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1.
Cogn Psychol ; 140: 101541, 2023 02.
Article in English | MEDLINE | ID: mdl-36587465

ABSTRACT

Face perception and recognition are important processes for social interaction and communication among humans, so understanding how faces are mentally represented and processed has major implications. At the same time, faces are just some of the many stimuli that we encounter in our everyday lives. Therefore, more general theories of how we represent objects might also apply to faces. Contemporary research on the mental representation of faces has centered on two competing theoretical frameworks that arose from more general categorization research: prototype-based face representation and exemplar-based face representation. Empirically distinguishing between these frameworks is difficult and neither one has been ruled out. In this paper, we advance this area of research in three ways. First, we introduce two additional frameworks for mental representation of categories, varying abstraction and ideal representation, which have not been applied to face perception and recognition before. Second, we fit formal computational models of all four of these theories to human perceptual judgments of the typicality and attractiveness (a strong correlate of typicality) of 100 young adult Caucasian female faces, with the models expressed within a face space derived from facial similarity judgments via multidimensional scaling. Third, we predict the perceived typicality and attractiveness of the faces using these models and compare the predictive performance of each to the empirical data. We found that of all four models, the ideal representation model provided the best account of perceived typicality and attractiveness for the present set of faces, although all models showed discrepancies from the empirical data. These findings demonstrate the relevance of mental categorization processes for representing faces.


Subject(s)
Face , Facial Recognition , Young Adult , Humans , Female , Recognition, Psychology
2.
J Biol Chem ; 295(2): 504-516, 2020 01 10.
Article in English | MEDLINE | ID: mdl-31771981

ABSTRACT

Peptidoglycan (PG) is a critical component of the bacterial cell wall and is composed of a repeating ß-1,4-linked disaccharide of N-acetylglucosamine and N-acetylmuramic acid appended with a highly conserved stem peptide. In Gram-negative bacteria, PG is assembled in the cytoplasm and exported into the periplasm where it undergoes considerable maturation, modification, or degradation depending on the growth phase or presence of environmental stressors. These modifications serve important functions in diverse processes, including PG turnover, cell elongation/division, and antibiotic resistance. Conventional methods for analyzing PG composition are complex and time-consuming. We present here a streamlined MS-based method that combines differential analysis with statistical 1D annotation approaches to quantitatively compare PGs produced in planktonic- and biofilm-cultured Pseudomonas aeruginosa We identified a core assembly of PG that is present in high abundance and that does not significantly differ between the two growth states. We also identified an adaptive PG assembly that is present in smaller amounts and fluctuates considerably between growth states in response to physiological changes. Biofilm-derived adaptive PG exhibited significant changes compared with planktonic-derived PG, including amino acid substitutions of the stem peptide and modifications that indicate changes in the activity of amidases, deacetylases, and lytic transglycosylases. The results of this work also provide first evidence of de-N-acetylated muropeptides from P. aeruginosa The method developed here offers a robust and reproducible workflow for accurately determining PG composition in samples that can be used to assess global PG fluctuations in response to changing growth conditions or external stimuli.


Subject(s)
Biofilms , Peptidoglycan/metabolism , Plankton/physiology , Pseudomonas aeruginosa/physiology , Biofilms/growth & development , Cell Wall/chemistry , Cell Wall/metabolism , Glycomics , Humans , Mass Spectrometry , Peptidoglycan/chemistry , Plankton/chemistry , Pseudomonas Infections/microbiology , Pseudomonas aeruginosa/chemistry
3.
Plant Cell ; 28(10): 2683-2696, 2016 10.
Article in English | MEDLINE | ID: mdl-27677881

ABSTRACT

To synthesize the cofactor thiamin diphosphate (ThDP), plants must first hydrolyze thiamin monophosphate (ThMP) to thiamin, but dedicated enzymes for this hydrolysis step were unknown and widely doubted to exist. The classical thiamin-requiring th2-1 mutation in Arabidopsis thaliana was shown to reduce ThDP levels by half and to increase ThMP levels 5-fold, implying that the THIAMIN REQUIRING2 (TH2) gene product could be a dedicated ThMP phosphatase. Genomic and transcriptomic data indicated that TH2 corresponds to At5g32470, encoding a HAD (haloacid dehalogenase) family phosphatase fused to a TenA (thiamin salvage) family protein. Like the th2-1 mutant, an insertional mutant of At5g32470 accumulated ThMP, and the thiamin requirement of the th2-1 mutant was complemented by wild-type At5g32470 Complementation tests in Escherichia coli and enzyme assays with recombinant proteins confirmed that At5g32470 and its maize (Zea mays) orthologs GRMZM2G148896 and GRMZM2G078283 are ThMP-selective phosphatases whose activity resides in the HAD domain and that the At5g32470 TenA domain has the expected thiamin salvage activity. In vitro and in vivo experiments showed that alternative translation start sites direct the At5g32470 protein to the cytosol and potentially also to mitochondria. Our findings establish that plants have a dedicated ThMP phosphatase and indicate that modest (50%) ThDP depletion can produce severe deficiency symptoms.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Thiamine Pyrophosphate/metabolism , Arabidopsis/enzymology , Arabidopsis Proteins/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Phosphoric Monoester Hydrolases/genetics , Phosphoric Monoester Hydrolases/metabolism
4.
Plant Cell ; 28(9): 2291-2311, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27495812

ABSTRACT

Recently, we described the ubiquitylation of PYL4 and PYR1 by the RING E3 ubiquitin ligase RSL1 at the plasma membrane of Arabidopsis thaliana This suggested that ubiquitylated abscisic acid (ABA) receptors might be targeted to the vacuolar degradation pathway because such ubiquitylation is usually an internalization signal for the endocytic route. Here, we show that FYVE1 (previously termed FREE1), a recently described component of the endosomal sorting complex required for transport (ESCRT) machinery, interacted with RSL1-receptor complexes and recruited PYL4 to endosomal compartments. Although the ESCRT pathway has been assumed to be reserved for integral membrane proteins, we show the involvement of this pathway in the degradation of ABA receptors, which can be associated with membranes but are not integral membrane proteins. Knockdown fyve1 alleles are hypersensitive to ABA, illustrating the biological relevance of the ESCRT pathway for the modulation of ABA signaling. In addition, fyve1 mutants are impaired in the targeting of ABA receptors for vacuolar degradation, leading to increased accumulation of PYL4 and an enhanced response to ABA Pharmacological and genetic approaches revealed a dynamic turnover of ABA receptors from the plasma membrane to the endosomal/vacuolar degradation pathway, which was mediated by FYVE1 and was dependent on RSL1. This process involves clathrin-mediated endocytosis and trafficking of PYL4 through the ESCRT pathway, which helps to regulate the turnover of ABA receptors and attenuate ABA signaling.

5.
Plant Physiol ; 174(2): 1012-1027, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28363991

ABSTRACT

Phosphoenolpyruvate carboxylase (PEPC) is a tightly controlled cytosolic enzyme situated at a crucial branch point of central plant metabolism. In developing castor oil seeds (Ricinus communis) a novel, allosterically desensitized 910-kD Class-2 PEPC hetero-octameric complex, arises from a tight interaction between 107-kD plant-type PEPC and 118-kD bacterial-type (BTPC) subunits. The native Ca2+-dependent protein kinase (CDPK) responsible for in vivo inhibitory phosphorylation of Class-2 PEPC's BTPC subunit's at Ser-451 was highly purified from COS and identified as RcCDPK1 (XP_002526815) by mass spectrometry. Heterologously expressed RcCDPK1 catalyzed Ca2+-dependent, inhibitory phosphorylation of BTPC at Ser-451 while exhibiting: (i) a pair of Ca2+ binding sites with identical dissociation constants of 5.03 µM, (ii) a Ca2+-dependent electrophoretic mobility shift, and (iii) a marked Ca2+-independent hydrophobicity. Pull-down experiments established the Ca2+-dependent interaction of N-terminal GST-tagged RcCDPK1 with BTPC. RcCDPK1-Cherry localized to the cytosol and nucleus of tobacco bright yellow-2 cells, but colocalized with mitochondrial-surface associated BTPC-enhanced yellow fluorescent protein when both fusion proteins were coexpressed. Deletion analyses demonstrated that although its N-terminal variable domain plays an essential role in optimizing Ca2+-dependent RcCDPK1 autophosphorylation and BTPC transphosphorylation activity, it is not critical for in vitro or in vivo target recognition. Arabidopsis (Arabidopsis thaliana) CPK4 and soybean (Glycine max) CDPKß are RcCDPK1 orthologs that effectively phosphorylated castor BTPC at Ser-451. Overall, the results highlight a potential link between cytosolic Ca2+ signaling and the posttranslational control of respiratory CO2 refixation and anaplerotic photosynthate partitioning in support of storage oil and protein biosynthesis in developing COS.


Subject(s)
Castor Oil/metabolism , Phosphoenolpyruvate Carboxylase/metabolism , Protein Kinases/metabolism , Ricinus/enzymology , Seeds/metabolism , Amino Acid Sequence , Antibody Formation , Binding Sites , Biocatalysis , Biophysical Phenomena , Calcium/metabolism , Cloning, Molecular , Gene Expression Regulation, Plant , Hydrophobic and Hydrophilic Interactions , Intrinsically Disordered Proteins/metabolism , Mitochondria/metabolism , Phosphorylation , Phosphoserine/metabolism , Protein Domains , Protein Interaction Domains and Motifs , Protein Kinases/chemistry , Ricinus/embryology , Ricinus/genetics , Sequence Alignment , Substrate Specificity
6.
J Vasc Res ; 54(5): 288-298, 2017.
Article in English | MEDLINE | ID: mdl-28930755

ABSTRACT

Biomaterial-based delivery of angiogenic growth factors restores perfusion more effectively than bolus delivery methods in rodent models of peripheral vascular disease, but the same success has not yet been demonstrated in clinically relevant studies of aged or large animals. These studies explore, in clinically relevant models, a therapeutic angiogenesis strategy for the treatment of peripheral vascular disease that overcomes the challenges encountered in previous clinical trials. Alginate hydrogels providing sustained release of vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF) were injected into ischemic hind limbs in middle-aged and old mice, and also in young rabbits, as a test of the scalability of this local growth factor treatment. Spontaneous perfusion recovery diminished with increasing age, and only the combination of VEGF and IGF delivery from gels significantly rescued perfusion in middle-aged (13 months) and old (20 months) mice. In rabbits, the delivery of VEGF alone or in combination with IGF from alginate hydrogels, at a dose 2 orders of magnitude lower than the typical doses used in past rabbit studies, enhanced perfusion recovery when given immediately after surgery, or as a treatment for chronic ischemia. Capillary density measurements and angiographic analysis demonstrated the benefit of gel delivery. These data together suggest that alginate hydrogels providing local delivery of low doses of VEGF and IGF constitute a safe and effective treatment for hind-limb ischemia in clinically relevant animal models, thereby supporting the potential clinical translation of this concept.


Subject(s)
Alginates/chemistry , Angiogenesis Inducing Agents/administration & dosage , Drug Carriers , Insulin-Like Growth Factor I/administration & dosage , Ischemia/drug therapy , Muscle, Skeletal/blood supply , Neovascularization, Physiologic/drug effects , Vascular Endothelial Growth Factor A/administration & dosage , Age Factors , Angiogenesis Inducing Agents/chemistry , Angiography, Digital Subtraction , Animals , Disease Models, Animal , Drug Compounding , Female , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Hindlimb , Hydrogels , Insulin-Like Growth Factor I/chemistry , Ischemia/diagnostic imaging , Ischemia/physiopathology , Mice, Inbred C57BL , Rabbits , Recovery of Function , Regional Blood Flow , Time Factors , Vascular Endothelial Growth Factor A/chemistry
7.
Biochem J ; 473(20): 3667-3682, 2016 Oct 15.
Article in English | MEDLINE | ID: mdl-27512054

ABSTRACT

Imported sucrose is cleaved by sucrose synthase (SUS) as a critical initial reaction in the biosynthesis of storage end-products by developing seeds. Although SUS is phosphorylated at a conserved seryl residue by an apparent CDPK (Ca2+-dependent protein kinase) in diverse plant tissues, the functions and mechanistic details of this process remain obscure. Thus, the native CDPK that phosphorylates RcSUS1 (Ricinus communis SUS1) at Ser11 in developing COS (castor oil seeds) was highly purified and identified as RcCDPK2 by MS/MS. Purified RcSUS1-K (-kinase) and heterologously expressed RcCDPK2 catalyzed Ca2+-dependent Ser11 phosphorylation of RcSUS1 and its corresponding dephosphopeptide, while exhibiting a high affinity for free Ca2+ ions [K0.5(Ca2+) < 0.4 µM]. RcSUS1-K activity, RcCDPK2 expression, and RcSUS1 Ser11 phosphorylation peaked during early COS development and then declined in parallel. The elimination of sucrose import via fruit excision triggered RcSUS1 dephosphorylation but did not alter RcSUS1-K activity, suggesting a link between sucrose signaling and posttranslational RcCDPK2 control. Both RcCDPK2-mCherry and RcSUS1-EYFP co-localized throughout the cytosol when transiently co-expressed in tobacco suspension cells, although RcCDPK2-mCherry was also partially localized to the nucleus. Subcellular fractionation revealed that ∼20% of RcSUS1-K activity associates with microsomal membranes in developing COS, as does RcSUS1. In contrast with RcCDPK1, which catalyzes inhibitory phosphorylation of COS bacterial-type phosphoenolpyruvate carboxylase at Ser451, RcCDPK2 exhibited broad substrate specificity, a wide pH-activity profile centered at pH 8.5, and insensitivity to metabolite effectors or thiol redox status. Our combined results indicate a possible link between cytosolic Ca2+-signaling and the control of photosynthate partitioning during COS development.


Subject(s)
Castor Oil/metabolism , Glucosyltransferases/metabolism , Plant Proteins/metabolism , Protein Kinases/metabolism , Seeds/enzymology , Seeds/metabolism , Hydrogen-Ion Concentration , Microsomes/metabolism , Phosphoenolpyruvate Carboxylase/metabolism , Phosphorylation
9.
Psychol Sci ; 27(2): 244-56, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26744069

ABSTRACT

Experience puts people in touch with nonsolid substances, such as water, blood, and milk, which are crucial to survival. People must be able to understand the behavior of these substances and to differentiate their properties from those of solid objects. We investigated whether infants represent nonsolid substances as a conceptual category distinct from solid objects on the basis of differences in cohesiveness. Experiment 1 established that infants can distinguish water from a perceptually matched solid and can correctly predict whether the item will pass through or be trapped by a grid. Experiments 2 and 3 showed that infants extend this knowledge to less familiar granular substances. These experiments indicate that concepts of cohesive and noncohesive material appear early in development, apply across several types of nonsolid substances, and may serve as the basis of later knowledge of physical phases.


Subject(s)
Child Development/physiology , Concept Formation , Pattern Recognition, Visual , Female , Habituation, Psychophysiologic , Humans , Infant , Male , Psychology, Child , Psychology, Developmental
10.
J Biol Chem ; 289(48): 33412-24, 2014 Nov 28.
Article in English | MEDLINE | ID: mdl-25313400

ABSTRACT

Sucrose synthase (SUS) catalyzes the UDP-dependent cleavage of sucrose into UDP-glucose and fructose and has become an important target for improving seed crops via metabolic engineering. A UDP-specific SUS homotetramer composed of 93-kDa subunits was purified to homogeneity from the triacylglyceride-rich endosperm of developing castor oil seeds (COS) and identified as RcSUS1 by mass spectrometry. RcSUS1 transcripts peaked during early development, whereas levels of SUS activity and immunoreactive 93-kDa SUS polypeptides maximized during mid-development, becoming undetectable in fully mature COS. The cytosolic location of the enzyme was established following transient expression of RcSUS1-enhanced YFP in tobacco suspension cells and fluorescence microscopy. Immunological studies using anti-phosphosite-specific antibodies revealed dynamic and high stoichiometric in vivo phosphorylation of RcSUS1 at its conserved Ser-11 residue during COS development. Incorporation of (32)P(i) from [γ-(32)P]ATP into a RcSUS1 peptide substrate, alongside a phosphosite-specific ELISA assay, established the presence of calcium-dependent RcSUS1 (Ser-11) kinase activity. Approximately 10% of RcSUS1 was associated with COS microsomal membranes and was hypophosphorylated relative to the remainder of RcSUS1 that partitioned into the soluble, cytosolic fraction. Elimination of sucrose supply caused by excision of intact pods of developing COS abolished RcSUS1 transcription while triggering the progressive dephosphorylation of RcSUS1 in planta. This did not influence the proportion of RcSUS1 associated with microsomal membranes but instead correlated with a subsequent marked decline in SUS activity and immunoreactive RcSUS1 polypeptides. Phosphorylation at Ser-11 appears to protect RcSUS1 from proteolysis, rather than influence its kinetic properties or partitioning between the soluble cytosol and microsomal membranes.


Subject(s)
Glucosyltransferases/metabolism , Intracellular Membranes/enzymology , Microsomes/enzymology , Plant Proteins/metabolism , Ricinus communis/enzymology , Seeds/enzymology , Phosphorylation/physiology , Proteolysis
11.
J Vasc Res ; 52(1): 62-9, 2015.
Article in English | MEDLINE | ID: mdl-26045306

ABSTRACT

Endothelial progenitor cells are being broadly explored for the treatment of ischemic cardiovascular diseases, but their response to molecules commonly used to promote the growth of new blood vessels has not been fully characterized. In this study, angiogenic sprout formation in a 3-dimensional, in vitro model by one type of endothelial progenitor, outgrowth endothelial cells (OECs), was characterized in response to exposure to stromal cell-derived factor (SDF) and vascular endothelial growth factor (VEGF) and then compared to mature endothelial cells. Exposure to SDF alone did not increase angiogenic sprouting in comparison to control media, while a combination of VEGF and SDF demonstrated greater potency than VEGF alone for all cell types. Together, VEGF and SDF reduced the sprout initiation time and maintained sprouting levels over time. In direct competition with mature endothelial cells, OECs preferentially localized to the tip cell position, suggesting an enhanced sprouting potential. Overall, these results reveal the impact of the combination of VEGF and SDF on endothelial cell sprouting, and support the enhanced potential of OECs, as opposed to mature endothelial cells, for treating ischemic diseases.


Subject(s)
Chemokine CXCL12/pharmacology , Endothelial Cells/drug effects , Neovascularization, Physiologic/drug effects , Vascular Endothelial Growth Factor A/pharmacology , Cell Culture Techniques , Cell Division , Chemokine CXCL12/physiology , Coculture Techniques , Endothelial Cells/cytology , Fetal Blood/cytology , Human Umbilical Vein Endothelial Cells , Humans , Microspheres , Microvessels/cytology , Recombinant Proteins/pharmacology , Time-Lapse Imaging , Vascular Endothelial Growth Factor A/physiology
12.
Plant Cell Environ ; 42(4): 1158-1166, 2019 04.
Article in English | MEDLINE | ID: mdl-30341950

ABSTRACT

Among 29 predicted Arabidopsis purple acid phosphatases (PAPs), AtPAP26 functions as the principle extracellular and intracellular PAP isozyme that is upregulated to recycle and scavenge Pi during Pi-deprivation or leaf senescence. Our companion paper documented the copurification of a secreted, high-mannose AtPAP26-S2 glycoform with AtGAL1 (At1g78850), a Pi starvation-inducible (PSI), and Galanthus nivalis agglutinin-related (mannose-binding) and apple domain lectin. This study tests the hypothesis that AtGAL1 binds AtPAP26-S2 to modify its enzymatic properties. Far-western immunodot blotting established that AtGAL1 readily associates with AtPAP26-S2 but not the low mannose AtPAP26-S1 glycoform nor other secreted PSI PAPs (i.e., AtPAP12 or AtPAP25). Analytical gel filtration indicated that 55-kDa AtGAL1 and AtPAP26-S2 polypeptides associate to form a 112-kDa heterodimer. Microscopic imaging of transiently expressed, fluorescent protein-tagged AtGAL1, and associated bimolecular fluorescence complementation assays demonstrated that (a) like AtPAP26, AtGAL1 also localizes to lytic vacuoles of Pi-deprived Arabidopsis and (b) both proteins interact in vivo. AtGAL1 preincubation significantly enhanced the acid phosphatase activity and thermal stability of AtPAP26-S2 but not AtPAP26-S1. We hypothesize that AtGAL1 plays an important role during Pi deprivation through its interaction with mannose-rich glycans of AtPAP26-S2 and consequent positive impact on AtPAP26-S2 activity and stability.


Subject(s)
Acid Phosphatase/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Galactokinase/metabolism , Phosphates/deficiency , Acid Phosphatase/isolation & purification , Arabidopsis Proteins/isolation & purification , Blotting, Western , Chromatography, Gel , Galactokinase/isolation & purification , Phosphates/metabolism , Protein Isoforms/isolation & purification , Protein Isoforms/metabolism
13.
Sci Adv ; 10(19): eadj8571, 2024 May 10.
Article in English | MEDLINE | ID: mdl-38728400

ABSTRACT

The development of sparse edge coding in the mammalian visual cortex depends on early visual experience. In humans, there are multiple indicators that the statistics of early visual experiences has unique properties that may support these developments. However, there are no direct measures of the edge statistics of infant daily-life experience. Using head-mounted cameras to capture egocentric images of young infants and adults in the home, we found infant images to have distinct edge statistics relative to adults. For infants, scenes with sparse edge patterns-few edges and few orientations-dominate. The findings implicate biased early input at the scale of daily life that is likely specific to the early months after birth and provide insights into the quality, amount, and timing of the visual experiences during the foundational developmental period for human vision.


Subject(s)
Visual Perception , Humans , Infant , Visual Perception/physiology , Female , Adult , Male , Visual Cortex/physiology , Photic Stimulation , Vision, Ocular/physiology
14.
Mol Microbiol ; 84(6): 1165-76, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22554073

ABSTRACT

Heteropolymeric B-band O-antigen (O-Ag) biosynthesis in Pseudomonas aeruginosa PAO1 follows the Wzy-dependent pathway, beginning with translocation of undecaprenyl pyrophosphate-linked anionic O-Ag subunits (O units) from the inner to the outer leaflets of the inner membrane (IM). This translocation is mediated by the integral IM flippase Wzx. Through experimentally based and unbiased topological mapping, our group previously observed that Wzx possesses many charged and aromatic amino acid residues within its 12 transmembrane segments (TMS). Herein, site-directed mutagenesis targeting 102 residues was carried out on the TMS and loops of Wzx, followed by assessment of each construct's ability to restore B-band O-Ag production, identifying eight residues important for flippase function. The importance of various charged and aromatic residues was highlighted, predominantly within the TMS of the protein, revealing functional 'hotspots' within the flippase, particularly within TMS2 and TMS8. Construction of a tertiary structure homology model for Wzx indicated that TMS2 and TMS8 line a central cationic lumen. This is the first report to describe a charged flippase lumen for mediating anionic O-unit translocation across the hydrophobic IM.


Subject(s)
Membrane Transport Proteins/metabolism , O Antigens/metabolism , Pseudomonas aeruginosa/enzymology , Amino Acid Substitution , Cell Membrane/metabolism , Membrane Transport Proteins/genetics , Models, Molecular , Mutagenesis, Site-Directed , Mutant Proteins/genetics , Mutant Proteins/metabolism , Protein Conformation , Pseudomonas aeruginosa/genetics
15.
Biofouling ; 29(10): 1169-80, 2013.
Article in English | MEDLINE | ID: mdl-24063626

ABSTRACT

Pseudomonas aeruginosa and Candida albicans are disparate microbial species, but both are known to be opportunistic pathogens frequently associated with nosocomial infections. The aim of this study was to provide a better understanding of the interactions between these microorganisms in dual-species biofilms. Several bacteriophage-resistant P. aeruginosa phenotypes have been isolated and were used in dual-species mixed-biofilm studies. Twenty-four and 48 h mixed-biofilms were formed using the isolated phenotypes of phage-resistant P. aeruginosa and these were compared with similar experiments using other P. aeruginosa strains with a defined lipopolysaccharide (LPS) deficiency based on chromosomal knockout of specific LPS biosynthetic genes. Overall, the results showed that the variants of phage-resistant P. aeruginosa and LPS mutants were both less effective in inhibiting the growth of C. albicans in mixed-biofilms compared to the wild-type strains of P. aeruginosa. Conversely, the proliferation of P. aeruginosa was not influenced by the presence of C. albicans. In conclusion, the ability of strains of P. aeruginosa to inhibit the formation of a biofilm of C. albicans appears to be correlated with the LPS chain lengths of phenotypes of P. aeruginosa, suggesting that LPS has a suppressive effect on the growth of C. albicans.


Subject(s)
Biofilms/growth & development , Biofouling/prevention & control , Candida albicans/physiology , Pseudomonas aeruginosa/virology , Candida albicans/ultrastructure , Microscopy, Electron, Scanning , Phenotype , Pseudomonas Phages/physiology , Pseudomonas aeruginosa/ultrastructure
16.
J Bacteriol ; 194(16): 4295-300, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22685285

ABSTRACT

migA and wapR are rhamnosyltransferase genes involved in the biosynthesis of Pseudomonas aeruginosa lipopolysaccharide core oligosaccharide. Here, we show that preferential expression of migA and wapR correlated with the levels of uncapped and O polysaccharide-capped core, respectively. wapR is negatively regulated, while migA is positively regulated by RhlR/RhlI quorum sensing.


Subject(s)
Bacterial Proteins/biosynthesis , Gene Expression Regulation, Bacterial , Lipopolysaccharides/metabolism , Phosphotransferases (Alcohol Group Acceptor)/biosynthesis , Pseudomonas aeruginosa/enzymology , Pseudomonas aeruginosa/genetics , Bacterial Proteins/metabolism , Base Sequence , Ligases/metabolism , Molecular Sequence Data , Promoter Regions, Genetic , Quorum Sensing , Transcription Factors/metabolism
17.
J Biol Chem ; 286(23): 20600-5, 2011 Jun 10.
Article in English | MEDLINE | ID: mdl-21498511

ABSTRACT

Heteropolymeric B-band lipopolysaccharide in Pseudomonas aeruginosa PAO1 is synthesized via the so-called Wzy-dependent pathway, requiring a functional Wzy for polymerization of O-antigen repeat units in the periplasm. Wzy is an integral inner membrane protein for which the detailed topology has been mapped in a recent investigation (Islam, S. T., Taylor, V. L., Qi, M., and Lam, J. S. (2010) mBio 1, e00189-10), revealing two principal periplasmic loops (PL), PL3 and PL5, each containing an RX(10)G motif. Despite considerable sequence conservation between the two loops, the isoelectric point for each peptide displayed marked differences, with PL3 exhibiting a net-positive charge and PL5 showing a net-negative charge. Data from site-directed mutagenesis of amino acids in each PL have led to the identification of several key Arg residues within the two RX(10)G motifs that are important for Wzy function, of which Arg(176), Arg(290), and Arg(291) could not be functionally substituted with Lys. These observations support the proposed role of each PL in a catch-and-release mechanism for Wzy-mediated O-antigen polymerization.


Subject(s)
Bacterial Proteins/metabolism , Carrier Proteins/metabolism , Membrane Proteins/metabolism , O Antigens/biosynthesis , Periplasm/metabolism , Pseudomonas aeruginosa/metabolism , Amino Acid Substitution , Bacterial Proteins/genetics , Carrier Proteins/genetics , Membrane Proteins/genetics , Mutagenesis, Site-Directed , O Antigens/genetics , Periplasm/genetics , Pseudomonas aeruginosa/genetics
18.
Environ Microbiol ; 14(8): 1995-2005, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22513190

ABSTRACT

Exopolysaccharide is a critical biofilm matrix component, yet little is known about how the synthesis of multiple exopolysaccharides is regulated. Pseudomonas aeruginosa can produce several biofilm matrix exopolysaccharides that include alginate, Psl and Pel. Here we demonstrated that AlgC, a key enzyme that provides sugar precursors for the synthesis of alginate and lipopolysaccharides (LPS) is also required for both Psl and Pel production. We showed that forced-synthesis of Psl in alginate-producing mucoid bacteria reduced alginate production but this was not due to transcription of the alginate biosynthesis-operon. Likewise, when either alginate or Psl were overproduced, levels of B-band LPS decreased. Induction of Pel resulted in a reduction of Psl levels. Because the effects of reduced exopolysaccharide synthesis when another is overproduced didn't appear to be regulated at the transcriptional level, this suggests that the biosynthesis pathways of Psl, Pel, alginate, and LPS compete for common sugar precursors. As AlgC is the only enzyme that provides precursors for each of these exopolysaccharides, we propose that AlgC is a key checkpoint enzyme that coordinates the total amount of exopolysaccharide biosynthesis by controlling sugar precursor pool. Our data also provide a plausible strategy that P.aeruginosa utilizes to modulate its biofilm matrix exopolysaccharides.


Subject(s)
Biofilms , Gene Expression Regulation, Bacterial , Polysaccharides, Bacterial/biosynthesis , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Operon , Polysaccharides, Bacterial/genetics , Pseudomonas aeruginosa/enzymology
19.
Cognition ; 229: 105256, 2022 12.
Article in English | MEDLINE | ID: mdl-35988453

ABSTRACT

Across the lifespan, humans are biased to look first at what is easy to see, with a handful of well-documented visual saliences shaping our attention (e.g., Itti & Koch, 2001). These attentional biases may emerge from the contexts in which moment-tomoment attention occurs, where perceivers and their social partners actively shape bottom-up saliences, moving their bodies and objects to make targets of interest more salient. The goal of the present study was to determine the bottom-up saliences present in infant egocentric images and to provide evidence on the role that infants and their mature social partners play in highlighting targets of interest via these saliences. We examined 968 unique scenes in which an object had purposefully been placed in the infant's egocentric view, drawn from videos created by one-year-old infants wearing a head camera during toy-play with a parent. To understand which saliences mattered in these scenes, we conducted a visual search task, asking participants (n = 156) to find objects in the egocentric images. To connect this to the behaviors of perceivers, we then characterized the saliences of objects placed by infants or parents compared to objects that were otherwise present in the scenes. Our results show that body-centric properties, such as increases in the centering and visual size of the object, as well as decreases in the number of competing objects immediately surrounding it, both predicted faster search time and distinguished placed and unplaced objects. The present results suggest that the bottom-up saliences that can be readily controlled by perceivers and their social partners may most strongly impact our attention. This finding has implications for the functional role of saliences in human vision, their origin, the social structure of perceptual environments, and how the relation between bottom-up and top-down control of attention in these environments may support infant learning.


Subject(s)
Attention , Parents , Humans , Infant , Learning , Visual Perception
20.
Infant Behav Dev ; 66: 101666, 2022 02.
Article in English | MEDLINE | ID: mdl-34837790

ABSTRACT

Recent studies have found that infants show relational learning in the first year. Like older children, they can abstract relations such as same or different across a series of exemplars. For older children, language has a major impact on relational learning: labeling a shared relation facilitates learning, while labeling component objects can disrupt learning. Here we ask: Does language influence relational learning at 12 months? Experiment 1 (n = 64) examined the influence of a relational label on learning. Prior to the study, the infants saw three pairs of objects, all labeled "These are same" or "These are different". Experiment 2 (n = 48) examined the influence of object labels prior to the study, with three objects labeled (e.g., "This is a cup, this is a tower."). We compared the present results with those of Ferry et al. (2015), where infants abstracted same and different relations after undergoing a similar paradigm without prior labels. If the effects of language mirror those in older children, we would expect that infants given relational labels (Experiment 1) will be helped in abstracting same and different compared to infants not given labels and that infants given object labels (Experiment 2) will be hindered relative to those not given labels. We found no evidence for either prediction. In Experiment 1, infants who had heard relational labels did not benefit compared to infants who had received no labels (Ferry et al., 2015). In Experiment 2, infants who had heard object labels showed the same patterns as those in Ferry et al. (2015), suggesting that object labels had no effect. This finding is important because it highlights a key difference between the relational learning abilities of infants and those seen in older children, pointing to a protracted process by which language and relational learning become entwined.


Subject(s)
Language , Learning , Adolescent , Child , Humans , Infant , Language Development
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