs studied were in agreement with HW and linkage equilibrium; however, an important HW departure for SE33 was found in the Mestizo population (p ≤ 0.0001). Regarding paternity and forensic statistical parameters, high values of combined power discrimination and mean power of exclusion were found using these seven markers. The principal co-ordinate analysis based on allele frequencies of three mini-STRs showed the complex genetic architecture of the Mestizo population.Conclusion: The results indicate that this set of loci is suitable to genetically identify individuals in the Mexican population, supporting its effectiveness in human identification casework. In addition, these findings add new statistical values and emphasise the importance of the use of non-CODIS markers in complex populations in order to avoid erroneous assumptions.

Estudio preliminar de la diversidad genética de perros con fenotipo Poodle en Colombia usando microsatélites

El objetivo de este estudio fue estimar la variabilidad genética de una muestra poblacional de perros con fenotipo Poodle en Bogotá - Colombia. La variabilidad genética de 81 perros con fenotipo Poodle se estudió mediante el genotipado de cinco loci de microsatélites. La heterocigosidad esperada (HE) para los loci: AHTK253, CXX279, INU030, INU055 y REN162C04 fue de 0,747, 0,788, 0,767, 0,679 y 0,751, respectivamente. La HE para el conjunto de la población fue de 0,746. La población de Poodles analizada tuvo una alta diversidad genética y se encontró que no existe una subestructuración poblacional, hecho que desvía el equilibrio HW; con lo que demuestra que la población de Poodle colombiana sea una perfecta candidata para estudios de asociación genética, y de conservación de la biodiversidad de los animales domésticos.

Abstract 18449: Genetic Variants for Katp Channels as Protective Factors in Ischemic Heart Disease Susceptibility: Gender Differences

Background: Coronary artery disease has been widely considered a “man’s disease” and not a major concern for women. However, scientists have been trying to unlock the female physiology of ischemic heart disease (IHD). In fact, women are more prone than men to microvascular coronary dysfunction. Recently, we proposed that genetic polymorphisms (SNP) of coronary ion channels may be involved in IHD susceptibility. Our published data showed that the genotype distribution of SNP rs5215 for Kir6.2 subunit of ATP-dependent potassium (KATP) channels moderately deviated from the HW equilibrium and that rs5215_GG is correlated with IHD susceptibility. In particular, we described that rs5215_GG genotype of Kir6.2 might have a ‘‘protective’’ role in IHD genesis. Aim and methods: In order to better describe the role of SNP rs5215_GG in IHD susceptibility in woman, we conducted a gender difference in term of prevalence of rs5215_GG on our previous population, i.e. 242 patients with acute coronary syndrome or stable angina, of which 155 with CAD, 46 with microvascular dysfunction and 41 with anatomically and functionally normal coronary arteries. Results: Among 242 patients, rs5215_GG was found to be more frequent in women (16/82; 19.51%; p=0.0450) compared to men. In particular, rs5215_GG was more frequent in female patients with anatomically and functionally normal coronary arteries (8/24; 33.3%; p=0.0428) compared with women with CAD, whereas there was no difference between men in normal and CAD groups. Discussion and conclusion: The polymorphism rs5215_GG is a missense SNP (ATC-GTC) that results in the substitution of isoleucine (I) residue with valine (V) in the Kir6.2 subunit of KATP channels. Our data validate the hypothesis that rs5215_GG could reduce susceptibility to IHD. In fact, rs5215_GG was confirmed to be present more frequently in woman presenting anatomically and functionally normal coronary arteries. Our results suggest that coronary KATP channels expressing rs5215_GG play a protective key role against IHD. Our data endorse the prospect of associations between SNP encoding coronary ion channels and IHD, validating the idea that ion channels play in coronary homeostasis and that their polymorphisms act in IHD genesis.

Abstract 19305: Diabetes Mellitus and Ischemic Heart Disease Susceptibility: Which is the Influence of Genetic Polymorphisms for Katp Channels?

Background: Evidences show that diabetes mellitus (DM) lead to higher risk for ischemic heart disease (IHD). Recently, we proposed that genetic polymorphisms (SNP) of coronary ion channels might be involved in IHD. We showed that the genotype distribution of SNP rs5215 for Kir6.2 subunit of ATP-dependent potassium (KATP) channels moderately deviated from the HW equilibrium and that rs5215_GG is linked to IHD susceptibility: it might have a ‘‘protective’’ role in IHD genesis. Moreover, previous studies described that SNP rs5215_GG is strongly correlated with DM and hyperinsulinism in human. Aim and methods: In the present study, we hypothesized that SNP rs5215_GG for Kir6.2 subunit for KATP channels had some influences in the susceptibility of DM to IHD. We performed a subgroup analysis about the influence of rs5215_GG in diabetic patients on our previous population, i.e. 242 patients candidates to elective or emergency coronary angiography for acute coronary syndrome or chronic angina, of which 155 with CAD, 46 with microvascular dysfunction and 41 with anatomically and functionally normal coronary arteries (control group). Results: Among 242 patients, comparing subjects with DM to those without, there was not any significant difference in the prevalence of rs5215_GG. However, among 33 subjects without DM with normal coronary arteries, SNP rs5215_GG had a higher prevalence compared to those in the CAD group (10/33; 30.3%; p= 0.0277) as well as non-diabetic patients in the microvascular dysfunction group (9/33; 27.2%; p= 0.0295). Discussion and conclusion: The variant rs5215_GG results in the substitution of isoleucine (I) residue with valine (V) in the Kir6.2 subunit of KATP channels. Our results confirm that rs5215_GG might decrease susceptibility to IHD in patients without DM. In fact, rs5215_GG was confirmed to be present more frequently in patients without DM presenting anatomically and functionally normal coronary arteries. We believe that absence of DM is protective against IHD genesis and it might be improved by coronary KATP channels expressing rs5215_GG. Taken together, our data ratify the possibility that ion channels play a key role in coronary homeostasis and that SNP encoding for ion channels are involved in IHD pathophysiology.

Population genetics of the Federally Threatened Miccosukee gooseberry (Ribes echinellum), an endemic North American species

Ribes echinellum (Coville) Rehder (Miccosukee gooseberry; Grossulariaceae) is a Federally Threatened species known from only two localities: Jefferson County (Florida, FL) and McCormick County (South Carolina, SC). This perennial shrub, ca. 1 m tall, is deciduous, and reproduces both vegetatively (clonal growth) and sexually (seed production). Recent surveys of the FL population revealed a dramatic decline in plant numbers. To assist in conservation and management of this species in FL and SC populations, microsatellite genetic markers were used to identify genotypes and assess the genetic structure of R. echinellum. We genotyped seven microsatellite loci in 102 individuals: 74 collected in FL and 28 in SC. Unbiased heterozygosity was between 0.28 and 0.53. All seven loci were polymorphic, showing a range of 1.52–2.13 effective number of alleles per locus (mean = 1.75). The two populations of R. echinellum show low genetic diversity, especially in SC. Clonality was not widespread, but was higher in the SC population. Both populations show signatures of bottlenecks but isolation by distance was not evident. We found significant deviation from HW equilibrium, with higher number of heterozygotes than expected. However when HW test was done for the combined populations as two separate groups, only FL showed a significant HW test and for SC the test was non-significant. Bayesian analysis and FST values suggest high genetic divergence between the populations. These results are important for developing a recovery plan and an ex situ and reintroduction conservation programs.

Microsatellite markers unravel the population genetic structure of the Azorean Leontodon: implications in conservation

The genus Leontodon L. (Asteraceae) comprises approximately 50 species with a natural distribution area covering North America, Europe, northern Africa, and western Asia. Two of these species are endemic to the Azores Archipelago: Leontodon filii and Leontodon rigens. Although both species were targeted with several taxonomic revisions, so far no studies into their genetic diversity have been carried out. In this research, the population genetic structure and diversity of both taxa were assessed using five newly developed SSR markers. Four hundred and thirty-seven individuals collected throughout the archipelago were included in the study. A total of 98 alleles (25–12 per locus, average = 19.6) and an overall excess of homozygotes (multilocus Fis = 0.37, range 0.16–0.53) were found for L. rigens populations. For L. filii, 52 alleles in total (8–13 per locus, average = 10.4) were found, overall near the HW equilibrium (multilocus Fis = 0.07, range −0.25 to 0.57). The two species showed an equivalent proportion of rare alleles (L. rigens 80.6 %; L. filii 76.9 %). Both a Principal Coordinate Analysis and a Bayesian analysis proposed the existence of two well-defined groups, but pooled L. filii populations from Faial Island with L. rigens populations. The largest proportion of genetic variability was found within populations (L. rigens 72.6%; L. filii 78.9 %). The highest values of gene flow were obtained for L. filii within the central group of islands. Our results update the current distribution given for the Azorean Leontodon taxa, clearly indicating that conservation measures should be applied to several populations. The results also reveal that a revision of the Azorean Leontodon should be carried out to clarify species delimitation.

Association of CYP1A1 A4889G and T6235C polymorphisms with the risk of breast cancer in Brazilian women.

e11551 Background: Estrogen, activated by cytochrome P450 enzymes (CYP1A1), participates in the breast cancer (BC) development. G and C variant alleles of CYP1A1 A4889G and T6235C polymorphisms encode enzymes with increased activity in activation of this compound when compared with the respective wild alleles. This study aimed to clarify the roles of the polymorphisms in the risk of BC. Methods: CYP1A1 A4889G and T6235C genotypes of 742 sporadic breast cancer (SBC) patients (median age: 52 years, 638 Caucasians, 104 non-Caucasians) and 742 healthy women (median age: 40 years, 638 Caucasians, 104 non-Caucasians) were obtained in genomic DNA by PCR and enzymatic digestion. Results: Patient’s and control’s samples were in HW equilibrium at CYP1A1 A4889G (χ2= 0.15, P= 0.70; χ2= 1.15, P= 0.28) and T6235C (χ2= 2.65, P= 0.10; χ2= 1.93, P= 0.16) loci. The frequency of CYP1A1 4889AG+GG genotypes was higher in patients than in controls (29.0% versus 23.2%, P= 0.004). Carriers of the G allele were under a 1.50 (95% CI: 1.14-1.97)-fold increased risk for SBC than those with the wild AA genotype. The frequencies of 4889AG+GG genotypes in Caucasian SBC patients were higher than those found in Non-Caucasian (30.4% vs 20.2%, P= 0.03), and also in controls (30.4% vs 23.2%, P= 0.002). Carriers of G allele were under a 1.61 (95% CI: 1.20-2.15)-fold increased risk for SBC than others. A higher frequency of the CYP1A1 4889AG+GG genotypes was also seen in patients with an earlier median age at first full-term pregnancy (FFTP), compared to those who had the first pregnancy with a later medium age (33.8% vs 26.1%, P=0.03), and also when compared to controls (33.8% vs 23.2%, P= 0.001). Women with the CYP1A1 4889AG+GG genotypes and with earlier FFTP were under a 1.87 (95% CI: 1.32-2.67)-fold increased risk for SBC than others. The CYP1A1 6235CC genotype was more common in pre-obese or obese SBC patients than in underweight or normal SBC patients (6.8% vs 4.0%, P= 0.04). Conclusions: Our data suggest that CYP1A1 A4889G and T6235C polymorphisms alter BC risk in Brazilian women. We believe that women with variant alleles of the referred polymorphisms should receive additional medical attention for disease prevention and early diagnosis.

Su2028 Pinaverium Bromide Plus Symeticone Interacts With the IL-8 +396 GG Variant in the Improvement of Abdominal Pain in IBS. A Report From the Mexican IBS Study Group

Background: IBS is a functional gastrointestinal disorder with a high placebo response; however, Pinaveriumbromide 100mg + Symeticone 300mg bid (PB+S) has shown to improve the severity of abdominal pain and bloating.1-2 Recently, increased IL-8 concentrations have been found in intestinal segments of IBD compared with non-inflamed IBS mucosa,3 and according to our findings, lower IL-8 concentrations are present in unstimulated-cultured PBMCs in IBS vs. controls.4 Additionally, IBS has been associated with the IL-8 intronic variant GG at +396 (rs2227307).5 These results suggest that this cytokine may be related with an inflammatory variant of IBS. Aim: To analyze the association between improvement of abdominal pain in IBS patients treated with PB+S and IL-8 variant rs2227307. Methods: Two-hundred and seventy nine patients with active IBS were randomly allocated to PB+S (n=104) or placebo (n=109), in a 3-month double-blind clinical trial. A 10-cm VAS was used to evaluate the severity of abdominal pain. The rs2227307 was analyzed by QPCR. MANOVA analysis for repeated measurements adjusted by gender, age, BMI and IBS subtype, was performed. A priori multiplicative interaction was considered significant if p,0.20 and for main effects if p,0.05. Results: Mean (+/-SD) age of the population was 36 (+/-9) years and mean BMI 26.5 (+/-5.3). Allele frequencies were T=0.63, G=0.37 and genotypes TT= 38.7%, TG= 48.4% and GG=12.9%. The sample was in HW equilibrium (X2= 0.38, p= 0.54). Abdominal pain improved in patients with the three genotypes with a size effect of 34% (favoring PB+S p=0.004). The larger size effect was in those with GG (70.5%, interaction with treatment p=0.18). Conclusions: This study shows that patients with the rs2227307 do not have the same magnitude of response to placebo compared to those with the other genotypes. We hypothesized that an underlying inflammatory-variant of IBS may be more responsive to PB+S. An effect on mucosal immunological mediators and/or epithelial permeability needs to be elucidated. This study was supported by Takeda-Mexico.

Su2029 Risk Factors for Development of Irritable Bowel Syndrome Six Months After Gastroenteritis With Campylobacter Concisus: Role of Psychometric Scores At Baseline

Background: IBS is a functional gastrointestinal disorder with a high placebo response; however, Pinaveriumbromide 100mg + Symeticone 300mg bid (PB+S) has shown to improve the severity of abdominal pain and bloating.1-2 Recently, increased IL-8 concentrations have been found in intestinal segments of IBD compared with non-inflamed IBS mucosa,3 and according to our findings, lower IL-8 concentrations are present in unstimulated-cultured PBMCs in IBS vs. controls.4 Additionally, IBS has been associated with the IL-8 intronic variant GG at +396 (rs2227307).5 These results suggest that this cytokine may be related with an inflammatory variant of IBS. Aim: To analyze the association between improvement of abdominal pain in IBS patients treated with PB+S and IL-8 variant rs2227307. Methods: Two-hundred and seventy nine patients with active IBS were randomly allocated to PB+S (n=104) or placebo (n=109), in a 3-month double-blind clinical trial. A 10-cm VAS was used to evaluate the severity of abdominal pain. The rs2227307 was analyzed by QPCR. MANOVA analysis for repeated measurements adjusted by gender, age, BMI and IBS subtype, was performed. A priori multiplicative interaction was considered significant if p,0.20 and for main effects if p,0.05. Results: Mean (+/-SD) age of the population was 36 (+/-9) years and mean BMI 26.5 (+/-5.3). Allele frequencies were T=0.63, G=0.37 and genotypes TT= 38.7%, TG= 48.4% and GG=12.9%. The sample was in HW equilibrium (X2= 0.38, p= 0.54). Abdominal pain improved in patients with the three genotypes with a size effect of 34% (favoring PB+S p=0.004). The larger size effect was in those with GG (70.5%, interaction with treatment p=0.18). Conclusions: This study shows that patients with the rs2227307 do not have the same magnitude of response to placebo compared to those with the other genotypes. We hypothesized that an underlying inflammatory-variant of IBS may be more responsive to PB+S. An effect on mucosal immunological mediators and/or epithelial permeability needs to be elucidated. This study was supported by Takeda-Mexico.

Genetic characterization of Libyan date palm resources by microsatellite markers.

Molecular typing of 377 female date palm trees belonging to 18 Libyan cultivars and representing common genotypes in the central Libyan oasis of Al Jufrah was performed using 16 highly polymorphic microsatellite or SSR loci. A total of 110 alleles with an average of 6.88 alleles per locus were scored indicating the high level of polymorphism existing among the cultivars thus allowing their genetic fingerprinting. Moreover 28 alleles out of 110 were fixed. All the cultivars were characterized by negative values of the Fixation Index (F) due to an excess of heterozygotes with respect to HW equilibrium. The pattern of genetic diversity among cultivars was estimated by codominant genetic distances and presented by principal coordinates analysis (PCoA). The observed pattern evidences the genetic diversity existing among cultivars that allow distinguishing them easily. The average dissimilarity internal to each cultivar ranged from 0 to 21. Seven cultivars showed value zero indicating no genetic difference within cultivar in agreement with their Fixation Index (F = 1). A varietal identification key was also built using multiloci genotyping with only three microsatellite loci that identified 23 alleles in total. The possibility to attribute the unknown male plant to a cultivar was also considered and male parentage analysis was performed. Fifty-five male plants out of 63 were assigned to a definite cultivar with high confidence level. The positive result obtained in identifying males confirmed the suitability of SSR for clone fingerprinting and cultivar identification, thus opening new prospects for date palm breeding.

Genetic Characterization of Colombian Brahman Cattle Using Microsatellites Markers

Genetic structure and diversity of 3789 animals of the Brahman breed from 23 Colombian regions were assessed. Considering the Brahman Zebu cattle as a single population, the multilocus test based on the HW equilibrium, shows significant differences (P < 0.001). Genetic characterization made on the cattle population allowed to examine the genetic variability, calculating a H(o) = 0.6621. Brahman population in Colombia was a small subdivision within populations (F(it) = 0.045), a geographic subdivision almost non-existent or low differentiation (F(st) = 0.003) and the F(is) calculated (0.042) indicates no detriment to the variability in the population, despite the narrow mating takes place or there is a force that causes the variability is sustained without inbreeding actually affect the cattle population. The outcomes of multivariate analyses, Bayesian inferences and interindividual genetic distances suggested that there is no genetic sub-structure in the population, because of the high rate of animal migration among regions.