Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel alpha-amylase inhibitory properties

Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9-kDa LTP from Capsicum annuum seeds that we call Ca-LTP(1). Ca-LTP(1) was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein`s biological activity. Bidimensional electrophoresis of the 9-kDa band, which corresponds to the purified Ca-LTP(1), showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of alpha-helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca-LTP(1) from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca-LTP(1) indicated that it was mainly localized within dense vesicles. In addition, isolated Ca-LTP(1) exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca-LTP(1) also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca-LTP(1) is able to inhibit mammalian alpha-amylase activity in vitro.

Molecular characterization of Arabidopsis thaliana PUF proteins - binding specificity and target candidates

PUF proteins regulate both stability and translation through sequence-specific binding to the 3` UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3` UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3` UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants.

A short proregion of trialysin, a pore-forming protein of Triatoma infestans salivary glands, controls activity by folding the N-terminal lytic motif

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect that transmits the protozoan parasite Trypanosoma cruzi, the etiological agent of Chagas` disease. Its saliva contains trialysin, a protein that forms pores in membranes. Peptides based on the N-terminus of trialysin lyse cells and fold into alpha-helical amphipathic segments resembling antimicrobial peptides. Using a specific antiserum against trialysin, we show here that trialysin is synthesized as a precursor that is less active than the protein released after saliva secretion. A synthetic peptide flanked by a fluorophore and a quencher including the acidic proregion and the lytic N-terminus of the protein is also less active against cells and liposomes, increasing activity upon proteolysis. Activation changes the peptide conformation as observed by fluorescence increase and CD spectroscopy. This mechanism of activation could provide a way to impair the toxic effects of trialysin inside the salivary glands, thus restricting damaging lytic activity to the bite site.

The Xanthomonas citri effector protein PthA interacts with citrus proteins involved in nuclear transport, protein folding and ubiquitination associated with DNA repair

P>Xanthomonas axonopodis pv. citri utilizes the type III effector protein PthA to modulate host transcription to promote citrus canker. PthA proteins belong to the AvrBs3/PthA family and carry a domain comprising tandem repeats of 34 amino acids that mediates protein-protein and protein-DNA interactions. We show here that variants of PthAs from a single bacterial strain localize to the nucleus of plant cells and form homo- and heterodimers through the association of their repeat regions. We hypothesize that the PthA variants might also interact with distinct host targets. Here, in addition to the interaction with alpha-importin, known to mediate the nuclear import of AvrBs3, we describe new interactions of PthAs with citrus proteins involved in protein folding and K63-linked ubiquitination. PthAs 2 and 3 preferentially interact with a citrus cyclophilin (Cyp) and with TDX, a tetratricopeptide domain-containing thioredoxin. In addition, PthAs 2 and 3, but not 1 and 4, interact with the ubiquitin-conjugating enzyme complex formed by Ubc13 and ubiquitin-conjugating enzyme variant (Uev), required for K63-linked ubiquitination and DNA repair. We show that Cyp, TDX and Uev interact with each other, and that Cyp and Uev localize to the nucleus of plant cells. Furthermore, the citrus Ubc13 and Uev proteins complement the DNA repair phenotype of the yeast Delta ubc13 and Delta mms2/uev1a mutants, strongly indicating that they are also involved in K63-linked ubiquitination and DNA repair. Notably, PthA 2 affects the growth of yeast cells in the presence of a DNA damage agent, suggesting that it inhibits K63-linked ubiquitination required for DNA repair.

Selective Oxidations of Organoboron Compounds Catalyzed by Baeyer-Villiger Monooxygenases

The applicability of Baeyer-Villiger monooxygenases (BVMOs) in organoboron chemistry has been explored through testing chemo-and enantioselective oxidations of a variety of boron-containing aromatic and vinylic compounds. Several BVMOs, namely: phenylacetone monooxygenase (PAMO), M446G PAMO mutant, 4-hydroxyacetophenone monooxygenase (HAPMO) and cyclohexanone monooxygenase (CHMO) were used in this study. The degree of chemoselectivity depends on the type of BVMO employed, in which the biocatalysts prefer boron-carbon oxidation over Baeyer-Villiger oxidation or epoxidation. Interestingly, it was discovered that PAMO can be used to perform kinetic resolution of boron-containing compounds with good enantioselectivities. These findings extend the known biocatalytic repertoire of BVMOs by showing a new family of compounds that can be oxidized by these enzymes.

Naturlig familjeplanering : Kvinnor och mäns upplevenser samt sjuksköterskans kunskap och användningsområden

Syftet med studien var att beskriva hur kvinnor och män upplevde naturlig familjeplanering (NFP) samt vilken kunskap sjuksköterskan hade om NFP och när hon kunde använda sig av den i sin profession. Artiklar valdes ut från databaserna Elin, Pub Med och Blackwell synergy. De sökord som användes var NFP, Catholic, Satisfaction, Awareness, Odeblad, Billing, Billings, Nurse, Pregnancy, Birthcontrol, Family, Mucus, Family planning, Natural family planning och Ovulation. Sammanlagt valdes 18 artiklar ut varav 14 användes i resultatet. Två artiklar till resultatet söktes manuellt och beställdes. Artiklarna kom från Nigeria, USA, Ungern, Peru, Filippinerna, Sri Lanka, Brasilien, Nya Zeeland, Tyskland, Canada och Iran. Några artiklar från Sverige eller de nordiska länderna hittades inte under sökningen. Resultatet visade att många var nöjda med NFP som metod. Det som var bland det mest positiva var att metoden inte gav några bieffekter och att kvinnorna lärde känna sin kropp. Det negativt med metoden var att användarna tyckte att det var svårt att avhålla sig från samlag under den fertila perioden samt att det fanns brister i metodens säkerhet. Många av dem som använde sig av metoden var katoliker. Många sjusköterskor var dåligt pålästa om metoden och skulle inte rekommendera den. Sjuksköterskan skulle kunna använda sig av metoden då kvinnan ville bli gravid, i fattiga länder och där religion sätter stopp för traditionella preventivmedel.

Palliativ vård : Anhörigvårdarens upplevelser av palliativ hemsjukvård.

Syftet med denna studie var att belysa anhörigvårdares upplevelser av den palliativa vården i hemmet utifrån aktuell vetenskaplig litteratur. Litteraturen som använts till resultatet har varit vetenskapliga artiklar av i huvudsak kvalitativ design. Artiklarna söktes i databaserna Blackwell Synergy, Elin och Pub Med. Sökorden som användes var Caring, Caregivers, Experiences, Family, Homecare, Needs, Palliative care, Perception. Sökorden kombinerades för att få artiklar som överensstämde med syftet och frågeställningen. Resultatet visade att anhörigvårdarna upplevde det som en självklarhet att den sjuke personen skulle vårdas hemma för att kunna behålla sitt normala liv och sin värdighet så långt som möjligt. Att få kunskap om hur de utförde omvårdnaden samt information om sjukdomen samt om risker och fördelar vid behandling upplevdes som viktigt och gav dem förståelse för situationen. Anhörigvårdarna levde med krav och förväntningar från flera håll, från den sjuke, andra anhöriga, från sjukvårdsteamet, samhället men också från dem själva. Periodvis var de både fysiskt och psykiskt utmattade, vilket gjorde dem sårbara. Perioden av palliativ hemsjukvård karaktäriserades av rädslor, osäkerhet, frustration och utmattning men även av positiva känslor och glädje över att kunna skapa livskvalité för den sjuke personen.

Cedric Gael Bryant, Lee Family Professor of English

Cedric Gael Bryant, Lee Family Professor of English, reading Beloved: A Novel, by Toni Morrison (PS3563 O8749 B4 1987)

Jorge Olivares, Allen Family Professor of Latin American Literature

Jorge Olivares, Allen Family Professor of Latin American Literature reading Sexual injustice : Supreme Court decisions from Griswold to Roe by Marc Stein

The preimaginal stages of the ensign wasp Evania appendigaster (Hymenoptera, Evaniidae), a cockroach egg predator

Evania appendigaster is a cosmopolitan wasp that deposits eggs in the oothecae of some species of cockroaches; its larvae then consume the cockroach eggs and embryos. It is a candidate for the biological control of cockroaches, but little is known about its basic biology. Here we describe the external morphology of all immature stages of E. appendigaster and compare them with the larvae of related species. The life cycle of E. appendigaster includes three larval instars, each with 13 body segments. Their mouthparts were generally reduced, except for the mandibles, which were always sclerotized and toothed, and were especially robust in second-instar larvae. Antennal and mouthpart sensilla were basiconic and difficult to observe. Larvae of E. appendigaster are similar in form to other described evaniid larvae, but quite different from the two available descriptions of larvae of gasteruptiid and aulacid wasps. Further descriptions of evaniid larvae will be useful in determining how widespread this morphology is within the family, and in understanding phylogenetic relationships within Hymenoptera.

Effects of hypercapnic hyperpnea on recovery from isoflurane or sevoflurane anesthesia in horses

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A dense GNSS meteorological network for observing deep convection in the Amazon

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Thermal and Rheological Properties of a Family of Botryosphaerans Produced by Botryosphaeria rhodina MAMB-05

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The stable archipelago in the region of the Pallas and Hansa dynamical families

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)