Towards a generic prototyping approach for therapeutically-relevant peptides and proteins in a cell-free translation system

Advances in peptide and protein therapeutics increased the need for rapid and cost-effective polypeptide prototyping. While in vitro translation systems are well suited for fast and multiplexed polypeptide prototyping, they suffer from misfolding, aggregation and disulfide-bond scrambling of the translated products. Here we propose that efficient folding of in vitro produced disulfide-rich peptides and proteins can be achieved if performed in an aggregation-free and thermodynamically controlled folding environment. To this end, we modify an E. coli-based in vitro translation system to allow co-translational capture of translated products by affinity matrix. This process reduces protein aggregation and enables productive oxidative folding and recycling of misfolded states under thermodynamic control. In this study we show that the developed approach is likely to be generally applicable for prototyping of a wide variety of disulfide-constrained peptides, macrocyclic peptides with non-native bonds and antibody fragments in amounts sufficient for interaction analysis and biological activity assessment.

Esquisitar pontes: Possíveis diálogos entre feministas queer chicanas e brasileiras para uma crítica literária de margem / Esquisiting Bridges: Possible dialogues between queer feminist Chicanas and Brazilians for a literary criticism from the margins

Resumo: A proposta deste artigo é esquisitar pontes e aproximar os feminismos contra- hegemônicos da América Latina, mais especificamente do Brasil, dos pensamentos produzidos pelas Chicanas, na fronteira entre os EUA e o México. Através dessa aproximação, buscamos potencializar a crítica literária feminista brasileira ao considerar processos de produção de conhecimento advindos de posicionamentos marginalizados. Enquanto a conceituação do esquisito vem do trabalho da pesquisadora brasileira Eliana Ávila (2015), a construção de pontes entre distintos grupos marginalizados emerge do pensamento fronteiriço de Gloria Anzaldúa e Cherríe Moraga (1981). Os trabalhos de Lélia Gonzalez (1984, 1988) e Larissa Pelúcio (2012) também se entrelaçam aos de autoras Chicanas para questionar relações de poder a apagamentos culturais históricos. Ao esquisitar pontes, desenvolve-se diálogos e articulações a partir de uma visão conscientemente parcial capazes de encontrar as potencialidades políticas para construções epistemológicas que levam em consideração os saberes localizados. É desse posicionamento que reforçamos a proposta de um queer esquisito e questionamos as relações geográficas de poder a partir de uma perspectiva brasileira.Palavras-chave: pontes; esquisito; queer; chicana; geopolítica; feminismo.Abstract: The objective of this article is to esquisitar (queer, in a free translation) bridges and to bring closer counter-hegemonic feminisms from Latin-America, more specifically from Brazil, and those developed by the Chicanas, in the borders between the USA and Mexico. Through this dialogue, we intend to potentialize the Brazilian feminist literary criticism by considering processes of knowledge production from marginalized positions. While the concept of esquisito comes from the works of the Brazilian researcher Eliana Ávila (2015), the construction of bridges between distinct groups emerges from the border thinking of Gloria Anzaldúa e Cherríe Moraga (1981). The works of Lélia Gonzalez (1984; 1988) and Larissa Pelúcio (2012) are also intertwined to the ones from Chicana authors to question power relations and historical cultural invisibilities. By esquisiting bridges, it is possible to develop political potentialities and epistemological constructions that take into consideration situated knowledges. From this perspective, we reinforce the proposal of an esquisito queer and question the geopolitics from a Brazilian point of view.Keywords: bridges; esquisito; queer; chicana; geopolitics; feminism.

Translation methods of English-Indonesian subtitles in Mariana Atencio’s “TEDx talk: what makes you special?”

This research aims at finding out methods used to translate connotative and denotative phrases in the video subtitle of Mariana Atencio’s “Tedx Talk: What Makes You Special?”. The video was translated to Indonesian by Badaruddin Saahe and has been reviewed by Santi Deviyanti Rahayu. This research method was qualitative descriptive, focusing on Newmark’s translation theory. The data of this research were taken from the transcript of the video and the provided subtitles. This study revealed twenty-eight phrases with connotative meaning, and the translation process employed both single and mixed methods. Single method types of translation employed were faithful translation, adaptation, free translation, and idiomatic translation, while mixed methods employed faithful and adaptation, faithful and semantic, faithful and communicative, adaptation and communicative, free and idiomatic, and idiomatic and adaptation. This study also revealed twelve phrases with denotative meaning, in which the translator chooses to translate them denotatively using various methods, despite the phrases conveying other meanings. This relates to the second study issue, which explains the connection between denotative meaningful sentences and Newmark (1988) translation approach. Following his observations, the researcher discovered that only one method was used: word-for-word and literal translation. To translate phrases with this denotative meaning, the translator employs two ways simultaneously: word-for-word and literal, literal and adaptation, literal & faithful, faithful & communicative, and Idiomatic & communicative. The researcher may deduce from the previous explanation that mixed techniques and denotative meaning are related in the sense that the combined methods share the same qualities, namely direct, contextual, and communicative. As stated by Fromkin (2001), this is also related to the properties of the denotation itself.

Intercâmbio intelectual, formação de professores e programas escolares: Alda Lodi, expert

Este artigo procura examinar a trajetória de Alda Lodi, enfocando o modo como sua atuação, no campo do ensino de matemática, consolidou seu papel de expert da educação em Minas Gerais. Formada na Escola Normal de Belo Horizonte, Lodi viajou aos Estados Unidos em 1927 como parte de uma missão pedagógica vinculada à reforma Francisco Campos. Sua viagem de estudos ao Teachers College funcionou como elemento de ascensão profissional, ao mesmo tempo em que fundamentou as discussões que ela desenvolveu com as professoras-alunas da Escola de Aperfeiçoamento de Belo Horizonte, onde foi professora de Metodologia da Aritmética. Neste artigo, procuramos explorar suas contribuições para o ensino de matemática a partir dos programas de Aritmética e Geometria do ensino primário – escritos por ela para a Secretaria de Educação de Minas Gerais entre os anos de 1941 e 1953. Com isso, procuramos contribuir para os estudos que investigam o papel que os experts desempenharam na sistematização de saberes a e para ensinar matemática no Brasil.Cultural exchange, teacher training and school programs: Alda Lodi, expertThis article aims to examine Alda Lodi’s career, focused on the way her work in the field of mathematics education helped to place her as an education expert in Minas Gerais. Alda Lodi graduated at the Belo Horizonte Normal School in 1915. In 1927, she travelled to the United States to study education as part of a pedagogical mission organized by Francisco Campos, who was reforming the education system of Minas Gerais state. Her time at the Teachers College, Columbia University was very important to her career. It also played a central role in the discussions developed by her at the Escola de Aperfeiçoamento de Belo Horizonte (Belo Horizonte Teacher Training School, in free translation), in the field of Methodology of Arithmetic. In this article we are going to explore her contributions to the teaching of mathematics using two Arithmetic and Geometry programs she wrote between the 1940’s and 1950’s for the Minas Gerais Department of Education. Thus, we aim to contribute to the studies that deal with the role played by experts in the systematization of the knowledge to teach and for teaching in the field of mathematics education in Brazil.Keywords: History of mathematics education; Alda Lodi; Experts; Teaching programs.

TRANSLATION METHOD USED IN DELI MALAY AND SERDANG MALAY FOLKLORES FROM INDONESIAN TO ENGLISH

This study aimed to determine the many types of translation methods, how they were employed in the translation of the folklores "Deli Malay" and "Serdang Malay" from Indonesian to English, and why the translators used the prevailing types in these folklores. The descriptive qualitative method was used to perform this research. The data was gathered from a folklore book and an interview with the folklores' translator. Descriptive qualitative research is the method used to analyze the data. The findings of this study revealed that literal translation accounted for 39 sentences (16%), faithful translation accounted for 23 sentences (9%), semantic translation accounted for four sentences (2%), adaptation translation accounted for seven sentences (3%), free translation accounted for 158 sentences (64%) and communicative translation accounted for 16 sentences (6%). A total of 247 sentences were extracted from the data. Because he did not follow any theories or methods of translation when translating the texts, the translator utilized free translation as the dominating method. Instead, he evaluated the translations' target audience, youngsters, and made them acceptable and simple to understand.

Stable m7G Cap-Distal 5'UTR Hairpin Structure Mediates Distinct 40S and 60S Binding Dynamics

The 5' untranslated region (UTR) of diverse mRNAs contains secondary structures that can influence protein synthesis by modulating the initiation step of translation. Studies support the ability of these structures to inhibit 40S subunit recruitment and scanning, but the dynamics of ribosomal subunit interactions with mRNA remain poorly understood. Here, we developed a reconstituted Saccharomyces cerevisiae cell-free translation system with fluorescently labeled ribosomal subunits. We applied this extract and single-molecule fluorescence microscopy to monitor, in real time, individual 40S and 60S interactions with mRNAs containing 5' UTR hairpin structures with varying thermostability. In comparison to mRNAs containing no or weak 5' UTR hairpins (ΔG >= -5.4 kcal/mol), mRNAs with stable hairpins (ΔG <= -16.5 kcal/mol) showed reduced numbers of 60S recruitment to mRNA, consistent with the expectation of reduced translation efficiency for such mRNAs. Interestingly, such mRNAs showed increased numbers of 40S recruitment events to individual mRNAs but with shortened duration on mRNA. Correlation analysis showed that these unstable 40S binding events were nonproductive for 60S recruitment. Furthermore, although the mRNA sequence is long enough to accommodate multiple 40S, individual mRNAs are predominantly observed to engage with a single 40S at a time, indicating the sequestering of mRNA 5' end by initiating 40S. Altogether, these observations suggest that stable cap-distal hairpins in 5' UTR reduce initiation and translation efficiency by destabilizing 40S-mRNA interactions and promoting 40S dissociation from mRNA. The premature 40S dissociation frees mRNA 5'-end accessibility for new initiation events, but the increased rate of 40S recruitment is insufficient to compensate for the reduction of initiation efficiency due to premature 40S dissociation. This study provides the first single-molecule kinetic characterization of 40S/60S interactions with mRNA during cap-dependent initiation and the modulation of such interactions by cap-distal 5' UTR hairpin structures.

Reconstitution of Prenyltransferase Activity on Nanodiscs by Components of the Rubber Synthesis Machinery of the Para Rubber Tree and Guayule

Natural rubber of the Para rubber tree (Hevea brasiliensis) is synthesized as a result of prenyltransferase activity. The proteins HRT1, HRT2, and HRBP have been identified as candidate components of the rubber biosynthetic machinery. To clarify the contribution of these proteins to prenyltransferase activity, we established a cell-free translation system for nanodisc-based protein reconstitution and measured the enzyme activity of the protein-nanodisc complexes. Co-expression of HRT1 and HRBP in the presence of nanodiscs yielded marked polyisoprene synthesis activity. By contrast, neither HRT1, HRT2, or HRBP alone nor a complex of HRT2 and HRBP manifested such activity. Similar analysis of guayule (Parthenium argentatum) proteins revealed that three HRT1 homologs (PaCPT1–3) manifested prenyltransferase activity only if co-expressed with PaCBP, the homolog of HRBP. Our results thus indicate that two heterologous subunits form the core prenyltransferase of the rubber biosynthetic machinery. A recently developed structure modeling program predicted the structure of such heterodimer complexes including HRT1/HRBP and PaCPT2/PaCBP. HRT and PaCPT proteins were also found to possess affinity for a lipid membrane in the absence of HRBP or PaCBP, and structure modeling implicated an amphipathic α-helical domain of HRT1 and PaCPT2 in membrane binding of these proteins.

Developing Culturally Effective Strategies for Chinese to English Geotourism Translation by Corpus-Based Interdisciplinary Translation Analysis

As a new field of translation with its own special genre, geotourism has not yet been firmly established because geotourism translations are currently not of a sufficient professional standard. This situation does not provide geotourists with the genre’s full target of enjoyment, learning and engagement through science popularisation tourism activities. In order to better meet these three definitive purposes in geotourism, this study analyses the three basic categories of geotourism—geological features (GFs), geological processes (GPs) and cultural elements (CEs)—to determine effective strategies of geotourism translation from Chinese into English. Challenges in translation include scientific jargon, language style and cultural gaps. In this article, the advantages of Hu’s Eco-translatology theory are explained and used for minimising translation problems; and the corpus linguistics method, superior for quantitative and qualitative analysis, is utilised. As well, digital auxiliary tools Tmxmall (2014) and Sketch Engine (2003) were employed to facilitate corpus research. Through analysis, effective strategies in each of the key geotourism categories, GFs, GPs and CEs, were identified, shaped and recommended for future translators’ attention. In the results, literal translation, transliteration and free translation, addition and use of official UNESCO names were recommended to render GFs. Division and shift translation, literal translation and shift and division were recommended for GPs. Literal translation, transliteration and free translation and addition were recommended for CEs. Since this is an initial investigation in the genre of geotourism, this study has attempted to build a model platform for future study and wider research in geotourism translation and translation pedagogy for the improvement of geotourism translation quality.

On the Prescription Translation of Traditional Chinese Medicine

Traditional Chinese Medicine (TCM) is an essential legacy of Chinese culture, which makes a great contribution to China and the world's medical fields. To make it accepted by more people from different countries, the translation of TCM becomes important. As an important branch of TCM, the study of prescription promotes the development of TCM, and its translation is also significant. However, there are various problems in TCM prescription translation and there are few standards in TCM translation. Thus, this research uses Christiane Nord's "Functionality plus Loyalty" Translation Theory to analyze the TCM prescription translation. Based on characteristics of prescription formation, the paper applies Christiane Nord's "Functionality plus Loyalty" theory to TCM prescription translation. It draws a conclusion that TCM prescription translation can be done according to "Functionality plus Loyalty" theory, and there are three ways of prescription names translation. They are literal translation, free translation and transliteration.