Chronic obstructive pulmonary disease (COPD) is a multifactorial disease of the respiratory system which develops as a result of a complex interaction of genetic and environmental factors closely related to lifestyle. We aimed to assess the combined effect of the PI3K/AKT/mTOR signaling pathway  and sirtuin  genes to COPD risk. SNPs of SIRT1 , SIRT3 , SIRT6 (rs107251), AKT1 (rs2494732), PIK3R1 , MTOR , PTEN  genes were genotyped by real-time polymerase chain reaction (PCR) among 1245 case and control samples. Logistic regression was used to detect the association of SNPs in different models. Linear regression analyses were performed to estimate the relationship between SNPs and lung function parameters and smoking pack-years. Significant associations with COPD were identified for SIRT1 (rs3818292) , SIRT3 (rs3782116)  and SIRT3 (rs536715)  under the dominant model, SIRT6 (rs107251) , PIK3R1: , rs831125 , rs3730089 ), PTEN: , and rs2735343 ). A significant genotype-dependent variation of lung function parameters was observed for SIRT1 (rs3818292), SIRT3 (rs3782116), PIK3R1 (rs3730089), and MTOR (rs2536). Gene-gene combinations that remained significantly associated with COPD were obtained; the highest risk of COPD was conferred by a combination of G allele of the PIK3R1 (rs831125) gene and GG of SIRT3 (rs536715) (OR = 3.45). The obtained results of polygenic analysis indicate the interaction of genes encoding sirtuins SIRT3, SIRT2, SIRT6 and PI3KR1, PTEN, MTOR and confirm the functional relationship between sirtuins and the PI3K/AKT/mTOR signaling pathway. Chronic obstructive pulmonary disease (COPD) is a multifactorialrespiratory system disease that affects the distalparts of the respiratory tract  and lungparenchyma with lung emphysema development . Сhronic obstructive pulmonary disease developsas a result of complex interaction between molecular geneticand environmental factors, closely related to lifestyle, andsmoking is considered the main cause of COPD . Published data suggest that the COPD pathogenesismay involve dysregulation of stress responses that inhibitcellular senescence, which includes a wide range of signalingcascades and their regulators .The PI3K/AKT/mTOR intracellular signaling pathway isa universal pathway controlling cell growth, metabolism, andproliferation . The key components of thissignaling pathway are phosphatidylinositol-3 kinase (PI3K),serine/threonine protein kinase (AKT), and serine/threoninekinase  . Signal transduction through the PI3K/AKT/mTOR signaling cascade is essential for cellular senescence.This signaling pathway is inhibited by the tyrosine phosphatasesPTEN (phosphatase and tensin homolog) and SHIP-1(inositol polyphosphate-5-phosphatase D). Both enzymeshave oxidation-sensitive cysteine residue in the active region.Oxidative stress is the main mechanism that causes acceleratedsenescence through its damaging effects on DNA and thePI3K/AKT/mTOR signaling pathway activation . In COPD and other age-associated diseases, the expressionof genes encoding endogenous antioxidant molecules isreduced, which leads to an increased level of oxidative stressand cellular senescence activation .NAD-dependent protein deacetylases from the sirtuins familyare considered as potential factors that decrease senescence.We aimed to assess the combined effect of the PI3K/AKT/mTOR signaling pathway  andsirtuins  genes on COPD risk.DNA samples were collected from unrelated subjects whowere Tatars in ethnicity and resided in the Republic of Bashkortostan.The study was approved by the Ethics Committeeat the Institute of Biochemistry and Genetics . All participants of this study providedwritten informed consent. The COPD group included 621patients (539 (86.79 %) males and 82 (13.21 %) females) witha mean age of 64.42 ± 10.71 years. There were 510 (82.13 %)smokers and former smokers and 111 (17.87 %) nonsmokersin the COPD group. The smoking index was 45.34 ± 23.84pack years in the smokers and former smokers. The controlgroup included 624 subjects (555 (88.94 %) males and 69(11.06 %) females) with a mean age of 59.67 ± 12.31. Therewere 526 (84.29 %) smokers and former smokers and 98(15.71 %) nonsmokers in the group; the smoking index was38.75 ± 24.87 pack years in the smokersIn all patients, spirometry was performed to assess lungfunction, including vital capacity (VC), forced vital capacity(FVC), forced expiration volume in the first second (FEV1),and the FEV1/FVC ratio. The group of patients had the followingparameters : FEV1 = 40.75 ± 18.33;FVC = 45.01 ± 18.22; VC = 49.32 ± 14.34; FEV1/FVC =59.5 ± 12.34. Inclusion and exclusion criteria for the COPDand control have been previously described .Genotyping. DNA was isolated from peripheral bloodleukocytes by phenol-chloroform extraction. The set includedSNPs of the following genes: SIRT1 ,SIRT3 , SIRT6 (rs107251), AKT1(rs2494732), PIK3R1 ,MTOR , PTEN 1.http://vavilov.elpub.ru/jour/manager/files/Suppl_Korytina_Engl_27_5.pdfSupplementary Materials are available in the online version of the paper:The SNPs were selected for the study based on the followingcriteria: functional effect of SNP on gene expressionor relation to non-synonymous substitutions, and/or associationswith complex human diseases; minor allele frequency (MAF) of ≥ 5 % in the European populations according tothe National Center for Biotechnology Information database(http://www.ncbi.nlm.nih.gov/projects/SNP/). The functionalsignificance of the SNPs was verified using RegulomeDBVersion 1.1 (https://regulomedb.org), SNPinfo Web Server(https://snpinfo.niehs.nih.gov), and HaploReg v3 . Data were presented in Supplementary Material 2.SNP genotyping was performed by real-time polymerasechain reaction (PCR) using commercial kits for fluorescencedetection  anda BioRad CFX96TM instrument . The methods of analysis were described in detailpreviously .Statistical analyses. Statistical analyses of the results wereperformed using the software packages IBM SPSS Statis-tics 22.0 and PLINK v. 1.07 . The methodswere described in detail previously .Association analyses of allele or genotype combinationswith COPD were carried out using the APSampler 3.6.1program (http://sourceforge.net/projects/apsampler/). TheBenjiamini–Hochberg correction for multiple testing wasperformed using special software (http://www.sdmproject.com/utilinies/?show=FDR) to decrease the false discoveryrate (FDR) and to obtain Рcor- FDR. The linkage disequilibriumstructure LD (D’) and haplotype frequencies were calculatedwith Haploview 4.2.Before analyzing the association of candidate gene alleleswith COPD, we verified whether the genotype frequencydistributions corresponded to the Hardy–Weinberg equilibriumand evaluated minor allele frequencies (MAF) both in thecombined group of patients and healthy subjects and in eithergroup individually . All studiedSNPs were in Hardy–Weinberg equilibrium in the controlgroup: SIRT1 (rs3758391) (PX-B = 0.24), SIRT1 (rs3818292)(PX-B = 0.47), SIRT3 (rs3782116) (PX-B = 0.5), SIRT3(rs536715) (PX-B = 0.75), SIRT6 (rs107251) (PX-B = 0.67),AKT1 (rs2494732) (PX-B = 0.2), PIK3R1 (rs10515070)(PX-B = 0.65), PIK3R1 (rs831125) (PX-B = 0.25), PIK3R1(rs3730089) (PX-B = 0.22), MTOR (rs2295080) (PX-B = 0.15),MTOR (rs2536) (PX-B = 0.24), PTEN (rs701848) (PX-B = 0.85),PTEN (rs2735343) (PX-B = 0.06).The groups of COPD patients and healthy controls differedsignificantly in the genotypes and/or alleles frequencydistributions of SIRT1 (rs3818292), SIRT3 , SIRT6 (rs107251), AKT1 (rs2494732), PIK3R1, and PTEN  (Table 1). Statistically significant results ofassociation analysis of the studied gene loci and COPD areshown in Table 2.An association of SIRT1 (rs3818292) with COPD was establishedin the dominant model .The risk of COPD was increased in heterozygous individuals. An association of COPD with theheterozygous genotype  and in thedominant model of SIRT3 (rs3782116) and in the dominant, log-additive  models and with the heterozygous genotype of SIRT3 (rs536715). It shouldbe noted that in both cases the COPD risk was associatedwith the frequent G allele (rs3782116 – OR = 1.21 95 % CI1.03–1.43 и rs536715 – OR = 1.58 95 % CI 1.32–1.91) andthe GG genotype (rs3782116 – OR = 1.44 95 % CI 1.16–1.81и rs536715 – OR = 1.99 95 % CI 1.58–2.51).We carried out a linkage disequilibrium analysis of thers3758391 and rs3818292 loci of the SIRT1, rs3782116 andrs536715 of the SIRT3, which showed the absence of linkagedisequilibrium between the loci of the SIRT1 gene  and the SIRT3 gene . Basedon the obtained data, haplotypes association analysis wasnot performed. Association of SIRT6 (rs107251) with thedevelopment of COPD was detected in the dominant model, but the association with the heterozygousCT genotype was more significant . The risk of COPD was associated with the CC ge-notype of SIRT6 (rs107251) (OR = 1.54 95 % CI 1.23–1.93).We have identified the association SNPs of the PIK3R1 gene with COPD. The risk ofCOPD was associated with heterozygous genotypes of thestudied SNPs of the PIK3R1 gene: rs10515070 , rs831125  and withthe GG genotype of rs3730089 .We showed the absence of linkage disequilibrium betweenrs10515070, rs831125, rs3730089 of the PIK3R1 gene:for rs10515070 and rs831125 , forrs10515070 and rs3730089 , forrs831125 and rs3730089 .Based on the obtained data, haplotypes association analysiswas not performed. Association of PTEN (rs701848)and COPD was established in the dominant , recessive  andlog-additive models Padj = 0.0015, OR = 1.35). We haveidentified the association of PTEN (rs2735343) with COPD inthe log-dominant model  and for theheterozygous genotype . Linkagedisequilibrium between the rs701848 and rs2735343 was notdetected , thus, haplotype associationanalysis was not performed.Association of the studied genes lociand quantitative phenotypes with lungfunction parameters and smoking indexLung function decline is a key clinical feature of airwayobstruction in COPD that indicates progression of the disease.We investigated the relationship between the studied genes lociand lung function parameters: Forced Vital Capacity (FVC),Forced Expiration Volume in 1 s (FEV1), and FEV1/FVCratio in COPD patients (Table 3). The heterozygous genotypeof PIK3R1 (rs3730089) (P = 0.013) and the TT genotypeof MTOR (rs2536) (P = 0.013) were associated with adecrease in the FVC value. Carriers of the SIRT3 (rs3782116)AA genotype exhibited a higher FVC value (P = 0.0015).Individuals that presented the AA genotype of SIRT1(rs3818292) (P = 0.017), the GG genotype of PIK3R1(rs3730089) (P = 0.025), and the AA genotype of SIRT3(rs3782116) (P = 0.0028) showed a significant increase in theirFVC. Meanwhile, carriers of the heterozygous genotypes ofSIRT1 (rs3818292) (P = 0.04), MTOR (rs2295080) (P = 0.025),and SIRT3 (rs3782116) (P = 0.016) exhibited a lower FVC value (see Table 3). The carriers of the heterozygous genotypeof PIK3R1 (rs831125) (P = 0.0082), the AA genotype ofSIRT1 (rs3818292) (P = 0.036) had a significantly highersmoking index.Analysis of gene-gene interactionsUsing the APSampler algorithm, we have identified gene-genecombinations significantly associated with сhronic obstructivepulmonary disease. In order to identify significant interactionsof functionally related sirtuins genes, SIRT2 (rs10410544) wasincluded in the analysis . We obtained2324 patterns associated with сhronic obstructive pulmonarydisease. Table 4 shows the results of the most significant genegenecombinations with PFDR less than 0.05 and OR morethan 2.0 for risk combinations) or less than 0.35 for protectivecombinations. A total of 19 gene-gene combinations fulfilledthe above-mentioned criteria. Nine patterns were associatedwith an increased risk of COPD; ten were protective. Allele Gand genotype GG of PIK3R1 (rs831125) contributed to themost significant combinations associated with COPD risk(four patters).The highest risk of COPD was conferred by the combinationof these variants of PIK3R1 (rs831125) with the GG genotypeof SIRT3 (rs536715) (OR = 3.45); and with the C alleleof PTEN (rs2735343) (OR = 3.06) and their combination:genotype GA of PIK3R1 (rs831125) together with the G alleleof SIRT3 (rs536715) and the C allele of PTEN (rs2735343)(OR = 2.86). The analysis on the gene-gene interaction of thestudied gene loci established an association of the T alleleof MTOR (rs2536) only in combinations with the G alleleof PIK3R1 (rs831125) (OR = 2.71). Three of the identifiedpatterns included the G allele of PIK3R1 (rs3730089) in combination with the GG genotype of SIRT3 (rs536715)(OR = 2.09), or with the CC genotype of SIRT6 (rs107251)(OR = 2.01).The most significant protective patterns included the A alleleor the AA genotype of PIK3R1 (rs831125) and the A alleleof PIK3R1 (rs3730089) in combination with the A allele orthe AG genotype of SIRT3 (rs3782116) and the A allele ofSIRT3 (rs536715) (see Table 4). Thus, the PIK3R1 (rs831125),PIK3R1 (rs3730089), and SIRT3 (rs536715) loci exhibited anallele-specific effect, when the G allele of PIK3R1 (rs831125),the G allele of PIK3R1 (rs3730089), and the G allele and theGG genotype of SIRT3 (rs536715) were part of risk patterns,and alternative alleles of the same polymorphic loci werepresent in protective patterns.As a result our study, significant associations between theSIRT1 (rs3818292), SIRT3 , SIRT6(rs107251) polymorphic variants and COPD were found.SIRT1 is the most studied member of the mammalian sirtuinfamily. It has been shown that SIRT1 plays an important rolein signaling pathways involved in cellular senescence andcell death . SIRT1 deacetylates manykey regulatory proteins and transcription factors involved inDNA repair, inflammation, expression of antioxidant genes,and cellular senescence, including the PI3K/AKT/mTORsignaling pathway genes, transcription factor FOXO3a, p21,p16, Klotho proteins .Increased expression of SIRT1 inhibits the TGF-β1/SMAD3signaling pathway and impairs epithelial-mesenchymal transformation,which leads to a decrease in COPD-associatedairway remodeling . SIRT1 levels arereduced in peripheral pulmonary and peripheral blood mononuclearcells of patients with COPD .We found that the COPD risk is higher in heterozygouscarriers of SIRT1 (rs3818292). Moreover, this polymorphicvariant demonstrates an association with a decrease in FVС1, which reflects the progression of the disease. Functionalanalysis showed that SIRT1 (rs3818292) is in linkage witha SNP in the 5′-untranslated DNA region (rs3740051) thatchanges the NFKB transcription factor binding site. We didnot associate SIRT1 (rs3758391) with COPD. The results ofour study are in agreement with the previously published datareported for the Han Chinese population .According to functional analysis, rs3758391 is located in thepromoter region of the SIRT1 gene, and the C variant disruptsbinding sites for several transcription factors and regulatoryproteins, affecting gene expression. The role of rs3758391in the development of age-associated diseases is well known.Mitochondrial dysfunction in respiratory epithelial cellsplays an important role in the pathogenesis of COPD . SIRT3 is the main mitochondrial deacetylaseregulating many enzymes involved in energy metabolism,respiratory chain components, the tricarboxylicacid cycle,ketogenesis, and fatty acid beta-oxidation .SIRT3 can directly control the production of reactive oxygenspecies (ROS) by deacetylating manganese superoxidedismutase (SOD2), the main mitochondrial antioxidant enzyme. SIRT3 is involved in regulatingthe activity of the DNA repair enzyme OGG1, which leadsto increased damage to mtDNA and apoptosis of alveolarepithelial cells . A number of studies haveshown the SIRT3 association with various complex diseases.We studied the association between two SIRT3 gene functionalpolymorphisms (rs3782116 and rs536715) and сhronicobstructive pulmonary disease. Functional analysis showedthat rs3782116 is located in the region of binding sites forhsa-miR-328; polymorphic loci rs3782116 and rs536715 arelocated in DNA regions that bind regulatory proteins. Bothpolymorphic loci were associated with COPD in our cohort;the disease development risk was associated with the G allelesof rs3782116 and rs536715. It should be noted that the carriersof the homozygous rare allele A of rs3782116 of the SIRT3gene had higher values of VC and FVC1. The contributionof SIRT3 variants to COPD has not been studied previously.At the same time, effects of the SIRT3 gene SNPs have beenfairly extensively investigated in age-associated diseases inwhich oxidative stress and cellular senescence play a key role.We obtained significant associations of SIRT6 (rs107251)with COPD; the frequent C allele is associated with COPDrisk, while the heterozygous genotype has a protective effecton the disease development. rs107251 is located in the DNAregion that binds to the SOX8 regulatory protein and it is inclose linkage with rs350846, localized in the 3-non-translationalregion of the SIRT6 gene – a binding site for severalmiRNAs . SIRT6 participates in the regulationof genome stability, NF-kB signaling, glucose homeostasis,exhibits ADP-ribosyltransferase and histone deacetylase activity,plays a role in DNA repair and maintenance of telomericchromatin integrity . In the studyby N. Takasaka et al. (2014), a decrease in SIRT6 levels wasshown in respiratory epithelial cells of COPD patients due tocigarette smoke exposure, leading to cellular senescence anddisruption of autophagy processes. Association of the SIRT6gene SNPs with COPD has not been studied previously, butthere is evidence of their association with cardiovasculardiseases, which are often comorbid pathologies in COPD.The PIK3R1 gene encodes regulatory subunit 1 of phosphoinositide3-kinase, a key element of the PI3K/AKT/mTORsignaling cascade . We investigated threePIK3R1 gene polymorphic loci , which showed a significant association withCOPD in our studied group. Carriers of rare alleles of thesepolymorphic loci had a high risk of COPD. In addition, weinvestigated the relationship between rs3730089 genotypesand VC and FVC1 values; thus, heterozygotes have lowervalues, and these results are in agreement with associationanalysis. Functional analysis showed that an intronic variantrs831125 is located in a binding site for regulatory proteins;rs3730089 is a missense variant with a “benign” effect accordingto the PolyPhen-2 database (http://genetics.bwh.harvard.edu/pph2/), located in a binding site for regulatory proteinsand affecting splice sites. SNPs of the PIK3R1 gene have notbeen evaluated for COPD before. Previously, an associationhas been observed between rs3730089 and type 2 diabetes.The phosphatase PTEN regulates the activity of phosphoinositide-3 kinase (PI3K) . Smokingas a major risk factor for COPD provokes oxidative stress,which, in turn, affects PTEN expression .We investigated two PTEN gene functional polymorphicloci; rs70184 is located in PTEN gene 3′ region and changesbinding sites for hsa-miR-1252 and hsa-miR-1304 miRNA;rs2735343, located in the intronic region, affects binding sitesfor several regulatory proteins.Significant associations with COPD in our sample werefound with PTEN gene loci; thus, homozygous carriers ofthe rare C allele of rs701848 and heterozygous carriers ofrs2735343 had a significant risk of COPD development.Published data have demonstrated an association betweenPTEN (rs701848) and COPD; the risk was significantly reducedfor homozygous T allele carriers, which is consistentwith the data obtained for our sample .PTEN participates in the regulation of various biologicalprocesses, including cell proliferation, apoptosis, inflammatoryreactions, transcription, and genomic stability . Decreased levels of PTEN lead to activation of PI3Ksignaling and increased cell senescence in COPD . It has been shown that decreased PTEN activity inCOPD increases the activity of matrix metalloprotease MMP9in bronchial epithelial cells, which consequently contributesto the progression of inflammation and extracellular matrixdegradation .The analysis of gene-gene interactions revealed significantsynergy between polymorphic loci of genes encodingphosphoinositide-3-kinase (PIK3R1) and mitochondrialdeacetylase (SIRT3), which were present in most significantcombinations associated with an increased risk of сhronicobstructive pulmonary disease. The C allele in the PTEN(rs2735343) was part of four informative combinations associatedwith a high risk of сhronic obstructive pulmonarydisease.The results of polygenic analysis indicate the interactionof genes encoding sirtuins SIRT3, SIRT2, SIRT6 and PI3KR1,PTEN, MTOR and confirm the functional relationship betweensirtuins and the PI3K/AKT/mTOR signaling pathway.The obtained results of single locus and polygenic analysis indicatethe contribution of SIRT3 , SIRT6(rs107251) and PIK3R1 polymorphisms to COPD and interaction of genes enco-ding the key components of the PI3K/AKT/mTOR signallingpathway and sirtuins, and confirm the involvement of cellularsenescence mechanisms with COPD developmentThe authors declare no conflict of interest.Barnes P.J., Baker J., Donnelly L.E. Cellular senescence asa mechanism and target in chronic lung diseases. Am. J.Respir. Crit. Care Med. 2019;200(5):556-564. 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Even though NoShow’s 52,825 bp genome is most similar to phages in cluster AB, its  Mycobacterium abscessus  revealed a siphovirus morphology , prepared for sequencing using the NEB Ultra II library kit, and sequenced on Illumina MiSeq (v3 reagents), yielding 59,426 single-end 150-base reads. Raw reads were assembled and checked for completeness using Newbler (v2.9) and Consed (v29) with default settings (MK279840), Muddy , and FF47  (Geneious Prime (2022.0.2)) and has the highest G + C content . NoShow possessed 11 bp 3′ single-stranded genome termini (5′-CGGAGAGGCTT-3′) (NoShow’s DNA was extracted by Phenol-Chloroform (settings . Within http://phages.wustl.edu/starterator), ARAGORN (v1.2.38) and tRNAscan-SE (v1.3) (The genome was annotated using PECAAN (v2021) , PhameraE (v1.3) , 11, HHPE (v1.3) , and BlaE (v1.3) , and theE (v1.3) . DefaultConsistent with cluster AB phages, NoShow lacks any identifiable immunity repressor or integrase functions and is, therefore, considered a lytic phage –18. Unli–21–


The name of the first author was incorrectly expressed in the Citation. The correct Citation is: Winck FV, Arvidsson S, Riaño-Pachón DM, Hempel S, Koseska A, et al. (2013) Genome-Wide Identification of Regulatory Elements and Reconstruction of Gene Regulatory Networks of the Green Alga Chlamydomonas reinhardtii under Carbon Deprivation. PLoS ONE 8(11): e79909. doi:10.1371/journal.pone.0079909.


AbstractDoryctopambolusgen. n. is erected to contain Doryctopambolus pilcomayensis , comb. n., which was previously placed within Pambolus (Pambolinae), as well as three new species, Doryctopambolus clebschisp. n., Doryctopambolus dominicanussp. n. and Doryctopambolus sarochensissp. n. Members PageBreakof this new genus are mainly characterised by the presence of at least one pair of conspicuous propodeal apico-lateral projections, which are similar to those present in all members of Pambolinae and in species of three Australasian doryctine genera. We generated DNA barcoding sequences for the three newly described species. We discuss the morphological similarity between species of the Australasian Echinodoryctes Belokobylskij, Iqbal & Austin and Doryctopambolus. A key for the described species of Doryctopambolus is provided.The new Neotropical doryctine genus  Eucharitidae, Figitidae, Ichneumonidae and Braconidae, among others. Within the cyclostome group of subfamilies belonging to Braconidae, the presence of acute tubercles or spines on the apical lateral region of propodeum has been considered as the main diagnostic character employed to distinguish members of the cosmopolitan Pambolinae , all belonging to the cyclostome subfamily Doryctinae, also have these propodeal projections, indicating that this feature has actually evolved on separate occasions within Braconidae.The presence of propodeal or scutellar spines is a common feature found in species of a number of hymenopteran parasitoid families, including mbolinae . HoweverPambolus described to date , comb. n. and three additional Neotropical species described herein. We also molecularly characterise the three newly described species with the DNA barcoding locus , Colección Nacional de Insectos, Instituto de Biología, Universidad Nacional Autónoma de México (CNIN-IB UNAM), Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”, Buenos Aires, Argentina (MACN), and Museo de La Plata, Argentina (MLP). Terminology for body structure, sculpture and wing venation features follows Digital colour photographs of all the species included in this work were taken with a Leica® Z16 APO-A stereoscopic microscope and a Leica® DFC295/DFC290 HD camera, and were edited using the Leica Application Suite® program. Digital scanning electron microscope (SEM) photographs of DNA sequences belonging to the barcoding locus [~650 bp of the cytochrome oxidase I (COI) mitochondrial DNA gene; Nunes & Zaldívar-Riveróngen. n.urn:lsid:zoobank.org:act:2C42D9FA-392C-4F6E-8A6E-CA00EAE711C0http://species-id.net/wiki/DoryctopambolusPambolus pilcomayensis van Achterberg & Braet, 2004Doryctopambolus can be distinguished from members of most doryctine genera except Concurtisella bidens, Echinodoryctes, Fijispathius and Ryukuspathius by having the propodeum with at least one pair of conspicuous apico-lateral projections. SpePageBreakPageBreakcies of Doryctopambolus and Concurtisella bidens are the only Neotropical doryctine taxa reported to have these projections, though they mainly differ by their first subdiscal cell  and ovipositor length . Doryctopambolus differs from the Australasian Fijispathius and Ryukyuspathius mainly by the fore wing first subdiscal cell open at apex (closed in the later two genera) and the first metasomal segment not petiolate . Doryctopambolus is morphologically similar to the Australian Echinodoryctes , propodeum evenly curved and strongly rugose-areolate (globose and mostly smooth in Echinodoryctes), hind coxa without basoventral tubercle and all femora without dorsal protuberances (both present in Echinodoryctes).Species ofdoryctes . HoweverHead: head globose; antennal sockets distinctly separated from each other by at least 0.5 times its diameter; frons almost flat, without median carina or furrows; ocelli arranged in equilateral triangle; eye with distinct and sparse setae; gena and temple smooth; malar suture absent; first flagellomere slightly shorter than scape and pedicel combined, slightly longer than second flagellomere; antenna with 16–28 antennomeres; occipital carina meeting hypostomal carina before mandible. Mesosoma: Length of mesosoma about two times its maximum width; neck of pronotum fairly long; pronotal crest conspicuous; mesoscutum declivous anteriorly; mesoscutal lobes smooth and polished medially; notauli complete and strongly impressed; scutellar sulcus deep, with its height 0.8–0.9 times height of scutellar disc; precoxal sulcus complete and scrobiculate, as long as mesopleuron; prepectal carina coarse and complete; propodeum evenly curved and strongly rugose-areolate, with at least one pair of conspicuous apico-lateral projections; propodeal bridge absent. Legs: fore tibia with a row of 7–8 stout spines; middle tibia without spines; femora without dorsal protuberances; hind coxa without basal tubercle. Wings: partially reduced to well-developed wings; fore wing veins r-m and 2RS present; m-cu arising interstitial or slightly antefurcal with vein 2RS, cu-a distinctly postfurcal with vein 1M; first subdiscal cell open at apex; hind wing vein M+CU equal length of vein 1M; cu-a present, m-cu absent; stigma present on male hind wing. Metasoma: length of first metasomal tergum 1.3–1.6 times its apical width, apical width about 2.0–2.3 times basal width; basal sternal plate (acrosternite) about 0.33–0.5 times length of tergum; suture between second and third metasomal tergites absent; second metasomal tergite at least sculptured basally; third metasomal tergite usually smooth, sometimes sculptured basally; remaining metasomal tergites entirely smooth and polished; ovipositor about same length of metasoma.Small size, 2.2–3.6 mm; black to light brown species. Neotropical. Known from central Argentina to northern Venezuela, and from Dominican Republic in the Caribbean.Doryctopambolus and Doryctopambolus pilcomayensis comb. nov. are described and redescribed in this study, respectively. Four additional speciePageBreaks belonging to this genus, two from Cerro Saroche, Lara, Venezuela, and two from Argentina, were also identified. The two Argentinian species could not be described due to their bad state of preservation.Of the two species from Venezuela, one was represented by a single male and the other one by an incomplete female, and their allospecificity was corroborated with DNA barcoding sequences . The Parque Nacional Cerro Saroche is a natural reserve of about 32,294 h mainly composed of xeric vegetation with deciduous and semideciduous shrubs comb. n.Pambolus pilcomayensis van Achterberg & Braet 2004: pp. 341–344.Doryctopambolus sarochensis sp. n. by having one pair of propodeal apico-lateral projections (two pairs in Doryctopambolus sarochensis sp. n.) and third metasomal tergite entirely smooth . Doryctopambolus pilcomayensis differs PageBreakfrom Doryctopambolus dominicanussp. n. and Doryctopambolus clebschi sp. n. by having the vertex striate (smooth in Doryctopambolus dominicanus sp. n. and Doryctopambolus clebschi sp. n.), first metasomal tergite not petiolate (at least 0.5 of tergum) and second metasomal tergite entirely sculptured (mostly smooth).This species distinguishes from Colour: head light to dark brown, antenna light brown, malar space light brown to yellowish, mandible and palpi light brown; propleuron and pronotum light brown, remainder of mesonotum brown to black; wings slightly infuscate, narrow banded with brown veins; metasoma brown to black, first two metasomal tergites darker; legs brown to dark brown, fore and middle coxae, trochanter, trochantellus and tarsi light brown to pale yellow; ovipositor and sheaths light brown with dark apex. Head: antero-dorsal surface of eyes bordered by groove; eyes setose; face costate-rugose laterally, central area slightly swollen and smooth; clypeus costate-rugose; malar space 0.8–1.0 times eye height; frons and vertex striate; temple in dorsal view 0.7 times eye width; antenna with 26 antennomeres. Mesosoma: length of mesosoma 2.0 times its maximum height; pronotum rugose; pronotal groove wide and scrobiculate laterally, dorsally smooth; propleuron costate-rugose; notauli deep and scrobiculate, meeting scutellum with parallel carinae; scutellum smooth and setose; scutellar sulcus with three parallel carinae; mesopleuron porcate dorsally, smooth near precoxal sulcus and ventrally; ventral portion of mesopleuron separated by shallow crenulate median groove with some rugosities near middle coxae; propodeum with one pair of conspicuous apico-lateral projections, longer than first flagellomere; median carina present only basally. Legs: hind coxa costate to costate-rugose dorsally. Wings: fore wing length 3.7–4.8 times its maximum width, r:3RSa:3RSb = 2:8-12:18-32; 2RS:3RSa:r-m = 6:8-12:2-3; m-cu arising interstitial or slightly antefurcal to vein 2RS; 1cu-a interstitial with 1M. Metasoma: length of first metasomal tergite 1.3 times its apical width, apical width about 2.0 times basal width costate-rugose, with two or three longitudinal carinae reaching apex of tergum; second metasomal tergite strongly costate; remaining metasomal tergites smooth and polished; basal sternal plate (acrosternite) about 0.33 times length of tergum; ovipositor nearly as long as metasoma.Females. Body length 3.0–3.6 mm, fore wing 2.0 mm, ovipositor 1.8 mm. Males. Essentially as females; body length 2.1–3.0 mm; body brown, with antennae, trochanter, trochantellus, tarsi, fore and middle coxae light brown to yellowish; face, frons and vertex slightly sculptured; stigma present in hind wing.Brazil: Goiás State (GO), Itumbiara, Malaise trap, C.H. Marchiori col.: one female 24-I-1998, three males 28-I-1998, one female and one male 31-I-1998, two males 7-II-1998, three females and one male 28-II-1998, one female and one male 7-III-1998, one female 1-IV-1998, one female and one male 15-IV-1998, one female 08-XI-1998 (DCBU); Mato Grosso do Sul State (MS), Bodoquena, Parque Nacional da Serra da Bodoquena, Faz. Pitangueiras, Bandeja Am. 21 (yellow pan trap), Crepaldi RA et al. col: one female V-2009 (DCBU). Argentina: Misiones Province, Estación Experimental Loreto, Dr. A. Ogloblin: one female 15-VI-1930 (MLP); Buenos Aires Province, Reserva Ecológica de Vicente López, 34.492°S, 58.480°W: one male 16-I-2011 (MACN); one male, no data (MLP).Twenty-two specimens . PageBreakSouthern Brazil to Argentina.Doryctopambolus pilcomayensis near Buenos Aires can be explained by the courses of the Parana and Uruguay basins, which carry downstream many plant and animal species from tropical areas to higher latitudes.The presence of Zaldívar-Riverón & Martínezsp. n.urn:lsid:zoobank.org:act:0992A72F-1163-48FB-A1BF-D5E59F7D117Fhttp://species-id.net/wiki/Doryctopambolus_clebschiDoryctopambolus clebschi is morphologically similar to Doryctopambolus dominicanus sp. n. However, Doryctopambolus clebschi differs from the latter species by having the face smooth medially and slightly rugose laterally (entirely rugose in Doryctopambolus dominicanus sp. n.), mesopleuron mostly smooth (mostly smooth-rugose), first metasomal tergite mostly smooth apically , and second metasomal tergite mostly smooth, slightly costate baso-laterally . These two species distinguish from the remaining two described species of Doryctopambolus, Doryctopambolus pilcomayensis comb. n. and Doryctopambolus sarochensis sp. n., by having the vertex smooth , first metasomal tergite distinctly petiolate (not petiolate), and second metasomal tergite partially smooth .. Body length 2.8 mm; fore wing 2.3 mm; ovipositor 1.7 mm. Colour: head brown, antennae brown, turning honey yellow to apex, palpi yellow; mesosoma and first metasomal tergites brown, second metasomal tergite honey yellow, remaining metasomal tergites light brown; wings hyaline; veins and stigma light brown; legs light brown to honey yellow; ovipositor and sheaths light brown to brown, apex strongly sclerotised and dark. Head: antenna with 16-17 antennomeres; eyes small, ovoid and setose; face smooth and glabrous medially, slightly rugose and pilose laterally, median area not swollen; clypeus slightly rugose; malar space 0.4 times eye height; frons and vertex smooth; temple in dorsal view 2.7 times eye width. Mesosoma: 1.8 times longer its maximum height; pronotum rugose laterally; pronotal groove wide and scrobiculate; propleuron slightly rugose; mesoscutal lobes entirely smooth and polished, sparsely pilose; notauli wide, deep and scrobiculate, meeting before scutellum in a longitudinally costate area; scutellum smooth, sparsely setose; scutellar sulcus with three parallel carinae; height of scutellar sulcus 0.8 times height of scutellar disc; subalar sulcus wide and scrobiculate, joining mesopleural sulcus, remaining area of mesopleuron mostly smooth and polished, slightly rugose near mesopleural sulcus; venter of mesopleuron smooth; propodeum with one pair of long and sharp apico-lateral projections, shorter than first flagellomere. Legs: hind coxa mostly smooth, slightly rugose ventrally. Wings: fore wing length 3.6 times its maximum width, r:3RS:3RSb = 2:3:10; 2RS:3RSa:r-m = 4:3:2; m-cu arising antefurcal with vein 2RS; 1cu-a interstitial with 1M; hind wing vein M + CU about equal length of vein 1M. Metasoma: length of first metasomal tergum 1.5 times its apical width, apical width about 2.3 times basal width, first metasomal tergite costate basally and medially, smooth apically; second metasomal tergite mostly smooth and polished, slightly costate baso-laterally; remaining metasomal tergites smooth and polished; basal sternal plate (acrosternite) about 0.5 times length of tergum; ovipositor about 1.1 times longer than metasoma.FemaleVariation. Females. Body length 2.6–2.8 mm.Males. Unknown.PageBreakHolotype. Female (CNIN IB-UNAM). Dominican Republic: A. Bermudez NP, La Ciénega, Telostablones, 19.066°N, 70.863°W, 15-16-IX-2009, sweep, H. Clebsch col. . Paratype. One female (CNIN IB-UNAM), same data as holotype.Dominican Republic.This species is named in honour to our good friend and colleague Hans Clebsch, who collected the specimens assigned to this species.Zaldívar-Riverón & Martínezsp. n.urn:lsid:zoobank.org:act:99C34338-DB4F-4AE2-A662-4DDA9957BA43http://species-id.net/wiki/Doryctopambolus_dominicanusD. Clebschi.See . Body length 2.7 mm; fore wing 1.8 mm; ovipositor 1.3 mm. Colour: head light brown, antennae light brown, turning honey yellow to apex, palpi yellow; mesosoma and first and basal two thirds of second metasomal tergites dark brown; wings hyaline; veins and stigma light brown; legs light brown to honey yellow; ovipositor and sheaths honey yellow, apex strongly sclerotised and dark. Head: PageBreak16 flagellomeres  about 0.5 times length of tergum; ovipositor 0.9 times as long as metasoma.FemaleHolotype.Female (CNIN IB-UNAM). Dominican Republic: Punta Cana , Malaise trap, Masner col. .Dominican Republic.The name of this species refers to the country where the holotype was collected, Dominican Republic.Nunes & Zaldívar-Riverónsp. n.urn:lsid:zoobank.org:act:9C8C7812-2519-4849-A2CB-D3EED92F7FFBhttp://species-id.net/wiki/Doryctopambolus_sarochensisDoryctopambolus sarochensis sp. n. distinguishes from the remaining species of the genus by having the vertex striate-rugose (striate in Doryctopambolus pilcomayensis comb. n. and smooth in the remaining two species), two pairs of propodeal apico-lateral projections, the basal ones small and blunt and the second pair long and distinctly truncate, and third metasomal tergite costate baso-laterally (entirely smooth in the other species).. Body length 3.0 mm; fore wing 2.0 mm; ovipositor 1.7 mm. Colour: head brown, malar space honey yellow, antenna honey yellow, turning brown at apex, mandible honey yellow, palpi yellow; mesosoma and metasoma dark brown to black; wings slightly infuscate; veins and stigma brown; fore leg honey yellow; middle leg honey yellow, with femur light brown; hind coxa and femur brown,PageBreak trochanter, trochantellus, tibia and tarsi honey yellow; ovipositor and sheaths honey yellow, apex strongly sclerotised and dark. Head: 23 antennomeres; eyes setose; face striate medially, striate-rugose laterally, with median area slightly swollen; clypeus costate-rugose; malar space 0.6 times eye height; frons and vertex striate-rugose; temple in dorsal view 0.8 times eye width. Mesosoma: two times longer its maximum height; pronotum rugose laterally; pronotal groove wide and scrobiculate; propleuron costate-rugose; mesoscutal lobes mostly smooth and shinning, rugose along notauli and at lateral edges; notauli narrow and scrobiculate, meeting before scutellum in a longitudinally costate-rugose area; scutellum smooth, with some setae; scutellar sulcus with three parallel carinae; height of scutellar sulcus 0.8 times height of scutellar disc; subalar sulcus wide, deep and scrobiculate, joining mesopleural sulcus in an inverted “V” shape; mesopleuron porcate dorsally, porcate-rugose laterally, smooth ventrally; venter of mesopleuron smooth; propodeum with two pairs of apico-lateral projections, the most basal one small and blunt, the apical one long and distinctly truncate, longer than first flagellomere. Legs: hind coxa strongly costate-rugose dorsally, poorly sculptured ventrally. Wings: fore wing length 3.8 times its maximum width, r:3RS:3RSb = 3:6:22; 2RS:3RSa:r-m = 8:6:5; m-cu arising antefurcal with vein 2RS; 1cu-a interstitial with 1M. Metasoma: length of first metasomal tergite 1.5 times its apical width, apical width about 2.0 times basal width, costate rugose with two dorsal carinae converging at apex; second metasomal tergite strongly costate; third metasomal tergite mostly smooth, costate baso-laterally, remaining metasomal tergites smooth and polished; basal sternal plate (acrosternite) about 0.33 times length of tergum; ovipositor as long as metasoma.FemaleVariation.Females. Body length 2.7–3.0 mm; antenna with 22–23 antennomeres. Males. Unknown.Holotype.Female (CNIN IB-UNAM). Venezuela: Lara, Parque Nacional Cerro Saroche, Cañaote #3, 10°11'083"N, -69°26'013"W; 1000 msnm; R. Briceño, R. Paz, W. Román, D. Torres colls.; . Paratype. Onefemale (CNIN IB-UNAM), same data as holotype .NortheastVenezuela.The name of this species refers to the place where the type material was collected..Echinodoryctes tetraspinosus Belokobylskij, lqbal & Austin. One specimen. Paratype. Female. Australia: Prevelly Park, W of Margaret River, 1 nov. 1984 W.A., I. & N. Lawrence, in marri nuts on ground. Doryctopambolus cf. pilcomayensis: Doryctopambolus sp. 1: one female (MLP): Argentina, Misiones, Estación Experimental Loreto, Dr. A. Ogloblin, 25-VI-1931; one male (MLP): Argentina, Misiones, Estación Experimental Loreto, Dr. A. Ogloblin, 8-VI-1933; Doryctopambolus sp. 2: one male (MACN): Argentina, Ciudad Autónoma de Buenos Aires, Reserva Ecológica Costanera Sur, trampa de caída, Mamani, Turienzo, Zapata, 19-I-2009; Doryctopambolus sp. 3 (CNIN IB-UNAM): Venezuela, Lara, Parque Nacional Cerro Saroche, Cañaote, 10°11'083"N, 69°26'013"W; 1000 m; R. Briceño, R. Paz, PageBreakW. Román, D. Torres colls. ; Doryctopambolus sp. 4 (CNIN IB-UNAM): Venezuela, Carabobo, Palmichal, 10.2859N, 68.2399W, 931 m, 30-31-iii-07, YPT/64 plates, shade coffee/Orange grove plantations, H. Clebsch col. .Doryctompambolus, Doryctopambolus clebschi (two specimens), Doryctopambolus dominicanus (one specimen), Doryctopambolus sarochensis (two specimens) and the two undescribed species from Venezuela (one specimen each), was congruent with the interspecific variation observed in other braconid genera, ranging from 4.2 to 14%. The lowest interspecific genetic distance occurred between specimens of the two Dominican species, Doryctopambolus clebschi and Doryctopambolus dominicanus.Interspecific variation of the barcoding locus among the examined species of


Postoperative Atrial Fibrillation (POAF) is the most common arrhythmia in cardiac surgery. Its incidence ranges between 20-40%. The objective of this study was to determine the predictive factors of POAF that occurred in patients undergoing coronary artery bypass grafting (CABG), and the impact of this event on morbidity and mortality.This bi-directional and longitudinal cohort study was made with 3,010 patients undergoing CABG between June/2009 and July/2010, aged ≥ 18 years. We excluded 382 patients for chronic or paroxysmal atrial fibrillation, atrial flutter, or association of the surgery evaluated with other procedures.The incidence of POAF was 12.4% ; rate female: male ratio of 1:3; CRI (p=0.001), creatinine >/= 2.2mg/dl (p=0.002); COPD (p=0.001), ICC (p=0.004), EuroSCORE (p<0.001), blood transfusion (p<0.001), Peripheral Arterial Insufficiency (PAI) ; emergency surgery/emergency (p= 0.006), postoperative stroke (p<0.001) and RI postoperatively (p <0.001). Patients with POAF had longer postoperative hospital stay (p<0.001), higher mortality at one year (p <0.001) and a higher rate of readmission within 30 days (p = 0.004).The age, male gender, CRI, COPD, peripheral arterial disease, creatinine >/= 2,2mg/dl, CHF, EuroSCORE, blood transfusion, emergency surgery/emergency (p= 0.006), postoperative stroke (p<0.001) and RI postoperatively (p<0.001) variables were independently predictive of POAF event. Due to the strong correlation with the increased length of hospitalization, mortality and re-hospitalization in these patients, we must implement strategies for POAF prophylaxis and prevention.


Computerized testing of neurocognitive function yields an accurate and reliable assessment . There iThe purpose of this study was to investigate neurocognitive function and recovery patterns in college students who incur migraine headaches compared to college students who do not.Volunteers (ages 18-29) completed computerized neurocognitive baseline (B) testing. Forty-four migraineurs incurring a migraine (M) were matched to 44 non-migraine (NM) controls for sex, age and education level. Verbal and visual memory, processing speed and reaction time were measured at 24 hours, 48 hours and 7 days post migraine.Repeated measures ANOVAs revealed declines in neurocognitive function of migraineurs in verbal memory , visual memory [md (24hr-B)M=-4.70 +15.61, NM=3.05+10.94; p=.041), and reaction time [md(24hr-B)M=.02±.09, NM=-.01±.04.


Fragility fractures are a common and increasing cause of morbidity and mortality in the general population; risk factors for fractures are commoner in HIV . This study aims to determine the prevalence and associations of low bone mineral density (BMD) and high fracture risk (FR) in an HIV cohort, suggest screening and management guidelines.A cross-sectional study of 223 randomly selected HIV-infected outpatients was undertaken. Recruitment was stratified by gender for age groups 30-39, 40-49 yrs and ≥50 yrs. Osteopenia and osteoporosis were diagnosed according to the WHO criteria. Patients completed a detailed questionnaire including combination HIV drugs (cART) history, & a dual-energy X-ray absorptiometry (DEXA) of Lumbar spine & Left Hip. Investigations included serum Ca, phosphate, 25OH vit D, alkaline phosphatase (Alk P) PTH, albumin, sex hormone binding globulin (SHBG), testosterone, CD4, HIV RNA. BMD risk factors were recorded including previous fractures, smoking, malabsorption, alcohol consumption, BMI, chronic diseases, physical activity index and past medication. FRAX score (10yr probability major fractures), and remaining lifetime fracture probability (RLFP) were calculated. Controls were from the Twin Research Unit at Kings College London. Data were analysed using multivariate logistic regression.Demographics: 133(60%) were male, 106(48%) were Caucasian, 71(33%) had AIDS at diagnosis. 190(85%) were taking cART, of whom 50(26%) were on their first line therapy. Osteoporosis/osteopenia were present in 13%/39% of males, 11%/29% females, and was approximately 2.4/3.0 fold greater than age-matched controls. The overall mean 10 yr fracture risk was 3.16%. RLFP exceeded 1.0 in 76% HIV patients, and <20% controls.Factors associated with low BMD after multivariate analysis: adjusted OR(95 % CI)/p-value BMI 0.90, p<0.001, Alk P 1.01, p 0.05, testosterone 1.04, p 0.01, Initiated cART 3.61, p 0.01. The lack of association with age is notable, adjusted OR 1.08, p 0.35Reduced bone mineral density and subsequent fracture risk is much commoner in patients with HIV compared to controls, especially, and those taking cART, and occurred across the age ranges. Hence screening for BMD and risk factors for fragility fractures is indicated in patients with HIV at a younger age than in the general population, especially if they are on cART.


Staphylococcus aureus Hospitalizations," by Matthew J Kuehnert et al., an error occurred. In Table 3, columns 3 and 5, the rates shown for hospitalization with S. aureus and MRSA-related discharge diagnoses were per 10,000 discharges, rather than per 1,000 discharges, as indicated.In “Methicillin-resistant–http://www.cdc.gov/eid/11/6/04-0831-t3.htmThe corrected table appears in the updated article at We regret any confusion this error may have caused.


The article title is incorrect. “The correct title is: MicroRNA-130b Suppresses Migration and Invasion of Colorectal Cancer Cells through Downregulation of Integrin β1.”The correct Citation is: Zhao Y, Miao G, Li Y, Isaji T, Gu J, et al. (2014) MicroRNA-130b Suppresses Migration and Invasion of Colorectal Cancer Cells through Downregulation of Integrin β1. PLoS ONE 9(2): e87938. doi:10.1371/journal.pone.0087938


The Sanger Mouse Genetics Project was incorrectly abbreviated in the citation. The correct citation is: Migdalska AM, van der Weyden L, Ismail O, White JK, Sanger Mouse Genetics Project, et al. (2012) Modeling Partial Monosomy for Human Chromosome 21q11.2-q21.1 Reveals Haploinsufficient Genes Influencing Behavior and Fat Deposition. PLoS ONE 7(1): e29681.


Granulicella mallensis MP5ACTX8T is a novel species of the genus Granulicella in subdivision 1of Acidobacteria. G. mallensis is of ecological interest being a member of the dominant soil bacterial community active at low temperatures and nutrient limiting conditions in Arctic alpine tundra. G. mallensis is a cold-adapted acidophile and a versatile heterotroph that hydrolyzes a suite of sugars and complex polysaccharides. Genome analysis revealed metabolic versatility with genes involved in metabolism and transport of carbohydrates. These include gene modules encoding the carbohydrate-active enzyme (CAZyme) family involved in breakdown, utilization and biosynthesis of diverse structural and storage polysaccharides including plant based carbon polymers. The genome of Granulicella mallensis MP5ACTX8T consists of a single replicon of 6,237,577 base pairs (bp) with 4,907 protein-coding genes and 53 RNA genes. T (= ATCC BAA-1857T = DSM 23137T), is the type strain of the species Granulicella mallensis [Granulicella, in subdivision 1 of Acidobacteria, was first described by Pankratov et al. in 2010 [Granulicella mallensis  was described along with other species of the genus Granulicella isolated from tundra soil [Granulicella species.Strain MP5ACTX8Acidobacteria is one of the most ubiquitous bacterial phyla found in diverse habitats and is abundant in most soil environments [Acidobacteria are phylogenetically and physiologically diverse [Acidobacterium [Terriglobus [Edaphobacter [Granulicella [Acidipila [Telmatobacter [Acidicapsa [Bryocella [Bryobacter [Blastocatella [Thermotomaculum [Holophaga [Geothrix [Acanthopleuribacter [Candidatus Koribacter versatilis’ [Candidatus Solibacter usitatus’ [Candidatus Chloracidobacterium thermophilum’ [Acidobacteria are relatively difficult to cultivate with slow growth rates and typically require up to several weeks to develop visible colonies on solid media. Nevertheless, the phylogenetic diversity, ubiquity and abundance of this group suggest that they play important ecological roles in soils. The abundance of Acidobacteria has been found to correlate with soil pH [Acidobacteria being most abundant in slightly acidic soils. Our previous studies have shown that Acidobacteria dominate in the acidic tundra heaths of northern Finland [Acidobacteria [T was classified as a novel species of the genus Granulicella [Granulicella mallensis MP5ACTX8T , G. sapmiensis S6CTX5AT (96.2%) and G. arctica MP5ACTX2T (96.1%) and 94.6 – 97.4% to G. rosea TPO1014T (94.6%), G. aggregans TPB6028T (96.0%), G. pectinivorans TPB6011T (96.1%), G. paludicola OB1010T (96.5%) and G. paludicola LCBR1 (97.4%). Phylogenetic analysis based on the 16S rRNA gene of taxonomically classified strains of family Acidobacteriaceae placed G. paludicola type strain OB1010 T as the closest taxonomically classified relative of G. mallensis MP5ACTX8T  at pH 3.5–6.5 (optimum pH 5) and at +4 to +28 °C (optimum 24–27 °C) [T forms opaque white mucoid colonies with a diameter of approximately 1 mm. Cells are Gram-negative, non-motile, aerobic rods, approximately 0.5–0.7 mm wide and 0.6–1.3 mm long. Growth observed with up to 1.5% NaCl (w/v) (T is shown in 4–27 °C) . On R2 aCl (w/v) . The celG. mallensis utilizes D-glucose, maltose, cellobiose, D-fructose, D-galactose, lactose, lactulose, D-mannose, D-ribose, raffinose, sucrose, trehalose, D-xylose, N-acetyl-D-glucosamine, glucuronate, glutamate, melezitose and salicin, but does not utilize D-arabinose, acetate, formate, pyruvate, malate, mannitol, D- or L-alanine, D-glycine, L-leucine, L-ornithine, gluconic acid, aspartate, dulcitol, butyrate, caproate, valerate, lactate, oxalate, propionate, fumarate, adonitol, methanol, ethanol, succinate, D-sorbitol or myoinositol, when grown using VL55 mineral medium with 100 mg yeast extract l-1. G. mallensis hydrolyzes aesculin, starch, pectin, laminarin and lichenan, but not gelatin, cellulose, xylan, sodium alginate, pullulan, chitosan or chitin on R2 medium. Strains show positive reaction for acid and alkaline phosphatases, leucine arylamidase, a-chymotrypsin, naphthol-AS-BI-phosphohydrolase, α- and β-galactosidases, α- and β-glucosidases, N-acetyl- β-glucosaminidase, β-glucuronidase, trypsin and valine arylamidase, but negative for α-fucosidase, α-mannosidase, esterase (C4 and C8), lipase (C14) and cystine arylamidase. Strain MP5ACTX8T reduces nitrate to nitrite. Strain MP5ACTX8T is resistant to the antibiotics erythromycin, chloramphenicol, neomycin, rifampicin, streptomycin, gentamicin, polymyxin B and penicillin, but susceptible to ampicillin, kanamycin, tetracycline, lincomycin, novobiocin and bacitracin [G. mallensis are iso-C15:0 (45.3%), C16:1ω7c (28.7%), iso-C13:0 (8.3%) and C16:0 (8.9%). The cellular fatty acid compositions of strain MP5ACTX8T were relatively similar to that of other Granulicella strains with fatty acids iso-C15:0 and C16:1ω7c being most abundant in all strains. Strain MP5ACTX8T contains MK-8 as the major quinone.The major cellular fatty acids in G. mallensis strain MP5ACTX8T was selected for sequencing in 2009 by the DOE Joint Genome Institute (JGI) community sequencing program. The Quality Draft (QD) assembly and annotation were completed on December 26, 2010. The complete genome was made available on Dec. 1, 2011. The genome project is deposited in the Genomes On-Line Database (GOLD) [T is deposited in GenBank (CP003130). e (GOLD)  and the G. mallensis MP5ACTX8T was cultivated on R2 medium as previously described [escribed . Genomicescribed .G. mallensis MP5ACTX8T (JGI ID 4088692) was generated at the DOE Joint genome Institute (JGI) using a combination of Illumina [The finished genome of Illumina  and 454 Illumina . The 454Illumina . The 454Illumina . The sofIllumina  was usedIllumina  was usedIllumina  or sequeGenes were identified using Prodigal  as part The genome consists of one circular chromosome of 6,211,694 bp in size with a GC content of 57.8 mol% and consists of 53 RNA genes . Of the Granulicella mallensis type strain MP5ACTX8T has the largest genome size of 6.2 Mbp. among the three tundra soil strains of subdivision 1 Acidobacteria [Granulicella mallensis identified a high abundance of genes assigned to COG functional categories for transport and metabolism of carbohydrates (9.1%) and amino acids (6.7%) and involved in cell envelope biogenesis (8.3%) and transcription (8.6%). Further genome analysis revealed an abundance of gene modules encoding for functional activities within the carbohydrate-active enzymes (CAZy) family [G. mallensis hydrolyzed complex carbon polymers, including CMC, pectin, lichenin, laminarin and starch, and utilized sugars such as cellobiose, D-mannose, D-xylose, D-trehalose. This parallels genome predictions for CDSs encoding for enzymes such as cellulases, pectinases, alginate lyases, trehalase and amylases. In addition, the G. mallensis genome contained a cluster of genes in the neighborhood of the cellulose synthase gene (bcsAB) which included cellulase (bscZ) (endoglucanase Y) of family GH8, cellulose synthase operon protein (bcsC) and a cellulose synthase operon protein (yhjQ) involved in cellulose biosynthesis. Detailed comparative genome analysis of G. mallensis MP5ACTX8T with other Acidobacteria strains for which finished genomes were available is reported in Rawat et al. [G. mallensis is involved in hydrolysis, the utilization of stored carbohydrates, and in the biosynthesis of exopolysaccharides from organic matter and plant based polymers in the soil. Therefore, we infer that strain G. mallensis may be central to carbon cycling processes in arctic and boreal soil ecosystems.bacteria . Genome ) family  involvedt et al. . The dat


Hepatic dysfunction has been associated with outcome of ICU patients. However, most scoring systems including APACHE II only marginally reflect acute liver dysfunction on admission. Indocyanine green (ICG) is eliminated by hepatobiliary excretion. Therefore, the ICG plasma disappearance rate (ICG-PDR) is used as a dynamic liver function test. ICG-PDR has been associated with mortality in several studies.A prospective study to compare prediction of 28-day mortality by ICG-PDR, other markers of liver function and scoring systems (primary endpoint). In the subgroup of patients with transpulmonary thermodilution (TPTD) monitoring , predictive capabilities of ICG-PDR were compared with cardiac index (CI), extravascular lung water index (EVLWI), global end-diastolic volume index (GEDVI) and pulmonary vascular permeability index (PVPI). ICG-PDR  and all other predictors were determined within 48 hours after admission. Statistics: IBM SPSS 20.P <0.001) and SOFA . Among markers of hepatic function on admission, ICG-PDR provided the largest AUC , which was larger than for GOT , bilirubin  and INR . Among TPTD parameters, only PVPI significantly predicted 28-day mortality , whereas CI, GEDVI and EVLWI were not predictive. Prediction of 28-day mortality by SOFA could not be improved by models including any hepatic parameter. By contrast, ICG-PDR was independently (P = 0.036) associated with mortality when included in a model (R = 0.58) with APACHE II. This model based on APACHE II and ICG-PDR provided the largest of all ROC-AUCs . ICG-PDR itself was independently associated with age (P = 0.025), but not with any other biometric parameter .A total of 154 patients , age 59 ± 13 years, APACHE II score 16.0 ± 5.7, SOFA score 7.6 ± 4.2. Etiology: sepsis 14.4%, cirrhosis 28.8%, GI bleeding 8.9%, ARDS 17.8%, cardiogenic shock 4.1%, acute renal failure 3.4%, various 22.6%. The 28-day mortality was significantly predicted by APACHE II (ROC-AUC: 0.762; ICG-PDR on admission is an independent predictor of 28-day mortality. Predictive capabilities particularly of APACHE II can be improved by combination with ICG-PDR. Among TPTD-derived parameters, only PVPI provides significant prediction of mortality.


The authors would like to add Matthias Gunther as the 6th author. The updated byline is: David A. Feinberg1,2,3*, Steen Moeller4, Stephen M. Smith5, Edward Auerbach4, Sudhir Ramanna1, Matthias Gunther1, Matt F. Glasser6, Karla L. Miller5, Kamil Ugurbil4, Essa Yacoub4The updated author contributions are: Conceived and designed the experiments: DAF SMS KU EY. Performed the experiments: DAF SM EA SR MG EY. Analyzed the data: DAF SM SMS EY KLM MFG. Contributed reagents/materials/analysis tools: DAF SM EA SR SMS MG MFG. Wrote the paper: DAF SM SMS KU EY KLM.


Plasmopara viticola, is one of the most severe diseases of grapevine and is commonly controlled by fungicide treatments. The beneficial microorganism Trichoderma harzianum T39 (T39) can induce resistance to downy mildew, although the molecular events associated with this process have not yet been elucidated in grapevine. A next generation RNA sequencing (RNA-Seq) approach was used to study global transcriptional changes associated with resistance induced by T39 in Vitis vinifera Pinot Noir leaves. The long-term aim was to develop strategies to optimize the use of this agent for downy mildew control.Downy mildew, caused by P. viticola inoculation. RNA-Seq analysis resulted in the identification of 7,024 differentially expressed genes, highlighting the complex transcriptional reprogramming of grapevine leaves during resistance induction and in response to pathogen inoculation. Our data show that T39 has a dual effect: it directly modulates genes related to the microbial recognition machinery, and it enhances the expression of defence-related processes after pathogen inoculation. Whereas several genes were commonly affected by P. viticola in control and T39-treated plants, opposing modulation of genes related to responses to stress and protein metabolism was found. T39-induced resistance partially inhibited some disease-related processes and specifically activated defence responses after P. viticola inoculation, causing a significant reduction of downy mildew symptoms.More than 14.8 million paired-end reads were obtained for each biological replicate of T39-treated and control leaf samples collected before and 24 h after The global transcriptional analysis revealed that defence processes known to be implicated in the reaction of resistant genotypes to downy mildew were partially activated by T39-induced resistance in susceptible grapevines. Genes identified in this work are an important source of markers for selecting novel resistance inducers and for the analysis of environmental conditions that might affect induced resistance mechanisms. Plasmopara viticola (Berk. and Curt.) Berl. and de Toni is a biotrophic oomycete that causes downy mildew in grapevine Release 3 [78] in control (C), Trichoderma harzianum T39-treated (T39), Plasmopara viticola-inoculated control (C+P.v.), and P. viticola-inoculated T39-treated (T39+P.v.) plants. Expression values in each biological replicate (numbered from 1 to 3) and each sequencing replicate (named A and B) are also listed. Click here for fileCorrelations between sequencing replicates of RNA-Seq analysis. Comparison of the expression levels of all grapevine genes, expressed as fragments per kilobase of transcript per million fragments mapped (FPKM), in the two sequencing replicates (named A and B) of (A) control (C) biological replicate no. 1; (B) C biological replicate no. 2 (B); (C) C biological replicate no. 3; (D) Trichoderma harzianum T39-treated (T39) biological replicate no. 1; (E) T39-treated biological replicate no. 2; (F) T39-treated biological replicate no. 3; (G) Plasmopara viticola-inoculated control (C+P.v.) biological replicate no. 1; (H) C+P.v. biological replicate no. 2; (I) C+P.v. biological replicate no. 3; (J) P. viticola-inoculated T39-treated plants (T39+P.v) biological replicate no. 1; (K) T39+P.v biological replicate no. 2; and (L) T39+P.v biological replicate no. 3. Click here for filePrincipal component analysis of grapevine treatments. Principal component analysis (PCA) is based on the expression values (FPKM) of all grapevine genes for each sequencing replicate (named A and B) of each biological replicate (numbered from 1 to 3) for control (C), Trichoderma harzianum T39-treated (T39), Plasmopara viticola-inoculated control (C+P.v.), and P. viticola-inoculated T39-treated (T39+P.v.) plants. Click here for filePearson’s correlation coefficients between sequencing and biological replicates of RNA-Seq analysis. Pearson’s correlation coefficients are based on the gene expression values (FPKM) of all grapevine genes in two sequencing replicates (A and B) of each biological replicate (numbered from 1 to 3) for control (C), Trichoderma harzianum T39-treated (T39), Plasmopara viticola-inoculated control (C+P.v.), and P. viticola-inoculated T39-treated (T39+P.v.) plants. Click here for fileExpression levels, clustering, and annotation results of differentially expressed genes. Gene expression values (FPKM) and standard errors are reported for differentially expressed  genes in control (C), Trichoderma harzianum T39-treated (T39), Plasmopara viticola-inoculated control (C+P.v.), and P. viticola-inoculated T39-treated (T39+P.v.) plants. Differentially expressed genes were identified by the DESeq package with a false discovery rate (FDR) of 5% and a fold-change greater than two in at least one pairwise comparison. Fold-changes, clustering results, and functional annotation are reported for each gene. Click here for filePrimer sequences of the grapevine genes analysed by real-time RT-PCR. Forward and reverse primer sequences of real-time RT-PCR analysis are reported for 24 selected grapevine genes. Grapevine Actin (TC81781)  and VATP16 (XM_002269086.1) [94] were used as constitutive genes for normalising the real-time RT-PCR data. Click here for fileExpression levels of grapevine genes that are members of the actin gene family. Gene expression values (FPKM) and standard errors are reported for grapevine genes belonging to the Actin gene family in control (C), Trichoderma harzianum T39-treated (T39), Plasmopara viticola-inoculated control (C+P.v.), and P. viticola-inoculated T39-treated (T39+P.v) plants. Click here for file


Corrigendum to Anthony L. Cunningham, Andrew N. Harman, Najla Nasr (2013) Initial HIV mucosal infection and dendritic cells. EMBO Mol Med 5 (658–660) DOI: 10.1002/emmm.201202763The name of the second author in this article, “Andrew N. Harman,” missed the middle initial N. The correct author names are listed above.


O-Sulfated Heparan Sulfate Recognized by the Antibody HS4C3 Contributes to the Differentiation of Mouse Embryonic Stem Cells via Fas Signaling.The word "Contributes" is misspelled in the article title. The correct title is: 3-O-Sulfated Heparan Sulfate Recognized by the Antibody HS4C3 Contribute to the Differentiation of Mouse Embryonic Stem Cells via Fas Signaling. PLoS ONE 7(8): e43440. doi:10.1371/journal.pone.0043440 The correct citation is: Hirano K, Sasaki N, Ichimiya T, Miura T, Van Kuppevelt TH, et al. (2012) 3-


Long-term potentiation (LTP) at thalamic input synapses to the lateral nucleus of the amygdala (LA) has been proposed as a cellular mechanism of the formation of auditory fear memories. We have previously shown that signaling via ERK/MAPK in both the LA and the medial division of the medial geniculate nucleus/posterior intralaminar nucleus (MGm/PIN) is critical for LTP at thalamo-LA synapses. Here, we show that LTP-inducing stimulation of thalamo-LA inputs regulates the activation of ERK and the expression of ERK-driven immediate early genes (IEGs) in both the LA and MGm/PIN. Further, we show that pharmacological blockade of NMDAR-driven synaptic plasticity, NOS activation, or PKG signaling in the LA significantly impairs high-frequency stimulation-(HFS-) induced ERK activation and IEG expression in both regions, while blockade of extracellular NO signaling in the LA impairs HFS-induced ERK activation and IEG expression exclusively in the MGm/PIN. These findings suggest that NMDAR-driven synaptic plasticity and NO-cGMP-PKG signaling within the LA coordinately regulate ERK-driven gene expression in both the LA and the MGm/PIN following LTP induction at thalamo-LA synapses, and that synaptic plasticity in the LA promotes ERK-driven transcription in MGm/PIN neurons via NO-driven “retrograde signaling”. Fear conditioning is a type of associative learning that is widely studied as a model of learning and memory across a variety of species. Fear conditioning has been extensively characterized at the behavioral level, particularly auditory fear conditioning, in which a tone  is paired with footshock . In brief, auditory fear conditioning is thought to involve transmission and integration of sensory information from CS and US pathways within the lateral nucleus of the amygdala (LA), where alterations in synaptic transmission are believed to encode key aspects of the learning –3. In suLong-term potentiation (LTP), an experimental model of synaptic plasticity, is widely believed to be a potential mechanism by which fear conditioning promotes synaptic alterations in the LA , 6. In sin vitro slice recording methods have pointed to a role for a number of intracellular signaling pathways in LTP at thalamic input synapses to the LA, including CaMKII  or 60 min , or 60 min (ERK1: t(5) = 1.847, P > .05; ERK2: t(5) = 0.434, P > .05) time points. Further, this effect was not observed in LFS controls  = 0.725, P > .05; pERK2: t(5) = 0.759, P > .05), or 30 min (pERK1: t(7) = 1.076, P > .05; pERK2: t(7) = 0.300, P > .05) time points following LFS. The findings for the LA are presented in Figures  Figures , left. I Figures , right; t(5) = 2.897, P < .05; pERK2: t(5) = 2.596, P < .05) and 30 min after stimulation (pERK1: t(3) = 5.655, P < .05; pERK2: t(4) = 3.747, P < .05). No significant differences were observed for the 60 min time point (pERK1: t(4) = 0.405, P > .05; pERK2: t(4) = 0.073, P > .05)  = 0.520, P > .05; ERK2: t(5) = 0.731, P > .05), 30 min (ERK1: t(4) = 0.661, P > .05; ERK2: t(4) = 0.648, P > .05), or 60 min (ERK1: t(4) = 1.131, P > .05; ERK2: t(4) = 0.195, P > .05) time points. Interestingly, LFS controls were also observed to have elevated levels of phospho-ERK1 and phospho-ERK2 in MGm/PIN at 5 minutes (pERK1: t(5) = 3.193, P < .05; pERK2: t(5) = 4.073, P < .05), but not at 30 min (pERK1: t(5) = 0.529, P > .05; pERK2: t(5) = 0.067, P > .05)  = 1.551, P > .05; ERK2: t(5) = 0.691, P > .05) or 30 min (ERK1: t(4) = 0.224, P > .05; ERK2: t(4) = 1.611, P > .05) time points following LFS. The fact that both HFS and LFS induced increases in ERK activation in the MGm/PIN at 5 min suggests that the elevated ERK phosphorylation in MGm/PIN at this time point was due to local electrical stimulation. Importantly, at the 30 min time point only HFS promoted significant ERK activation in the MGm/PIN, suggesting that the enhanced ERK phosphorylation in MGm/PIN at this later time point is specifically associated with LTP. The findings for the MGm/PIN are presented in Figures  Figures , left. I Figures , right. In our first series of experiments, we observed significant increases in ERK activation in the LA and MGm/PIN following HFS of the thalamo-LA pathway. In the present experiments, we used a combination of Western blotting and immunohistochemistry to examine whether LTP-inducing stimulation of thalamic input synapses to the LA regulate the ERK-driven IEGs Arc/Arg3.1, EGR-1 and c-Fos in the LA and the MGm/PIN. As before, anesthetized rats received either HFS or LFS of thalamic inputs to the LA and were sacrificed by decapitation 2 hours later Figures , a time t(8) = 6.502, P < .05; c-Fos: t(8) = 3.901, P < .05; EGR-1: t(8) = 5.273, P < .05; t(8) = 1.294, P > .05; c-Fos: t(8) = 0.241, P > .05; EGR-1: t(8) = 2.822, P > .05; t(8) = 3.642, P < .05; c-Fos: t(5) = 3.403, P < .05; EGR-1: t(6) = 2.59, P < .05; t(8) = 1.163. P > .05; c-Fos: t(5) = 0.094, P > .05; EGR-1: t(6) = 1.720, P > .05; The results of our Western blotting experiments are depicted in Figures t(5) = 6.894, P < .05; t(5) = 0.124, P > .05; t(5) = 3.822, P < .05; c-Fos: t(5) = 4.144, P < .05; Figures t(5) = 0.709, P > .05; c-Fos: t(5) = 0.121, P > .05; Figures Immunohistochemical localization of the three IEGs after HFS and LFS in both the LA and MGm/PIN can be seen in Figures t(6) = 22.556, P < .05], EGR-1 , and c-Fos  proteins  = 1.913, P > .05; EGR-1: t(5) = 0.227, P > .05; c-Fos: t(5) = 0.395, P > .05]. Rats receiving HFS exhibited Arc/Arg3.1, EGR-1, and c-Fos labeled cells throughout the MGm and PIN, while very few labeled cells were observed in the MGv  = 4.075, P < .05; pERK2: t(4) = 3.904, P < .05; t(4) = 0.173, P > .05; pERK2: t(4) = 1.273, P > .05] or 7-Ni , however, significantly impaired HFS-induced ERK activation in the LA (t(4) = 5.026, P < .05; pERK2: t(4) = 3.640, P < .05; The findings are depicted in Figures n the LA . Interest(4) = 3.035, P < .05; pERK2: t(4) = 3.864, P < .05; t(5) = 1.405, P > .05; pERK2: t(5) = 1.348, P > .05], 7-Ni , or c-PTIO . In the MGm/PIN, we observed a significant increase in phospho-ERK1/2 activation 30 min after HFS in vehicle-infused controls [pERK1: Thus, blockade of NMDAR-driven synaptic plasticity and NO signaling at the level of the LA impairs HFS-induced ERK activation not only in the LA, but also in the MGm/PIN. Further, extracellular release of NO in the LA is required for HFS-induced ERK activation in the MGm/PIN, but not in the LA. In this series of experiments, we examined whether blockade of NMDAR-driven synaptic plasticity and NO signaling in the LA impairs HFS-induced expression of the IEGs Arc/Arg3.1, c-Fos, and EGR-1 in both the LA and the MGm/PIN. As in the previous experiment, anesthetized rats were given intra-LA infusion of ifenprodil, 7-Ni, or c-PTIO prior to LTP-inducing stimulation of the thalamo-LA pathway . In addit(7) = 3.440. P < .05; c-Fos: t(7) = 2.405, P < .05; EGR-1: t(7) = 5.009, P < .05; t(7) = 0.377. P > .05; c-Fos: t(7) = 0.708, P > .05; EGR-1: t(7) = 0.306, P > .05; t(7) = 2.629. P < .05; c-Fos: t(7) = 3.783, P < .05; EGR-1: t(7) = 3.440, P < .05; t(7) = 0.211. P > .05; c-Fos: t(7) = 0.592, P > .05; EGR-1: t(7) = 0.139, P > .05; The findings for rats infused with ifenprodil are depicted in Figures t(7) = 2.374. P < .05; c-Fos: t(7) = 2.462, P < .05; EGR-1: t(7) = 2.402, P < .05; t(7) = 0.672. P > .05; c-Fos: t(7) = 1.101, P > .05; EGR-1: t(7) = 1.499, P > .05; t(7) = 3.066. P < .05; c-Fos: t(7) = 2.383, P < .05; EGR-1: t(7) = 2.687, P < .05; t(7) = 0.065. P > .05; c-Fos: t(7) = 0.025, P > .05; EGR-1: t(7) = 0.460, P > .05; The findings for rats infused with 7-Ni are depicted in Figures t(7) = 2.374. P < .05; c-Fos: t(7) = 2.462, P < .05; EGR-1: t(7) = 2.402, P < .05; t(7) = 2.545. P < .05; c-Fos: t(7) = 2.406, P < .05; EGR-1: t(7) = 2.509, P < .05; t(7) = 3.066. P < .05; c-Fos: t(7) = 2.383, P < .05; EGR-1: t(7) = 2.687, P < .05; t(7) = 1.241. P > .05; c-Fos: t(7) = 0.696, P > .05; EGR-1: t(7) = 1.600, P > .05; The findings for rats infused with c-PTIO are depicted in Figures t(7) = 7.972. P < .05; c-Fos: t(7) = 3.686, P < .05; EGR-1: t(7) = 4.599, P < .05; t(6) = 1.688. P > .05; c-Fos: t(7) = 0.631, P > .05; EGR-1: t(7) = 1.287, P > .05; t(7) = 7.972. P < .05; c-Fos: t(7) = 4.064, P < .05; EGR-1: t(7) = 4.901, P < .05; t(6) = 1.688. P > .05; c-Fos: t(7) = 1.101, P > .05; EGR-1: t(7) = 0.401, P > .05; The findings for rats infused with PKG inhibitor Rp-8-Br-PET-cGMPS are depicted in Thus, similar to the findings of our ERK experiments, blockade of NMDAR-driven synaptic plasticity and NO signaling at the level of the LA impairs HFS-induced IEG expression not only in the LA but also in the MGm/PIN. Further, extracellular release of NO in the LA appears to be required for HFS-induced IEG expression in the MGm/PIN, but not in the LALong-term potentiation (LTP) at thalamo-LA synapses has been proposed as a candidate cellular mechanism of the formation of auditory fear memories, yet little is known about the molecular mechanisms underlying LTP at this synapse. In the present study, we have examined the regulation of ERK and that of three different ERK-driven IEGs at both sides of the thalamo-LA synapse after LTP-inducing stimulation. We found that LTP-inducing stimulation at thalamo-LA synapses is accompanied by ERK activation and ERK-driven gene expression not only in the LA, but also in regions of the MGm/PIN that are presynaptic to the LA. Further, pharmacological disruption of either NMDAR-driven synaptic plasticity or NO-cGMP-PKG signaling at the level of the LA impairs ERK activation and IEG expression in each region. Collectively, these findings suggest that NMDAR-driven synaptic plasticity and NO signaling within the LA coordinately regulate ERK activation and ERK-driven gene expression in both the LA and the MGm/PIN following LTP induction at thalamo-LA synapses.A recent study in our lab showed that LTP-inducing stimulation of thalamo-LA inputs induces ERK activation in the LA, and that intra-LA infusion of ERK/MAPK inhibitor impairs LTP at thalamo-LA synapses . HoweverOur findings also revealed that LTP-inducing stimulation of thalamo-LA synapses regulates the expression of the ERK-driven IEGs Arc/Arg3.1, c-Fos, and EGR-1 in both the LA and MGm/PIN. Previous studies have extensively documented the role of Arc/Arg3.1 –32 and EIn contrast to Arc/Arg3.1, both EGR-1 and c-Fos are thought to behave as transcription factors, regulating the expression of late-response genes that are critical for long-term synaptic plasticity. Importantly, several studies have observed associative increases in the expression of c-Fos and EGR-1 in the LA after cued fear conditioning –40. FurtOur findings of enhanced activation of ERK-driven transcriptional regulation in both LA and MGm/PIN neurons is consistent with previous work that has shown that infusion of a MEK inhibitor into either the LA or the MGm/PIN impairs LTP in the LA , 23 and in vitro [While most widely studied in the hippocampus , 58–63 ain vitro , 18.∼2-3 mm distance in such a rapid manner. However, previous reports have suggested that retrograde transport can occur very rapidly in neurons, between ∼4–8 mm/hr [In the present study, we used pharmacological methods to ask whether NMDAR-driven synaptic plasticity and NO-cGMP-PKG signaling may be regulating both ERK and ERK-driven transcription within the LA and the MGm/PIN. We showed that blockade of NR2B (via ifenprodil), NOS (via 7-Ni), or PKG (via Rp-8-Br-PET-cGMPS) significantly impaired HFS-induced activation of ERK and ERK-driven gene expression in both the LA and the MGm/PIN. Remarkably, however, blockade of extracellular NO signaling (via c-PTIO) significantly impaired HFS-induced activation of ERK and ERK-driven gene expression in the MGm/PIN, but not in the LA. These findings suggest that the ERK activation and downstream IEG expression in MGm/PIN following LTP is driven by NO “retrograde signaling” at the level of the LA. The identity of the signal that links NO release at the level of the LA with ERK activation and ERK-driven gene expression at the level of the MGm/PIN is currently unknown, as is how such a signal may propagate in a retrograde manner from synapse to nucleus across a –8 mm/hr . Further–8 mm/hr , or inhi–8 mm/hr , which m–8 mm/hr . The present findings are consistent with a revised model of the molecular events underlying synaptic plasticity at thalamo-LA synapses in which NMDAR-driven synaptic plasticity and NO signaling in LA neurons promotes pre- and postsynaptic alterations at thalamo-LA synapses via regulation of ERK-driven gene expression in MGm/PIN and LA neurons, respectively, . In thatIn support of this model, recent studies from our lab and others have shown that ERK-driven transcription in the MGm/PIN is required not only for fear memory consolidation , 67, 68 In summary, our findings suggest that NMDAR-driven synaptic plasticity and NO signaling within the LA coordinately regulate ERK activation and ERK-driven gene expression in both the LA and the MGm/PIN following LTP induction at thalamo-LA synapses, and further suggest that synaptic plasticity in the LA promotes ERK-driven transcription in MGm/PIN neurons via NO-driven “retrograde signaling”. These findings further extend what is known about the molecular basis of LTP within the LA, and provide additional evidence that studying LTP at thalamo-LA synapses may inform us about the molecular basis of fear memory formation in the amygdala.


These brain and renal injuries were associated with increased gene expression of NADPH oxidase components, NADPH oxidase activity and oxidative stress in brain and kidney tissues as well as systemic oxidative stress. Treatment with the ARB, olmesartan (10 mg/kg/day) reduced blood pressure in saline-drinking KK-Ay mice and attenuated cognitive decline, BBB disruption, glomerular injury and albuminuria, which were associated with a reduction of NADPH oxidase activity and oxidative stress in brain and kidney tissues as well as systemic oxidative stress. Furthermore, a suppressive dose of azelnidipine (3 mg/kg/day) exaggerated these beneficial effects of olmesartan. These data support the hypothesis that a CCB enhances ARB-associated cerebrovascular-renal protective effects through suppression of NADPH oxidase-dependent oxidative stress in type 2 diabetes.Recent clinical trials have demonstrated that combination therapy with renin-angiotensin system inhibitors plus calcium channel blockers (CCBs) elicits beneficial effects on cardiovascular and renal events in hypertensive patients with high cardiovascular risks. In the present study, we hypothesized that CCB enhances the protective effects of an angiotensin II type 1 receptor blocker (ARB) against diabetic cerebrovascular-renal injury. Saline-drinking type 2 diabetic KK-A The beneficial effects of renin-angiotensin system (RAS) inhibition with angiotensin converting enzyme inhibitors (ACEIs) and angiotensin II type 1 receptor blockers (ARBs) have been demonstrated in hypertensive patients with high cardiovascular-renal risks including heart failure , diabetePrevious large-scale epidemiological studies have indicated that CKD is not related to the incidence of stroke . Howevery mice treated with the ARB, olmesartan [It has been highlighted that type 2 diabetes and hypertension are risk factors for cerebrorenal injury including cognitive decline ,22, BBB mesartan .y mice [Experimental protocols and animal care were performed according to the guidelines for the care and use of animals established by Kagawa University, Japan. The experiments were approved by the Animal Experimentation Ethics Committee at Kagawa University (No. 112). At the end of the experiment, organs were dissected under sodium pentobarbital anesthesia . Six-week-old male type 2 diabetic KK-Ay mice  and contn = 11) and saline-drinking C57BL6 mice . Type 2 diabetic KK-Ay mice were divided into four groups: tap water drinking KK-Ay mice ; saline-drinking KK-Ay mice ; saline-drinking KK-Ay mice treated with olmesartan  ; saline-drinking KK-Ay mice treated with olmesartan plus azelnidipine  . It has previously been reported that azelnidipine at 3 mg/kg body weight/day did not change blood pressure in KK-Ay mice [y mice .After a two week period of acclimatization and measuring basal parameters, 8-week-old mice were underwent combination treatments for 16 weeks. Control C57BL6 mice were divided into two groups: tap water drinking C57BL6 mice  was monitored in conscious mice by tail-cuff plethysmography . Urinary albumin and creatinine concentrations were measured by using commercially available assay kits  . PostpraAt the end of the experiment blood, brain and kidney samples were harvested under anesthesia with sodium pentobarbital . The brain and kidney tissues were harvested and fixed in 10% buffered paraformaldehyde or embedded in Tissue-Tek OCT compound , and remaining tissues were snap-frozen in liquid nitrogen. Small amounts of brain and renal cortical tissues were collected in RNAlater and stored overnight at 4°C RNAlater-treated samples were subsequently snap-frozen in liquid nitrogen and stored at -80°C until processing for RNA extraction and reverse transcription-polymerase chain reaction (RT-PCR) analysis.A passive avoidance test was performed to evaluate cognitive function as described previously . Detailsn = 6 per group) as previously described [BBB permeability was determined using the EB extravasation technique  and Mallory-Azan reagents to evaluate glomerular sclerosis and tubulointerstitial fibrosis, respectively ,32. The Glomerular podocyte injury was evaluated by immunohistochemical analysis of desmin and was performed as previously described ,31. To investigate oxidative stress in the brain and kidney tissues, DHE immunofluorescence staining was performed as previously described . DetailsNADPH oxidase-derived superoxide anion  generation was measured using lucigenin-enhanced chemiluminescence, as described previously . Details The mRNA expression in brain and renal cortical tissues were analyzed by RT-PCR using a LightCycler FastStart DNA Master SYBR Green I kit and an ABI Prism 7000 Sequence Detection System  as previously described ,31. The Plasma and urine 8-hydroxy-2’-deoxyguanosine (8-OHdG) , plasma level of non-esterified fatty acid (NEFA), triglyceride (TG), total cholesterol (TCho) , and insulin  were measured using commercially available kits.post hoc test. Values of P < 0.05 were considered statistically significant. All values are presented as means ± S.E.M.. Statistical comparisons of differences among groups were performed using one-way repeated-measures analysis of variance (ANOVA), followed by the Newman-Keuls y mice showed elevated SBP compared with that in age-matched C57BL6 or C57BL6 + 0.9% NaCl mice   . In addi tissues . These dy mice, DHE staining was significantly increased in kidney tissues compared with that in C57BL6 mice Click here for additional data file.Table S1(DOCX)Click here for additional data file.Figure S1Body weight changes during the experimental period. KK-Ay mice showed higher body weight compared to C57BL mice. However, none of the treatments affected body weight gains in KK-Ay + 0.9% NaCl mice (n=11). aP < 0.05 vs. C57BL/6, bP < 0.05 vs. C57BL/6 + 0.9% NaCl.(TIF)Click here for additional data file.Figure S2NADPH oxidase subunits gene expression in brain tissues analyzed by RT-PCR. NADPH oxidase subunits gp91phox (A) and p47phox (B) mRNA levels in whole brain tissues. Saline-dinking KK-Ay mice showed upregulation of NADPH oxidase subunit mRNA levels in brain tissues, which were attenuated by treatment with olmesartan. Furthermore, the combination of olmesartan plus azelnidipine completely prevented these changes resulting in levels similar to that in C57BL6 mice (n=8). aP < 0.05 vs. C57BL/6, bP < 0.05 vs. C57BL/6 + 0.9% NaCl, cP < 0.05 vs. KK-Ay, dP < 0.05 vs. KK-Ay + 0.9% NaCl, eP < 0.05 vs. KK-Ay + 0.9% NaCl + olmesartan.(TIF)Click here for additional data file.Figure S3NADPH oxidase subunits gene expression in kidney tissues analyzed by RT-PCR. NADPH oxidase subunits gp91phox (A) and p22phox (B) mRNA levels in renal cortical tissues. In saline-dinking KK-Ay mice, superoxide production in renal tissues was associated with upregulation of NADPH oxidsase subunit genes expression. Treatment with olmesartan markedly attenuated these changes. Furthermore, the combination of olmesartan plus azelnidipine completely prevented these changes resulting in levels similar to that in C57BL6 mice (n=8). aP<0.05 vs. C57BL/6, bP<0.05 vs. C57BL/6 + 0.9% NaCl, cP<0.05 vs. KK-Ay, dP<0.05 vs. KK-Ay + 0.9% NaCl, eP<0.05 vs. KK-Ay + 0.9% NaCl + olmesartan.(TIF)Click here for additional data file.


Salmonella enterica strains that are representatives of the S. enterica serovar Typhimurium complex in reference collection A (SARA) are closely related but exhibit differences in antibiotic resistance, which could have public health consequences. To better understand the mechanisms behind these resistances, we sequenced the genomes of two multidrug-resistant strains: SARA64 (Muenchen) and SARA33 (Heidelberg).The  Salmonella enterica is one of the most important bacterial enteric pathogens and has been implicated in food-borne illnesses worldwide .orldwide . Emergenorldwide , 3. Explorldwide , 5, consintI1) . Two of these strains, SARA64 and SARA33, exhibited resistance to ampicillin, chloramphenicol, tetracycline, streptomycin, sulfisoxazole, and kanamycin. SARA33 also showed resistance to gentamicin. Both strains were positive for the integrase found in integrons class I (intI1) ; howeverintI1) . In ordede novo assembled using CLC Genomics Workbench version 5.5.1 . The G+C mol% values of SARA64 and SARA33 were 52.0 and 52.1%, respectively, which are similar to the reported GC content for other Salmonella strains . The genomes were sequenced using an Ion Torrent (PGM) sequencing system with the 200-bp reads chemistry  at 30 to 40× coverage, using an Ion PGM 200 sequencing kit, according to the manufacturer’s instructions. Genomic sequence contigs for each strain were  strains . Strain http://www.ncbi.nlm.nih.gov/genomes/static/Pipeline.html) (in silico multilocus sequence typing (MLST) (http://cge.cbs.dtu.dk/services/) -laa, aph(3′)-la, strA, strB, and aadA1], sulfonamides (sul1 and sul2), beta-lactams (blaOXA), tetracycline (tetB), and phenicol (catA1), which explain its MDR phenotype. Some of these resistance genes were on a genomic island similar to GI-DT12 in Salmonella enterica serovar Typhimurium T000240 (intI1-blaOXA-aadA1-qacEΔ1-sul1). The genes strA, strB, and sul2 were located on a plasmid.These draft genome sequences were annotated using the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP) (ne.html) . Identitrvices/)  using thdatabase ; strain database . Antibiocreening . SARA64  T000240 , which caac(6′)-ly, aadA5, aadB, aa(6′)-33, and aadA1], sulfonamides (sul1 and sul2), beta-lactams (blaOXA-2 and blaTEM), and tetracycline (tetD). A novel integron cassette was identified that contained one hypothetical protein (hp) with unknown function, followed by aadA1 and aa(6′)-33 genes [intI1-hp-aac(6′)-33-aadA1].In contrast, SARA33 carried resistance genes for aminoglycosides [Salmonella enterica strains are now available in GenBank under accession numbers AUQD00000000 for strain SARA33 (2213-Heidelberg) and AUQE00000000 for strain SARA64 (2244-Muenchen).The draft genome sequences of these two


Immunohistochemical studies suggest that the rheumatoid nodule (RN) is a Th1 granuloma , yet theWe measured circulating cytokines of DMARD-naïve early RA patients, and compared profiles of patients with RN to those without RN.A cross-sectional cohort of 149 DMARD-naïve adults with early RA (symptom duration ≤ 2 years). Assessments included a smoking history, the presence of subcutaneous RN, the Simplified Disease Activity Index (SDAI), and X-rays scored using the modified Larsen method, Rheumatoid Factor (RF) and anti cyclic citrullinated peptides (aCCP) tests. Serum cytokines were measured using the Bio-Plex® suspension array system. Differences between patients with RN and those without RN were explored using univariate and multivariate techniques, correcting for SDAI by regression analysis.Of 149 patients , the majority were Black Africans (93%) and 34 (22.8%) had RN. Patients with RN were more likely to be males, but neither smoking history nor symptom duration was significantly different between the two groups. RN patients had severe RA with higher SDAI (p=0.04) and X-ray scores (p=0.004), and higher RF (p=0.005) and aCCP (p=0.04) titers. Cytokine profiles in the RN patients showed significant elevation of predominantly Th1 cytokines, with higher interferon-gamma (IFN-G) (p=0.05), interleukin (IL)-12 (p=0.02), and IL-2 (p=0.05). In addition, cytokines of macrophage  and fibroblast origin  were significantly higher in these patients. Multivariate analysis showed IL-7 (p<0.001) and IFN-G (p=0.001) to be independently significant.DMARD-naïve early RA patients with RN have a unique serum cytokine profile, with significant elevation of predominantly Th1 and fibroblast cytokines.


Diastolic dysfunction (DD) is common in patients with cardiovascular disease. Current cardiac magnetic resonance techniques for assessment of DD require the acquisition of tagged imaging sequences, and complex post-processing. A new post-processing software allows for rapid strain assessment using feature tracking analysis (FT)  based on conventional steady-state free precession (SSFP)-Cine sequences. The aim of this study was to compare tagging (TAG) with FT for assessment of diastolic function in patients with DD and healthy controls.20 healthy volunteers and 10 patients with echocardiographic diagnosed DD Grade II-III were investigated by the 2 techniques. Cardiovascular magnetic resonance imaging (Philips Intera 1.5T) using CSPAMM and SSFP-Cine sequences was performed for matched mid-ventricular short-axis slices. Each modality was analyzed offline using dedicated post-processing software  and early-diastolic strain rate (EDSr) was calculated from both datasets. EDSr derived from CSPAMM and SSFP-Cine data were compared and inter-observer and intra-observer variability assessed using Bland-Altman analysis and Pearson correlation coefficient.-1) was highly correlated with TAG EDSr (76.71+/- 17.45s-1), Bland-Altman analyses for method comparison yielded a mean difference of 1.4 (95% CI: 0.0 to 2.9). Limits of agreement (+/- 1.96 SD) were -14.9 and 12.0. In patients with DD FT EDSr (61.40+/- 15.60s-1) was not significantly different to TAG EDSr (57.60+/- 12.30s-1) (p=ns). The same was found in the control group FT EDSr (86.52+/- 10.55s-1) and TAG (86.27+/- 10.31s-1) (p=ns). Intra-observer variability yielded a mean difference of 1.2+/- 4.9 (FT) and 0.9+/- 3.5 (TAG). Inter-observer variability displayed a mean difference of 2.8+/- 5.84 (FT) and 0.3+/- 3.7 (TAG). Pearson correlation coefficient was (0.96 (FT); 0.97 (TAG)) for intra-observer variability and (0.94 (FT); 0.97 (TAG)) for inter-observer variability.For all measurements FT EDSr (78.15+/- 17.17 sAssessment of EDSr using a novel feature tracking technique shows good agreement with tagging derived data and may thus be used for rapid clinical determination of diastolic dysfunction. However, compared to feature tracking, tagging revealed lower intra - and inter-observer variability and may therefore remain the preferred technique for research applications.None.