FindingNemo Extraction 2: Phenol-free Method v2

This is a sub-protocol designed to extract/isolate ultra-high molecular weight (UHMW) DNA to obtain ultra-long (UL) reads on Nanopore sequencers using a phenol-free extraction method. A DNA extraction protocol that yields clean and homogeneous UHMW DNA is important for a good UL sequencing output. The choice of protocol should be based on achieving these parameters. Kit-free, phenol-free protocol is a modification of NEB's Monarch HMW DNA Extraction Kit for Cells & Blood, with the option to use SDS or CTAB in the lysis buffer. This protocol also uses glass beads for DNA precipitation matrix. We tested this sub-protocol in human cell line, with input cells of 3 millions but can be varied from 1-5 millions. As a rule of thumb, a million cells will suffice for one load on a MinION.

Microbial analysis and enrichment of anaerobic phenol and p-cresol degrading consortia with addition of AQDS

Quinones and humus are ubiquitous in the biosphere and play an important role in the anaerobic biodegradation and biotransformation of organic acids, poisonous compounds as well as inorganic compounds. The impact of humic model compound, anthraquinone-2, 6-disulfonate (AQDS) on anaerobic phenol and p-cresol degradation were studied. Four methanogenic AQDS-free phenol and p-cresol enrichments and two phenol-AQDS enrichments were obtained using two sludges with potential biodegradability of phenol and cresol isomers as inoculum. 16S rRNA gene-cloning analysis combined with fluorescence in situ hybridization revealed that syntrophic aromatic compound degrading bacterium Syntrophorhabdus aromaticivorans was dominant in four AQDS-free enrichments, whereas phenol degrading Cryptanaerobacter phenolicus was dominant in two phenol-AQDS enrichments. Neither co-culture of Syntrophorhabdus aromaticivorans with Methanospirillum hungatei nor two phenol-AQDS enrichments could metabolize phenol using AQDS as the terminal electron acceptor. Further degradation experiments suggested that C. phenolicus related microbes in two phenol-AQDS enrichments were responsible for the conversion of phenol to benzoate, and benzoate was further degraded by benzoate degraders of Syntrophus aciditrophicus or Sporotomaculum syntrophicum to acetate.

Prenylated Trans-Cinnamic Esters and Ethers against Clinical Fusarium spp.: Repositioning of Natural Compounds in Antimicrobial Discovery

Onychomycosis is a common nail infection mainly caused by species belonging to the F. oxysporum, F. solani, and F. fujikuroi species complexes. The aim of this study was to evaluate the in vitro susceptibility of six representative strains of clinically relevant Fusarium spp. toward a set of natural-occurring hydroxycinnamic acids and their derivatives with the purpose to develop naturally occurring products in order to cope with emerging resistance phenomena. By introducing a prenylated chain at one of the hydroxy groups of trans-cinnamic acids 1–3, ten prenylated derivatives (coded 4–13) were preliminarily investigated in solid Fusarium minimal medium (FMM). Minimal inhibitory concentration (MIC) and lethal dose 50 (LD50) values were then determined in liquid FMM for the most active selected antifungal p-coumaric acid 3,3′-dimethyl allyl ester 13, in comparison with the conventional fungicides terbinafine (TRB) and amphotericin B (AmB), through the quantification of the fungal growth. Significant growth inhibition was observed for prenylated derivatives 4–13, evidencing ester 13 as the most active. This compound presented MIC and LD50 values (62–250 µM and 7.8–125 µM, respectively) comparable to those determined for TRB and AmB in the majority of the tested pathogenic strains. The position and size of the prenylated chain and the presence of a free phenol OH group appear crucial for the antifungal activity. This work represents the first report on the activity of prenylated cinnamic esters and ethers against clinical Fusarium spp. and opens new avenues in the development of alternative antifungal compounds based on a drug repositioning strategy.

Sinapic Acid Esters: Octinoxate Substitutes Combining Suitable UV Protection and Antioxidant Activity

In 2021, Hawaii will permanently ban the use and sale of octinoxate-based sunscreens as studies have shown serious impacts of such UV filters on the coral reef. This ban, which could be generalized to other countries, highlights the extreme need to offer alternative UV filters that are not only effective in terms of sun protection, but also healthy with regards to human health and the environment. In this context, a wide library of p-hydroxycinnamic esters deriving from naturally occurring sinapic acid has been synthesized using a Knoevenagel–Doebner condensation. The UV filtering activities as well as the antioxidant properties of these sinapic acid esters were then investigated. The results showed promising UVB protection and antioxidant efficacy. A Structure–Activity Relationship (SAR) study on the sinapic acid esters highlighted the need of a free phenol to, as expected, observe antioxidant activity, but also to obtain a higher intensity of protection. Moreover, the nature of the ester moiety also proved to be a key structural feature for the UV absorbance, as higher steric hindrance on the ester moiety leads to more active compounds. The judicious structural design of sinapic esters thus provides promising compounds combining UV protection and antioxidant activity.

Effect of hydrothermal treatment on the carbon structure of Inner Mongolia lignite

Understanding the structural properties of lignite during hydrothermal treatment would aid in predicting the subsequent behavior of coal during the pyrolysis, liquefaction, and gasification processes. Here, hydrothermal treatment of Inner Mongolia lignite (IM) was carried out in a lab autoclave. The distribution of carbon in the lignite was monitored via solid 13C nuclear magnetic resonance spectroscopy, and the functional groups of oxygen in lignite were determined by Fourier transform infrared spectroscopy. The curve-fitting method was used to calculate the content of the functional groups quantitatively. The results show that hydrothermal treatment is an effective method for upgrading the lignite. The side chains of the aromatic ring in lignite are altered, while the main macromolecular structure remains nearly the same. The hydrothermal treatment of IM could be divided into three temperature-dependent stages. The first stage (< 493 K) is the decomposition reaction of oxygen functional groups, where the O/C ratio decreases from 0.203 in raw IM to 0.185 for the IM treated at 493 K. In the second stage (493–533 K), hydrolysis of functional groups and hydrogen transfer between water and lignite occur. Here, the ratio of methylene to methyl increases from 0.871 in IM-493 to 1.241 for IM-533, and the content of quinone generates from the condensation of free phenol increased. The third stage (> 533 K) involves breakage of the covalent bond, and the content of CH4 and CO in the emission gas clearly increase.

THE RESEARCH OF CONDITIONS FOR THE FORMATION OF PHENOL FORMALDEHYDE RESINS MODIFIED WITH HYDROXYL CONTAING POLYETHER POLYOL

The aim of the study was to study the conditions of copolycondensation of phenol alcohol with hydroxyl-containing polyether polyol in the presence of maleic anhydride chain extender in order to obtain modified thermoset phenolic-formaldehyde oligomers. Investigation of the dependence of the content of free phenol, formaldehyde and methylol groups on time during the synthesis of the modified phenol-formaldehyde resin with the ratios hydroxyl-containing polyether polyol:phenol alcohol 1:60 mol/mol and 1:100 mol/mol. It was revealed that maleic anhydride, present in the reaction medium as a copolycondensation catalyst, also participates in copolycondensation reactions as a chain extender. Moreover, as the molar fraction of phenol alcohol increases in the synthesis, an increase in the amount of maleic anhydride involved as a chain extender and an increase in the molecular weight of the resulting modified phenol-formaldehyde oligomer are observed.

The study of powdered phenol-formaldehyde resins for the manufacture of plywood

В настоящее время перед отечественным производителем древесных композиционных материалов стоят задачи по увеличению объемов производства, повышению качества и конкурентоспособности, снижению токсичности выпускаемой продукции. Решение этих задач основано на разработке новых и совершенствовании имеющихся технологий современного производства клеёв и композиционных материалов на их основе. Цель исследования – повышение эффективности производства фанеры путём применения клеев на основе порошкообразных термореактивных полимеров поликонденсационного типа. Для достижения поставленной цели в работе необходимо исследовать влияние наполнителя на свойства клеевой композиции и готовой продукции. Исходными компонентами для проведения экспериментов являлись порошковая фенолоформальдегидная смола и аэросил технический. Установлено, что введение аэросила технического в клеящие составы на основе поликонденсационной порошковой фенолоформальдегидной смолы СФЖ-3013 увеличивает прочность фанеры, также с увеличением количества наполнителя уменьшается содержание свободных продуктов в смоле: содержание свободных фенола и формальдегида уменьшается до 0,05 – 0,06% и 0,02 – 0,03% соответственно. Предполагается, что при горячем способе склеивания происходит гелеобразование, частицы аэросила образуют разветвленные цепочки ~Si-O-Si~, которые целиком пронизывают объем клея, этот процесс сопровождается связыванием формальдегида в процессе структурирования. Полученные результаты исследований могут быть использованы в работе специалистов деревообрабатывающих производств при управлении технологическими процессами склеивания. Разработанные составы клеевой композиции на основе фенолоформальдегидных смол, модифицированных (наполненных) техническим аэросилом позволяют сократить расход клеевых материалов; повысить прочность и качество склеиваемой продукции; ускорить процесс отверждения связующего; сократить продолжительность склеивания; снизить энергозатраты путем уменьшения времени склеивания; уменьшить себестоимость клея, за счёт замещения основных компонентов наполнителем. At present, the domestic manufacturer of wood composite materials is faced with tasks to increase production volumes, improve quality and competitiveness, and reduce the toxicity of manufactured products. The solution to these problems is based on the development of new and improvement of existing technologies for the modern production of adhesives and composite materials based on them. The purpose of the study is to increase the efficiency of plywood production by using adhesives based on powdered thermoset polymers of the polycondensation type. To achieve this goal in the work, it is necessary to study the effect of the filler on the properties of the adhesive composition and finished products. The initial components for the experiments were powder phenol-formaldehyde resin and technical aerosil. It was found that the introduction of technical aerosil in adhesives based on polycondensation powder phenol-formaldehyde resin SFZh-3013 increases the strength of plywood, as the amount of filler increases, the content of free products in the resin decreases: the content of free phenol and formaldehyde decreases to 0.05 – 0.06% and 0.02 – 0.03%, respectively. It is assumed that during the hot bonding method, gel formation occurs, so that aerosil particles form branched chains ~ Si-O-Si ~ that completely penetrate the glue volume, this process is accompanied by formaldehyde bonding during structuring. The obtained research results can be used in the work of specialists in woodworking industries in the management of gluing processes. The developed compositions of the adhesive composition based on phenol-formaldehyde resins modified (filled) with technical aerosil allow you to: reduce the consumption of adhesive materials; to increase the strength and quality of glued products; speed up the curing process of the binder; shorten bonding time; reduce energy costs by reducing the bonding time; reduce the cost of glue, due to the replacement of the main components with filler.

Functional Components, Antioxidant Activity and Hypoglycemic Ability Following Simulated Gastro-Intestinal Digestion of Pigments from Walnut Brown Shell and Green Husk

To assess the effects of digestion on the functional components of walnut pigment and their bioactivities, we developed an in vitro model simulating gastro-intestinal digestion. Results showed an increase in the contents of flavonoids and conjugated phenols (with retention rates higher than 100%) in husk pigment after digestion. The lowest of the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging abilities was reached in the group with the minimum flavonoid content after digestion. Close correlation was observed between free phenol content and total reducing power, as the reducing power among different groups of husk pigment was in consistent with free phenols changes. The inhibitory effect of walnut pigment on α-amylase with/without digestion enzyme was similar. However, shell pigment showed improved inhibitory effect on α-glucosidase activity, with an increased inhibitory rate of 5.42%. In general, the antioxidant activity and hypoglycemic ability of walnut pigment were prone to chemical and enzymatic changes during simulated digestion, which were also related to the alteration of flavonoids and phenols.