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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. Dual catalysis is widely employed by natural metalloenzymes to functionalize challenging substrates. Now, this concept is applied to artificial metalloenzymes by designing a hydroaminase with two biotinylated gold cofactors enabling an unnatural σ,π-activation mechanism of terminal alkynes.

Background There is an urgent need for a vaccine with efficacy against SARS-CoV-2. We hypothesize that peptide vaccines containing epitope regions optimized for concurrent B cell, CD4+ T cell, and CD8+ T cell stimulation would drive both humoral and cellular immunity with high specificity, potentially avoiding undesired effects such as antibody-dependent enhancement (ADE) (figure 1). Leveraging methods initially developed for prediction of tumor-specific antigen targets, we combine computational prediction of T cell epitopes, recently published B cell epitope mapping studies, and epitope accessibility to select candidate peptide vaccines for SARS-CoV-2 (figure 2). Methods SARS-CoV-2 HLA-I and HLA-II ligands were predicted using multiple MHC binding prediction software. T cell vaccine candidates were further refined by predicted immunogenicity, viral source protein abundance, sequence conservation, coverage of high frequency HLA alleles, and co-localization of CD4+/CD8+ T cell epitopes. B cell epitope regions were chosen from linear epitope mapping studies of convalescent patient serum, filtering to select regions with surface accessibility, high sequence conservation, spatial localization near functional domains of the spike glycoprotein, and avoidance of glycosylation sites. Using murine compatible T/B cell epitopes, vaccine studies were performed with downstream ELISA/ELISpot to monitor immunogenicity. Results We observed distribution of HLA-I (n = 2486) and -II (n = 3138) ligands evenly across the SARS-CoV-2 proteome, with significant overlap between predicted human and murine ligands (figure 3). Applying a multivariable immunogenicity model trained from IEDB viral tetramer data (AUC 0.7 and 0.9 for HLA-I and -II, respectively), alongside filters for entropy and protein expression resulted in 292 CD8+ and 616 CD4+ epitopes (figure 4). From an initial pool of 58 B cell epitope candidates, three epitope regions were identified (figure 5). Combining B cell and T cell analyses, alongside manufacturability heuristic, we propose a set of SARS-CoV-2 vaccine peptides for use in subsequent murine studies and clinical trials (figure 6). Preliminary murine studies demonstrate evidence of T and B cell activation (figure 7). Conclusions A peptide vaccine targeting B cells, CD4+ T cells, and CD8+ T cells in parallel may prove an important part of a multifaceted response to the COVID-19 pandemic. Adapting methods for predicting tumor-specific antigens, we presented a set of peptide candidates with high overlap for T and B cell epitopes and broad haplotype population coverage, with validation of immunogenicity in murine vaccine studies. Acknowledgements The authors appreciate funding support from University of North Carolina University Cancer Research Fund (AR and BGV), the Susan G. Komen Foundation (BGV), the V Foundation for Cancer Research (BGV), and the National Institutes of Health (CCS, 1F30CA225136). We would like to thank members of the #DownWithTheCrown Slack channel for helpful discussion and feedback.

The Philippines is a primarily agricultural nation in Southeast Asia, despite rapid industrialization in many areas of the archipelago...

The purpose of this research is to get information on the value of defectives, water content, taste, and the overall acceptance of Arabica coffee derived from 20 coffee producers in West Sumatera. Taste testing involves trained q grader. The Data of research results analyzed descriptively. The results showed that the quality defectives of coffee is at a quality level from 3 to 5 and 70% quality of coffee in accordance with SNI (Indonesian National Standards) 01-2907-2008. The type of defective in coffee beans is the rupture seeds, brown seeds, black seeds then followed by perforated seeds. Taste of coffee Brew are Excellent Aroma and complex coffee flavor, good acidity and thickness of seeds. Vanilla flavor and taste of fruits and spices. The color of the Arabica coffee powder varies from brown, black and color like cinnamon, As well as overall acceptance kinda like. This indicates the need to implement standard operational procedures to improve the quality of coffee in West Sumatera Framework

Microalgae Chlorella vulgaris contains micronutrients such as mineral zinc and selenium having role to produce antibody and improve body immunity. However, the impact of Chlorella vulgaris supplementation to goat milks’s chemical compositions has not being well documented. This research’s objective is to identify Chlorella vulgaris suppmentation as zinc and selenium source to improve goat milk’s quality as health drink in COVID-19 pandemy. Fifteen Etawah cross breed or Peranakan Etawa (PE) goats groupped into 5 treatments dan 3 repetitions. The goats were fed with grass, tofu waste and tempe waste. Chlorella supplement is fed to treated goat feed every day with level 0g (P0 = control), 2, 5g (P1), 5g (P2), 7, 5g (P3), and 10g (P4). The research’s result shows that zinc content in goat milk increase 2, 12 ppm or 24, 67%, while selenium increase 0, 08 mcg or 5, 5% after getting supplementation of Chlorella 10g/goat/day (P4). The provision of Chlorella also increase as solids total, protein and milk fat. Milk production increased by 28, 36% with Chlorella vulgaris supplements (P4). Based on the results of this study it can be stated that Chlorella vulgaris supplementation in goat PE feed can be used as a source of natural zinc and selenium minerals to improve the quality of goat milk as a health drink to increase body immunity, especially during the Covid-19 pandemic.

Research has been conducted to determine the Minang domain arabica coffee sensory quality profile, which can be used as a reference for specialty coffee. Seed processing is carried out at the research location (Solok Regency, South Solok Regency, Pasaman, Agam, and Fifty Cities) until the grain stage is wet (washed). Fianda Coffee Roastery carries out the coffee roasting process. Coffee uses W600i Roaster with a long roast method with two treatments, namely temperature 1800C for 15 minutes, temperature B 2000C for 10 minutes. The taste test was conducted at a two-door cafe and the Indonesian Coffee and Cocoa Research Institute Laboratory, Jember, East Java, following the SCAA. The test results showed that dry coffee beans' moisture content for all treatments was

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&rsquo, 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.

Nuclear distribution protein C (NudC) is a mitotic regulator that plays a role in cytokinesis. However, how NudC is regulated during cytokinesis remains unclear. Here, we show that NudC is phosphorylated by Aurora B, a kinase critical for cell abscission. NudC is co-localized with Aurora B at the midbody and co-immunoprecipitated with Aurora B in mitosis. Inhibition of Aurora B by ZM447439 reduced NudC phosphorylation, suggesting that NudC is an Aurora B substrate in vivo. We identified T40 on NudC as an Aurora B phosphorylation site. NudC depletion resulted in cytokinesis failure with a dramatic elongation of the intercellular bridge between daughter cells, sustained Aurora B activity at the midbody, and reduced cell abscission. These cytokinetic defects can be rescued by the ectopic expression of wild-type NudC. Reconstitution with T40A phospho-defective NudC was found to rescue the cytokinesis defect. In contrast, reconstitution with the T40D phospho-mimetic NudC was inefficient in supporting the completion of cytokinesis. These results suggest that that dynamic phosphorylation of NudC by Aurora B regulates cytokinesis.

Phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P2] is a key second messenger that regulates actin and membrane dynamics, as well as other cellular processes. Many of the effects of PtdIns(4,5)P2are mediated by binding to effector proteins that contain a pleckstrin homology (PH) domain. Here, we identify two novel effectors of PtdIns(4,5)P2in the budding yeast Saccharomyces cerevisiae: the PH domain containing protein Slm1 and its homolog Slm2. Slm1 and Slm2 serve redundant roles essential for cell growth and actin cytoskeleton polarization. Slm1 and Slm2 bind PtdIns(4,5)P2through their PH domains. In addition, Slm1 and Slm2 physically interact with Avo2 and Bit61, two components of the TORC2 signaling complex, which mediates Tor2 signaling to the actin cytoskeleton. Together, these interactions coordinately regulate Slm1 targeting to the plasma membrane. Our results thus identify two novel effectors of PtdIns(4,5)P2regulating cell growth and actin organization and suggest that Slm1 and Slm2 integrate inputs from the PtdIns(4,5)P2and TORC2 to modulate polarized actin assembly and growth.

Telomeres are areas of heterochromatin composed of TTAGGG repeats located at the ends of linear chromosomes. They play a critical role in keeping genome stable and preventing premature aging diseases and the development of cancer. Characterizing mechanisms of telomere maintenance and understanding how their deregulation contributes to human diseases are therefore important for developing novel therapies. A key mechanism driving telomere maintenance and replicative immortality in cancer cells is telomere elongation by telomerase, and many emerging potential telomere-based therapies have focused on targeting telomerase components. By contrast, recent studies on telomere maintenance mechanism suggest that disrupting telomere stability by interfering with alternative mechanisms of telomere synthesis or protection may also yield new strategies for the treatment of cancer. This review will focus on emerging regulators of telomere synthesis or maintenance, such as G4 telomeric DNA, the CST complex, the t-loop, and shelterins, and discuss their potential as targets for anti-cancer chemotherapeutic intervention in the future.

Protein acetylation has been implicated in playing an important role during mitotic progression. Aurora B kinase is known to play a critical role in mitosis. However, whether Aurora B is regulated by acetylation is not known. Using IP with an anti-acetyl lysine antibody, we identified Aurora B as an acetylated protein in PC3 prostate cancer cells. Knockdown of HDAC3 or inhibiting HDAC3 deacetylase activity led to a significant increase (P

IbM of Mangosteen UKM group is one of Agricultural Polytechnic State of Payakumbuh’s proposal program which has aim to increase the productivity value and competitiveness also to dig up the mangosteen local food potention in line with functional food paradigm development. Prebiotic and probiotic mangosteen food technology as the food quality for life safety is one of future plan effort to dig up local food potention that is Safe, Healthy, Intact and Halal (Allowed in Islamic way). By this IbM program is expected to change people’s mindset so they have willingness, be able and can to process the mangosteen to be product which about to add economical value and various functional food using technology in processing. Mangosteen (Garcinia mangostana.L) aka Queen of tropical fruit is a fruit that has many superiorities compared to another fruits like natural colour source that can be used as food coloring, also to be utilized as anti-oxidant, anti-diarrhea, anti-cancer and others. The Methods done are field survey, Focus Group Discussion (FGD), Demonstration, illumination, training that focused to the products and methods with guidance system also effective monitoring in creating functional food products and always to be preferred. In its implementation Mitra’s group in society contributing as the business doer in IbM activity. College has role giving innovation and technology transfer and evaluation and guidance to ‘Mitra’. The result expected is prebiotic and probiotic mangosteen food products like mangosteen yogurt, Mangosteen syrup, Mangosteen-dried extract and mangosteen cake, service, journal, and book.

There are so many ways to do in reducing cholesterol concentration in blood. Yogurt and strawberry juice have ability to repair blood lipid profile at hypercholesterolemia. Research and lactobacillus development potention as the cholesterol killing probiotic that to be mixed with strawberry juice is necessary to do to support community health. The purpose of this reasearch was measuring the mixture of yogurt and strawberry juice effectiveness to repair blood lipid profile. The design of this research that to be used was experimental by pretest and posted randomized control design progam. The sample of this research was thirty white mice (Rattus Norvegicus). Cholesterol Total, HDL and LDL to be analyzed by WI-M-KK2 BLK-SB methode (Clinical Chemstry Auto Analyzer) using tool Selectra E Auto Analyzer. The data was analyzed by Anova test, continued by LSD test using computer program . The result after giving mixture of yogurt and strawberry juice with the dosage 2ml and 3ml per day could obviously reduce LDL Level (p 0,05). The differentiation in dosis giving showed unmeaning reduction result (p>0,05). Conclusion, the mixture of yogurt and strawberry juice could repair blood lipid profile, where it’s proved to reduce drastically choleterol Total Level, LDL in 30 days.

The telomeric CTC1/STN1/TEN1 (CST) complex has recently been implicated in promoting replication recovery under replications stress at genomic regions, yet its precise role is unclear. Here we report that STN1 is enriched at GC-rich repetitive sequences genome-wide in response to hydroxyurea (HU)-induced replication stress. STN1 deficiency exacerbates fragility of these sequences under replication stress, resulting in chromosome fragmentation. We find that upon fork stalling, CST proteins form distinct nuclear foci that colocalize with RAD51. Furthermore, replication stress induces physical association between CST with RAD51 in an ATR-dependent manner. Strikingly, CST deficiency diminishes HU-induced RAD51 foci formation and reduces RAD51 recruitment to telomeres and non-telomeric GC-rich fragile sequences. Collectively, our findings establish that CST promotes RAD51 recruitment to GC-rich repetitive sequences in response to replication stress to facilitate replication restart, thereby providing insights into the mechanism underlying genome stability maintenance.