Covert Genetic Selections to Optimize Phenotypes

In many high complexity systems (cells, organisms, institutions, societies, economies, etc.), it is unclear which components should be regulated to affect overall performance. To identify and prioritize molecular targets which impact cellular phenotypes, we have developed a selection procedure (“SPI”–single promoting/inhibiting target identification) which monitors the abundance of ectopic cDNAs. We have used this approach to identify growth regulators. For this purpose, complex pools of S. cerevisiae cDNA transformants were established and we quantitated the evolution of the spectrum of cDNAs which was initially present. These data emphasized the importance of translation initiation and ER-Golgi traffic for growth. SPI provides functional insight into the stability of cellular phenotypes under circumstances in which established genetic approaches cannot be implemented. It provides a functional “synthetic genetic signature” for each state of the cell (i.e. genotype and environment) by surveying complex genetic libraries, and does not require specialized arrays of cDNAs/shRNAs, deletion strains, direct assessment of clonal growth or even a conditional phenotype. Moreover, it establishes a hierarchy of importance of those targets which can contribute, either positively or negatively, to modify the prevailing phenotype. Extensions of these proof-of-principle experiments to other cell types should provide a novel and powerful approach to analyze multiple aspects of the basic biology of yeast and animal cells as well as clinically-relevant issues

ALASAN TEMPAT WARUNG PERJUDIAN KOA TETAP EKSIS DI PAINAN TIMUR KECAMATAN IV JURAI KABUPATEN PESISIR SELATAN

Skripsi ini membahas tentang alasan tempat warung perjudian koa tetap eksis di Painan Timur. Koa/ceki merupakan permainan kartu yang ada di Minangkabau. Awalnya Permainan koa ini hanya merupakan permainan biasa yang dianggap sebagai hiburan ketika salah satu dari warga yang akan melakukan resepsi pernikahan. Namun pada akhirnya permainan ini diajadikan ajang taruhan atau perjudian, dengan tujuan mendeskripsikan alasan tempat warung perjudian koa tetap eksis di Painan Timur. Dalam penelitian ini teori yang digunakan adalah teori fenomenologi Alfred Schutz, yang meliputi Because Motive dan In Order to Motive. Pendekatan yang dilakukan adalah pendekatan kualitatif dengan tipe penelitian deskriptif. Penentuan informan dalam penelitian ini dengan menggunakan teknik purposive sampling. Untuk mendapatkan data, penulis menggunakan teknik wawancara mendalam kepada 12 informan masyarakat sekitar warung. Hasil penelitian menunjukkan bahwa yang melatarbelakangi motif masyarakat membiarkan warung perjudian koa tetap eksis di Painan Timur dari 12 informan. Meliputi Because Motive diantaranya (1) adanya sikap pro dan kontra antar tetangga, (2) perubahan permainan dari generasi ke generasi, (3) minimnya tenggang rasa antar pemain dengan warga sekitar (1) tidak ada sanksi hukum yang tegas dari pemerintah daerah, (2) pemilik warung tergiur mendapatkan keuntungan besar, (3) tidak adanya tanggapan baik dari pemilik warung dan pemain, (4) harapan merubah perilaku berjudi koa. Berdasarkan penelitian yang dilakukan, diperoleh kesimpulan bahwa adanya because motive dan in order to motive yang dilakukan oleh para pemain perjudian koa dapat merugikan dirinya sendiri, keluarga, pemilik warung dan orang lain yang berada disekitarnya, serta perilaku menyimpang dari ajaran agama Islam yang mengarah keperbuatan haram

Systematic engineering of artificial metalloenzymes for new-to-nature reactions

Artificial metalloenzymes (ArMs) catalyzing new-to-nature reactions could play an important role in transitioning toward a sustainable economy. While ArMs have been created for various transformations, attempts at their genetic optimization have been case specific and resulted mostly in modest improvements. To realize their full potential, methods to rapidly discover active ArM variants for ideally any reaction of interest are required. Here, we introduce a reaction-independent, automation-compatible platform, which relies on periplasmic compartmentalization in Escherichia coli to rapidly and reliably engineer ArMs based on the biotin-streptavidin technology. We systematically assess 400 ArM mutants for five bioorthogonal transformations involving different metals, reaction mechanisms, and reactants, which include novel ArMs for gold-catalyzed hydroamination and hydroarylation. Activity enhancements up to 15-fold highlight the potential of the systematic approach. Furthermore, we suggest smart screening strategies and build machine learning models that accurately predict ArM activity from sequence, which has crucial implications for future ArM development.ISSN:2375-254

A Bioclimate-Based Maximum Entropy Model for Comperiella calauanica Barrion, Almarinez and Amalin (Hymenoptera: Encyrtidae) in the Philippines

Comperiella calauanica is a host-specific endoparasitoid and effective biological control agent of the diaspidid Aspidiotus rigidus, whose outbreak from 2010 to 2015 severely threatened the coconut industry in the Philippines. Using the maximum entropy (Maxent) algorithm, we developed a species distribution model (SDM) for C. calauanica based on 19 bioclimatic variables, using occurrence data obtained mostly from field surveys conducted in A. rigidus-infested areas in Luzon Island from 2014 to 2016. The calculated the area under the ROC curve (AUC) values for the model were very high (0.966, standard deviation = 0.005), indicating the model’s high predictive power. Precipitation seasonality was found to have the highest relative contribution to model development. Response curves produced by Maxent suggested the positive influence of mean temperature of the driest quarter, and negative influence of precipitation of the driest and coldest quarters on habitat suitability. Given that C. calauanica has been found to always occur with A. rigidus in Luzon Island due to high host-specificity, the SDM for the parasitoid may also be considered and used as a predictive model for its host. This was confirmed through field surveys conducted between late 2016 and early 2018, which found and confirmed the occurrence of A. rigidus in three areas predicted by the SDM to have moderate to high habitat suitability or probability of occurrence of C. calauanica: Zamboanga City in Mindanao; Isabela City in Basilan Island; and Tablas Island in Romblon. This validation in the field demonstrated the utility of the bioclimate-based SDM for C. calauanica in predicting habitat suitability or probability of occurrence of A. rigidus in the Philippines

A bioclimate-based maximum entropy model for Comperiella calauanica barrion, Almarinez and Amalin (Hymenoptera: Encyrtidae) in the Philippines

Comperiella calauanica is a host-specific endoparasitoid and effective biological control agent of the diaspidid Aspidiotus rigidus, whose outbreak from 2010 to 2015 severely threatened the coconut industry in the Philippines. Using the maximum entropy (Maxent) algorithm, we developed a species distribution model (SDM) for C. calauanica based on 19 bioclimatic variables, using occurrence data obtained mostly from field surveys conducted in A. rigidus-infested areas in Luzon Island from 2014 to 2016. The calculated the area under the ROC curve (AUC) values for the model were very high (0.966, standard deviation = 0.005), indicating the model’s high predictive power. Precipitation seasonality was found to have the highest relative contribution to model development. Response curves produced by Maxent suggested the positive influence of mean temperature of the driest quarter, and negative influence of precipitation of the driest and coldest quarters on habitat suitability. Given that C. calauanica has been found to always occur with A. rigidus in Luzon Island due to high host-specificity, the SDM for the parasitoid may also be considered and used as a predictive model for its host. This was confirmed through field surveys conducted between late 2016 and early 2018, which found and confirmed the occurrence of A. rigidus in three areas predicted by the SDM to have moderate to high habitat suitability or probability of occurrence of C. calauanica: Zamboanga City in Mindanao; Isabela City in Basilan Island; and Tablas Island in Romblon. This validation in the field demonstrated the utility of the bioclimate-based SDM for C. calauanica in predicting habitat suitability or probability of occurrence of A. rigidus in the Philippines

Design and evolution of chimeric streptavidin for protein-enabled dual gold catalysis

Artificial metalloenzymes result from anchoring an organometallic catalyst within an evolvable protein scaffold. Thanks to its dimer of dimers quaternary structure, streptavidin allows the precise positioning of two metal cofactors to activate a single substrate, thus expanding the reaction scope accessible to artificial metalloenzymes. To validate this concept, we report herein on our efforts to engineer and evolve an artificial hydroaminase based on dual gold activation of alkynes. Guided by modelling, we designed a chimeric streptavidin equipped with a hydrophobic lid shielding its active site, which enforces the advantageous positioning of two synergistic biotinylated gold cofactors. Three rounds of directed evolution using Escherichia coli cell-free extracts led to the identification of mutants favouring either the anti-Markovnikov product (an indole carboxamide with 96% regioselectivity, 51 turnover numbers), resulting from a dual gold σ,π-activation of an ethynylphenylurea substrate, or the Markovnikov product (a phenyl-dihydroquinazolinone with 99% regioselectivity, 333 turnover numbers), resulting from the π-activation of the alkyne by gold

Minor word formation processes in Michael Scott`s the alchemyst: the secret of the immortal Nicholas Flamel novel

The aim of this paper is to describe the process and challenges in building exposure scenarios for engineered nanomaterials (ENM), using an exposure scenario format similar to that used for the European Chemicals regulation (REACH). Over 60 exposure scenarios were developed based on information from publicly available sources (literature, books, and reports), publicly available exposure estimation models, occupational sampling campaign data from partnering institutions, and industrial partners regarding their own facilities. The primary focus was on carbon-based nanomaterials, nano-silver (nano-Ag) and nano-titanium dioxide (nano-TiO2), and included occupational and consumer uses of these materials with consideration of the associated environmental release. The process of building exposure scenarios illustrated the availability and limitations of existing information and exposure assessment tools for characterizing exposure to ENM, particularly as it relates to risk assessment. This article describes the gaps in the information reviewed, recommends future areas of ENM exposure research, and proposes types of information that should, at a minimum, be included when reporting the results of such research, so that the information is useful in a wider context

Protor-1 is required for efficient mTORC2-mediated activation of SGK1 in the kidney

International audienceThe mTOR (mammalian target of rapamycin) protein kinase is an important regulator of cell growth and is a key target for therapeutic intervention in cancer. Two complexes of mTOR have been identified: complex 1 (mTORC1), consisting of mTOR, Raptor and mLST8 and complex 2 (mTORC2) consisting of mTOR, Rictor, Sin1, mLST8 and Protor-1 or Protor-2. Both complexes phosphorylate the hydrophobic motifs of AGC kinase family members: mTORC1 phosphorylates S6K, while mTORC2 regulates phosphorylation of Akt, PKC and SGK1. To investigate the roles of the Protor isoforms, we generated single as well as double Protor-1 and Protor-2 knockout mice and studied how activation of known mTORC2 substrates was affected. We observed that loss of Protor-1 and/or Protor-2 did not affect the expression of the other mTORC2 components, nor their ability to assemble into an active complex. Moreover, Protor knockout mice display no defects in the phosphorylation of Akt and PKC at their hydrophobic or turn motifs. Strikingly, we observed that Protor-1 knockout mice displayed markedly reduced hydrophobic motif phosphorylation of SGK1 and its physiological substrate NDRG1 in the kidney. These data suggest that Protor-1 may play a role in enabling mTORC2 to efficiently activate SGK1, at least in the kidney