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  • 1. Adomas, Aleksandra
    et al.
    Heller, Gregory
    Olson, Åke
    Osborne, Jason
    Karlsson, Magnus
    Nahalkova, Jarmila
    Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences P.O. Box 7026, Uppsala, Sweden.
    Van Zyl, Len
    Sederoff, Ron
    Stenlid, Jan
    Finlay, Roger
    Asiegbu, Frederick O.
    Comparative analysis of transcript abundance in Pinus sylvestris after challenge with a saprotrophic, pathogenic or mutualistic fungus2008In: Tree Physiology, ISSN 0829-318X, E-ISSN 1758-4469, Vol. 28, no 6, p. 885-897Article in journal (Refereed)
    Abstract [en]

    To investigate functional differences in the recognition and response mechanisms of conifer roots to fungi with different trophic strategies, Pinus sylvestris L. was challenged with a saprotrophic fungus Trichoderma aureoviride Rifai. The results were compared with separate studies investigating pine interactions with a pathogen, Heterobasidion annosum (Fr.) Bref. sensu stricto and an ectomycorrhizal symbiont, Laccaria bicolor Maire (Orton). Global changes in the expression of 2109 conifer genes were assayed 1, 5 and 15 days after inoculation. Gene expression data from a cDNA microarray were analyzed by the 2-interconnected mixed linear model statistical approach. The total number of genes differentially expressed compared with the uninfected control was similar after challenge with the pathogen and the ectomycorrhizal symbiont, but the number of differentially expressed genes increased over time, for H. annosum, and decreased for L. bicolor. Inoculation of pine roots with T aureoviride resulted overall in a much lower number of genes with changed transcript levels compared with inoculation with H. annosum or L. bicolor. Functional classification of the differentially expressed genes revealed that the ectomycorrhizal fungus triggered transient induction of defence-related genes. The response and induction of defence against the pathogen was delayed and the magnitude increased over time. Thus, there were specific transcriptional responses depending on whether the conifer roots were challenged with mutualistic, saprotrophic or pathogenic fungi. This suggests that pine trees are able to recognize diverse fungal species and specifically distinguish whether they are pathogenic, neutral or beneficial microbial agents.

  • 2. Asiegbu, F O
    et al.
    Choi, W B
    Li, G S
    Nahalkova, Jarmila
    Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, Box 7026, Uppsala, Sweden.
    Dean, R A
    Isolation of a novel antimicrobial peptide gene (Sp-AMP) homologue from Pinus sylvestris (Scots pine) following infection with the root root fungus Heterobasidion annosum2003In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 228, no 1, p. 27-31Article in journal (Refereed)
    Abstract [en]

    A new family of antimicrobial peptide homologues termed Sp-Amp has been discovered in Pinus sylvestris (Scots pine). This is the first report of such proteins to be characterized in a conifer species. Sp-AMP1 was identified in a substructured cDNA library of root tissue infected with the root rot fungus Heterobasidion annosum and encodes a mature peptide of 79 amino acid residues. Three additional members of the Sp-AMP family (Sp-AMPs 2-4) encode cysteine-rich proteins of 105 amino acids, each containing an N-terminal region with a probable cleavage signal sequence. Northern analysis confirmed that Sp-AMP expression is elevated in Scots pine roots upon infection with H. annosum. These peptides share 64% amino acid identity with a mature protein from Macadamia integrifolia (MiAMP1), which allowed us to build a homology model for preliminary analysis. Southern analyses further confirmed that several copies of the gene are present in the Scots pine genome. The potential significance of Sp-AMP in the H. annosum-conifer pathosystem is discussed.

  • 3. Asiegbu, F O
    et al.
    Nahalkova, Jarmila
    Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Li, G S
    Pathogen-inducible cDNAs from the interaction of the root rot fungus Heterobasidion annosum with Scots pine (Pinus sylvestris L.)2005In: Plant Science, ISSN 0168-9452, E-ISSN 1873-2259, Vol. 168, no 2, p. 365-372Article in journal (Refereed)
    Abstract [en]

    Subtractive hybridization was used to select cDNAs representing genes that are differentially expressed during interaction of the necrotroph Heterobasidion annosum and its conifer host (Pinus sylvestris). We obtained 966 ESTs from the subtraction cDNA library, which included 509 singletons and 147 contigs. The sequences of 492 clones (51%) significantly matched National Centre for Biotechnology Information Database entries. Four hundred and seventy-four ESTs (49%) had not been previously described. The ESTs with moderate to high similarity scores based on BlastX were organized into categories based on their putative function. Among the genes identified, 16% were associated with metabolism and other cellular functions, 14% with cell rescue and defence and 39% were classified as unknown. Seven of the genes shared significant homology to fungal genes. A cDNA encoding an antimicrobial peptide (AMP) was the most abundant transcript representing 2% of the total sequenced clones. The expression pattern of five ESTs (peroxidase, anti-microbial peptide, resistance gene analogue, unknown protein, thaumatin) were analysed by virtual Northern blot, and confirmed elevated levels of the gene transcripts upon pathogen infection. These ESTs provide insight into the host-pathogen interaction and also represent a resource for future research on H. annosum-conifer pathosystems.

  • 4. Asiegbu, F.O.
    et al.
    Choi, W.
    Li, G.
    Nahalkova, Jarmila
    Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, Box 7026, Uppsala, Sweden.
    Dean, R.A.
    Isolation of a novel antimicrobal peptide gene (Sp-AMP) homologue from Pinus sylvestris (Scots pine) following infection with the root rot fungus Heterobasidion annosum2003In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 228, no 1, p. 27-31Article in journal (Refereed)
    Abstract [en]

    A new family of antimicrobial peptide homologues termed Sp-Amp has been discovered in Pinus sylvestris (Scots pine). This is the first report of such proteins to be characterized in a conifer species. Sp-AMP1 was identified in a substructured cDNA library of root tissue infected with the root rot fungus Heterobasidion annosum and encodes a mature peptide of 79 amino acid residues. Three additional members of the Sp-AMP family (Sp-AMPs 2–4) encode cysteine-rich proteins of 105 amino acids, each containing an N-terminal region with a probable cleavage signal sequence. Northern analysis confirmed that Sp-AMP expression is elevated in Scots pine roots upon infection with H. annosum. These peptides share 64% amino acid identity with a mature protein from Macadamia integrifolia (MiAMP1), which allowed us to build a homology model for preliminary analysis. Southern analyses further confirmed that several copies of the gene are present in the Scots pine genome. The potential significance of Sp-AMP in the H. annosum–conifer pathosystem is discussed.

  • 5. Frykman, Susanne
    et al.
    Hur, Ji-Yeun
    Frånberg, Jenny
    Aoki, Mikio
    Winblad, Bengt
    Nahalkova, Jarmila
    Department of Neurobiology, Care Sciences and Society (NVS), Karolinska Institutet Dainippon Sumitomo Pharma Alzheimer Center (KASPAC), Novum, Huddinge, Sweden.
    Behbahani, Homira
    Tjernberg, Lars O.
    Synaptic and Endosomal Localization of Active gamma-Secretase in Rat Brain2010In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 5, no 1, p. e8948-Article in journal (Refereed)
    Abstract [en]

    Background

    A key player in the development of Alzheimer's disease (AD) is the gamma-secretase complex consisting of at least four components: presenilin, nicastrin, Aph-1 and Pen-2. gamma-Secretase is crucial for the generation of the neurotoxic amyloid beta-peptide (A beta) but also takes part in the processing of many other substrates. In cell lines, active gamma-secretase has been found to localize primarily to the Golgi apparatus, endosomes and plasma membranes. However, no thorough studies have been performed to show the subcellular localization of the active gamma-secretase in the affected organ of AD, namely the brain.

    Principal Findings

    We show by subcellular fractionation of rat brain that high gamma-secretase activity, as assessed by production of A beta 40, is present in an endosome-and plasma membrane-enriched fraction of an iodixanol gradient. We also prepared crude synaptic vesicles as well as synaptic membranes and both fractions showed high A beta 40 production and contained high amounts of the gamma-secretase components. Further purification of the synaptic vesicles verified the presence of the gamma-secretase components in these compartments. The localization of an active gamma-secretase in synapses and endosomes was confirmed in rat brain sections and neuronal cultures by using a biotinylated gamma-secretase inhibitor together with confocal microscopy.

    Significance

    The information about the subcellular localization of gamma-secretase in brain is important for the understanding of the molecular mechanisms of AD. Furthermore, the identified fractions can be used as sources for highly active gamma-secretase.

  • 6. Gemeiner, P.
    et al.
    Nahalka, J.
    Vikartovska, A.
    Nahalkova, J.
    Tomaska, M.
    Sturdik, E.
    Markovic, O.
    Malovikova, A.
    Zatkova, I.
    Ilavsky, M.
    Calcium pectate gel could be a better alternative to calcium alginate gel in multiple applications of immobilized cells1996In: Immobilized cells: basic and applications / [ed] Wijffels R.H., Buitelaar R.M., Bucke C., Tramper J., Netherlands: Elsevier, 1996, p. 76-83Chapter in book (Other academic)
  • 7. Gemeiner, P
    et al.
    Nahalka, J
    Vikartovska, A
    Nahalkova, J
    Tomaska, M
    Sturdik, E
    Markovic, O
    Malovikova, A
    Zatkova, I
    Ilavsky, M
    Calcium pectate gel could be a better alternative to calcium alginate gel in multiple applications of immobilized cells1996In: IMMOBILIZED CELLS: BASICS AND APPLICATIONS, 1996, p. 76-83Conference paper (Refereed)
  • 8. Hajduch, M
    et al.
    Nahalkova, Jarmila
    Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia.
    Hrib, J
    Vookova, B
    Gemeiner, P
    An electrophoretic analysis of the seed protein body proteins from Pinus nigra2001In: Biologia plantarum, ISSN 0006-3134, E-ISSN 1573-8264, Vol. 44, no 1, p. 137-140Article in journal (Refereed)
    Abstract [en]

    Protein bodies (PBs) of European black pine (Pirus nigra Am.) were isolated from mature seeds. Extracted soluble matrix proteins and crystalloid proteins PBs proteins were investigated by SDS-PAGE electrophoresis in presence and absence of 2-mercaptoethanol. The proteins of molecular masses 16, 17, 18, 61 and 65 kDa were presented only in crystalloid protein samples. Only 15 kDa protein was present in soluble matrix proteins and not in crystalloid proteins. Another protein bands were present in both soluble matrix and crystalloid proteins. 20, 37, 38, 39 and 48 kDa proteins were strongly visible among crystalloid proteins. Bands of 23 and 32 kDa were more visible in soluble matrix protein samples. Different composition in crystalloid proteins was found in absence of 2-mercaptoethanol: no proteins with molecular mass 71 kDa and more proteins in soluble matrix. In case of crystalloid proteins we detected 7 protein bands in interval from 71 to 212 kDa.

  • 9. Hajduch, Martin
    et al.
    Nahalkova, Jarmila
    Hrib, Jiri
    Vookova, Bozena
    Gemeiner, Peter
    Soluble and insoluble seed protein body proteins from Pinus nigra1999In: Biologia (Bratislava), ISSN 0006-3088, E-ISSN 1336-9563, Vol. 54, no Suppl. 7, p. 61-61Article in journal (Refereed)
  • 10. Hrib, J
    et al.
    Janisch, R
    Vookova, B
    Nahalkova, Jarmila
    Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia .
    Gemeiner, P
    Hajduch, M
    Protein bodies in the pine megagametophyte in vitro culture: ultrastructural, histochemical and electrophoretic observations2000In: Biologia plantarum, ISSN 0006-3134, E-ISSN 1573-8264, Vol. 43, no 3, p. 329-336Article in journal (Refereed)
    Abstract [en]

    The megagametophytes of the European black pine (Pinus nigra Am.) were cultured on modified MS medium. After 10 d, protein bodies showed well-marked degradation on freeze-etched replicas and in preparations observed by scanning electron microscopy. After 20 d of cultivation, the megagametophyte cells were completely empty. Proteins secreted into the agar medium were determined by electrophoresis and 15 different proteins, in the range of 6.5 to 71 kDa, were identified.

  • 11. Hrib, Jiri
    et al.
    Vookova, Bozena
    Nahalkova, Jarmila
    Hajduch, Martin
    Kotacka, Libor
    Janisch, Roman
    Gemeiner, Peter
    Komrska, Jiri
    Structure and function of protein bodies: A model study on pine cells1999In: Biologia (Bratislava), ISSN 0006-3088, E-ISSN 1336-9563, Vol. 54, no Suppl. 7, p. 60-60Article in journal (Refereed)
  • 12. Nahalka, J
    et al.
    Nahalkova, Jarmila
    Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta, Bratislava, Slovak-Republic.
    Gemeiner, P
    Blanarik, P
    Elicitation of plumbagin by chitin and its release into the medium in Drosophyllum lusitanicum Link. suspension cultures1998In: Biotechnology letters, ISSN 0141-5492, E-ISSN 1573-6776, Vol. 20, no 9, p. 841-845Article in journal (Refereed)
    Abstract [en]

    Polysaccharides (chitin/pectin) that are involved in the interactions between plants and microorganisms were applied to the cultured cells of Drosophyllum lusitanicum. In the case of chitin addition, elicitation and crystallization of plumbagin in the medium were observed. N-Acetylchitooligosaccharides smaller than heptamers [(GlcNAc)(n) (n<7)] elicited the biosynthesis of plumbagin but did not increase the hypersensitive response (HR). On the other hand, carboxymethylchitin (DP similar to 200) led to the accumulation of plumbagin in cells and to HR death as well as to the lysis of the cells and release of plumbagin into the medium. The response of cultured cells to the N-Acetylchitosaccharides varied depending on the chemo/physiological conditions of the cells. Addition of pectin (1 g/l) resulted in enhanced HR and decreased biosynthesis of plumbagin.

  • 13. Nahalkova, J
    et al.
    Chrtiansky, J
    Hrib, J
    Vookova, B
    Gemeiner, P
    Hajduch, M
    Lectin-like and antifungal activity of protein body proteins from Pinus nigra seeds1996In: Chemické listy (Print), ISSN 0009-2770, E-ISSN 1213-7103, Vol. 90, no 9, p. 698-699Article in journal (Refereed)
  • 14.
    Nahalkova, Jarmila
    et al.
    Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Asiegbu, F O
    Daniel, G
    Hrib, J
    Vookova, B
    Pribulova, B
    Gemeiner, P
    Isolation and immunolocalization of a Pinus nigra lectin (PNL) during interaction with the necrotrophs: Heterobasidion annosum and Fusarium avenaceum2001In: Physiological and molecular plant pathology, ISSN 0885-5765, E-ISSN 1096-1178, Vol. 59, no 3, p. 153-163Article in journal (Refereed)
    Abstract [en]

    Pinus nigra ARN. lectin (PNL) was isolated from protein bodies of European black pine seeds by affinity chromatography of protein bodies extract on GIcNAc-C-glycosylated Spheron 300 beads. In vitro tests confirmed that PNL has the ability to agglutinate rat erythrocytes, GIcNAc-C-glycosylated HEMA BIO 1000 microbeads and Trichoderma viride spores. Western blot immuno-assays using antibodies against PNL antigen demonstrated specificity of the antisera. Immuno dot assays with polyclonal antibody against the purified PNL exhibited strong cross reaction to both Scots pine and Norway spruce seed extracts but not to fungal cell wall extracts from either Heterobasidion annosum or Fusarium avenaceum or Phlebiopsis gigantea. The indirect immunofluorescence assay showed a strong interaction of PNL with hyphal materials of the necrotrophs (H. annosum and F avenaceum). Results from immunolocalization experiments suggest that PNL is not degraded during seed germination and development since it was localized in all parts of juvenile pine seedlings. The protein was mainly observed on the cytoplasmic membranes and on the primary cell walls. In infected seedlings, a strong labelling of hyphal materials with PNL antisera was recorded only at the early stages of infection but not at the later stages of hyphal invasion. At advanced stages of the infection, most of the fungal hyphae were covered by host phenolic-like substances which probably masked the interaction of the antiserum with pathogen cell wall materials; hence the low labelling. The potential function of PNL as a recognition molecule during interaction of conifer trees with necrotrophic parasites is discussed.

  • 15.
    Nahalkova, Jarmila
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolutionary Biology, Molecular Evolution.
    Fatehi, Jamshid
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolutionary Biology, Molecular Evolution.
    Red fluorescent protein (DsRed2) as a novel reporter in Fusarium oxysporum f. sp. Lycopersici2003In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 225, no 2, p. 305-309Article in journal (Refereed)
    Abstract [en]

    pAn-DsRed2 vector was constructed for constitutive cytoplasmic expression of the red fluorescent protein (DsRed2) under control of the glyceraldehyde-3-phosphate dehydrogenase gene promoter from Aspergillus nidulans. DsRed2-transformation of two Fusarium oxysporum f. sp. lycopersici strains pathogenic against tomato host resulted in bright red cytoplamic fluorescence of the fungus. The transformants were screened based on the hygromycin B resistance, brightness, stability and rate of appearance of the DsRed2 fluorescence. The transormed fungi were growing normally and their pathogenicity did not change after transformation procedure. The function of novel DsRed2 marker was verified by fluorescence microscopy of the infected tomato seedlings. The results indicate that DsRed2 can be used as a efficient novel reporter gene for monitoring of the F. oxysporum within the host tissues.

  • 16.
    Nahalkova, Jarmila
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolutionary Biology, Molecular Evolution.
    Fatehi, Jamshid
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolutionary Biology, Molecular Evolution.
    Red fluorescent protein (DsRed2) as a novel reporter in Fusarium oxysporum f. sp lycopersiei2003In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 225, no 2, p. 305-309Article in journal (Refereed)
    Abstract [en]

    pAn-DsRed2 vector was constructed for constitutive cytoplasmic expression of the red fluorescent protein (DsRed2) under control of the glyceraldehyde-3-phosphate dehydrogenase gene promoter from Aspergillus nidulans. DsRed2-transformation of two Fusarium oxysporum f. sp. lycopersici strains pathogenic against tomato host resulted in bright red cytoplamic fluorescence of the fungus. The transformants were screened based on the hygromycin B resistance, brightness, stability and rate of appearance of the DsRed2 fluorescence. The transormed fungi were growing normally and their pathogenicity did not change after transformation procedure. The function of novel DsRed2 marker was verified by fluorescence microscopy of the infected tomato seedlings. The results indicate that DsRed2 can be used as a efficient novel reporter gene for monitoring of the F oxysporum within the host tissues.

  • 17.
    Nahalkova, Jarmila
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolution, Genomics and Systematics, Molecular Evolution.
    Fatehi, Jamshid
    Olivain, Chantal
    Alabouvette, Claude
    Tomato root colonization by fluorescent-tagged pathogenic and protective strains of Fusarium oxysporum in hydroponic culture differs from root colonization in soil2008In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 286, no 2, p. 152-157Article in journal (Refereed)
    Abstract [en]

    The colonization process of tomato roots inoculated separately or/and simultaneously by a pathogenic Fusarium oxysporum f. sp. lycopersici strain Fol8 and the protective F. oxysporum strain Fo47, genetically tagged with the red and green fluorescent protein genes, respectively, was studied in a hydroponic culture. Plants were coinoculated with Fol8 and Fo47 at two conidial concentration ratios of 1/1 and 1/100, in which biological control was not effective or effective, respectively. First observation of fungi on root was possible 48 h after inoculation at a high inoculum level and 5 days post inoculation at the lower concentration of inoculum. The pattern of root colonization was similar for both strains with the initial development of hyphal network on the upper part of taproot, followed by the growth of hyphae towards the elongation zone, lateral roots and root apices. Finally, the whole elongation zone and root apex were invaded by both strains but no specific infection sites were observed. When coinoculated, both strains could grow very closely or even at the same spot on the root surface. At the nonprotective ratio, Fol8 was the successful colonizer, but application of Fo47 at a concentration 100 times > , Fol8 delayed vessel colonization by the pathogen.

  • 18.
    Nahalkova, Jarmila
    et al.
    Slovak Acad Sci, Inst Chem, Dubravska Cesta 9, SK-84238 Bratislava, Slovakia.
    Hrib, J
    Gemeiner, P
    Vookova, B
    Chrtiansky, J
    Lectin-like activity in European black pine (Pinus nigra) seed protein bodies1999In: Biologia (Bratislava), ISSN 0006-3088, E-ISSN 1336-9563, Vol. 54, no 1, p. 113-117Article in journal (Refereed)
    Abstract [en]

    A study of black pine seed protein bodies with regard to the presence of defense-related proteins was performed. Interaction of the intact protein bodies with saccharides revealed specificity of surface membrane binding sites for chitin, cellulose, laminarin and the respective mono- or disaccharides. Crude protein fraction isolated from protein bodies possessed an ability to agglutinate fungal spores. Inhibiton of the fungal spores agglutination confirmed the presence of lectin-like proteins specific to basic fungal cell wall components.

  • 19.
    Nahalkova, Jarmila
    et al.
    nstitute of Chemistry, Slovak Academy of Sciences, SK-842 38 Bratislava.
    Pribulova, B
    Svitel, J
    Konigstein, K
    Petrusova, M
    Gemeiner, P
    Petrus, L
    Glycosylmethylamines as a new tool in the lectin research1999In: Chemické zvesti, ISSN 0366-6352, E-ISSN 1336-9075, Vol. 53, no 5, p. 340-343Article in journal (Refereed)
  • 20. Nahalkova, Jarmila
    et al.
    Volkmann, Inga
    Aoki, Mikio
    Winblad, Bengt
    Bogdanovic, Nenad
    Tjernberg, Lars O.
    Behbahani, Homira
    CD147, a gamma-secretase associated protein is upregulated in Alzheimer's disease brain and its cellular trafficking is affected by presenilin-22010In: Neurochemistry International, ISSN 0197-0186, E-ISSN 1872-9754, Vol. 56, no 1, p. 67-76Article in journal (Refereed)
    Abstract [en]

    gamma-Secretase activity has been extensively investigated due to its role in Alzheimer's disease. Here, we studied the association of CD147, a transmembrane glycoprotein belonging to the immunoglobulin family, with gamma-secretase and its expression in Alzheimer's disease and control tissues. Subcellular fractionation of postmitochondrial supernatant from Fat brain on step iodixanol gradient in combination with co-immunoprecipitation using an anti-nicastrin antibody showed association of limited amount of CD147 to gamma-secretase. By immunoblotting of postnuclear pellets from Alzheimer's disease and control human brain tissues we showed that CD147 with molecular weight 75 kDa is upregulated in frontal cortex and thalamus of the Alzheimer's disease brains. Immunohistochemistry of brain tissues from Alzheimer's disease and control revealed specific Upregulation of CD147 in neurons, axons and capillaries of Alzheimer's disease frontal cortex and thalamus. The effect of presenilin-1 and -2, which are the catalytic subunits of gamma-secretase, on CD147 expression and subcellular localization was analyzed by confocal microscopy in combination with flow cytometry and showed that PS2 affected the subcellular localization of CD147 in Mouse embryonic fibroblast cells. We suggest that a small fraction of CD147 present in the brain is associated with the gamma-secretase, and can be involved in mechanisms dysregulated in Alzheimer's disease brain.

  • 21.
    Nahálková, Jarmila
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    The protein-interaction network with functional roles in tumorigenesis, neurodegeneration, and aging2016In: Molecular and Cellular Biochemistry, ISSN 0300-8177, E-ISSN 1573-4919, Vol. 423, no 1-2, p. 187-196Article, review/survey (Refereed)
    Abstract [en]

    The present review summarizes the knowledge about a protein-interaction network, which includes proteins with significant functions in the mechanisms of aging and age-related diseases. All the detected interacting proteins TPPII, p53, MYBBP1A, CDK2 and SIRT7, SIRT6, and CD147 are suitable for the development of antitumor therapeutics and treatments for diseases of aging. TPPII and SIRT6 directly affect glucose metabolism which drive malignant growth. In addition, SIRT6 activators are attractive candidates for Alzheimer's disease (AD) due to the protection effect of SIRT6 overexpression from DNA damage. TPPII activity exhibits a decreasing effect on mTOR signaling, and its requirement for the degradation of A beta peptides in the human fibroblasts suggests that it has dual functions in tumorigenesis and AD-related pathology. Likewise, the direct promotion of the invasiveness of breast epithelial cells and the contribution to the A beta degradation by stimulating the matrix metalloproteinases production suggest a double functional role for CD147. An association of the partial portion of cellular CD147 to gamma-secretase further supports the functional relation to AD pathology. The animal and cellular models with downregulated or knockout TPPII, p53, SIRT6, SIRT7, and MYBBP1A expression levels illustrate similar functions of the interacting proteins. They demonstrate similar effects on the length of life span, premature aging, and lipid metabolism. The presented protein-interaction network is relevant to the discoveries of the mechanisms of tumorigenesis, aging, and neurodegeneration.

  • 22.
    Nahálková, Jarmila
    et al.
    Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84238, Bratislava, Slovak Republic.
    Svitel, Juraj
    Gemeiner, Peter
    Danielsson, Bengt
    Pribulová, Bozena
    Petrus, Ladislav
    Affinity analysis of lectin interaction with immobilized C- and O- gylcosides studied by surface plasmon resonance assay2002In: Journal of Biochemical and Biophysical Methods, ISSN 0165-022X, E-ISSN 1872-857X, Vol. 52, no 1, p. 11-18Article in journal (Refereed)
    Abstract [en]

    A biosensor based on the surface plasmon resonance (SPR) principle was used for kinetic analysis of lectin interactions with different immobilized saccharide structures. A novel affinity ligands beta-D-glycopyranosylmethylamines derived from common D-aldohexoses linked to the carboxymethyl dextran layer of the SPR sensor surface served for interactions with a wide range of lectins. The method of preparation and use of the beta-D-mannopyranosyl glycosylated sensor surface was described. The results of affinity analysis of lectin-ligand interactions were evaluated and compared with data obtained from measurements using commercially available p-aminophenyl alpha-D-glycopyranosides. Possible applications and advantages of C- and O-glycosylated SPR biosensors are discussed.

  • 23.
    Nahálková, Jarmila
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Tomkinson, Birgitta
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    TPPII, MYBBP1A and CDK2 form a protein–protein interaction network2014In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 564, p. 128-135Article in journal (Refereed)
    Abstract [en]

    Tripeptidyl-peptidase II (TPPII) is an aminopeptidase with suggested regulatory effects on cell cycle, apoptosis and senescence. A protein–protein interaction study revealed that TPPII physically interacts with the tumor suppressor MYBBP1A and the cell cycle regulator protein CDK2. Mutual protein–protein interaction was detected between MYBBP1A and CDK2 as well. In situ Proximity Ligation Assay (PLA) using HEK293 cells overexpressing TPPII forming highly enzymatically active oligomeric complexes showed that the cytoplasmic interaction frequency of TPPII with MYBBP1A increased with the protein expression of TPPII and using serum-free cell growth conditions. A specific reversible inhibitor of TPPII, butabindide, suppressed the cytoplasmic interactions of TPPII and MYBBP1A both in control HEK293 and the cells overexpressing murine TPPII. The interaction of MYBBP1A with CDK2 was confirmed by in situPLA in two different mammalian cell lines. Functional link between TPPII and MYBBP1A has been verified by gene expression study during anoikis, where overexpression of TPP II decreased mRNA expression level of MYBBP1A at the cell detachment conditions. All three interacting proteins TPPII, MYBBP1A and CDK2 have been previously implicated in the research for development of tumor-suppressing agents. This is the first report presenting mutual protein–protein interaction network of these proteins.

  • 24. Olivain, C
    et al.
    Humbert, C
    Nahalkova, Jarmila
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolutionary Biology, Molecular Evolution.
    Fatehi, J
    L'Haridon, F
    Alabouvette, C
    Colonization of tomato root by pathogenic and nonpathogenic Fusarium oxysporum strains inoculated together and separately into the soil2006In: Applied and Environmental Microbiology, ISSN 0099-2240, E-ISSN 1098-5336, Vol. 72, no 2, p. 1523-1531Article in journal (Refereed)
    Abstract [en]

    In soil, fungal colonization of plant roots has been traditionally studied by indirect methods such as microbial isolation that do not enable direct observation of infection sites or of interactions between fungal pathogens and their antagonists. Confocal laser scanning microscopy was used to visualize the colonization of tomato roots in heat-treated soil and to observe the interactions between a nonpathogenic strain, Fo47, and a pathogenic strain, Fo18, inoculated onto tomato roots in soil. When inoculated separately, both fungi colonized the entire root surface, with the exception of the apical zone. When both strains were introduced together, they both colonized the root surface and were observed at the same locations. When Fo47 was introduced at a higher concentration than Fo18, it colonized much of the root surface, but hyphae of Fo18 could still be observed at the same location on the root. There was no exclusion of the pathogenic strain by the presence of the nonpathogenic strain. These results are not consistent with the hypothesis that specific infection sites exist on the root for Fusarium oxysporum and instead support the hypothesis that competition occurs for nutrients rather than for infection sites.

  • 25. Petrus, L
    et al.
    Nahalkova, Jarmila
    Chem Slovenskej Akad Vied, Bratislava 84238, Slovakia.
    Gemeiner, P
    Petrusova, M
    Research of carbohydrates as the diversity factor of the living matter2000In: Chemické listy (Print), ISSN 0009-2770, E-ISSN 1213-7103, Vol. 94, no 9, p. 764-766Article in journal (Refereed)
    Abstract [en]

    An overview of the biological roles of carbohydrate chains in glycoconjugates is given with respect to their immediate impact on the diversity of the living matter. Some space is devoted also to new applications of glycosylmethylamines in the lectin research.

  • 26. Vookova, B
    et al.
    Hrib, J
    Gregova, E
    Nahalkova, Jarmila
    Gemeiner, P
    Defense reaction of pine seeds: distribution of protein bodies extract from Pinus nigra seeds in agar medium plates1999In: Biologia (Bratislava), ISSN 0006-3088, E-ISSN 1336-9563, Vol. 54, no 1, p. 107-111Article in journal (Refereed)
    Abstract [en]

    Diffusion and distribution of protein bodies (PB) extract from Pinus nigra seeds in agar medium plates was studied. After application of PB extract on filter paper discs it was placed on agar plates for 20 days. Proteins diffused from the discs were distributed in the plates. Identical protein pattern (8-54 kDa molecules) of PB extract was defined by SDS-PAGE analysis in agar strips at different distances from the discs. The predominant peptides and proteins were 8, 9, 13 and 18 kDa. The 32-36 kDa proteins were absent. These results suggest that the proteins during the maintenance on agar medium plates were separated. Relative quantity of individual proteins was different in certain distances from the discs. It is probable that these proteins can be involved in defense of pine megagametophyte. We assume that proportion of individual proteins is also of importance in their defense function.

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