Their immunologic effects following oral administration is less clear. The purpose of this systematic review was to consolidate and evaluate the available data regarding the specific immunologic effects of dietary polysaccharides.A large body of literature suggests that certain polysaccharides affect immune system function. Much of this literature, however, consists of Studies were identified by conducting PubMed and Google Scholar electronic searches and through reviews of polysaccharide article bibliographies. Only articles published in English were included in this review. Two researchers reviewed data on study design, control, sample size, results, and nature of outcome measures. Subsequent searches were conducted to gather information about polysaccharide safety, structure and composition, and disposition.We found 62 publications reporting statistically significant effects of orally ingested glucans, pectins, heteroglycans, glucomannans, fucoidans, galactomannans, arabinogalactans and mixed polysaccharide products in rodents. Fifteen controlled human studies reported that oral glucans, arabinogalactans, heteroglycans, and fucoidans exerted significant effects. Although some studies investigated anti-inflammatory effects, most studies investigated the ability of oral polysaccharides to stimulate the immune system. These studies, as well as safety and toxicity studies, suggest that these polysaccharide products appear to be largely well-tolerated.Trametes versicolor mushroom improved survival and immune function in human RCTs of cancer patients; glucans, arabinogalactans and fucoidans elicited immunomodulatory effects in controlled studies of healthy adults and patients with canker sores and seasonal allergies. This review provides a foundation that can serve to guide future research on immune modulation by well-characterized polysaccharide compounds.Taken as a whole, the oral polysaccharide literature is highly heterogenous and is not sufficient to support broad product structure/function generalizations. Numerous dietary polysaccharides, particularly glucans, appear to elicit diverse immunomodulatory effects in numerous animal tissues, including the blood, GI tract and spleen. Glucan extracts from the  Oftenin vitro -16, but injected . For clin orally . We thusStudies were identified by conducting electronic searches of PubMed and Google Scholar from their inception to the end of October 2009. The reference lists of the selected articles were checked for additional studies that were not originally found in the search.The following search terms were combined with the term polysaccharide: dietary AND immune, or oral AND immune, or dietary AND inflammation, or oral AND inflammation. When specific polysaccharides or polysaccharide-rich plants and fungi were identified, further searches were conducted using their names with the same search terms. Studies were selected based on the following inclusion criteria:1. Rodent or human studies2. The presence of test group and control group 3. Studies reporting statistically significant immunomodulatory effects4. English language5. Studies published up to October 2009.50), composition and structure, and disposition.Two researchers  reviewed the list of unique articles for studies that fit the inclusion criteria. Uncertainties over study inclusion were discussed between the researchers and resolved through consensus. Searches were then conducted to obtain specific polysaccharide product information: safety  were shown in RCTs to increase lymphocyte proliferation and the number of CD8+ lymphocytes [Panax quiquefolium (North American ginseng) was shown in an RCT of healthy older adults to decrease the incidence of acute respiratory illness and symptom duration [Undaria pinnatifida (wakame) fucoidans found both immune stimulating and suppressing effects, including increased stromal-derived factor-1, IFN-g, CD34+ cells and CXCR4-expressing CD34+ cells and decreased blood leukocytes and lymphocytes [A number of studies in healthy human adults demonstrated immune stimulating effects of oral polysaccharides. Arabinogalactans from phocytes  and to iphocytes . A furanduration . Finallyphocytes .Agaricus subrufescens (A. blazei) (aqueous extracts [Lentinula edodes (shiitake)  spp. have also been shown to enhance immune function in healthy young animals [Cyamopsis tetragonolobus galactomannan (guar gum) or highly methoxylated pectin feeding exerted numerous stimulating effects on antibody production in older animals [Studies in healthy animals showed a number of immune stimulating effects of various glucan products from extracts , aqueousextracts , α-1,6 aextracts , and whoextracts ); Lentinlentinan  and β-gllentinan ); Saccha-glucans ,28); Lamaminarin ); Sclerohosphate ); Sclerorum (SSG ); and Ph (powder  and aque (powder ). A fura animals -35. Cyam animals .Avena (oat) spp. soluble glucans [E. vermiformis and fed Avena spp. particulate glucans [E. coli injections fed L. digitata glucans (laminarin) [U. pinnatifida fucoidans [Staphylococcus aureus or Candida albicans injections fed S. cerevisiae glucans (scleroglucan) [A. subrufescens (A. blazei Murrill) [Evidence for the effectiveness of oral polysaccharides against infection and immune challenges has been mainly demonstrated in animals. Immune stimulating effects have been shown in resting and exercise-stressed animals with thioglycollate, clodronate, or HSV-1 injections fed  glucans -41; anim glucans ,43; animminarin) ; animalsucoidans ; animalsoglucan) ; and aniMurrill) .S. cerevisiae β-1,3;1-6 glucans decreased IL-4, IL-5 and percent eosinophils, and increased IL-12 in nasal fluid [Additional controlled human and animal studies have shown anti-inflammatory and anti-allergy effects of some polysaccharide products. In an RCT of adults with seasonal allergic rhinitis, al fluid , while aal fluid .Cladosiphon okamuranus Tokida fucoidans [Cyamopsis tetragonolobus galactomannans [Malus spp. pectins [A. subrufescens aqueous extracts [of Ganoderma tsugae [Pyrus pyrifolia pectins [Pholiota nameko heteroglycan (PNPS-1) [Animal models of inflammatory bowel disease have shown anti-inflammatory effects of ucoidans , Cyamopsomannans , Malus s pectins , and mix pectins . Animalsextracts , an aquea tsugae , and Pyr pectins . Anti-in(PNPS-1) .Trametes versicolor glucans have demonstrated anti-cancer effects in humans. In two RCTs and five controlled trials, PSK from T. versicolor mycelia increased survival of advanced stage gastric, colon and colorectal cancer patients [T. versicolor fruit bodies found increased IgG and IgM antibodies and total leukocyte and neutrophil counts, along with a decrease in the number of patients withdrawing from the study due to disease progression [A. subrufescens glucans showed increased NK cell activity and fewer chemotherapy side effects [patients -62 with gression . An RCT  effects .A. subrufescens demonstrating anti-cancer activities in animal models include an aqueous extract [G. lucidum [G. frondosa [Hordeum vulgare β-glucans [Laminaria angustata powder [Lentinula edodes products  [L. edodes (KS-2) improved survival of animals with cancer cell injections [Lycium barbarum (LBP3p), Lentinus lepidus (PG101) and A. subrufescens (ATOM) demonstrated a number of immune stimulating effects in animal cancer models [T. versicolor (PSP): animals with cancer cell implantations showed decreased tumor growth and vascular density [In numerous animal models of cancer, a wide range of polysaccharides have shown anti-tumorogenic effects. Glucan products sourced from  extract , an aque extract , and an  extract . Anti-ca lucidum -69; the frondosa -72; Hord-glucans -76; Lamia powder ; Lentinu; Ple; PleA. ss powder , Sacchar,3glucans,73; and um (SSG) ,83. A gljections ; apple ajections -90. Finar models -93. Inte density .A. subrufescens aqueous extract [A. subrufescens extracts in healthy animals or animals with cancers found no evidence of toxicity  or extract  , effects can be unequivocally attributed to polysaccharides. Such associations are certainly more tenuous when considering product powders or products obtained by extraction methods designed to isolate polysaccharides, but without complete compositional analyses.® products) have been shown to be metabolized by human colonic bacteria. Orally ingested fucoidans, glucans and mannans (or their fragments) have been detected in numerous tissues and organs throughout the body [Dietary polysaccharides are known to impact gut microbial ecology ,102, andthe body ,108,109,One can only speculate upon the mechanisms by which the polysaccharides discussed in this review influence immunologic function, particularly when one considers the exceedingly complex environment of the GI tract. It is possible that fragments of polysaccharides partially hydrolyzed by gut bacteria may either bind to gut epithelia and exert localized and/or systemic immune system effects, or be absorbed into the bloodstream, with the potential to exert systemic effects. Current studies investigating the link between the bioconversion of dietary polysaccharides, their bioavailability and their downstream effects on the host metabolism and physiology are utilizing metabolomic and metagenomic approaches that can detect and track diverse microbial metabolites from immunomodulatory polysaccharides . These aAdditional RCTs of well-characterized products are needed to more completely understand the immunomodulatory effects and specific applications of oral polysaccharides. Such studies will need to better investigate the optimal timing and duration for polysaccharide ingestion. That is, should they be consumed continuously, before, at the time of, or after exposure to a pathogen or environmental insult? Only a few studies have actually investigated the impact of timing of polysaccharide intake to achieve optimal benefits. Daily feeding with some polysaccharides appears to result in tolerance (and diminished benefits); this has been demonstrated for some mushroom β-glucans ,26. For The dietary polysaccharides included in this review have been shown to elicit diverse immunomodulatory effects in animal tissues, including the blood, GI tract, and spleen. In controlled human trials, polysaccharide intake stimulated the immune system in the blood of healthy adults, dampened the allergic response to a respiratory inflammatory agent, and improved survival in cancer patients. Additional RCTs of well-characterized products are needed to more completely understand the immunomodulatory effects and specific applications of oral polysaccharides(a)anthracene; DMH: N-N'-dimethylhydrazine; DMN: dimethylhydrazine; DSS: dextran sulfate sodium; EBV: Epstein-Barr virus; GALT: gut-associated lymphoid tissue; GI: gastrointestinal; H202: hydrogen peroxide; HSV: herpes simplex virus; ICR: imprinting control region; ID: intradermal; IEL: intraepithelial lymphocytes; IFN-λ: interferon gamma; IG: intragastric; IgA: immunoglobulin A; IgE: immunoglobulin E; IgG: immunoglobulin G; IgM: immunoglobulin M; IL: interleukin; IMC: invasive micropapillary carcinoma; IN: intranasally; IP: intraperitoneal; IV: intravenous; LPS: lipopolysaccharide; Mø: macrophage; mAb: monoclonal antibody; 3-MCA: methylcholanthrene; MLN: mesenteric lymph nodes; MM-46 carcinoma: mouse mammary carcinoma; MW: molecular weight; NK: natural killer; NOAEL: no observable adverse effect level; OVA: ovalbumin; PBL: peripheral blood leukocytes; PBMC: peripheral blood mononuclear cells; PHA: phytohaemagglutinin; PMA: phorbol 12-myristate 13-acetate; PML: polymorphonuclear lymphocyte; RCT: randomized, controlled trial; RNA: ribonucleic acid; SC: subcutaneous; SD rats: Sprague Dawley; TCR: T cell receptor; TLR: toll like receptor; TNF-α: tumor necrosis factor alpha; UC: ulcerative colitis; WT: wild type.♀: female; ♂: male; Ab: antibody; AIDS: autoimmune deficiency syndrome; AOM: azoxymethane; BBN: N-butyl-N'-butanolnitrosamine; BLCL: Burkitt's Lymphoma Cell Line; BW: body weight; CBC: complete blood count; CD: cluster of differentiation; CFU: colony forming unit; ConA: concanavalin A; CXCR: CXC chemokine receptor; DMBA: 7,12-dimethylbenz® powder and Advanced Ambrotose® powder) discussed in this review.The authors are employees of the Research & Development Department at Mannatech, Incorporated, which sells two of the polysaccharide products (AmbrotoseJER and EDN conducted literature searches and wrote the manuscript. RAS provided technical guidance. All authors read and approved the final manuscript.


Yarrowia lipolytica, Debaryomyces hansenii, Kluyveromyces lactis and Candida glabrata. Y. lipolytica is a dimorphic yeast and has good capacities to secrete proteins. The translocation of nascent protein through the endoplasmic reticulum membrane was well studied in Y. lipolytica and is largely co-translational as in the mammalian protein secretion pathway.Protein secretion is a universal cellular process involving vesicles which bud and fuse between organelles to bring proteins to their final destination. Vesicle budding is mediated by protein coats; vesicle targeting and fusion depend on Rab GTPase, tethering factors and SNARE complexes. The Génolevures II sequencing project made available entire genome sequences of four hemiascomycetous yeasts, S. cerevisiae proteins involved in vesicular secretion and these protein sequences were used for the BLAST searches against Génolevures protein database . These proteins are well conserved between these yeasts and Saccharomyces cerevisiae. We note several specificities of Y. lipolytica which may be related to its good protein secretion capacities and to its dimorphic aspect. An expansion of the Y. lipolytica Rab protein family was observed with autoBLAST and the Rab2- and Rab4-related members were identified with BLAST against NCBI protein database. An expansion of this family is also found in filamentous fungi and may reflect the greater complexity of the Y. lipolytica secretion pathway. The Rab4p-related protein may play a role in membrane recycling as rab4 deleted strain shows a modification of colony morphology, dimorphic transition and permeability. Similarly, we find three copies of the gene (SSO) encoding the plasma membrane SNARE protein. Quantification of the percentages of proteins with the greatest homology between S. cerevisiae, Y. lipolytica and animal homologues involved in vesicular transport shows that 40% of Y. lipolytica proteins are closer to animal ones, whereas they are only 13% in the case of S. cerevisiae.We identified Y. lipolytica is more representative of vesicular secretion of animals and other fungi than is S. cerevisiae.These results provide further support for the idea, previously noted about the endoplasmic reticulum translocation pathway, that  Yarrowia lipolytica is a hemiascomycetous dimorphic yeast, generally regarded as safe (GRAS), which has been used for biotechnological applications. It is able to produce large amounts of several metabolites such as citric acid and to secrete a variety of extracellular proteins  . Y. lipboratory . We focuboratory -8, on thboratory  and on tboratory . SeveralY. lipolytica and we compared them to the proteins of the three other yeasts, Candida glabrata, Kluyveromyces lactis and Debaryomyces hansenii. C. glabrata has become the second most common cause of candidiasis after Candida albicans. C. glabrata is not dimorphic, in contrast to other Candida species, and is phylogenetically closer to Saccharomyces cerevisiae . VesEndoplasmic reticulum to Golgi transport is mediated by the action respectively of the COPII and COPI coat complexes -23 for r for r23 S. cerevisiae, there are one Sec23p, one Sec23p-related protein, one Sec24p, two Sec24p-related proteins (Sfb2p and Sfb3p) and one Sec12p homologue (Sed4p). In Y. lipolytica and D. hansenii, all the COP II coat components are well conserved and we find two Sec23p-homologues, two Sec24p-homologues but no Sed4p proteins. In C. glabrata, there are two Sec23p-homologues, three Sec24p-homologues and two Sec13p-homologues and in K. lactis we found the same proteins as in S. cerevisiae with the exception of Sfb2p and Sed4p.Vesicle budding is initiated by the activation of the GTPase Sar1p by the endoplasmic reticulum integral membrane guanine exchange factor Sec12p ,28. Sar1S. cerevisiae components are conserved in Y. lipolytica, though the Y. lopolytica Sec28p is only weakly related to the S. cerevisiae protein. In K. lactis, the Arf1 protein homologue was not identified but another Arf protein could play the role of Arf1p .The COP I coat assembles by the same process as COP II complex involving an Arfp-GTPase .S. cerevisiae, by homology to the mammalian adaptor protein (AP) subunit sequences, three potential heterotetrameric adaptor protein complexes have been identified . TheS. cerevisiae, Ypt1p, Ypt31p/32p and Sec4p are the essential Rab GTPases which regulate the exocytic pathway and Ypt6p, Ypt7p and Ypt51p/52p/53p are involved in the endocytic pathway. S. cerevisiae also has two other Rab GTPase, Ypt10p and Ypt11p which are also present in C. glabrata and we can find Ypt11p in K. lactis. Ypt10p seems to be involved in endocytic function and Ypt11p is required for endoplasmic reticulum inheritance . The Us complex . In Y. ls is low . Cog2p wY. lipolytica does not align with the S. cerevisiae sequence. Nevertheless the Y. lipolytica sequence also contains potential coiled-coil regions  and Tip20p ,106. Dslnalysis, ). The TiS. cerevisiae TRAPP II complex  is composed of ten subunits and could have a role in retrograde transport of Golgi vesicles  areeckpoint ; Imh1p, eckpoint ,120] andeckpoint  [for a reckpoint ]; Coy1p,RE Gos1p  and Rud3RE Gos1p .The TRAPP II subunit Trs120p is required for vesicle traffic from early endosome to the late Golgi .The Vps Class C/HOPS complex . Twentyer fungi , Vam3p aormation ,143. Vam mutants . In Y. lunctions . The mulunctions . Sft1p wS. cerevisiae, Sly1p acting between the endoplasmic reticulum and Golgi .S. cerevisiae sequences were collected from Saccharomyces Genome Database [Hemiascomycetous yeast genome sequences, BLAST searches of vesicular secretion proteins and BLAST results  were obtained from the Génolevures web site . S. cereDatabase . BLASTs Database  and InfoDatabase . ProteinDatabase , ExPASy Database  and CBS Database .S. cerevisiae vesicular secretion was made from literature. These S. cerevisiae protein sequences were used for BLAST searches with the Génolevures web site. For protein families such as Rab protein, autoBLAST, which means BLAST of a sequence against its own genome, were made to identify all the members of the family. The protein sequences of the new members were identified by BLAST searches against the NCBI eukaryotic protein sequences.A list of proteins implicated in The percentages of proteins with the greatest homology Fig.  and 8 weY. lipolytica,Yl: Yarrowia lipolytica; C. glabrata, Cg: Candida glabrata; K. lactis, Kl: Kluyveromyces lactis; D. hansenii, Dl: Debaryomyces hansenii; S. cerevisiae, Sc: Saccharomyces cerevisiae; N. crassa, Nc: Neurospora crassa; SNARE: Soluble N-ethylmaleimide-sensitive factor Attachment protein Receptor.DS conceived the study, carried out the molecular genetic studies, the sequence analyses and drafted the manuscript. JMB participated in the sequence analyses. All authors read and approved the final manuscript.Yarrowia lipolytica identified proteins coatsDrawing of . PM: plasma membrane, ER: endoplasmic reticulum, RE: recycling endosome, EE: early endosome, LE: late endosome, MVB: multi-vesicular bodies, SV: secretory vesicle.Click here for fileYarrowia lipolytica identified Ypt/Rab GTPasesDrawing of . PM: plasma membrane, ER: endoplasmic reticulum, RE: recycling endosome, EE: early endosome, LE: late endosome, MVB: multi-vesicular bodies, SV: secretory vesicle.Click here for fileFull image of Figure 2Click here for fileYarrowia lipolytica identified tethering factorsDrawing of . PM: plasma membrane, ER: endoplasmic reticulum, RE: recycling endosome, EE: early endosome, LE: late endosome, MVB: multi-vesicular bodies, SV: secretory vesicle.Click here for fileDrawing of Yarrowia lipolytica identified SNARE and SM proteins. PM: plasma membrane, ER: endoplasmic reticulum, RE: recycling endosome, EE: early endosome, LE: late endosome, MVB: multi-vesicular bodies, SV: secretory vesicle.Click here for fileE-values. E-values found for BLAST of Yarrowia lipolytica proteins against Saccharomyces cerevisiae, Candida glabrata, Kluyveromyces lactis, Debaryomyces hansenii, Schizosaccharomyces pombe (Sp),Neurospora crassa, other fungi, animals, plants, obtained with NCBI web site. Numbers between brackets indicate the order of best BLAST hits. Fungi: Ashbya gossypii (Ag), Aspergillus clavatus (Ac), Aspergillus fumigatus (Af), Aspergillus nidulans (Asn), Aspergillus niger (An), Aspergillus orizae (Ao), Aspergillus parasiticus (Ap), Aspergillus terreus (Ast), Chaetomium globosum (Chg), Coccidioides immitis (Ci), Coprinopsis cinerea (Cc), Cryptococus neoformans (Cn), Gibberzlla zeae (Gz), Hypocrea lixii (Hl), Magnaporthe grisea (Mg), Neosartorya fischeri (Nf), Neurospora crassa (Nc), Paracoccidioides brasiliensis (Pb), Phaeosphaeria nodorum (Pn), Ustilago maydis (Um). Animals: Aedes aegypti (Aa), Aiptasia pulchella (Ap), Anopheles gambiae (Ang), Apis mellifera (Am), Bombyx mori (Bm), Bos taurus (Bt), Caenorhabditis briggsae (Cb), Caenorhabditis elegans (Ce), Canis familiaris (Cf), Danio rerio (Dr), Drosophila grimshawi (Dg), Drosophila melanogaster (Dm), Drosophila pseudoobscura (Dp), Gallus gallus (Gg), Homo sapiens (Hs), Macaca mulatta (Mam), Mus musculus (Mm), Oryzias latipes (Ol), Pan troglodytes (Pt), Pongo pygmaeus (Pp), Rattus norvegicus (Rn), Strongylocentrus purpuratus (Stp), Xenopus laevis (Xl), Xenopustropicalis (Xt). Plants: Arabidopsis thaliana (At), Brassica oleracea (Bo), Brassica rapa (Br), Hyacinthus orientalis (Ho), Lotus japonicus (Lj), Medicago truncatula (Mt), Nicotiana tabacum (Nt), Oenothera odorata (Oo), Oriza sativa (Os), Pisum sativum (Ps), Solanum chacoense (Soc), Solanum tuberosum (St), Zea mays (Zm). .Click here for fileE-values. E-values found for NCBI BLAST of Saccharomyces cerevisiae proteins against Yarrowia lipolytica and animal proteins .Click here for fileCandida. glabrata, Kluyveromyces lactis, Debaryomyces hansenii genes coding for the proteins potentially implicated in vesicular transport.List of Click here for file


The last word in the title is misspelled. The correct spelling is "onychodystrophy." The title should therefore read: DLA Class II Alleles Are Associated with Risk for Canine Symmetrical Lupoid Onychodystrophy (SLO). The corrected citation is: Wilbe M, Ziener ML, Aronsson A, Harlos C, Sundberg K, et al. (2010) DLA Class II Alleles Are Associated with Risk for Canine Symmetrical Lupoid Onychodystrophy (SLO). PLoS ONE 5(8): e12332. doi:10.1371/journal.pone.0012332


Gonium is intermediate in organizational complexity between its unicellular relative, Chlamydomonas, and its multicellular relatives with differentiated cell types, such as Volvox. Gonium pectorale consists of ~16 biflagellate cells arranged in a flat plate. The detailed molecular analysis of any species necessitates its accessibility to genetic manipulation, but, in volvocine algae, transformation procedures have so far only been established for Chlamydomonas reinhardtii and Volvox carteri.Green algae of the family Volvocaceae are a model lineage for studying the molecular evolution of multicellularity and cellular differentiation. The volvocine alga G. pectorale was achieved using a heterologous dominant antibiotic resistance gene, the aminoglycoside 3'-phosphotransferase VIII gene (aphVIII) of Streptomyces rimosus, as a selectable marker. Heterologous 3'- and 5'-untranslated flanking sequences, including promoters, were from Chlamydomonas reinhardtii or from Volvox carteri. After particle gun bombardment of wild type Gonium cells with plasmid-coated gold particles, transformants were recovered. The transformants were able to grow in the presence of the antibiotic paromomycin and produced a detectable level of the AphVIII protein. The plasmids integrated into the genome, and stable integration was verified after propagation for over 1400 colony generations. Co-transformants were recovered with a frequency of ~30–50% when cells were co-bombarded with aphVIII-based selectable marker plasmids along with unselectable plasmids containing heterologous genes. The transcription of the co-transformed, unselectable genes was confirmed. After heterologous expression of the luciferase gene from the marine copepod Gaussia princeps, which was previously engineered to match the codon usage in C. reinhardtii, Gonium transformants show luciferase activity through light emission in bioluminescence assays.Stable nuclear transformation of C. reinhardtii and from V. carteri work in G. pectorale and allow the functional expression of heterologous genes, such as the selectable marker gene aphVIII of S. rimosus or the co-transformed, codon-optimized G. princeps luciferase gene, which turned out to be a suitable reporter gene in Gonium. The availability of a method for transformation of Gonium makes genetic engineering of this species possible and allows for detailed studies in molecular evolution using the unicellular Chlamydomonas, the 16-celled Gonium, and the multicellular Volvox.Flanking sequences that include promoters from  Chlamydomonas reinhardtii . Unfortunately, no molecular tools and hardly any nucleotide sequence data are available for genera that are intermediate in organizational complexity between Chlamydomonas and Volvox. Because a molecular analysis of species with intermediate organizational complexity is important for the understanding of molecular evolution, we planned to establish a transformation technique in Gonium pectorale to allow for its genetic manipulation. This coenobial  volvocine green alga builds a slightly convex plate, which typically contains 16 cells in a rather square or rhomboidal arrangement, with four cells in the center and 12 cells in the periphery  of Streptomyces rimosus, which confers resistance to paromomycin  and cells were disrupted using a Sonopuls™ HD2070 sonicator (Bandelin Electronic) and the lysate was transferred to a 24-well plate. After addition of 150 μl 0.05 mM coelenterazine  in assay buffer, the 24-well plate was exposed to a chemiluminescence-sensitive film  for 30 s at 20°C , and frozen at -20°C for at least 20 min. After thawing, 20 μl samples were added to 125 μl of the assay buffer . Following incubation for 15 min at 20°C in the dark, samples were transferred to clear polystyrene vials , 50 μl 0.01 mM coelenterazine was added, and bioluminescence was assayed at 20°C using a MiniLumat LB9506 luminometer . The luminescence was recorded as relative light units.Quantitation of bioluminescence was performed as described by Shao and Bock . For it,For analysis of induction of luciferase activity in heat-shocked transformants, organisms were subject to a temperature shift from 23 to 36°C for 1 h, because in preliminary tests, shifts to 36° resulted in the strongest induction in comparison to lower or higher temperatures (data not shown). After a 1 h recovery phase at 23°C, cells were lysed by freezing and thawing and luciferase activity was assayed at 20°C as described above . As a reAlignment of sequences was done using the MUltiple Sequence Comparison by Log-Expectation program (MUSCLE) . Minor mThe novel sequences that are described in this study have been deposited under the following accession numbers:Gonium pectorale SAG 12.85: rbcL [GenBank: FJ793553], psaA [GenBank: FJ793556], psaB [GenBank: FJ793559], ITS [GenBank: FJ793562]; Gonium pectorale CCAP 32/14: rbcL [GenBank: FJ793554], psaA [GenBank: FJ793557], psaB [GenBank: FJ793560], ITS [GenBank: FJ793563]; Gonium pectorale NIES-1710: rbcL [GenBank: FJ793555], psaA [GenBank: FJ793558], psaB [GenBank: FJ793561], ITS [GenBank: FJ793564].The accession numbers of other cited sequences are:Gonium pectorale NIES-569: rbcL [GenBank: D63437], psaA [GenBank: AB044242], psaB [GenBank: AB044463]; Gonium pectorale UTEX 2570: ITS [GenBank: AF054425]; Gonium pectorale AWCAf2–3: ITS [GenBank: AF054431]; Gonium pectorale AWC-Laos: ITS [GenBank: AF182429]; Gonium pectorale Coleman 16-1: ITS [GenBank: U66969]; Gonium pectorale UTEX 2075: ITS [GenBank: AF054434]; Gonium pectorale UTEX 2581: ITS [GenBank: AF054433]; Gonium octonarium GO-LC-1+: rbcL [GenBank: D63436], psaA [GenBank: AB044241], psaB [GenBank: AB044462]; Gonium octonarium UTEX 842: ITS [GenBank: U66968]; Gonium quadratum NIES-653: rbcL [GenBank: D63438], psaA [GenBank: AB044243], psaB [GenBank: AB044464]; Gonium quadratum AWC-Cal3-3: ITS [GenBank: AF182430]; Gonium quadratum AWC-Cat: ITS [GenBank: AF182431]; Gonium multicoccum UTEX 2580: rbcL [GenBank: D63435], psaA [GenBank: AB044240], psaB [GenBank: AB044461]; Gonium multicoccum UTEX 783: ITS [GenBank: U66967]; Gonium viridistellatum UTEX 2519: rbcL [GenBank: D86831], psaA [GenBank: AB044244], psaB [GenBank: AB044465]; Gonium viridistellatum UTEX 2520: ITS [GenBank: AF182432]; Tetrabaena socialis  NIES-571: rbcL [GenBank: D63443], psaA [GenBank: AB044415], psaB [GenBank: AB044466]; Tetrabaena socialis  UTEX 14: ITS [GenBank: U66976]; Basichlamys sacculifera (= Gonium sacculiferum) NIES-566: rbcL [GenBank: D63430], psaA [GenBank: AB044416], psaB ; Basichlamys sacculifera (= Gonium sacculiferum) UTEX 822: ITS [GenBank: U66972]; Astrephomene gubernaculifera NIES-418: rbcL [GenBank: D63428], psaA [GenBank: AB044234], psaB [GenBank: AB044458]; Astrephomene gubernaculifera UTEX 1393: ITS [GenBank: AF054422]; Astrephomene perforata NIES-564: rbcL [GenBank: D63429], psaA [GenBank: AB044238], psaB [GenBank: AB044460]; Astrephomene perforata UTEX 2475: ITS [GenBank: U66939]; Pandorina morum NIES-574: rbcL [GenBank: D63442], psaA [GenBank: AB044226], psaB [GenBank: AB044452]; Pandorina morum Poona: ITS [GenBank: AF182433]; Eudorina unicocca UTEX 1215: rbcL [GenBank: D63434], psaA [GenBank: AB044209], psaB [GenBank: AB044440]; Eudorina elegans NIES-456: rbcL [GenBank: D63432], psaA [GenBank: AB044199], psaB [GenBank: AB044435]; Pleodorina californica UTEX 809: rbcL [GenBank: D63439], psaA [GenBank: AB044192], psaB [GenBank: AB044430]; Volvox aureus NIES-541: rbcL [GenBank: D63445], psaA [GenBank: AB044182], psaB [GenBank: AB044424]; Volvox carteri NIES-732: rbcL [GenBank: D63446], psaA [GenBank: AB044185], psaB [GenBank: AB044425]; Volvox globator UTEX 955: rbcL [GenBank: D86836], psaA [GenBank: AB044187], psaB [GenBank: AB044428]; Chlamydomonas reinhardtii 137C: rbcL [GenBank: J01399], psaA [GenBank: AB044419], psaB [GenBank: AB044470]; plasmid pPmr3 [GenBank: AY429514].aphVIII: aminoglycoside 3'-phosphotransferase VIII gene; ars: arylsulfatase gene; gPCR: genomic PCR; hsp70A: heat shock protein 70A gene; ITS: internal transcribed spacer; JM: Jaworski's Medium; MMLV: Moloney murine leukemia virus; PCR: polymerase chain reaction; psaA: photosystem I P700 chlorophyll a apoprotein A1 gene; psaB: photosystem I P700 chlorophyll a apoprotein A2 gene; psaD: photosystem I reaction center subunit II (chloroplastic) gene; rbcL: ribulose-1,5-bisphosphate carboxylase (large subunit) gene; rbcS: ribulose-1,5-bisphosphate carboxylase  gene; rRNA: ribosomal RNA; RT-PCR: reverse transcription PCR; UTR: untranslated region.KL conducted the experiments and analyzed the data. AH (corresponding author) conceived and coordinated the study, critically evaluated the data, and wrote the manuscript. All authors read and approved the final manuscript.Gonium pectorale strainsDescription of the phylogenetic analysis of utilized . The identity of the utilized Gonium pectorale strains SAG 12.85, CCAP 32/14 and NIES-1710 was verified in a phylogenetic analysis. Therefore, we cloned and sequenced certain DNA fragments that have been used in phylogenetic analyses of other volvocine algae. These include fragments of chloroplast genes encoding photosystem I P700 chlorophyll a apoprotein A1 (psaA), photosystem I P700 chlorophyll a apoprotein A2 (psaB) and ribulose bisphosphate carboxylase (rbcL), as well as the internal transcribed spacer sequences, ITS 1 and ITS 2, that flank the 5.8S ribosomal RNA (rRNA) nuclear gene.Click here for filepsaA cDNA fragments from several volvocine species.Sequence alignment of Click here for filepsaB cDNA fragments from several volvocine species.Sequence alignment of Click here for filerbcL cDNA fragments from several volvocine species.Sequence alignment of Click here for fileSequence alignment of ITS sequences flanking the 5.8S rRNA gene from several volvocine species.Click here for filepsaA, psaB and rbcL from several volvocine species.Sequence comparison of Click here for fileSequence comparison of ITS sequences flanking the 5.8S rRNA gene from several volvocine speciesClick here for filepsaA cDNA fragments from several volvocine speciesPhylogeny based on . Relationships among psaA cDNA fragments from several volvocine species. The unrooted tree was calculated using the neighbor-joining method of PHYLIP. Numbers indicate bootstrap analysis values obtained using 10000 resampled data sets. The analysis is based on the alignment given in Additional File Gonium pectorale strains are highlighted in light blue. Gonium pectorale strains used in this study are indicated by a dark blue arrow.Click here for filepsaB cDNA fragments from several volvocine speciesPhylogeny based on . Relationships among psaB cDNA fragments from several volvocine species. The unrooted tree was calculated using the neighbor-joining method of PHYLIP. Numbers indicate bootstrap analysis values obtained using 10000 resampled data sets. The analysis is based on the alignment given in Additional File Gonium pectorale strains are highlighted in light blue. Gonium pectorale strains used in this study are indicated by a dark blue arrow.Click here for filerbcL cDNA fragments from several volvocine speciesPhylogeny based on . Relationships among rbcL cDNA fragments from several volvocine species. The unrooted tree was calculated using the neighbor-joining method of PHYLIP. Numbers indicate bootstrap analysis values obtained using 10000 resampled data sets. The analysis is based on the alignment given in Additional File Gonium pectorale strains are highlighted in light blue. Gonium pectorale strains used in this study are indicated by a dark blue arrow.Click here for filepsaA, psaB and rbcL cDNA fragments from several volvocine speciesPhylogeny based on a combined data set of . Relationships within a combined data set generated from the psaA, psaB and rbcL sequences from several volvocine species. The unrooted tree was calculated using the neighbor-joining method of PHYLIP. Numbers indicate bootstrap analysis values obtained using 30000 resampled data sets. The analysis is based on the alignments given in Additional Files Gonium pectorale strains are highlighted in light blue. Gonium pectorale strains used in this study are indicated by a dark blue arrow.Click here for filePhylogeny based on ITS 1, ITS 2 and 5.8S rRNA sequences from several volvocine species. Relationships among ITS 1/5.8S rRNA/ITS 2 sequences from several volvocine species. The unrooted tree was calculated using the neighbor-joining method of PHYLIP. Numbers indicate bootstrap analysis values obtained using 10000 resampled data sets. The analysis is based on the alignment given in Additional File Gonium pectorale strains are highlighted in light blue. Gonium pectorale strains used in this study are indicated by a dark blue arrow.Click here for file


The wrong file was inadvertently uploaded for Table S17. The authors regret this error. The file should contain the following table:Criteria Number of genes======================================Total representative genes: 20,905Have at least one GO annotation: 12,515Have at least one GO process annotation: 8,058Have at least one GO component annotation: 7,797Have at least one GO function annotation: 10,170The correct Supplementary Table file can be obtained at:Click here for additional data file.


Because the author Tuncay Delibasi was added to the author byline after publication, the Author Contributions section does not correctly reflect the new authorship. The new Author Contributions should read:Conceived and designed the experiments: GPF TB RA TD KE MLW JL. Performed the experiments: GPF TB RA TD KE HKE MLW JL. Analyzed the data: GPF TB RA TD KE MLW JL. Contributed reagents/materials/analysis tools: RA MLW JL. Wrote the paper: GPF TB RA TD KE MLW JL.


There are two Co atoms in the asymmetric unit, each lying on an inversion centre and adopting a distorted octahedral coordination. Classical and non-classical hydrogen bonds are responsible for formation of a three-dimensional polymeric network in the crystal.The title cobalt(II) complex, [CoCl DOI: 10.1107/S1600536809042263/dn2499Isup2.hkl            Structure factors: contains datablocks I. DOI:  crystallographic information; 3D view; checkCIF report            Additional supplementary materials:


The ZnII ion is also four-coordinated by the two phenolate O atoms of the Schiff base ligand and by two cis-coordinated chloride anions.In the title compound, [CuZnCl DOI: 10.1107/S1600536809006928/hg2482Isup2.hkl            Structure factors: contains datablocks I. DOI:  crystallographic information; 3D view; checkCIF report            Additional supplementary materials:


The two crystallographically independent oxalate ligands, each lying on an inversion center, act as bridging ligands, linking Nd atoms into an extended zigzag chain. Neighboring chains are linked by the pyrazine-2-carboxylate ligands into a two-dimensional layerlike network in the (10In the title complex, [Nd(C DOI: 10.1107/S1600536809031250/hy2217Isup2.hkl            Structure factors: contains datablocks I. DOI:  crystallographic information; 3D view; checkCIF report            Additional supplementary materials:


Correction to: Isr J Health Policy Reshttps://doi.org/10.1186/s13584-019-0338-0Incorrect: Hierarchy of hair loss stigma: media portrayals of cancer, alopecia areata, and cancer in Israeli newspapersCorrect: Hierarchy of hair loss stigma: media portrayals of cancer, alopecia areata, and ringworm in Israeli NewspapersThe original publication of this article  containeThe original publication has been updated.


Affiliation 1 and affiliation 2 are incomplete and incorrectly switched. The complete, correct affiliation 1 is: Research Centre for Olive, Citrus and Tree Fruit—Council for Agricultural Research and Economics, 87036 Rende (CS) IT. The complete, correct affiliation 2 is: Department of Biology, Ecology and Earth Science, University of Calabria, 87036 Arcavacata Rende (CS) IT.Affiliation 3 is incomplete. The complete affiliation 3 is: Agenzia Nazionale per le Nuove Tecnologie, l’Energia e lo Sviluppo Economico Sostenibile (ENEA), TRISAIA Research Center, S.S. 106 Jonica, 75026 Rotondella (MT) IT.


Corrigendum: After publication, a reader notified the authors that there are taxonomic and spelling issues throughout the article and in the reference list. The following changes have been made: A type species has been identified for Fimbriaphyllia, which is now assigned to the family Euphylliidae. The publication is now registered with ZooBank (urn:lsid:zoobank.org:pub:EBF69BA0-897E-4AC8-ADF5-4A7115CA1353).Physogyra lichtensteni, Plerogyra sinuosa , Plesiastrea versipora and Blastomussa wellsi are now classified as incertae sedis.Faviidae is no longer listed as a family. Favia rotumana is now referred to as Dipsastraea rotumana, and Favia pallida is now referred to as Dipsastraea pallida.The taxonomic status of Trachyphyllidae [sic] has been removed.F. divisa is now described as having both flabello-meandroid and phacelo-flabellate or phaceloid corallite structures.Euphyllidae is now spelled as Euphylliidae, Caryophyllidae as Caryophylliidae and Lobophyllidae as Lobophylliidae.Figure 4 has been revised to reflect that Fimbriaphyllia divisa and Fimbriaphyllia yaeyamaensis both have phaceloid and phacelo-flabellate colony formations. These traits are indicated in the branches next to the photos of F. divisa and F. yaeyamaensis in the dichotomous tree based on phylogeny.New Figures 5 and 6 have been added to demonstrate the skeletal morphology of the Euphyllia and Fimbriaphyllia species, and height comparisons between F. paradivisa and E. glabrescens, respectively.New references Alloiteau (1952), Benzoni et al. (2014), Budd et al. (2012), Chamisso & Eysenhardt (1821), Chen et al. (2005), Eyal et al. (2016), Gravier (1910), Griffith (2004), Huang et al. (2016), Khodzori et al. (2015), Matthai (1928), Mazlan et al. (2005), Milne & Haime (1848), Milne & Haime (1849), Milne & Haime (1851), Nemenzo (1960), Oken (1815-1816), Pillai (1971), Quoy & Gaimard (1824), Richards & Beger (2013), Saville-Kent (1893), Shirai (1980), Spengler (1799), Umbgrove (1939), Veron & Hodgson (1989), Veron (1990), Veron (1992) and Wells (1971) have been added. Additionally, titles for references Akmal et al. (2017), Arrigoni et al. (2016) and Veron & Pichon (1980) have been corrected.Lastly, the authors have added an Acknowledgements and a New Species Registration section and have updated the Funding statement, Grant Disclosure, and Field Study Permissions to reflect additional work done post-publication. Publisher’s Note: On July 8, 2018, this corrigendum was published with an incomplete LSID. This has now been corrected to: urn:lsid:zoobank.org:pub:EBF69BA0-897E-4AC8-ADF5-4A7115CA1353


AbstractColeoptera: Coccinellidae) known in Algeria now contains 75 species belonging in ten tribes. New country records include the European species Oenopiaconglobata and the invasive Asian species Harmoniaaxyridis. Sampling data is provided for 14 species found during a faunistic survey performed mostly in agroecosystems, together with host plant and prey species.An updated and corrected checklist of species of ladybird beetles ( Coccinellidae (ladybirds) is the most species-rich family in the recently recognized beetle superfamily Coccinelloidea (Family elloidea  with appelloidea . They arelloidea .Coccinellidae) of individual countries are relatively well known, and the fauna of Algeria is also relatively well documented   used by Exochomusquadripustulatus       Rhyzobiuslophantae    Tetrabrachyscribratellus   Tetrabrachysvolkonskyi   CoccinelliniAdaliabipunctata       undecimnotata used by Cheilomenespropinqua   Coccinellaseptempunctata Linnaeus, 1758  (new record) *PageBreakHarmoniaquadripunctata   Hippodamiatredecimpunctata   (as H. (Adonia) variegata by Myrrhaoctodecimguttata   Oenopiaconglobata  (new record) *Oenopiadoublieri         Henosepilachnaargus        Nephus (Bipunctatus) bicinctus   Nephus (Bipunctatus) bipunctatus   Nephus (Bipunctatus) conjunctus   Nephus (Sidis) hiekei   Nephus (Sidis) levaillanti  (syn. Scymnuslevaillanti used by Nephus (Nephus) ludyi   Nephus (Bipunctatus) peyerimhoffi  (source:  source:  *Nephus (Nephus) quadrimaculatus  (source:  source: Nephus (Nephus) redtenbacheri   Scymniscussplendidulus   Scymnus (Scymnus) apetzi Mulsant, 1846 (source:  source: Scymnus (Scymnus) bivulnerus Baudi di Selve, 1894 (source:  source: Scymnus (Mimopullus) fulvicollis Mulsant, 1846 (syn. Pullusfulvicollis used by Scymnus (Scymnus) interruptus  (source:  source: Scymnus (Scymnus) laetificus Weise, 1879  marginalis   Scymnus (Mimopullus) marinus   nubilus   Scymnus (Scymnus) pavesii Canepari, 1983  rufipes  (source:  source: Scymnus (Pullus) subvillosus  (syn Pullussubvillosus used by PageBreakScymnus (Scymnus) suffrianioides Sahlberg, 1913  suturalis Thunberg, 1795   used by SticholotidiniCoelopterussalinus Mulsant & Rey, 1852     Chilocoruscacti was introduced in Algeria but probably did not establish itself  using a CLIMEX model. Although meanwhile it has been found in a few countries with wet tropical climate (Kenya: H.axyridis in Algeria. The first specimen found was a male (Fig. succinea. The establishment of the species was confirmed by occurrence of many larvae and pupae in 2018. All adults found in 2018 were of form succinea, which is the most common colour form in the native Chinese as well as in most invasive populations (The occurrence of the invasive alien species e Kenya: , and in  Poutsma 008 usingulations .Oenopiaconglobata is a common tree inhabiting predatory ladybird living in most European countries and as a subspecies in large parts of Asia. The specimen collected in Algeria has yellow elytral background (Fig. und Fig. , while i