Characterization of KPAP, a Novel Pokeweed Antiviral Protein Isoform

Phytolacca americana is a perennial plant that harbors the pokeweed antiviral protein (PAP) genes which belong to the N-glycosidase family. These proteins are called ribosome inactivating proteins (RIPs) which remove an adenine base from the sarcin/ricin loop (SRL) of the 28S rRNA in eukaryotes. This depurination results in protein translation inhibition leading to a concentration-dependent cytotoxic consequence. Many RIPs in the plant kingdom are being discovered with new genomic sequencing technologies but, many of them remain uncharacterized. Recently, our lab discovered a new RIP called Novel PAP/KPAP in the sequenced pokeweed genome. KPAP transcript levels did not behave the same way as other isoforms under phytohormone stimulated biotic stress, however, they showed a slight decrease in drought. In this study, I provide preliminary groundwork on the KPAP gene model, expression in abiotic stresses, protein structure, depurination activity and potential binding arrangement on the SRL. My results show the presence of predicted upstream open reading frames (uORFs) in the long leader intron, perhaps belonging to retrotransposons. The enzymatic activity recorded from KPAP in an in vitro translation assay resembles the activity of PAP-I. Moreover, the exclusive expression of KPAP in leaves compared to other isoforms and its downregulation in drought may indicate a function related to photosynthesis, carbon metabolism, plant growth or leaf expansion. Structurally the predicted protein was docked in silico to an adenine base and showed a similar disposition of the active site compared to PAP-I. Lastly, KPAP was docked in silico to the E.coli SRL and residue interactions are discussed in the context of other RIPs. This early research shows KPAP’s similarities and differences compared to the other isoforms, however, much remains to be learned from this isoform and RIPs in general

Characterization of KPAP, a Novel Pokeweed Antiviral Protein Isoform

Phytolacca americana is a perennial plant that harbors the pokeweed antiviral protein (PAP) genes which belong to the N-glycosidase family. These proteins are called ribosome inactivating proteins (RIPs) which remove an adenine base from the sarcin/ricin loop (SRL) of the 28S rRNA in eukaryotes. This depurination results in protein translation inhibition leading to a concentration-dependent cytotoxic consequence. Many RIPs in the plant kingdom are being discovered with new genomic sequencing technologies but, many of them remain uncharacterized. Recently, our lab discovered a new RIP called Novel PAP/KPAP in the sequenced pokeweed genome. KPAP transcript levels did not behave the same way as other isoforms under phytohormone stimulated biotic stress, however, they showed a slight decrease in drought. In this study, I provide preliminary groundwork on the KPAP gene model, expression in abiotic stresses, protein structure, depurination activity and potential binding arrangement on the SRL. My results show the presence of predicted upstream open reading frames (uORFs) in the long leader intron, perhaps belonging to retrotransposons. The enzymatic activity recorded from KPAP in an in vitro translation assay resembles the activity of PAP-I. Moreover, the exclusive expression of KPAP in leaves compared to other isoforms and its downregulation in drought may indicate a function related to photosynthesis, carbon metabolism, plant growth or leaf expansion. Structurally the predicted protein was docked in silico to an adenine base and showed a similar disposition of the active site compared to PAP-I. Lastly, KPAP was docked in silico to the E.coli SRL and residue interactions are discussed in the context of other RIPs. This early research shows KPAPs similarities and differences compared to the other isoforms, however, much remains to be learned from this isoform and RIPs in general

Une exposition drôlement juste aux musées Gadagne

Réalisée par les musées Gadagne, l’exposition Lyon sur le divan propose un regard décalé sur l’architecture et les métamorphoses de la ville grâce à un dispositif scénographique mêlant références sérieuses, parodie et humour. Les responsables du projet montrent comment ce choix audacieux peut décontenancer le visiteur mais aussi l’aider à s’approprier des contenus pour finalement mieux appréhender la ville

Temperature-frequency evolution during 20 kHz cyclic loading of Dual- Phase 780 steel

Ultrasonic machine, which operates at around 20 kHz, is used to quickly get the fatigue behavior in the very high cycle fatigue domain. Cyclic loading at such high frequencies is accompanied by selfheating of the test specimen [1]–[3]. The self-heating magnitude depends on the material and cooling system. It is also accompanied by changes in loading frequency of testing. In contrast to thetemperature evolution and to our knowledge, the frequency evolution during an ultrasonic fatigue test has been very little studied. In this paper, the changes in both specimen temperature and loading frequency during ultrasonic cycling for the Dual-Phase 780 steel are investigated

PREVALÊNCIA DE DOR EM ADOLESCENTES ESTUDANTES DO ENSINO MÉDIO DIURNO DO MUNICÍPIO DE GARIBALDI/RS

Este estudo objetivou identificar a prevalência de dor em adolescentes estudantes do ensino médio diurno das escolas estaduais do município de Garibaldi/RS. Utilizou-se o questionário (Instrumento para avaliação da dor nas costas - IDC) composto de questões fechadas para avaliar a dor. O IDC foi aplicado em 688 adolescentes de ambos os sexos de 13 a 20 anos (média de idade de 15,6 anos), de duas escolas estaduais de Garibaldi/RS. Verificou-se que as regiões com maior prevalência de dor entre os escolares foram a lombar (46,6 %), os ombros (38,8 %) e os membros inferiores (37,1 %), sendo que a intensidade leve predominou nas três regiões. Houve maior frequência de dor na região lombar (46,9 %), com 17% dos adolescentes reportando referindo 1 a 4 vezes, seguido dos ombros (40,4 %), com 16,6 % dos adolescentes referindo dor de 1 a 4 vezes. Este tipo de informação é relevante, pois contribui no planejamento de ações educacionais e preventivas.  Descritores: Adolescente; Estudantes; Dor nas Costas.

Neonatal umbilical cord blood transplantation halts skeletal disease progression in the murine model of MPS-I

Umbilical cord blood (UCB) is a promising source of stem cells to use in early haematopoietic stem cell transplantation (HSCT) approaches for several genetic diseases that can be diagnosed at birth. Mucopolysaccharidosis type I (MPS-I) is a progressive multi-system disorder caused by deficiency of lysosomal enzyme α-L-iduronidase, and patients treated with allogeneic HSCT at the onset have improved outcome, suggesting to administer such therapy as early as possible. Given that the best characterized MPS-I murine model is an immunocompetent mouse, we here developed a transplantation system based on murine UCB. With the final aim of testing the therapeutic efficacy of UCB in MPS-I mice transplanted at birth, we first defined the features of murine UCB cells and demonstrated that they are capable of multi-lineage haematopoietic repopulation of myeloablated adult mice similarly to bone marrow cells. We then assessed the effectiveness of murine UCB cells transplantation in busulfan-conditioned newborn MPS-I mice. Twenty weeks after treatment, iduronidase activity was increased in visceral organs of MPS-I animals, glycosaminoglycans storage was reduced, and skeletal phenotype was ameliorated. This study explores a potential therapy for MPS-I at a very early stage in life and represents a novel model to test UCB-based transplantation approaches for various diseases

The Use of Baclofen as a Treatment for Alcohol Use Disorder: A Clinical Practice Perspective

Alcohol use disorder (AUD) is a brain disorder associated with high rates of mortality and morbidity worldwide. Baclofen, a selective gamma-aminobutyric acid-B (GABA-B) receptor agonist, has emerged as a promising drug for AUD. The use of this drug remains controversial, in part due to uncertainty regarding dosing and efficacy, alongside concerns about safety. To date there have been 15 randomized controlled trials (RCTs) investigating the use of baclofen in AUD; three using doses over 100 mg/day. Two additional RCTs have been completed but have not yet been published. Most trials used fixed dosing of 30–80 mg/day. The other approach involved titration until the desired clinical effect was achieved, or unwanted effects emerged. The maintenance dose varies widely from 30 to more than 300 mg/day. Baclofen may be particularly advantageous in those with liver disease, due to its limited hepatic metabolism and safe profile in this population. Patients should be informed that the use of baclofen for AUD is as an “off-label” prescription, that no optimal fixed dose has been established, and that existing clinical evidence on efficacy is inconsistent. Baclofen therapy requires careful medical monitoring due to safety considerations, particularly at higher doses and in those with comorbid physical and/or psychiatric conditions. Baclofen is mostly used in some European countries and Australia, and in particular, for patients who have not benefitted from the currently used and approved medications for AUD

Synthetic biology to access and expand nature's chemical diversity

Bacterial genomes encode the biosynthetic potential to produce hundreds of thousands of complex molecules with diverse applications, from medicine to agriculture and materials. Accessing these natural products promises to reinvigorate drug discovery pipelines and provide novel routes to synthesize complex chemicals. The pathways leading to the production of these molecules often comprise dozens of genes spanning large areas of the genome and are controlled by complex regulatory networks with some of the most interesting molecules being produced by non-model organisms. In this Review, we discuss how advances in synthetic biology — including novel DNA construction technologies, the use of genetic parts for the precise control of expression and for synthetic regulatory circuits — and multiplexed genome engineering can be used to optimize the design and synthesis of pathways that produce natural products