Kinome profiling of sugar signaling in plants using multiple platforms

Although the primary sequence of kinases shows substantial divergence between unrelated eukaryotes, variation in the motifs that are actually phosphorylated by eukaryotic kinases is much smaller. Hence arrays developed for kinome profiling of mammalian cells are useful for kinome profiling of plant tissues as well, facilitating the study of plant signal transduction. We recently employed the Pepscan kinomics chip to reveal the small GTPases in plant sucrose signaling. Here we show that employing a different peptide library (the Pepscan kinase chip) largely similar results are obtained, confirming these earlier data, but such a different library also contributes new insights into the molecular details mediating plant cell responses to a sugar stimulus. Thus when studying plant signal transduction employing peptide arrays, using multiple platforms both increases the confidence of results and provides additional information.Key words: sucrose, arabidopsis, kinome profiling, kinase, MAPK, SnRK, casein kinase, tyrosine phosphorylation, CDC2, AGC kinaseIn our recently published paper1 we analyze the changes in the plant kinome after sucrose feeding compared to control sorbitol feeding of Arabidopsis. We employed kinomics chips (Pepscan Presto, The Netherlands) containing 960 different kinase consensus peptides selected for their importance in mammalian signal transduction. In addition we used kinase chips (Pepscan Presto, The Netherlands), containing 1,152 peptides covering the majority of peptides available through Phosphobase (version 2.0). These chips contain peptides derived from phosphorylation events described in many kingdoms and are taken from animals, plants, fungi and even bacteria.^2,3 Full details as to the peptides spotted can be found at www. pepscanpresto.com. Results of the cluster analysis of three independent biological replicas of kinome profiling after treatment with water, sorbitol, sucrose and glucose are depicted in Figure 3A of our recent paper.¹ A further analysis of this set of experiments was not given, due to a lower than expected biological reproducibility. In contrast to the >0.8 correlation observed for the set described in our recent paper, correlations of below 0.5 were seen for the experimental set analyzed on the kinase1 chips. However, we feel that some observation made based on these kinome profiles contain valuable information, if only to suggest follow-up experiments.Comparison of the kinome profiles of Arabidopsis treated with sucrose or sorbitol for 1 h revealed a set of 93 differentially phosphorylated consensus peptides, with the majority of 59 peptides showing reduced phosphorylation after sucrose treatment (

Mechanisms and Evidence of Genital Coevolution: The Roles of Natural Selection,Mate Choice,and Sexual Conflict

Genital coevolution between the sexes is expected to be common because of the direct interaction between male and female genitalia during copulation. Here we review the diverse mechanisms of genital coevolution that include natural selection, female mate choice, male–male competition, and how their interactions generate sexual conflict that can lead to sexually antagonistic coevolution. Natural selection on genital morphology will result in size coevolution to allow for copulation to be mechanically possible, even as other features of genitalia may reflect the action of other mechanisms of selection. Genital coevolution is explicitly predicted by at least three mechanisms of genital evolution: lock and key to prevent hybridization, female choice, and sexual conflict. Although some good examples exist in support of each of these mechanisms, more data on quantitative female genital variation and studies of functional morphology during copulation are needed to understand more general patterns. A combination of different approaches is required to continue to advance our understanding of genital coevolution. Knowledge of the ecology and behavior of the studied species combined with functional morphology, quantitative morphological tools, experimental manipulation, and experimental evolution have been provided in the best-studied species, all of which are invertebrates. Therefore, attention to vertebrates in any of these areas is badly needed.Of all the evolutionary interactions between the sexes, the mechanical interaction of genitalia during copulation in species with internal fertilization is perhaps the most direct. For this reason alone, coevolution between genital morphologies of males and females is expected. Morphological and genetic components of male and female genitalia have been shown to covary in many taxa (Sota and Kubota 1998; Ilango and Lane 2000; Arnqvist and Rowe 2002; Brennan et al. 2007; Rönn et al. 2007; Kuntner et al. 2009; Tatarnic and Cassis 2010; Cayetano et al. 2011; Evans et al. 2011, 2013; Simmons and García-González 2011; Yassin and Orgogozo 2013; and see examples in TaxaMale structuresFemale structuresEvidenceLikely mechanismReferencesMollusks Land snails (Xerocrassa)Spermatophore-producing organsSpermatophore-receiving organsComparative among speciesSAC or female choiceSauder and Hausdorf 2009 SatsumaPenis lengthVagina lengthCharacter displacementLock and keyKameda et al. 2009Arthropods Arachnids (Nephilid spiders)MultipleMultipleComparative among speciesSACKuntner et al. 2009 Pholcidae spidersCheliceral apophysisEpigynal pocketsComparative (no phylogenetic analysis)Female choiceHuber 1999 Harvestmen (Opiliones)Hardened penes and loss of nuptial giftsSclerotized pregenital barriersComparative among speciesSACBurns et al. 2013Millipedes Parafontaria tonomineaGonopod sizeGenital segment sizeComparative in species complexMechanical incompatibility resulting from Intersexual selectionSota and Tanabe 2010 Antichiropus variabilisGonopod shape and sizeAccesory lobe of the vulva and distal projectionFunctional copulatory morphologyLock and keyWojcieszek and Simmons 2012Crustacean Fiddler crabs, UcaGonopodeVulva, vagina, and spermathecaTwo-species comparison, shape correspondenceNatural selection against fluid loss, lock and key, and sexual selectionLautenschlager et al. 2010Hexapodes OdonatesClasping appendagesAbdominal shape and sensory hairsFunctional morphology, comparative among speciesLock and key via female sensory systemRobertson and Paterson 1982; McPeek et al. 2009Insects Coleoptera: seed beetlesSpiny aedagusThickened walls of copulatory ductComparative among speciesSACRönn et al. 2007 Callosobruchus: Callosobruchus maculatusDamage inflictedSusceptibility to damageFull sib/half sib mating experimentsSACGay et al. 2011Reduced spinesNo correlated responseExperimental evolutionSACCayetano et al. 2011 Carabid beetles (Ohomopterus)Apophysis of the endophallusVaginal appendix (pocket attached to the vaginal apophysis)Cross-species matingsLock and keySota and Kubota 1998; Sasabi et al. 2010 Dung beetle: Onthophagus taurusShape of the parameres in the aedagusSize and location of genital pitsExperimental evolutionFemale choiceSimmons and García-González 2011 Diptera: Drosophila santomea and D. yakubaSclerotized spikes on the aedagusCavities with sclerotized plateletsCross-species matingsSACKamimura 2012 Drosophila melanogaster species complexEpandrial posterior lobes
Oviscapt pouchesComparative among speciesSAC or female choiceYassin and Orgogozo 2013Phallic spikesOviscapt furrowsCercal teeth, phallic hook, and spinesUterine, vulval, and vaginal shields D. mauritiana and D. secheliaPosterior lobe of the genital archWounding of the female abdomenMating with introgressed linesSACMasly and Kamimura 2014 Stalk-eyed flies (Diopsidae)Genital processCommon spermathecal ductComparative among species and morphologicalFemale choiceKotrba et al. 2014 Tse-tse flies: Glossina pallidipesCercal teethFemale-sensing structuresExperimental copulatory functionFemale choiceBriceño and Eberhard 2009a,b Phelebotomine: sand fliesAedagal filaments, aedagal sheathsSpermathecal ducts length, base of the ductComparative among speciesNone specifiedIlango and Lane 2000 Heteroptera: Bed bugs (Cimiciidae)Piercing genitaliaSpermalege (thickened exosqueleton)Comparative among speciesSACCarayon 1966; Morrow and Arnqvist 2003 Plant bugs (Coridromius)Changes in male genital shapeExternal female paragenitaliaComparative among speciesSACTatarnic and Cassis 2010 Waterstriders (Gerris sp.)Grasping appendagesAntigrasping appendagesComparative among speciesSACArnqvist and Rowe 2002 Gerris incognitusGrasping appendagesAntigrasping appendagesComparative among populationsSACPerry and Rowe 2012 Bee assassins (Apiomerus)AedagusBursa copulatrixComparative among speciesNoneForero et al. 2013 Cave insects (Psocodea), NeotroglaMale genital chamberPenis-like gynosomeComparative among speciesFemale competition (role reversal), coevolution SACYoshizawa et al. 2014 Butterflies (Heliconiinae)Thickness of spermatophore wallSigna: Sclerotized structure to break spermatophoresComparative among speciesSACSánchez and Cordero 2014Fish Basking shark: Cetorhinus maximusClasper clawThick vaginal padsMorphological observationNoneMatthews 1950 GambusiaGonopodial tipsGenital papillae within openingsComparative among speciesStrong character displacementLangerhans 2011 Poecilia reticulataGonopodium tip shapeFemale gonopore shapeComparative among populationsSACEvans et al. 2011Reptiles AnolesHemipene shapeVagina shapeShape correspondence, two speciesSexual selectionKöhler et al. 2012 Several speciesHemipene shapeVagina shapeShape correspondenceLock and key, female choice, and SACPope 1941; Böhme and Ziegler 2009; King et al. 2009 Asiatic pit vipersSpininess in hemipenesThickness of vagina wallTwo-species comparisonNonePope 1941 Garter snake: Thamnophis sirtalisBasal hempene spineVaginal muscular controlExperimental manipulationSACFriesen et al. 2014Birds WaterfowlPenis lengthVaginal elaborationComparative among speciesSACBrennan et al. 2007 TinamousPenis length/presenceVaginal elaborationComparative among speciesFemale choice/natural selectionPLR Brennan, K Zyscowski, and RO Prum, unpubl.Mammals MarsupialsBifid penisTwo lateral vaginaeShape correspondenceNoneRenfree 1987 EquidnaBifid penis with four rosettesSingle vagina splits into two uteriShape correspondenceNoneAugee et al. 2006; Johnston et al. 2007 Insectivores: Short-tailed shrew: Blarina brevicaudaS-shaped curve of the erect penisCoincident curve in the vaginaShape correspondenceNoneBedford et al. 2004 Common tenrec: Tenrec caudatusFiliform penis (up to 70% of the male’s body length)Internal circular folds in the vaginaLength correspondenceNoneBedford et al. 2004 Rodents: Cape dune mole: Bathyergus suillusPenis and baculum lengthVaginal lengthAllometric relationships within speciesNoneKinahan et al. 2007 Australian hopping mice (Notomys)Spiny penisDerived distal region in the vaginaMorphological observation and two-species comparisonCopulatory lockBreed et al. 2013 Pig: Sus domesticusFiliform penis endCervical ridgesArtificial inseminationFemale choiceBonet et al. 2013 Primates: Macaca arctoidesLong and filamentous glansVestibular colliculus (fleshy fold) that partially obstructs the entrance to the vaginaShape correspondence and comparison with close relativesNoneFooden 1967Open in a separate windowThe likely mechanism is that suggested by the authors, and it includes sexually antagonistic coevolution (SAC), natural selection, sexual selection, female choice, or none specified. The evidence provided by the studies can be comparative among species or among populations, experimental evolution, cross-species matings, full-sibling (sib)/half-sib matings, shape, and length correspondence. Shape correspondence is often taken as evidence of coevolution, although it is not as conclusive as other approaches.Male genitalia are among the most variable structures in nature (Eberhard 1985). In contrast, female genitalia have typically been found not to be as interspecifically variable as male genitalia in several studies that specifically examined and described them (Eberhard 1985, 2010a,b). Female genitalia are not studied as often as male genitalia, perhaps because of a male-biased view of evolutionary processes by researchers (Ah-King et al. 2014). However, studying female genitalia is undeniably challenging. Male genitalia are generally kept inside of the body cavity, but are everted before, or during copulation, so their functional morphology can be more easily studied than the internal genitalia of females. Female genitalia also tend to be softer than male genitalia and thus their morphology may be more difficult to describe, and can more easily be distorted on dissection and preservation. Female adaptations to sense or oppose features of male genitalia can be subtle, requiring careful study. Female genital tracts are under multiple sources of selection: not just mating, but also storing sperm, egg laying, birthing, and often interfacing with the terminal portion of the digestive tract. Therefore, selection balancing multiple functions may further constrain morphological evolution in female genitalia. However, even small morphological changes in female genitalia, for example, increases in vaginal muscle, may change a female’s ability to choose or reject a male during mating, or to manage the costs of mating. Thus, the functional consequences to male and female genital morphology are hard to predict unless one knows how genitalia function during intromission. Despite these challenges, recent studies have examined variation of female genitalia and evidence is accumulating that features of female genitalia are variable enough to support coevolutionary processes (Polihronakis 2006; Puniamoorthy et al. 2010; Siegel et al. 2011; Showalter et al. 2013; and see additional references in Ah-King et al. 2014).In this article, we will discuss different hypotheses of genital evolution that predict coevolution; however, this is not a review of that entire subject (but see Eberhard et al. 2010b; Simmons 2013). Rather, we discuss the various mechanisms of genital coevolution differentiating the potentially independent or overlapping roles of natural selection, female choice, and male–male competition (Fig. 1). This classification allows us to distinguish specifically those mechanisms of genital coevolution that involve sexual conflict (i.e., when the evolutionary interests of individuals of different sexes, particularly over mating, are different). We then highlight examples in different taxa organisms with particular emphasis on those that provide evidence of sexual conflict.Open in a separate windowFigure 1.Graphical classification of mechanisms of genital evolution and coevolution. Three circles depict the independent and co-occurring actions of natural selection, female choice, and male–male competition. Different specific versions of genital coevolution can occur depending on which of the three broader evolutionary mechanisms are occurring. Sexual conflict (hatched lines) occurs through the simultaneous action of male–male competition and female choice, or male–male competition and natural selection. SAC, sexually antagonistic coevolution. See text for explanation.     

Comparative Pathobiology of Kaposi Sarcoma-associated Herpesvirus and Related Primate Rhadinoviruses

With the emergence of the AIDS epidemic over the last 2 decades and the more recent identification of Kaposi sarcoma-associated herpesvirus (KSHV, Human herpesvirus 8), the genera of rhadinoviruses have gained importance as a family of viruses with oncogenic potential. First recognized in New World primates more than 30 y ago, the rhadinoviruses Saimiriine herpesvirus 2 and Ateline herpesvirus 2 have well-described transforming capabilities. Recently several new species-specific rhadinoviruses of Old World primates have been described, including retroperitoneal fibromatosis herpesvirus and rhesus rhadinovirus (Cercopithecine herpesvirus 17). Molecular analysis of these viruses has elucidated several functionally conserved genes and properties shared with KSHV involved in cellular proliferation, transformation, and immune evasion that facilitate the oncogenic potential of these viruses. This review examines the comparative pathobiology of KSHV, discusses the role of macaque rhadinoviruses as models of human disease, and outlines the derivation of specific pathogen-free animals.Abbreviations: CCL, cellular chemokine ligand; IRF, interferon regulatory factors; KSHV, Kaposi sarcoma-associated herpesvirus; LANA, latent nuclear antigen; MCD, multicentric Castleman disease; MCP1, monocyte chemotactic protein 1; miRNA, microRNA; ORF, open reading frame; PEL, primary effusion lymphoma; RFHV, retroperitoneal fibromatosis herpesvirus; RVV, rhesus rhadinovirus; SaHV2, Saimiriine herpesvirus 2; SPF, specific pathogen-free; SRV2, simian retrovirus type 2; THBS1, thrombospondinMembers of the herpesviridae are enveloped DNA viral agents that can infect a variety of host species, resulting in lifelong infection. The family is divided into Alphaherpesvirinae, Betaherpesvirinae, and Gammaherpesvirinae, according to biologic behavior and phylogenetic relationship. As a group, synthesis of viral DNA occurs in the nucleus, and production of infectious virions is associated with destruction of the cell. Herpesviruses have large complex genomes and often have acquired host genes that allow these viruses to modulate and persist in the face of host immune responses.^25,71 This condition (termed ‘latency’) is characteristic of all herpesviral infections of the natural host. Although most members of the herpesviridae are of relatively low virulence in their respective hosts, some lack strict host specificity, and cross-species transmission to an inadvertent host can be associated with severe and fatal disease.The gammaherpesvirinae subfamily is characterized by in vitro and in vivo infection of lymphoblastoid cells and is further divided into the lymphocryptovirus (γ1 herpesviruses) and rhadinovirus (γ2 herpesviruses) genera. Rhadinoviruses have taken on increased importance with the identification of the novel Kaposi sarcoma-associated herpesvirus (KSHV, Human herpesvirus 8) in association with Kaposi sarcoma, an inflammatory and neoplastic condition seen in many HIV-infected patients with AIDS.^20,22 Until the recognition of KSHV more than a decade ago, rhadinovirus infection of primates was thought to be restricted to the New World primate lineages, but subsequent investigation revealed a number of novel species-specific viruses in a variety of Old World primates (28 As discussed later, based largely on phylogenetic analysis, it is now believed that the rhadinoviruses are subdivided into 2 distinct groupings (rhadinovirus [RV] 1 and 2).⁷⁷ This review will examine 2 recently recognized rhadinoviruses of macaques (retroperitoneal fibromatosis virus [RFHV] and rhesus rhadinovirus [RRV, Cercopethecine herpesvirus 17]), focusing on their comparative pathobiology with KSHV, their impact on naturally occurring disease entities, and their roles as animal models of human disease.

Table 1.

Nomenclature of primate rhadinoviruses (RV)

Endoplasmic Reticulum Targeting and Insertion of Tail-Anchored Membrane Proteins by the GET Pathway

Hundreds of eukaryotic membrane proteins are anchored to membranes by a single transmembrane domain at their carboxyl terminus. Many of these tail-anchored (TA) proteins are posttranslationally targeted to the endoplasmic reticulum (ER) membrane for insertion by the guided-entry of TA protein insertion (GET) pathway. In recent years, most of the components of this conserved pathway have been biochemically and structurally characterized. Get3 is the pathway-targeting factor that uses nucleotide-linked conformational changes to mediate the delivery of TA proteins between the GET pretargeting machinery in the cytosol and the transmembrane pathway components in the ER. Here we focus on the mechanism of the yeast GET pathway and make a speculative analogy between its membrane insertion step and the ATPase-driven cycle of ABC transporters.The mechanism of membrane protein insertion into the endoplasmic reticulum (ER) has been extensively studied for many years (Shao and Hegde 2011). From this work, the signal recognition particle (SRP)/Sec61 pathway has emerged as a textbook example of a cotranslational membrane insertion mechanism (Grudnik et al. 2009). The SRP binds a hydrophobic segment (either a cleavable amino-terminal signal sequence or a transmembrane domain) immediately after it emerges from the ribosomal exit tunnel. This results in a translational pause that persists until SRP engages its receptor in the ER and delivers the ribosome-nascent chain complex to the Sec61 channel. Last, the Sec61 channel enables protein translocation into the ER lumen along with partitioning of hydrophobic transmembrane domains into the lipid bilayer through the Sec61 lateral gate (Rapoport 2007).Approximately 5% of all eukaryotic membrane proteins have an ER targeting signal in a single carboxy-terminal transmembrane domain that emerges from the ribosome exit tunnel following completion of protein synthesis and is not recognized by the SRP (Stefanovic and Hegde 2007). Nonetheless, because hydrophobic peptides in the cytoplasm are prone to aggregation and subject to degradation by quality control systems (Hessa et al. 2011), these tail-anchored (TA) proteins still have to be specifically recognized, shielded from the aqueous environment, and guided to the ER membrane for insertion. In the past five years, the guided-entry of TA proteins (GET) pathway has come to prominence as the major machinery for performing these tasks and the enabler of many key cellular processes mediated by TA proteins including vesicle fusion, membrane protein insertion, and apoptosis. This research has rapidly yielded biochemical and structural insights (​and2)2) into many of the GET pathway components (Hegde and Keenan 2011; Chartron et al. 2012a; Denic 2012). In particular, Get3 is an ATPase that uses metabolic energy to bridge recognition of TA proteins by upstream pathway components with TA protein recruitment to the ER for membrane insertion. However, the precise mechanisms of nucleotide-dependent TA protein binding to Get3 and how the GET pathway inserts tail anchors into the membrane are still poorly understood. Here, we provide an overview of the budding yeast GET pathway with emphasis on mechanistic insights that have come from structural studies of its membrane-associated steps and make a speculative juxtaposition with the ABC transporter mechanism.

Table 1.

A catalog of GET pathway component structures

Diminished Susceptibility to Cefoperazone/Sulbactam and Piperacillin/Tazobactam in Enterobacteriaceae Due to Narrow-Spectrum β-Lactamases as Well as Omp Mutation

Cefoperazone/sulbactam (CSL) and piperacillin/tazobactam (TZP) are commonly used in clinical practice in China because of their excellent antimicrobial activity. CSL and TZP-nonsusceptible Enterobacteriaceae are typically resistant to extended-spectrum cephalosporins such as ceftriaxone (CRO). However, 11 nonrepetitive Enterobacteriaceae strains, which were resistant to CSL and TZP yet susceptible to CRO, were collected from January to December 2020. Antibiotic susceptibility tests and whole-genome sequencing were conducted to elucidate the mechanism for this rare phenotype. Antibiotic susceptibility tests showed that all isolates were amoxicillin/clavulanic-acid resistant and sensitive to ceftazidime, cefepime, cefepime/tazobactam, cefepime/zidebactam, ceftazidime/avibactam, and ceftolozane/tazobactam. Whole-genome sequencing revealed three of seven Klebsiella pneumoniae strains harbored bla_SHV-1 only, and four harbored bla_SHV-1 and bla_TEM-1B. Two Escherichia coli strains carried bla_TEM-1B only, while two Klebsiella oxytoca isolates harbored bla_OXY-1-3 and bla_OXY-1-1, respectively. No mutation in the β-lactamase gene and promoter sequence was found. Outer membrane protein (Omp) gene detection revealed that numerous missense mutations of OmpK36 and OmpK37 were found in all strains of K. pneumoniae. Numerous missense mutations of OmpK36 and OmpK35 and OmpK37 deficiency were found in one K. oxytoca strain, and no OmpK gene was found in the other. No Omp mutations were found in E. coli isolates. These results indicated that narrow spectrum β-lactamases, TEM-1, SHV-1, and OXY-1, alone or in combination with Omp mutation, contributed to the resistance to CSL and TZP in CRO-susceptible Enterobacteriaceae.Antibiotic susceptibility tests

Cognitive Manic Symptoms in Bipolar Disorder Associated with Polymorphisms in the DAOA and COMT Genes

Introduction

Bipolar disorder is characterized by severe mood symptoms including major depressive and manic episodes. During manic episodes, many patients show cognitive dysfunction. Dopamine and glutamate are important for cognitive processing, thus the COMT and DAOA genes that modulate the expression of these neurotransmitters are of interest for studies of cognitive function.

Methodology

Focusing on the most severe episode of mania, a factor was found with the combined symptoms of talkativeness, distractibility, and thought disorder, considered a cognitive manic symptoms (CMS) factor. 488 patients were genotyped, out of which 373 (76%) had talkativeness, 269 (55%) distractibility, and 372 (76%) thought disorder. 215 (44%) patients were positive for all three symptoms, thus showing CMS (Bipolar disorder type 1 [n]488Men [n (%)]209 (43)Talkativeness [n (%)]373 (76)Distracibility [n (%)]269 (55)Thought disorder [n (%)]372 (76)Cognitive manic symptoms^* [n (%)]215 (44)Men [n (%)]81 (39)Non-Cognitive manic symptoms [n (%)]248 (51)Men [n (%)]117 (56)Unknown [n (%)]25 (5)Men [n (%)]11 (44)Anonymous blood donors (ABD)1044Men [n (%)]616 (59)Open in a separate window^*having all three symptoms: talkativeness, distractibility, and tought disorder.

Results

The finding of this study was that cognitive manic symptoms in patients with bipolar 1 disorder was associated with genetic variants in the DAOA and COMT genes. Nominal association for DAOA SNPs and COMT SNPs to cognitive symptoms factor in bipolar 1 disorder was found in both allelic (BP1 CMSBP1 non-CMSABDBP1 CMS vs. non-CMS^b BP1 CMS vs. ABD controls^b GeneSNP^a aa/ab/bbaa/ab/bbaa/ab/bbpEMP1^c EMP2^d OR [95% CI] ^e pEMP1^c EMP2^d OR [95% CI] ^e DAOA rs3916967 (C/T)32/88/8950/118/77177/494/3610.0180.0180.210.72 [0.55–0.93]0.0290.0260.280.78 [0.66–1.0] DAOA rs2391191 (A/C)28/75/7939/111/70179/487/3570.0550.0390.500.75 [0.57–1.0]0.0200.0190.210.75 [0.63–1.0] DAOA rs1935062 (C/A)26/67/8935/102/86146/460/4050.120.120.780.80 [0.58–1.0]0.0690.0660.520.80 [0.65–1.0] COMT rs5993883 (T/G)33/120/5371/112/57269/510/2230.0250.0300.270.73 [0.56–0.95]0.0017^* 1.0E^{−4 *} 0.021^* 0.68 [0.91–1.4] COMT rs165599 (G/A)29/94/8725/93/12687/443/5010.0930.0940.691.27 [1.0–1.8]0.0140.0170.161.34 [1.1–1.7]Open in a separate window^aSNP (minor allele(a)/major allele(b)).^bgender and rs1718119 as covariate.^cpoint-wise p-value from 10,000 pemutations with no covarite (EMP1).^dcorrected empirical p-value by max (T) permutation.^eodds ratio (OR), the proportion of minor versus major allele affected (cognitive manic symptoms factor)/proportion of minor versus major allele unaffected (non-cognitive manic symptoms factor or ABD controls).^*significant after correction for multiple testing by max (T) permutation.

Table 3

Haplotype association of haplotype group 1 in bipolar 1 patients with cognitive manic symptoms (CMS) compared with non-CMS patients or ABD controls in the DAOA gene.

Ophthalmic artery chemosurgery for less advanced intraocular retinoblastoma: five year review

Background

Ophthalmic artery chemosurgery (OAC) for retinoblastoma was introduced by us 5 years ago for advanced intraocular retinoblastoma. Because the success was higher than with existing alternatives and systemic side effects limited we have now treated less advanced intraocular retinoblastoma (Reese-Ellsworth (RE) I-III and International Classification Retinoblastoma (ICRB) B and C).

Methodology/Principal Findings

Retrospective review of 5 year experience in eyes with Reese Ellsworth (Reese-Ellsworth (RE) Classification For Intraocular Retinoblastoma GROUP I a. Solitary tumor, less than 4 disc diameters in size, at or behind the equator b. Multiple tumors, none over 4 disc diameters in size, all at or behind the equator GROUP II a. Solitary tumor, less than 4 to 10 disc diameters in size, at or behind the equator b. Multiple tumors, none over 4 to 10 disc diameters in size, all at or behind the equator GROUP III a. Any lesion anterior to the equator b. Solitary tumors larger than 10 disc diameters behind the equator GROUP IV a. Multiple tumors, some larger than 10 disc diameters b. Any lesion extending anteriorly to the ora serrata GROUP V a . Massive tumors involving over half the retina b . Vitreous seeding Open in a separate window

Table 2

International Classification for Retinoblastoma (ICRB) Scheme.

Use of methotrexate therapy is not associated with decreased prevalence of metabolic syndrome

With great interest, we read the article by Toms and colleagues [1] in the previous issue of Arthritis Research & Therapy, in which they assessed prevalences of metabolic syndrome (MetS) in rheumatoid arthritis (RA) patients. Moreover, they identified demographic and clinical factors that may be associated with MetS. Toms and colleagues found prevalences of up to 45% of MetS and demonstrated older age and health status (health assessment questionnaire) to be associated with MetS irrespectively of the definition used. Of most interest, an association between methotrexate (MTX) use and decreased presence of MetS was observed in patients more than 60 years of age. The investigators hypothesized that this may be attributed to a drug-specific effect (and not to an anti-inflammatory effect) either by changing levels of adenosine, which is known to interact with glucose and lipid metabolism, or by an indirect effect mediated through concomitant folic acid administration, thereby decreasing homocysteine levels.Recently, we also examined the prevalence of MetS in (a subgroup of) RA patients in the CARRÉ investigation, a prospective cohort study on prevalent and incident cardiovascular disease and its underlying cardiovascular risk factors [2]. The findings of Toms and colleagues stimulated us to perform additional analyses in our total study population (n = 353).The prevalences of MetS were 35% and 25% (Table ​(Table1)1) according to criteria of National Cholesterol Education Program (NCEP) 2004 and NCEP 2001, respectively. In multivariate backward regression analyses, we found significant associations between body mass index, pulse rate, creatinine levels, hypothyroidism and diabetes mellitus and the presence of MetS independently of the criteria used (Table ​(Table2).2). However, an independent association between single use of MTX or use of MTX in combination with other disease-modifying antirheumatic drugs, on the one hand, and a decreased prevalence of MetS, on the other hand, could not be demonstrated (even in the subgroup of patients over the age of 60).

Table 1

Characteristics of the study population

Stress-induced flowering

Many plant species can be induced to flower by responding to stress factors. The short-day plants Pharbitis nil and Perilla frutescens var. crispa flower under long days in response to the stress of poor nutrition or low-intensity light. Grafting experiments using two varieties of P. nil revealed that a transmissible flowering stimulus is involved in stress-induced flowering. The P. nil and P. frutescens plants that were induced to flower by stress reached anthesis, fruited and produced seeds. These seeds germinated, and the progeny of the stressed plants developed normally. Phenylalanine ammonialyase inhibitors inhibited this stress-induced flowering, and the inhibition was overcome by salicylic acid (SA), suggesting that there is an involvement of SA in stress-induced flowering. PnFT2, a P. nil ortholog of the flowering gene FLOWERING LOCUS T (FT) of Arabidopsis thaliana, was expressed when the P. nil plants were induced to flower under poor-nutrition stress conditions, but expression of PnFT1, another ortholog of FT, was not induced, suggesting that PnFT2 is involved in stress-induced flowering.Key words: flowering, stress, phenylalanine ammonia-lyase, salicylic acid, FLOWERING LOCUS T, Pharbitis nil, Perilla frutescensFlowering in many plant species is regulated by environmental factors, such as night-length in photoperiodic flowering and temperature in vernalization. On the other hand, a short-day (SD) plant such as Pharbitis nil (synonym Ipomoea nil) can be induced to flower under long days (LD) when grown under poor-nutrition, low-temperature or high-intensity light conditions.^1–9 The flowering induced by these conditions is accompanied by an increase in phenylalanine ammonia-lyase (PAL) activity.¹⁰ Taken together, these facts suggest that the flowering induced by these conditions might be regulated by a common mechanism. Poor nutrition, low temperature and high-intensity light can be regarded as stress factors, and PAL activity increases under these stress conditions.¹¹ Accordingly, we assumed that such LD flowering in P. nil might be induced by stress. Non-photoperiodic flowering has also been sporadically reported in several plant species other than P. nil, and a review of these studies suggested that most of the factors responsible for flowering could be regarded as stress. Some examples of these factors are summarized in 12^–14

Table 1

Some cases of stress-induced flowering

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Up to 60 different proteins are recruited to the site of clathrin-mediated endocytosis in an ordered sequence. These accessory proteins have roles during all the different stages of clathrin-mediated endocytosis. First, they participate in the initiation of the endocytic event, thereby determining when and where endocytic vesicles are made; later they are involved in the maturation of the clathrin coat, recruitment of specific cargo molecules, bending of the membrane, and finally in scission and uncoating of the nascent vesicle. In addition, many of the accessory components are involved in regulating and coupling the actin cytoskeleton to the endocytic membrane. We will discuss the different accessory components and their various roles. Most of the data comes from studies performed with cultured mammalian cells or yeast cells. The process of endocytosis is well conserved between these different organisms, but there are also many interesting differences that may shed light on the mechanistic principles of endocytosis.Receptor-mediated endocytosis is the process by which eukaryotic cells concentrate and internalize cell surface receptors from the plasma membrane into small (∼50 nm– ∼100 nm diameter) membrane vesicles (Chen et al. 2011; McMahon and Boucrot 2011; Weinberg and Drubin 2012). This mechanism has been studied extensively in mammalian tissue culture cells and in yeast, and despite the evolutionary distance between yeast and mammalian cells the mechanism of receptor-mediated endocytosis in the respective cell types show remarkable similarities. Indeed many of the ∼60 endocytic accessory proteins (EAPs) found in yeast have homologs in mammalian cells, although both cell types also have unique EAPs (McMahon and Boucrot 2011; Weinberg and Drubin 2012).In the following, we briefly describe known yeast and mammalian EAPs (Sigismund et al. 2012; see also Bökel and Brand 2013; Cosker and Segal 2014; Di Fiore and von Zastrow 2014).

Table 1.

Key endocytic proteins in mammals and in yeast

Aluminum induced proteome changes in tomato cotyledons

Cotyledons of tomato seedlings that germinated in a 20 µM AlK(SO₄)₂ solution remained chlorotic while those germinated in an aluminum free medium were normal (green) in color. Previously, we have reported the effect of aluminum toxicity on root proteome in tomato seedlings (Zhou et al.¹). Two dimensional DIGE protein analysis demonstrated that Al stress affected three major processes in the chlorotic cotyledons: antioxidant and detoxification metabolism (induced), glyoxylate and glycolytic processes (enhanced), and the photosynthetic and carbon fixation machinery (suppressed).Key words: aluminum, cotyledons, proteome, tomatoDifferent biochemical processes occur depending on the developmental stages of cotyledons. During early seed germination, before the greening of the cotyledons, glyoxysomes enzymes are very active. Fatty acids are converted to glucose via the gluconeogenesis pathway.^2,3 In greening cotyledons, chloroplast proteins for photosynthesis and leaf peroxisomal enzymes in the glycolate pathway for photorespiration are metabolized.^2–4 Enzymes involved in regulatory mechanisms such as protein kinases, protein phosphatases, and mitochondrial enzymes are highly expressed.^3,5,6The chlorotic cotyledons are similar to other chlorotic counterparts in that both contains lower levels of chlorophyll, thus the photosynthetic activities are not as active. In order to understand the impact of Al on tomato cotyledon development, a comparative proteome analysis was performed using 2D-DIGE following the as previously described procedure.¹ Some proteins accumulated differentially in Al-treated (chlorotic) and untreated cotyledons (Fig. 1). Mass spectrometry of tryptic digestion fragments of the proteins followed by database search has identified some of the differentially expressed proteins (Open in a separate windowFigure 1Image of protein spots generated by Samspot analysis of Al treated and untreated tomato cotyledons proteomes separated on 2D-DIGE.

Table 1

Proteins identified from tomato cotyledons of seeds germinating in Al-solution