Methods for isolating, identifying and quantifying anthocyanin metabolites in clinical samples

The metabolic fate of anthocyanins until recently was relatively unknown, primarily as a result of their instability at physiological pH and a lack of published methods for isolating and identifying their metabolites from biological samples. The aim of the present work was to establish methods for the extraction and quantification of anthocyanin metabolites present in urine, serum and fecal samples. 35 commercial and 10 synthetic analytes, including both known and predicted human and microbial metabolites of anthocyanins were obtained as reference standards. HPLC and MS/MS conditions were optimized for organic modifier, ionic modifier, mobile phase gradient, flow rate, column type and MS source and compound dependent parameters. The impact of sorbent, solvent, acid, preservative, elution and evaporation on SPE extraction efficiency was also explored. The HPLC-MS/MS method validation demonstrated acceptable linearity (r2, 0.997 ± 0.002) and sensitivity (LODs: urine, 100 ± 375 nM; serum, 104 ± 358 nM and feces 138 ± 344 nM) and the final SPE methods provided recoveries of 88.3 ± 17.8% for urine, 86.5 ± 11.1% for serum and 80.6 ± 20.9% for feces. Final methods were applied to clinical samples derived from an anthocyanin intervention study, where 36 of the 45 modeled metabolites were detected within urine, plasma or faecal samples. The described methods provide suitable versatility for the identification and quantification of an extensive series of anthocyanin metabolites for use in future clinical studies exploring absorption, distribution, metabolism and elimination

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Nucleotides and nucleosides are not only involved in cellular metabolism but also act extracellularly via P1 and P2 receptors, to elicit a wide variety of physiological and pathophysiological responses through paracrine and autocrine signalling pathways. For the first time, we have used an ion-pair reversed-phase high-performance liquid chromatography ultraviolet (UV)-coupled method to rapidly and simultaneously quantify 12 different nucleotides and nucleosides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine, uridine triphosphate, uridine diphosphate, uridine monophosphate, uridine, guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, guanosine): (1) released from a mouse renal cell line (M1 cortical collecting duct) and (2) in human biological samples (i.e., urine). To facilitate analysis of urine samples, a solid-phase extraction step was incorporated (overall recovery rate ? 98 %). All samples were analyzed following injection (100 ?l) into a Synergi Polar-RP 80 Å (250 × 4.6 mm) reversed-phase column with a particle size of 10 ?m, protected with a guard column. A gradient elution profile was run with a mobile phase (phosphate buffer plus ion-pairing agent tetrabutylammonium hydrogen sulfate; pH 6) in 2-30 % acetonitrile (v/v) for 35 min (including equilibration time) at 1 ml min(-1) flow rate. Eluted compounds were detected by UV absorbance at 254 nm and quantified using standard curves for nucleotide and nucleoside mixtures of known concentration. Following validation (specificity, linearity, limits of detection and quantitation, system precision, accuracy, and intermediate precision parameters), this protocol was successfully and reproducibly used to quantify picomolar to nanomolar concentrations of nucleosides and nucleotides in isotonic and hypotonic cell buffers that transiently bathed M1 cells, and urine samples from normal subjects and overactive bladder patients

Trypsin inhibition by macrocyclic and open-chain variants of the squash inhibitor MCoTI-II

MCoTl-I and MCoTl-II from the seeds of Momordica cochinchinensis are inhibitors of trypsin-like proteases and the only known members of the large family of squash inhibitors that are cyclic and contain an additional loop connecting the amino- and the carboxy-terminus. To investigate the contribution of macrocycle formation to biological activity, we synthesized a set of open-chain variants of MCoTl-II that lack the cyclization loop and contain various natural and non-natural amino acid substitutions in the reactive-site loop. Upon replacement of P1 lysine residue \#10 within the open-chain variant of MCoTl-II by the non-natural isosteric nucleo amino acid AlaG{[}beta-(guanin-9-yl)-L-alanine], a conformationally restricted arginine mimetic, residual inhibitory activity was detected, albeit reduced by four orders of magnitude. While the cyclic inhibitors MCoTl-I and MCoTl-II were found to be very potent trypsin inhibitors, with picomolar inhibition constants, the open-chain variants displayed an approximately 10-fold lower affinity. These data suggest that the formation of a circular backbone in the MCoTI squash inhibitors results in enhanced affinity and therefore is a determinant of biological activity

PENINGKATAN MINAT DAN HASIL BELAJAR PESERTA DIDIK PADA MATERI SUMBER DAYA DAN PERUBAHAN ENERGI DENGAN MEDIA GAME ANDROID SYNERGI

Penelitian tindakan ini berawal dari permasalahan yang terdapat di SDN Sukabumi 2, yaitu rendahnya minat dan hasil belajar siswa kelas IV dalam mengikuti sistem Belajar Dari Rumah (BDR) yang diterapkan selama pandemi Covid–19 sehingga 28% siswa memperoleh nilai di bawah KKM dikarenakan merasa jenuh. Tujuan dari penelitian ini adalah untuk meningkatkan minat siswa dan mendeskripsikan peningkatan hasil belajar siswa setelah menggunakan media Game Android Synergi pada pembelajaran kelas IV Tema 2 Subtema 2 tentang Pemanfaatan Sumber Daya Alam dan Perubahan Energi. Subjek penelitian adalah siswa kelas IV SDN Sukabumi 2 Kecamatan Mayangan Kota Probolinggo berjumlah 35 orang. Penggunaan game ini terbukti berhasil memberikan hasil yang signifikan dalam meningkatkan minat, keaktifan dan hasil belajar siswa. Hasil angket yang diberikan kepada 35 siswa adalah 34 siswa (97%) memberikan jawaban sangat setuju, 1 siswa (0,3%) memberikan jawaban setuju, dan tidak ada siswa yang memberikan jawaban tidak setuju terhadap penggunaan media Game Android Synergi dalam pembelajaran, sedangkan ketuntasan belajar siswa mencapai 97% atau hanya 1 siswa yang memperoleh nilai di bawah KKM. Kata Kunci: Game synergi, minat dan hasil belajar. IMPROVING STUDENTS’ INTEREST AND LEARNING OUTCOMEON MATERIAL OF RESOURCES AND ENERGY CHANGESUSING ANDROID SYNERGY GAME MEDIA Abstract: This action research began from the problems met at SDN Sukabumi 2, they are unmotivated students in learning process and learning outcomes of the fourth grade students in joining the online learning implemented during the Covid-19 pandemic, so that there were 28% of students got below the passing grade due to feeling bored. The purpose of this research was to increase students’ interest and to describe the improvement of students learning outcomes after using the Android Synergy Game media in the fourth graders discussing Sub-theme 2 about Utilization of Natural Resources and Energy Change. The research subjects were the fourth graders of SDN Sukabumi 2, Mayangan District, Probolinggo City, totaling 35 students. The use of this game has proven to be successful in providing significant results in increasing students’ interest, activity and learning outcomes. The results of the questionnaire given to 35 students showed that 34 students (97%) said strongly agree, 1 student (0.3%) said agree, and no student said disagree to the use of Synergy Android Game media, while students learning completeness reached 97% or only 1 got score under the passing grade. Keywords: Synergy game; interest and learning outcomes

Synergi: A Mixed-Initiative System for Scholarly Synthesis and Sensemaking

Efficiently reviewing scholarly literature and synthesizing prior art are crucial for scientific progress. Yet, the growing scale of publications and the burden of knowledge make synthesis of research threads more challenging than ever. While significant research has been devoted to helping scholars interact with individual papers, building research threads scattered across multiple papers remains a challenge. Most top-down synthesis (and LLMs) make it difficult to personalize and iterate on the output, while bottom-up synthesis is costly in time and effort. Here, we explore a new design space of mixed-initiative workflows. In doing so we develop a novel computational pipeline, Synergi, that ties together user input of relevant seed threads with citation graphs and LLMs, to expand and structure them, respectively. Synergi allows scholars to start with an entire threads-and-subthreads structure generated from papers relevant to their interests, and to iterate and customize on it as they wish. In our evaluation, we find that Synergi helps scholars efficiently make sense of relevant threads, broaden their perspectives, and increases their curiosity. We discuss future design implications for thread-based, mixed-initiative scholarly synthesis support tools.Comment: ACM UIST'2

Evaluation of the effectiveness of smart inverters in mitigating voltage variations and fluctuations

With the increasing integration level of photovoltaics (PV) generation into the distribution grid, there would be detrimental impacts on residential customers in the form of rapid voltage variations and overvoltages. These voltage disturbances are caused by intermittent and varying power injections of PV generation. They can be reduced with the installation of smart inverters alongside PV with appropriate control settings. The smart inverters monitor and manage the reactive power exchange between the distribution circuit and the PV system so as to mitigate the adversity of voltage impacts. This thesis analyzes the effects of PV generation in regards to range and variation of voltage in a real-world distribution system. It also evaluates the effectiveness of PV generation equipped with smart inverters in mitigating these issues. A detailed study is conducted on few control strategies that would mitigate increasing range and variability of voltages. This study helps determine suitable control settings that increase the effectiveness of the discussed control functions and improve the overall performance of the distribution systemElectrical and Computer Engineerin

A qualitative study of corporate environmental reporting through the reporting software, Synergi Life

The term sustainability is infiltrating every corner of the society these days, and the corporate world is no exception. The focus among stakeholders and the society at large is on corporations’ responsibility towards society. More information about and insight in a company’s business is becoming more demanded by stakeholders, in order for them to get a better overview of the corporate world’s sustainability. Being transparent, by reporting environmental, social, and governmental (ESG) information, is a relevant measure for companies to give the stakeholder and the society the information they need. However, such reporting is not universally mandatory and there are no universal rules, requirements nor standards that tell companies exactly how ESG reporting should be done and what information such reports must include. Therefore, the way companies report today is often very different from each other which makes it difficult for stakeholders to evaluate and compare similar companies. The latter issue is very broad and can be interoperated and research from many perspectives. This thesis is however, mainly focusing on the environmental aspect of ESG reporting. This is done by using a reporting software called Synergi Life as an example. Synergi Life is an established reporting software owned by DNV (Det Norske Vertias) that makes it possible for companies across sectors to report information on different matter such as, environmental activities. This qualitative study is using collected data from the software as well as participants with different roles to create an example that shows the main challenges of environmental reporting, from the companies’ perspective as well as the stakeholder’s perspective. Relevant theories such as stakeholder theory, shareholder theory and legitimacy theory are used together with other relevant studies that presents the guidelines, initiatives, and requirements of ESG reporting to give a broad foundation for the discussion and analysis

Potential of Grid-Scale PV and Storage Facilities to Participate in Capacity Markets

The recent FERC Order 841 has provided an opportunity for grid-scale PV and battery facilities to participate in the capacity market providing ancillary services. Order 841 combined with steadily decreasing PVand battery costs and increasing demand for renewable energy, gives high potential for a PV and battery facility to be economically viable. In this study, an analysis tool is developed as an auxiliary to the PV + Storage + Control + Grid (PSCG) simulation to estimate the megawatts (MW) of dispatchable capacity that can be reliably offered by a PV and storage facility. Limits are based on grid connection, power electronics, and battery energy capacity, and are impacted by variable insolation, load, and system state. The analysis tool visualizes these limitations and economic potential using a Weibull distribution confidence for the results. The study presents a solution that optimizes the PV and battery sizing along with state of charge reserve to give the highest yield given the adjustable parameters with a modular design that can be fitted to look at numerous other parameters that would affect the outcome

Identification and Characterization of AES-135, a Hydroxamic Acid-Based HDAC Inhibitor That Prolongs Survival in an Orthotopic Mouse Model of Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive, incurable cancer with a 20% 1 year survival rate. While standard-of-care therapy can prolong life in a small fraction of cases, PDAC is inherently resistant to current treatments, and novel therapies are urgently required. Histone deacetylase (HDAC) inhibitors are effective in killing pancreatic cancer cells in in vitro PDAC studies, and although there are a few clinical studies investigating combination therapy including HDAC inhibitors, no HDAC drug or combination therapy with an HDAC drug has been approved for the treatment of PDAC. We developed an inhibitor of HDACs, AES-135, that exhibits nanomolar inhibitory activity against HDAC3, HDAC6, and HDAC11 in biochemical assays. In a three-dimensional coculture model, AES-135 kills low-passage patient-derived tumor spheroids selectively over surrounding cancer-associated fibroblasts and has excellent pharmacokinetic properties in vivo. In an orthotopic murine model of pancreatic cancer, AES-135 prolongs survival significantly, therefore representing a candidate for further preclinical testing