ScRpb4, Encoding an RNA Polymerase Subunit from Sugarcane, Is Ubiquitously Expressed and Resilient to Changes in Response to Stress Conditions

RNA polymerase II is an essential multiprotein complex that transcribes thousands of genes, being a fundamental component of the transcription initiation complex. In eukaryotes, RNA polymerase II is formed by a 10-multisubunit conserved core complex, and two additional peripheral subunits, Rpb4 and Rpb7, form the Rpb4/7 subcomplex. Although transcription is vital for cell and organismal viability, little is known about the transcription initiation complex in sugarcane. An initial characterization of the sugarcane RNA polymerase subunit IV (ScRpb4) was performed. Our results demonstrate that ScRpb4 is evolutionarily conserved across kingdoms. At the molecular level, ScRpb4 expression was found in vegetative and reproductive tissues. Furthermore, the expression of ScRpb4 remained stable under various stress conditions, most likely to ensure a proper transcriptional response. Optimal conditions to express ScRpb4 in vitro for further studies were also identified. In this study, an initial characterization of the sugarcane polymerase II subunit IV is presented. Our results open the window to more specific experiments to study ScRpb4 function, for instance, crystal structure determination and pull-down assays as well as their function under biotic and abiotic stresses.

Influence of Sand Casting Waste as Substitutor of Quartz Sand in Mortar

Sand casting waste has the potential to replace quartz sand in mortar manufacture because it contains high silica. This study uses sand casting waste from the steel industry in Gresik, Indonesia to observe how it affects the quality of the mortar. Initial characterization were carried out to determine the properties of the material, including; magnetic test which results are not attracted by magnets, moisture content test with a value of 0.328%, XRD test to determine the crystallinity content which results contain 99.52% Silica Quartz, and XRF test to determine the content of the compound in which results are 81.25% Silica dominant. Then observations were made by making mortar with the replacement of quartz sand by sand casting with variations of 0% wt, 25% wt, 50% wt, and 100% wt and then tested its compressive strength at 3 days, 7 days, and 28 days. Based on the research that has been done, the optimum result using sand casting is at 25% wt with a compressive strength of 251.15 kgf/cm2 at 28 days of age which is higher than the standard.

Cell-Penetrating Antimicrobial Peptides Derived from an Atypical Staphylococcal δ-Toxin

The continued emergence and global distribution of infections caused by antimicrobial-resistant pathogens fuel our perpetual need for new or alternative therapies. Here, we present the discovery and initial characterization of bacterial cell-penetrating AMPs that were based on a family of virulence factors.

Endogenous protein tagging in medaka using a simplified CRISPR/Cas9 knock-in approach

The CRISPR/Cas9 system has been used to generate fluorescently labelled fusion proteins by homology directed repair in a variety of species. Despite its revolutionary success, there remains an urgent need for increased simplicity and efficiency of genome editing in research organisms. Here, we establish a simplified, highly efficient and precise strategy for CRISPR/Cas9 mediated endogenous protein tagging in medaka (Oryzias latipes). We use a cloning-free approach that relies on PCR amplified donor fragments containing the fluorescent reporter sequences flanked by short homology arms (30-40bp), a synthetic sgRNA and Cas9 mRNA. We generate eight novel knock-in lines with high efficiency of F0 targeting and germline transmission. Whole Genome Sequencing (WGS) results reveal single-copy integration events only at the targeted loci. We provide an initial characterization of these fusion-protein lines, significantly expanding the repertoire of genetic tools available in medaka. In particular, we show that the mScarlet-pcna line has the potential to serve as an organismal-wide label for proliferative zones and an endogenous cell cycle reporter.

Discovery and initial characterization of YloC, a novel endoribonuclease in Bacillus subtilis

Bacillus subtilis genome is predicted to encode numerous ribonucleases, including four 3’ exoribonucleases that have been characterized to some extent. A strain containing gene knockouts of all four known 3’ exoribonucleases is viable, suggesting that one or more additional RNases remain to be discovered. A protein extract from the quadruple RNase mutant strain was fractionated and RNase activity was followed, resulting in identification of an enzyme activity catalyzed by the YloC protein. YloC is an endoribonuclease and is a member of the highly conserved “YicC family” of proteins that is widespread in bacteria. YloC is a metal-dependent enzyme that catalyzes cleavage of single-stranded RNA, preferentially at U residues, and exists in an oligomeric form, most likely a hexamer. As such, YloC shares some characteristics with the SARS-CoV Nsp15 endoribonuclease. While the in vivo function of YloC in B. subtilis is yet to be determined, YloC was found to act similarly to YicC in an Escherichia coli in vivo assay that assesses decay of the small RNA, RyhB. Thus, YloC may play a role in small RNA regulation.

Initial Characterization of Bio-Slurry as Liquid Fertilizer

The demand for organic fertilizers in Indonesia is currently in a critical state. In 2015, it was noticed that the demand of Indonesia for NPK fertilizers reached over 6.5 million tons per year. Given the current state of demand for organic fertilizers in Indonesia, it is imperative to have alternative organic fertilizers that are environmentally friendly and readily available raw materials. One alternative is to use biogas waste that contains essential plant-based nutrients such as NPK (nitrogen, phosphorus, potassium) and additional nutrients such as magnesium (Mg), calcium (Ca) and sulfur (S). The composition of Bio-Slurry after fermentation consists of 25% ± 5 dry matter and 75% ± 5 water. If the dry substance is decomposed, it contains 18- 27% organic matter. The production of liquid organic fertilizers from BioSlurry requires some anaerobic fermentation process lasting 20 days. During the fermentation process it is necessary to additional raw materials such as cow urine, banana stalk and banana peel to add the N, P and K values to BioSlurry. In the initial characterization test, BioSlurry liquid (after pressing) showed an NPK value of <1% and a C-Organic value of 0.04.

Inhibitory Activity of Proanthocyanidins Against Escherichia coli 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase

1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is a key enzyme in the methylerythritol phosphate pathway for terpenoid biosynthesis. Furthermore, it is an ideal target for the screening of novel antibiotics because it is present in causative organisms, but absent from humans. To identify more lipophilic DXR inhibitors from natural resources, we tested the DXR inhibitory activity of five proanthocyanidins in this study. The results indicated that all these compounds specifically restrained the activity of DXR, with procyanid B2 exhibiting a relatively low effect against DXR (IC50 ∼ 305 μM) and procyanid C1 displaying moderate activity (IC50 75.1 μM). The other three compounds cinnamtannin A2, cinnamtannin B1, and cinnamtannin D1 (IC50 ∼ 89.3, 105.0, and 97.8 μM, respectively) showed DXR inhibitory effects that were slightly weaker than that of procyanid C1. In addition, based on the initial characterization, the structure–activity relationship of this series of compounds against DXR is discussed.

Module-Level TIM Degradation in HALT

In this study, TIM degradation is driven through HALT using temperature cycling and random vibration for two commercially available materials providing thermal conductivities of 6.5 and 8.0 W/m-K. HALT specimen were prepared by applying TIM through a 4-mil stencil over AlSiC baseplates in the shape of those used in Wolfspeed CAS325M12HM2 power electronics modules. Baseplates were mounted onto aluminum carrier blocks with embedded thermocouples to characterize the thermal resistance across the baseplate and TIM layer. Thermal dissipation into the top of the baseplates was provided by a custom heating block, which mimics the size and placement of the die junctions in CAS325 modules, applying power loads of 200, 300, and 400W. After initial characterization, samples were transferred to the HALT chamber with one set of samples exposed to temperature cycling only (TCO) and the other temperature cycling and vibration (TCV). Both sample sets were cycled between temperature extremes of −40 and 180 °C with random vibrations applied at a peak acceleration of 3.21 Grms. After hundreds of cycles, samples were reevaluated to assess changes in thermal resistance to provide an accelerated measure of TIM degradation. This will allow for reliability calculations of useful lifetime, provide a basis for developing accelerated testing method to related temperature cycling to faster methods of degradation, and additionally provide a means by which to develop a maintenance schedule for servicing the power modules which will enhance cooling and lifetime operation.