Candidate genes involved in cardiovascular risk factors by a family-based association study on the island of Kosrae,Federated States of Micronesia

Altered plasma levels of lipids and lipoproteins, obesity, hypertension, and diabetes are major risk factors for atherosclerotic cardiovascular disease. To identify genes that affect these traits and disorders, we looked for association between markers in candidate genes (apolipoprotein AII (apo AII), apolipoprotein AI-CIII-AIV gene cluster (apo AI-CIII-AIV), apolipoprotein E (apo E), cholesteryl ester transfer protein (CETP), cholesterol 7alpha-hydroxylase (CYP7a), hepatic lipase (HL), and microsomal triglyceride transfer protein (MTP)) and known risk factors (triglycerides (Tg), total cholesterol (TC), apolipoprotein AI (apo AI), apolipoprotein AII (apo AII), apolipoprotein B (apo B), body mass index (BMI), blood pressure (BP), leptin, and fasting blood sugar (FBS) levels.) A total of 1,102 individuals from the Pacific island of Kosrae were genotyped for the following markers: Apo AII/MspI, Apo CIII/SstI, Apo AI/XmnI, Apo E/HhaI, CETP/TaqIB, CYP7a/BsaI, HL/DraI, and MTP/HhpI. After testing for population stratification, family-based association analysis was carried out. Novel associations found were: 1) the apo AII/MspI with apo AI and BP levels, 2) the CYP7a/BsaI with apo AI and BMI levels. We also confirmed the following associations: 1) the apo AII/MspI with Tg level; 2) the apo CIII/SstI with Tg, TC, and apo B levels; 3) the Apo E/HhaI E2, E3, and E4 alleles with TC, apo AI, and apo B levels; and 4) the CETP/TaqIB with apo AI level. We further confirmed the connection between the apo AII gene and Tg level by a nonparametric linkage analysis. We therefore conclude that many of these candidate genes may play a significant role in susceptibility to heart disease.     

Genetic study of common variants at the Apo E,Apo AI,Apo CIII,Apo B,lipoprotein lipase (LPL) and hepatic lipase (LIPC) genes and coronary artery disease (CAD): variation in LIPC gene associates with clinical outcomes in patients with established CAD

Background  

Current evidence demonstrates that positive family history and several alterations in lipid metabolism are all important risk factors for coronary artery disease (CAD). All lipid abnormalities themselves have genetic determinants. Thus, objective of this study was to determine whether 6 genetic variants potentially related to altered lipid metabolism were associated with CAD and with lipid abnormalities in an Italian population. These genetic variables were: apolipoprotein E (Apo E), Apo AI, Apo CIII, Apo B, lipoprotein lipase (LPL) and the hepatic lipase (LIPC) genes. Furthermore, an 8 years prospective analysis of clinical cardiovascular events was related to the various genetic markers.     

Apolipoprotein AI and CIII gene polymorphisms and their association with lipid levels in Italian, Greek and Anglo-Irish populations of Australia

PRIMARY OBJECTIVE: The apolipoprotein (apo) AI-CIII-AIV gene cluster on chromosome 11 has been identified as a candidate region for hyperlipidaemia and in particular for hypertriglyceridaemia. Our aim was to detect associations between the apo AI and CIII polymorphisms and the plasma lipids, total cholesterol, triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol in normal, healthy, adults from three ethnic groups of Australia--Italian, Greek and Anglo-Irish, separately by gender. METHODS AND PROCEDURES: The SstI restriction fragment length polymorphisms (RFLP) in the 3' untranslated region of the apo CIII gene and the MspI RFLP in the third intron of the apo AI gene were scored and the lipid concentrations were ascertained using standard methodologies. t-tests were used to compare lipid levels between sexes and between populations, and multivariate ANOVA was used to detect if the two RFLPs had an effect on any of the lipid concentrations. MAIN OUTCOMES AND RESULTS: The two RFLPs exhibit strong linkage disequilibrium in all three populations (p < 0.001). There were some significant differences in allele frequencies among the populations: the minor S2 allele was more frequent in Italians (0.12) than Greeks (0.03) (p = 0.003), and the minor M2 allele was more common in Greeks (0.14) than Anglo-Irish (0.05) (p = 0.026). We found no significant association between either of the RFLPs and any of the lipid concentrations in either sex of all three populations. However, Kruskal-Wallis tests detected associations of borderline significance between apo AI MspI genotypes and triglycerides (p = 0.04) and between apo AI MspI genotypes and cholesterol levels (p = 0.03) in Anglo-Irish females. CONCLUSIONS: Because only two statistically significant associations were detected among a number of comparisons, our data suggest that the apo AI and CIII polymorphisms play only a very limited role in mediating variation in lipid concentrations in these three ethnic groups.     

A novel apolipoprotein E5 variant with a 24-bp insertion causing hyperlipidemia

A tandem 24-bp insertion in the apolipoprotein E (apo E) gene was detected in a patient with elevated triglyceride, apolipoprotein (apo) CII, and apo CIII levels. This novel variant, apo E5ss, showed in position apo E5 by isoelectric focusing and was of larger molecular weight than apo E3 during two-dimensional gel electrophoresis. Polymerase chain reaction-single strand conformation polymorphism analysis using the primer pairs that cover all the coding regions was useful for rapid detection of the variant of the apo E allele. Apo E5ss may have a 24-bp insertion caused by slipped mispairing, resulting in a tandem duplication of amino acid residues 135–142 [APOE, 24-BP INS, DUP CODONS 135–142]. The proband was the only person with apo E5ss among the 806 Japanese males that we examined. We inspected six other reported apo E5 variants in the literature. Received: July 10, 2001 / Accepted: August 8, 2001     

Amyloid substance within stenotic aortic valves promotes mineralization

Audet A, Côté N, Couture C, Bossé Y, Després J‐P, Pibarot P & Mathieu P
(2012) Histopathology  61, 610–619 Amyloid substance within stenotic aortic valves promotes mineralization Aims: Accumulation of apolipoproteins may play an important role in the pathobiology of calcific aortic valve disease (CAVD). We aimed to explore the hypothesis that apolipoprotein‐derived amyloid could play a role in the development of CAVD. Methods and results: In 70 explanted CAVD valves and 15 control non‐calcified aortic valves, we assessed the presence of amyloid by using Congo red staining. Immunohistochemistry was performed to document the presence of apolipoprotein AI (Apo‐AI). Apoptosis was documented by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) studies performed in control and CAVD valves. Control valves were free of amyloid. Deposition of amyloid was detected in all CAVD valves, and the amount was positively correlated with plasma high‐density lipoprotein and Apo‐AI levels. Apo‐AI within CAVD valves co‐localized with intense staining of fibrillar amyloid. In turn, deposition of amyloid co‐localized with apoptosis near mineralized areas. Isolation of amyloid fibrils confirmed that Apo‐AI is a major component of amyloid deposits in CAVD. In vitro, CAVD‐derived amyloid extracts increased apoptosis and mineralization of isolated aortic valvular interstitial cells. Conclusions: Apo‐AI is a major component of amyloid substance present within CAVD valves. Furthermore, amyloid deposits participate in mineralization in CAVD by promoting apoptosis of valvular interstitial cells.     

Molecular typing of Clostridioides difficile isolates from clinical and non‐clinical samples in Iran

Clostridioides difficile is a major cause of nosocomial infectious diarrhea in hospitalized patients throughout the world. We aimed to characterize C. difficile isolates among hospitalized patients, hospital staffs, and hospital environment samples obtained in three tertiary care hospitals of Iran with regard to their molecular types between June 2016 and November 2017. The toxigenicity of C. difficile isolates was determined by toxigenic culture and multiplex‐PCR. Toxigenic C. difficile isolates collected were ribotyped using capillary gel electrophoresis‐based PCR and the database of WEBRIBO ( http://webribo.ages.at ). Of 500 clinical and non‐clinical samples, toxigenic C. difficile were identified in 35 of 250 stool samples (14%) and in 3 of 250 swabs (1.2%). The most frequently found ribotypes (RTs) were 039, AI‐12, and AI‐21 (15.8, 10.52, and 10.52% of all isolates, respectively). Further RTs were: 017, 001, AI‐3, AI‐15, AI‐18, AI‐10, AI‐4, and PR21195 (as new ribotype). The epidemic RTs (027 and 078) seen in the Europe, North America, and Asia were completely absent in this study.     

Italian familial defective apolipoprotein B patients share a unique haplotype with other Caucasian patients

Familial defective apolipoprotein (apo) B-100 together with familial hypercholesterolemia are the two common genetic conditions that cause hypercholesterolemia. familial defective apolipoprotein B-100 is due to mutations around codon 3500 of the apo B gene. The most-characterized mutation is a G>A transition at nucleotide 10,708 that results in the substitution of arginine by glutamine at codon 3500 (Apo B Arg3500Gln). Two other mutations are caused by a C>T transition, one at nucleotide 10,800 (Apo B Arg3531Cys) and the other at nucleotide 10,707 (apo B Arg3500Trp). In the present study we describe three new Italian cases of familial defective apolipoprotein B-100 (Apo B Arg3500Gln), one from the Liguria region and two from Sicily, and the haplotype of the apo B gene co-segregating with the mutation. By screening two groups of probands, clinically diagnosed as having Familial Hypercholesterolemia (700 from mainland Italy and 305 from Sicily), the prevalence of familial defective apolipoprotein B-100 due to Arg3500Gln was found to be very low (0.28% and 0.65%, respectively). The Arg3531Cys mutation was not detected in any proband. In the three new families with Arg3500Gln mutation in the present study and in one previously described in Italy, the mutation was associated with a unique apo B haplotype, which is consistent with data previously reported for Caucasian patients [XbaI-, MspI+, EcoRI-, presence of the 5' signal peptide insertion (Ins)allele, and the 49-repeat allele of the 3'-VNTR]. Received: 27 February 2001 / Accepted: 24 September 2001     

Effects of apoE gene polymorphism on Lp(a) concentrations depend on the size of apo(a): a study of 466 white men

 Polymorphisms in the genes for the low-density lipoprotein (LDL) receptor ligands, apolipoprotein E (apoE), and apolipoprotein B (apoB) are associated with variation in plasma levels of LDL cholesterol. Lp(a) lipoprotein(a) [Lp(a)] is LDL in which apoB is attached to a glycoprotein called apolipoprotein(a) [apo(a)]. Apo(a) has several genetically determined isoforms differing in molecular weight, which are inversely correlated with Lp(a) concentrations in blood. The interaction of apo(a) with triglyceride-rich lipoproteins differs with the size of apo(a), and therefore the effects of apoE gene polymorphism on Lp(a) levels could also depend on apo(a) size. We have investigated the possible effect of genetic variation in the apoE and apoB genes on plasma Lp(a) concentrations in 466 white men with different apo(a) phenotypes. Overall there was no significant association between the common apoE polymorphism and Lp(a), but in the subgroup with apo(a)-S4, concentrations of Lp(a) differed significantly among the apoE genotypes (P=0.05). Lp(a) was highest in the apoE genotypes ɛ₂ɛ₃ and ɛ₃ɛ₃ and lowest in genotype ɛ₃ɛ₄, and the apoE polymorphism was estimated to account for about 2.4% of the variation in Lp(a). In contrast, in the subgroup with apo(a)-S2 Lp(a) was significantly lower (P=0.04) in apoE genotype ɛ₂ɛ₃ than in genotype ɛ₃ɛ₃. Lp(a) concentrations did not differ among the XbaI (P=0.65) or SP 24/27 (P=0.26) polymorphisms of the apoB gene. The expected effects of both apoE and apoB polymorphism on LDL levels were significant in the whole population sample and in subjects with large-sized apo(a) isoforms (P<0.01), whereas no effect was seen in those with low molecular weight apo(a) isoforms. We conclude that the influence of apoE genotypes on Lp(a) concentrations depends on the size of the apo(a) molecule in Lp(a), possibly because both apo(a)-S4 and apoE4 have high affinity for triglyceride-rich lipoproteins and may be taken up and degraded rapidly by remnant receptors. Received: 12 January 1995 / Accepted: 12 July 1996     

Frequencies of five genetic polymorphisms in coronarographed patients and effects on lipid levels in a supposedly healthy population

Allele frequencies of genetic polymorphisms were compared between supposedly healthy subjects and angiographically proven coronary artery disease patients. The polymorphic candidate loci investigated were the apolipoprotein (apo) B signal peptide and Xba I polymorphisms, the apo E polymorphism and two polymorphisms of lipoprotein lipase (LPL) gene: Hind/ III and PvuII. Apo B signal peptide and Hind III/LPL polymorphisms showed significant differences in allele partition between cases and controls; the rare alleles of both polymorphisms were less frequent (p<0.05) in cases. We looked for associations between the polymorphisms and lipid concentration variability in a supposedly healthy population (145 men and 144 women). Apo B signal peptide, apo E and Pvul II/LPL polymorphisms seem to influence some lipid metabolism parameters significantly. Apo AI and LpCIII levels were significantly different among apo B signal peptide genotypes: Del homozygotes had the highest concentrations of both variables. The e4 allele of apo E polymorphism was associated with increased concentrations of total cholesterol, LDL cholesterol and apo B. Increased LpAI:AII levels observed in E3 homozygotes (p<0.01) have not previously been reported. LpAI:AII concentration was also influenced by Pvu II/LPL polymorphisms.     

Antimicrobial activity of different Finnish monofloral honeys against human pathogenic bacteria

The antimicrobial activity and phenolic compounds of five Finnish honey products against important human pathogens Streptococcus pneumoniae, S. pyogenes, Staphylococcus aureus, and methicillin‐resistant S. aureus were analyzed. Microbroth dilution method and HPLC‐DAD were used in antimicrobial testing and phenolic compound determination, respectively. Significant antimicrobial activity (p < 0.01) against all the tested pathogens was found from willow herb (Epilobium angustifolium), heather (Calluna vulgaris), and buckwheat (Fagopyrum esculentum) honeys. This is the first report on antimicrobial activity of Finnish monofloral honeys against streptococcal and staphylococcal bacteria. To our knowledge this is also the first report on the antimicrobial effect of honey against S. pneumoniae.     

Effects of MASP2 haplotypes and MASP‐2 levels in hepatitis C‐infected patients

Mannan‐binding lectin (MBL) and MBL‐associated serine protease 2 (MASP‐2) are components of the lectin pathway, which activate the complement system after binding to the HCV structural proteins E1 and E2. We haplotyped 11 MASP2 polymorphisms in 103 HCV patients and 205 controls and measured MASP‐2 levels in 67 HCV patients and 77 controls to better understand the role of MASP‐2 in hepatitis C susceptibility and disease severity according to viral genotype and fibrosis levels. The haplotype block MASP2*ARDP was associated with protection against HCV infection (OR = 0.49, p = .044) and lower MASP‐2 levels in controls (p = .021), while haplotype block AGTDVRC was significantly increased in patients (OR = 7.58, p = .003). MASP‐2 levels were lower in patients than in controls (p < .001) and in patients with viral genotype 1 or 4 (poor responders to treatment) than genotype 3 (p = .022) and correlated inversely with the levels of alkaline phosphatase, especially in individuals with fibrosis 3 or 4 (R = ?.7, p = .005). MASP2 gene polymorphisms modulate basal gene expression, which may influence the quality of complement response against HCV. MASP‐2 levels decrease during chronic disease, independently of MASP2 genotypes, most probably due to consumption and attenuation mechanisms of viral origin and by the reduced liver function, the site of MASP‐2 production.     

Apolipoprotein AI and CIII gene polymorphisms and their association with lipid levels in Italian,Greek and Anglo-Irish populations of Australia

Primary objective: The apolipoprotein (apo) AI-CIII-AIV gene cluster on chromosome 11 has been identified as a candidate region for hyperlipidaemia and in particular for hypertriglyceridaemia. Our aim was to detect associations between the apo AI and CIII polymorphisms and the plasma lipids, total cholesterol, triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol in normal, healthy, adults from three ethnic groups of Australia-Italian, Greek and Anglo-Irish, separately by gender.

Methods and procedures: The Sst I restriction fragment length polymorphisms (RFLP) in the 3' untranslated region of the apo CIII gene and the MspI RFLP in the third intron of the apo AI gene were scored and the lipid concentrations were ascertained using standard methodologies. t-tests were used to compare lipid levels between sexes and between populations, and multivariate ANOVA was used to detect if the two RFLPs had an effect on any of the lipid concentrations.

Main outcomes and results: The two RFLPs exhibit strong linkage disequilibrium in all three populations (p &lt; 0.001). There were some significant differences in allele frequencies among the populations: the minor S2 allele was more frequent in Italians (0.12) than Greeks (0.03) (p = 0.003), and the minor M2 allele was more common in Greeks (0.14) than Anglo-Irish (0.05) (p = 0.026). We found no significant association between either of the RFLPs and any of the lipid concentrations in either sex of all three populations. However, Kruskal-Wallis tests detected associations of borderline significance between apo AI MspI genotypes and triglycerides (p = 0.04) and between apo AI MspI genotypes and cholesterol levels (p = 0.03) in Anglo-Irish females.

Conclusions: Because only two statistically significant associations were detected among a number of comparisons, our data suggest that the apo AI and CIII polymorphisms play only a very limited role in mediating variation in lipid concentrations in these three ethnic groups.

Das Apolipoprotein (Apo)-Gencluster AI-CIII-AIV auf Chromosom 11 ist als eine Kandidatenregion für Hyperlipidämie und im speziellen für Hypertriglyceridämie identifiziert worden. Unser Ziel war es, die Beziehungen zwischen dem Apo AI und CIII-Polymorphismus und PlasmaLipiden: Gesamt-Cholesterol, Triglyceride, HDL (High Density Lipoprotein) und LDL (Low Density Lipoprotein) bei normalen, gesunden Erwachsenen von 3 ethnischen Gruppen: australisch-italienisch; griechisch und anglo-irisch getrennt nach den Geschlechtern festzustellen. Der SstI Registriktions-Fragment-Längen-Polymorphismus in der 3' UTR (untranslated region) auf dem Apo CIII-Gen und der MspI RFLP im 3. Intron auf dem Apo AI-Gen wurden bewertet und die LipidKonzentrationen wurden mittels Standard-Methoden ermittelt. Zum Vergleich der Lipid-Konzentrationen zwischen den Geschlechtern und den Populationen wurde der t-Test verwendet, ANOVA wurde dann verwendet, wenn untersucht werden sollte, ob die 2 RFLPs einen Effekt auf irgendeine Lipid-Konzentration. Die 2 RFLPs zeigen ein starkes Bindungsungleichgewicht in allen 3 Populationen (p &lt; 0.001). Es gab einige signifikante Differenzen in den Allel-Frequenzen zwischen den Populationen: Das minor S2-Allel war häufiger bei der italienischen (0.12) als bei der griechischen (0.03) Gruppe. Das minor M2-Allel war häufiger bei der griechischen (0.14) als bei der anglo-irischen (0.05) Population (p = 0.026). Wir fanden weder eine signifikante Beziehung für eine der beiden RFLPs und irgendeiner Lipid-Konzentration noch zwischen den Geschlechtern bei allen 3 Populationen. Dennoch deckte der Kruskal-Wallis-Test Beziehungen im Signikanz-Grenzbereich zwischen Apo AI MspI-Genotypen und Triglyceriden (p = 0.04) und zwischen Apo AI Msp I-Genotypen und den Cholesterolspiegeln (p = 0.03) bei angloirischen Frauen auf. Weil nur 2 statistisch signifikante Beziehungen bei einer Reihe von Vergleichen nachgewiesen werden konnten, legen unsere Daten nahe, dass Apo AI und CIII-Polymorphismen nur eine sehr begrenzte Rolle in der Variation der Lipid-Konzentrationen zwischen diesen 3 ethnischen Gruppen spielen.

Objectif premier: le groupe génique de l'alipoprotéine (apo) AI-CIII-AIV sur le chromosome 11 a été identifié comme région candidate pour l'hyperlipidémie, en particulier pour l'hypertrigycéridémie. Notre but a été de détecter des associations entre les polymorphisme de l'apo AI et CIII et les liquides plasmatiques; cholestérol total, triglycérides, lipoprotéines de haute densité (LHD) et lipoprotéines de faible densité (LFD) du cholestérol, chez des adultes en bonne santé de trois groupes ethniques d'Australie: italiens, grecs et anglo-irlandais étudiés séparément par sexe. Procédure et méthode: les polymorphismes Sst I de longueur du fragment de restriction (RFLP) dans la région 3' non traduite du gène apo CIII et le RFLP MspI dans le troisième intron du gène apo AI ont été comptés et les concentrations lipidiques déterminées au moyen de méthodologies standart. Les niveaux de lipides entre sexes et entre populations ont été comparés par des tests-t et une ANOVA multivariée utilisée afin de détecter si les deux RFLP avaient un effet sur une quelconque des concentrations lipidiques. Résultats principaux: les deux RFLP présentent un fort déséquilibre de linkage dans les trois populations(p &lt; 0,001). Il y a quelques différences significatives dans les fréquences alléliques parmi les populations: l'allèle mineur S2 est plus fréquent chez les italiens (0,12) que chez les grecs (0,03) (p = 0, 003) et l'allèle mineur M2 est plus commun chez les grecs (0,14) que chez les anglo-irlandais (0,05) (p = 0, 026). On ne trouve pas d'association significative entre l'un des deux RFLP et l'une quelconque des concentrations lipidiques de l'un des deux sexes dans les trois populations. Cependant, les tests de Kruskal-Wallis détectent des associations à la limite d'être significatives entre les génotypes apo AI MspI et les triglycérides (p = 0, 04) ainsi qu'entre génotypes apo AI MspI et niveaux de cholestérol (p = 0, 03) chez les femmes anglo-irlandaises. Conclusions: étant donné que seulement deux associations significatives parmi un grand nombre de comparaisons ont été trouvées, nos données suggèrent que les polymorphismes apo AI et CIII jouent un rôle très limité dans la régulation de la variation en concentrations lipidiques dans ces trois groupes ethniques.     

Amomum tsao‐ko fruit extract suppresses lipopolysaccharide‐induced inducible nitric oxide synthase by inducing heme oxygenase‐1 in macrophages and in septic mice

Amomum tsao‐ko Crevost et Lemarié (Zingiberaceae) has traditionally been used to treat inflammatory and infectious diseases, such as throat infections, malaria, abdominal pain and diarrhoea. This study was designed to assess the anti‐inflammatory effects and the molecular mechanisms of the methanol extract of A. tsao‐ko (AOM) in lipopolysaccharide (LPS)‐induced RAW 264.7 macrophages and in a murine model of sepsis. In LPS‐induced RAW 264.7 macrophages, AOM reduced the production of nitric oxide (NO) by inhibiting inducible nitric oxide synthase (iNOS) expression, and increased heme oxygenase‐1 (HO‐1) expression at the protein and mRNA levels. Pretreatment with SnPP (a selective inhibitor of HO‐1) and silencing HO‐1 using siRNA prevented the AOM‐mediated inhibition of NO production and iNOS expression. Furthermore, AOM increased the expression and nuclear accumulation of NF‐E2‐related factor 2 (Nrf2), which enhanced Nrf2 binding to antioxidant response element (ARE). In addition, AOM induced the phosphorylation of extracellular regulated kinase (ERK) and c‐Jun N‐terminal kinase (JNK) and generated reactive oxygen species (ROS). Furthermore, pretreatment with N‐acetyl‐l ‐cysteine (NAC; a ROS scavenger) diminished the AOM‐induced phosphorylation of ERK and JNK and AOM‐induced HO‐1 expression, suggesting that ERK and JNK are downstream mediators of ROS during the AOM‐induced signalling of HO‐1 expression. In LPS‐induced endotoxaemic mice, pretreatment with AOM reduced NO serum levels and liver iNOS expression and increased HO‐1 expression and survival rates. These results indicate that AOM strongly inhibits LPS‐induced NO production by activating the ROS/MAPKs/Nrf2‐mediated HO‐1 signalling pathway, and supports its pharmacological effects on inflammatory diseases.     

Significant impact of the highly informative (CA)n repeat polymorphism of the APOA‐II gene on the plasma APOA‐II concentrations and HDL subfractions: The ECTIM study

High density lipoproteins (HDL) are heterogeneous in their apolipoprotein composition and the role of apolipoprotein A‐II (APOA‐II) in HDL structure and metabolism is poorly understood. Yet, studies of naturally occurring variations of APOA‐II in mice and experiments in transgenic mice overexpressing the APOA‐II gene (APOA‐II) have shown that APOA‐II expression influences APOA‐II plasma levels and HDL size and composition. In humans, two RFLPs (BstNI and MspI) have been described in the APOA‐II gene. These RFLPs, however, have been inconstantly associated with variations in APOA‐II plasma levels. In particular, the large multicentric ECTIM Study did not show any significant effect of the two RFLPs. Other polymorphisms consisting of repetitive sequences have been proposed as more informative markers than RFLPs. Thus, data from the ECTIM Study were reconsidered by integrating the additional information obtained from a highly informative multiallelic (CA)_n‐repeat polymorphism located in the second intron of the gene. The population study was composed of 763 non‐treated male controls and 594 cases of myocardial infarction. In controls, the (CA)₁₉ allele was associated with significantly decreased APOA‐II (P < 0.0009) and LpA‐II:A‐I (P < 0.02) plasma levels. Although the APOA‐I plasma levels were not affected by the polymorphism, the (CA)₁₉ allele was associated with an increased LpA‐I/LpA‐II:A‐I ratio (P < 0.004). No effect, however, could be detected on myocardial infarction. Study of the linkage disequilibrium and the estimation of haplotype frequencies indicated that the impact of the APOA‐II locus could hardly be detected by using the BstNI and MspI RFLPs. These data revive interest in evaluating the role of the APOA‐II locus in the control of APOA‐II plasma levels and HDL composition. © 2002 Wiley‐Liss, Inc.     

PKC,ERK/p38 MAP kinases and NF‐κB targeted signalling play a role in the expression and release of IL‐1β and CXCL8 in Porphyromonas gingivalis‐infected THP1 cells

Porphyromonas gingivalis is a keystone pathogen in periodontitis and is gaining importance in cardiovascular pathogenesis. Protease‐activated receptors (PARs), toll‐like receptors (TLRs) and nucleotide‐binding oligomerization domain (NOD) on monocytes recognize the structural components on P. gingivalis, inducing inflammatory intermediates. Here, we elucidate the modulation of PARs, TLRs, NODs, and the role of MAPK and NF‐κB in IL‐1β and CXCL8 release. THP1 cells were stimulated with P. gingivalis wild‐type W50 and its isogenic gingipain mutants: Rgp mutant E8 and Kgp mutant K1A. We observed modulation of PARs, TLRs, NOD, IL‐1β and CXCL8 expression by P. gingivalis. Gingipains hydrolyse IL‐1β and CXCL8, which is more evident for IL‐1β accumulation at 24 h. Inhibition of PKC (protein kinase C), p38 and ERK (extracellular signal‐regulated kinases) partially reduced P. gingivalis‐induced IL‐1β at 6 h, whereas PKC and ERK reduced CXCL8 at both 6 and 24 h. Following NF‐κB inhibition, P. gingivalis‐induced IL‐1β and CXCL8 were completely suppressed to basal levels. Overall, TLRs, PARs and NOD possibly act in synergy with PKC, MAPK ERK/p38 and NF‐κB in P. gingivalis‐induced IL‐1β and CXCL8 release from THP1 cells. These pro‐inflammatory cytokines could affect leucocytes in circulation and exacerbate other vascular inflammatory conditions such as atherosclerosis.     

Recently characterized molecular events in uncommon gynaecological neoplasms and their clinical importance

The introduction of new sequencing technologies has resulted in the discovery of commonly mutated genes in uncommon cancers, including non‐epithelial ovarian neoplasms and other rare gynaecological tumours, such as cervical embryonal rhabdomyosarcoma. In some of these neoplasms, mutations in certain genes are both frequent and specific enough for the genomic mutations and sometimes their associated protein loss or overexpression to be used as an aid to diagnosis. In this review, we contrast previous gene identification methods with newer ones, and discuss how the new sequencing technologies (collectively referred to as ‘next‐generation sequencing’) have permitted the identification of specific molecular events that characterize several rare gynaecological neoplasms. We highlight the value of using sequencing to complement traditional pathological methods when diagnosing certain tumours, and provide practical advice to pathologists dealing with these neoplasms. We focus on adult granulosa cell tumours (somatic monoallelic mutations in FOXL2), Sertoli–Leydig cell tumours, gynaecological embryonal rhabdomyosarcomas (germline and somatic mutations in DICER1), and small‐cell carcinoma of the ovary, hypercalcaemic type (biallelic mutations in SMARCA4). The new genetic findings provided by next‐generation sequencing in these uncommon neoplasms have brought these disorders back into focus, and point the way towards new diagnostic, preventive and therapeutic avenues.     

X‐linked duplication copy number variation in a familial overgrowth condition

We describe an overgrowth condition associated with X‐linked copy number variation. Three brothers displayed an overgrowth pattern at birth that continued postnatally. Clinical findings included macrocephaly, distinctive facial features, developmental delay and variable clubfoot. Normal fetal growth was noted until the third trimester by Hadlock standards, revealing a late gestational overgrowth pattern. Microarray analysis in the family showed a maternally inherited 680 kb copy number duplication at Xq26.1‐q26.2 in all three brothers. Molecular sequencing for known overgrowth conditions including GPC3, Sotos 1 (NSD1), Malan (NFIX), Perlman (DIS3L2), Weaver (EZH2), Opitz–Kaveggia (MED12) loci were negative. BWS IC1 and IC2 methylation and CDKN1C testing was also negative. Normal IGF1 levels excluded X‐linked acrogiantism. The duplicated region Xq26.1‐q26.2 contained IGSF1 and at least part of the lncRNA FIRRE. IGSF1, a highly expressed pituitary immunoglobulin superfamily gene, was recently implicated in a genome‐wide association study of canine size. IGSF1 variants were associated with large canine breeds compared to smaller breeds. Our findings support the hypothesis that an X‐linked variant encompassing the IGSF1 region may be associated with body size. Although IGSF1 loss has been noted in human hypothyroidism, this is the first reported phenotype in a family with copy number duplication in the region. Our findings suggest that prenatal evaluation, cross‐species evaluation, Mendelian, and GWAS studies may describe a distinctive familial condition and its corresponding phenotypic features.     

Dengue virus up‐regulates expression of notch ligands Dll1 and Dll4 through interferon‐β signalling pathway

The Notch signalling pathway is involved in multiple cellular processes and has been recently indicated to modulate the host immune response. However, the role of the Notch pathway in dengue virus (DENV) infection remains unknown. Our study has screened the expression profile of Notch receptors, ligands and target genes in human monocytes, macrophages and dendritic cells in response to DENV infection. The real‐time PCR data showed that Notch ligand Dll1 was significantly induced in DENV‐infected monocytes; and receptor Notch4, ligands Dll1 and Dll4, and target Hes1 were dramatically enhanced in DENV‐infected macrophages and dendritic cells. In macrophages, induction of Dll1 and Dll4 mediated by DENV2 was increased by treatment with interferon‐β (IFN‐β), and was impaired by neutralization of IFN‐β. The DENV‐induced Dll1 and Dll4 expression level was decreased by silencing key innate immune molecules, including Toll‐like receptor 3 (TLR3), MyD88, RIG‐I and IPS‐I. In IFN‐receptor‐depleted macrophages, the Dll1 and Dll4 induction was significantly alleviated. Functionally, activation of Notch signalling by Dll1 in CD4⁺ T cells enhanced the expression of a T helper type 1 (Th1) cytokine IFN‐γ, while Notch activation in macrophages had no direct effect on replication of DENV. Our data revealed that the expressions of Notch ligands in antigen‐presenting cells were differentially induced by DENV via innate immune signalling, which is important for Th1/Th2 differentiation during adaptive immune response.     

Respiratory sinus arrhythmia and serotonin transporter promoter gene polymorphisms: Taking a triallelic approach makes a difference

It has been recently reported in Psychophysiology that carriers of the short allele of an insertion/deletion (ins/del) functional polymorphism in the promoter region (5‐HTTLPR) of the serotonin transporter gene may display decreased resting respiratory sinus arrhythmia (RSA). However, this region hosts another functionally connected single‐nucleotide polymorphism (rs25531), which should also be genotyped in order to correctly categorize the low‐ and high‐expressing alleles of 5‐HTTLPR. The present study investigated resting RSA in an ethnically homogenous sample (N = 143) of participants genotyped for both the ins/del and rs25531 polymorphisms in 5‐HTTLPR. In contrast with the biallelic genotypes, based only on the ins/del alleles, the triallelic 5‐HTTLPR genotypes (i.e., including rs25531) showed no association with resting RSA. Taking a triallelic approach to 5‐HTTLPR is thus necessary in order to avoid false positive results.