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Although recent years have seen considerable interest in stimuli‐responsive hydrogels, their strict preparation conditions and narrow applicability limit their use as diverse sensors and soft robots. Herein, a versatile Agar‐Zwirrions hybrid hydrogel actuator (Agar/PSBMA) integrated with simultaneous temperature self‐sensing and wide‐range electrical response is developed. To prepare the Agar/PSBMA hydrogel, a simple and controllable preforming post‐enhancing and mechanical pressing method is used by introducing zwitterions materials into a temperature‐sensitive Agar matrix. Owing to the design, the compact multiplex complementary structure generated by this method and the materials can facilitate the improvement of flexibility, stretchability, and toughness while providing mechanical dissipation and adhesion properties. Importantly, the visible detected temperature self‐sensing ability during 10–40 °C, and quick and wide‐range bending responses of both high‐voltage and low‐voltage electric fields make it unique over other actuators. Furthermore, the electrical response behavior of the hydrogel is found to be impacted by mechanical characteristics and charge polarization based on the finite element Abaqus simulations analysis. The prepared versatile hydrogels show the potential for applications as soft robotics and controlled transportation of adhered substances while simultaneously monitoring their working temperature, which expands the response range of hydrogel actuators and broadens the scope of application. [ABSTRACT FROM AUTHOR]

This paper devised a novel Christmas‐tree structure for flexible neural probes, which features recording sites on side‐branches that can be folded along the shank of the probe by temporary encapsulation before implantation. This design reduces the size of the implant and minimizes the surgical footprint. Upon implantation, the temporary encapsulation dissolves in vivo, leading to self‐stretching of the branches, and an expansion of the lateral detecting range of the probe. We conducted in vitro experiment with our probe in 0.6% agar that mimics the brain tissue environment. The results demonstrate that the neural probe we have developed exhibits self‐stretching ability and provides an increased lateral recording range without causing additional damage to the brain tissue. © 2024 Institute of Electrical Engineer of Japan and Wiley Periodicals LLC. [ABSTRACT FROM AUTHOR]

A good organic single crystal of 2‐Amino‐1‐methyl‐4‐oxo‐4, 5‐di‐hydro‐1H‐imidazol‐3‐ium 4‐methyl‐benzene‐sulfonate (2ADPTS) is grown by slow evaporation technique, and the orthorhombic system with noncentrosymmetric space group (P212121) is corroborated by single‐crystal X‐ray diffraction analysis. The structural parameters of the 2ADPTS crystal are investigated and weighted up with the theoretical interpretation. The Hydrogen–Hydrogen interaction with (43.6%) in 2ADPTS promotes crystal structure packing stabilization. The presence of various functional groups and chemical composition of the 2ADPTS compound is identified by FTIR and FT‐Raman spectroscopy. The natural bond orbital evaluation interprets all the possible hydrogen bonding interactions and charge delocalizations in the crystal structure. Thermal analysis confirmed the thermal stability of the 2ADPTS crystal. The absorption spectrum of the 2ADPTS crystal is recorded using UV–Vis spectroscopy. The third‐order nonlinear property of the 2ADPTS crystal is confirmed by using the Z‐scan technique. The higher significant antibacterial activity of 2ADPTS is carried out by the standard Kirby‐Bauer test against four pathogenic bacteria commonly called the agar well diffusion process. [ABSTRACT FROM AUTHOR]

This article in the Journal of Fish Diseases examines the effects of the fungicide dimethomorph on various species of oomycetes, which are harmful to plants and animals, including humans. The study specifically focuses on three oomycete species that cause significant economic losses in agriculture and aquaculture. Through in vitro tests and scanning electron microscopy, the researchers found that dimethomorph inhibited the growth of the oomycetes and caused morphological changes in one species. The study suggests that more research is needed to fully understand these pathogens and develop effective treatments. [Extracted from the article]

TiO2 nanoparticles are effective alternative to the antibiotics. The quick recombination of the photogenerated charges and the inefficient exploitation of visible light remain the major problems limiting their large‐scale applications. Modifying the TiO2 with Silver that acting as an electron trap, plays an important role in reducing the recombination of TiO2 charges and shift its photo‐response to the visible light region. The present work aims to synthesize AgTiO2 nanoparticles using sol‐gel and coprecipitation methods and evaluating the antimicrobial behavior of them. Also, to find the optimal conditions based on the anatase phase of TiO2 due to its higher antimicrobial activity than the rutile form. Among Ag‐TiO2 nanoparticles obtained, the one prepared by sol‐gel method at 450 °C and PH=8–8.5 exhibits the best antimicrobial property. AgTiO2 nanoparticles have been characterized by means X‐ray Diffraction (XRD), Fourier transform infrared spectroscopy (FTIR),scanning electron microscope (SEM), ζ‐potential, dynamic light scattering (DLS) that showing promising results. Zeta potential values −103 mV has been reported that is considered as a stable colloidal suspension system that prevents nanoparticles aggregation. Antimicrobial behavior of them were evaluated using Diffusion (Agar disk‐diffusion), Dilution (MIC) and Bacterial colony counting methods and diameter of the inhibitory zone 15 mm and minimum inhibitory concentration 5 mg/ml was observed.Colony count results indicated that AgTiO2 able to kill bacteria at the lowest concentration of 5 mg/ml. Outcomes of MTT assay (measure cell cytoxicity) suggesting the absence of toxicity in living Hfff cells (as a normal human fibroblast) while surviving L929 cells (as a cancer cell line originated from mouse fibroblast cells). After 24 and 48 h incubation from 100 % to 77.3 % and 57.2 % was reduced respectively. This indicates that Ag/TiO2 nanoparticles act as a anticancer drug and have inherent selective toxicity nature towards cancer cells while posing no effect to normal cells. AgTiO2 nanoparticles are synthesized using sol‐gel and coprecipitation methods. The sample prepared by the sol‐gel method at a temperature of 450 °C and pH=8.5, which corresponds to the anatase phase of TiO2 synthesis, shows the best antimicrobial properties and acts as an anticancer drug with inherent selective toxicity nature towards cancer cells while posing no effect on normal cells. [ABSTRACT FROM AUTHOR]

The fig‐milk dessert, a traditional and nutritionally rich treat infused with bioactive compounds, was subjected to a comprehensive analysis in this study. The novelty of this research lies in the investigation of the in vitro antioxidant, anticancer, and antimicrobial potential of the fig‐milk dessert. This was accomplished through the utilization of the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assay, Annexin/propidium iodide staining, microtiter plate‐based assay and agar well diffusion, respectively, for the first time. Additionally, the study assessed the total phenols and flavonoid content of the extract using the Folin–Ciocalteu assay and the aluminum chloride method, respectively. The findings revealed that the cooking method exerted a significant influence on the bioactive properties and nutritional composition of the dessert. Among the samples analyzed, CM1, consisting of figs steamed for 2 min and milk heated to 70°C, exhibited remarkable characteristics. This sample demonstrated the highest peptide concentration (1290 mg/L), superior antioxidant and anticancer activities, and favorable sensory attributes. Specifically, CM1 induced apoptosis in 84% of AGS cells and inhibited 68% of free radicals in the DPPH assay. It is noteworthy that the fig‐milk dessert did not exhibit any antibacterial properties. These discerning results carry substantial implications for the development of functional dairy products endowed with both nutritional and potential therapeutic properties. [ABSTRACT FROM AUTHOR]

Hyrcanian forests comprise a long strip along the southern coast of the Caspian Sea in northern Iran. Oriental beech (Fagus orientalis) is one of the most valuable forest tree species in the Hyrcanian forests. Bark necrosis and bleeding spots of oriental beech were observed in Savadkuh forests in Mazandaran province during summer 2021. Ten samples were collected from symptomatic oriental beech trees. The bark canker symptoms included stem tissue necrosis, dark brown to black spots on the trunk, bleeding with dark exudates and dark lesions in the inner bark. Twenty‐six strains of culturable bacteria were isolated from the samples which in culture were predominantly round, convex, smooth with entire margins on nutrient agar and a metallic green sheen pigment on eosin‐methylene blue (EMB) agar. Fourteen strains induced necrosis and browning of the tissues around the inoculation sites on acorns after 2 weeks. Five representative strains also caused dieback in twigs of oriental beech 3 weeks after inoculation. These bacteria were Gram‐negative, catalase positive and oxidase negative. All strains produced colonies with a green metallic sheen on EMB agar. Strains were negative in tests for starch and gelatine hydrolysis or production of levan. A phylogenetic tree based on concatenated sequences of gyrB and infB genes, of representative strains well as in each of the single gene‐based phylogenetic trees, indicated that these strains clustered with Brenneria alni NCPPB 3934T with high bootstrap values. To the best of our knowledge, this is the first report of B. alni associated with bark necrosis and bleeding spots on oriental beech. [ABSTRACT FROM AUTHOR]

The macroalgae residue from the industrial agar extraction process contains a significant amount of carbon and has potential as a renewable feedstock. Unfortunately, it is often overlooked and is poorly utilized. This study aims to valorize this macroalgae residue through pyrolysis to produce silica‐rich biochar and other value‐added products in the form of biocrude oil (BCO) and biopyrolysis gas. The macroalgae residue was pyrolyzed at 300–700 °C with a heating rate of 20–40 °C/min. Yields of biochar, BCO, and gas of 62%, 25%, and 13% were obtained at a temperature of 700 °C and a heating rate of 20 °C/min. Biochar has a porous structure, a surface area exceeding 15 m2/g, and is dominated by amorphous silica of up to 13%. This silica‐rich biochar also contains Na and K, which hold potential benefits in agriculture, serving as soil ameliorants and playing a crucial role in enhancing soil fertility and promoting plant growth. In the meantime, BCO contains 29.3% carboxylic acid group as the most important chemical component. Other than that, the biopyrolysis gas contains mainly CH4 and H2 (up to 24–32%), which can act as chemical building blocks. Finally, a simple business scenario of silica‐rich biochar production reveals that it has a specific cost of 0.37 US$/kg. It could be economically viable as a soil ameliorant or fertilizer. However, challenges persist in scaling up production to an industrial scale. [ABSTRACT FROM AUTHOR]

Encapsulating drugs in poly(lactic‐co‐glycolic acid) (PLGA) nanoparticles can provide a gradual and consistent drug release. Herein, diclofenac sodium‐encapsulated PLGA nanoparticles (DS‐PLGA NPs) were synthesized, characterized, and their effects on bacterial proteolysis and elastolytic activity were evaluated using skim milk agar plate, elastin‐congo red, and staphylolysin disk susceptibility assays. Quantitative PCR was employed to examine the effect of nanoparticles on the expression of lasA/B genes. The DS‐PLGA NPs displayed spherical morphology with an average size of 86 nm, zeta potential of ‐15.2 mV, and DLS size of 124.6 nm. At a sub‐inhibitory concentration, DS‐PLGA NPs inhibited bacterial elastolytic activity by 41.6 to 62.1% during 30–180 min of incubation. The elastolytic curve slopes of samples from PLGA and DS‐PLGA treated groups were respectively 0.0714 and 0.0380 activity/min, indicating a significant reduction of bacterial extracellular elastases by DS‐PLGA NPs. Exposure of bacteria to a sub‐inhibitory concentration of the nanoparticles attenuated the expression of the lasA and lasB genes by 0.21 and 0.31 folds, respectively. Attenuation of the proteolytic activity of P. aeruginosa suggests that the antivirulence potential of the DS‐PLGA NPs along with the anti‐inflammatory properties of diclofenac sodium, can be considered for the treatment of acute and chronic P. aeruginosa infections. [ABSTRACT FROM AUTHOR]

In this research, the biosynthesis method was used for Fe3O4/CuO core‐shell nanocomposite using by aqueous Alcea medicinal plant extract, which aimed to synthesize antibacterial substances against common resistant bacteria. Antibacterial effects of various samples synthesized in different concentrations were evaluated against gram‐positive and negative bacteria in Mueller Hilton agar culture medium. The photocatalytic properties were carried out in a period of 140 minutes, and the maximum effect of the sample was 76 % in 100 minutes. FTIR, SEM, EDX and XRD tests confirmed the correct to design the experiment. synthesis of the nanocomposite with an average size of 38 nm and a surface charge of 23.1 mV. To obtain the best and most effective conditions for the synthesis of antibacterial nanoparticles, the CCD method was used. The main purpose of this research work is to use a medicinal plant as a reducer in the synthesis of metal composite nanoparticles, which plant itself has healing and antimicrobial properties in traditional medical science. [ABSTRACT FROM AUTHOR]

We prepared 81 samples of sweetened viscous solutions and gels with agar concentrations ranging from 0.025% to 0.60% and compared their sweetness intensities to reference solutions without agar. Sweetened viscous solutions containing agar showed significantly higher sweetness intensity in six solutions containing D‐glucose, three solutions containing sucrose, and four solutions containing D‐sorbitol compared to the reference solutions. Notably, some sweetened viscous solutions with extremely low viscosity showed higher sweetness intensity than the reference solution, which is a novel finding. Sweet samples containing agar showed significantly lower sweetness intensity than the reference solution in 15 samples containing D‐glucose, 17 samples containing sucrose, and 15 samples containing D‐sorbitol, supporting previous theories. We performed a simple regression analysis on the sweetness intensity of viscous solutions and gels containing agar and their physical properties. Our results showed a strong correlation between sweetness intensity and physical properties. These findings suggest that as sweetness intensity decreases, physical properties tend to increase. Practical Applications: To prevent noncommunicable diseases such as diabetes, it is recommended to practice carbohydrate restriction and reduce the consumption of sweet foods. In this study, upon being offered extremely low‐viscosity sweetened solutions, we anticipate perceiving a slightly stronger sweetness and an increased sense of satisfaction in contrast to tasting sweetened aqueous solutions without viscosity. These findings hold the potential to provide pioneering insights into the interplay between taste and texture for entities such as food manufacturers. They could facilitate applications designed to elevate the quality of health‐conscious food production. [ABSTRACT FROM AUTHOR]

Research on molecular crystals exhibiting light‐driven actuation has made remarkable progress through the development of various molecules and the identification of driving mechanisms. However, crystals developed to date have been driven mainly by ultraviolet (UV) or blue light irradiation, and driving by red or near‐infrared (NIR) light has not been attempted yet. Herein, a broad‐wavelength light‐driven molecular crystals that exhibit high‐speed bending by photothermal effect is developed. Titanium carbide (Ti3C2Tx) MXene nanosheets are integrated into salicylideneaniline crystals to extend the wavelength range that causes photothermally driven bending to UV, visible, and NIR light. In addition, unlike the thin pristine molecular crystals that show slow photoisomerization‐induced bending only under UV light, the MXene layer enables the molecular crystals to be actuated rapidly regardless of their thickness over a wide range of wavelengths. The hybridization of molecular crystals with MXene, which exhibits strong biocompatibility as well as NIR light‐driven photothermal effect, allows for the bending of the hybrid crystals inside agar phantoms mimicking biological tissue. Last, it is confirmed that MXene hybridization can be extended to common molecular crystals including various salicylideneaniline and anisole derivatives. [ABSTRACT FROM AUTHOR]

The article focuses on identifying a novel splicing mutation in the Ectodysplasin A (EDA) gene within a hypohidrotic ectodermal dysplasia (HED) pedigree, exploring the molecular pathogenesis of HED through genetic testing technologies. Topics covered include the characterization of HED, involvement of specific genes like EDA, and the identification of various mutation types leading to HED, emphasizing the significance of this novel splicing mutation in the EDA gene within a Chinese family.

Adding gelling agents to convert the liquid state of the semen extender to a solid state allows for an increased sperm life span. Gelatin and alginate have been used to study the effects of gelling agents on sperm quality. However, there are other gelling agents that have not been studied, such as agar. In addition, studying different sources of gelling agents or the effect of mixing more than one gelling agent with semen extenders on sperm fertility has received little attention. Therefore, the objective of this study was to evaluate the effect of adding agar and a mixture of gelling agents from different sources to semen extender on ram sperm traits and fertility. The first trial evaluated the effect of the addition of 2.5–3 mg mL−1 of gelatin mixed with 0.5–20 mg mL−1 of agar or alginate to ram semen extender on sperm (motility, progressive motility, live/dead, membrane integrity) and semen (pH) characteristics. The response variables were evaluated 1, 72 and 144 h after storage at 4°C. In the second trial, two sources (feed grade and bacteriological) of gelatin and agar were evaluated on the response variables as in Trial 1. In trial 3, a total of 34 ewes were inseminated with doses supplemented (n = 17) with or without (n = 17) agar and gelatin. The pregnancy rate was diagnosed 40 days after insemination. In general, adding agar and gelatin improves (p <.05) sperm motility, membrane integrity and the ratio of live sperm after 144 h of storage compared to the Control group, regardless of the source (bacteriological or feed grade). However, the pregnancy rate in ewes was not influenced (p ≥.05) by semen doses stored with agar and gelatin. In conclusion, the addition of agar and gelatin preserves ram sperm motility and membrane integrity after 144 of storage at 4°C without affecting the pregnancy rate in inseminated ewes. [ABSTRACT FROM AUTHOR]

A series of novel 1,2‐bissubstituted disulfanes bearing beta‐amino acid, dihydropyrimidine‐2,4‐(1H,3H)‐dione, hydrazide, hydrazone and azole moieties were synthesized. The antibacterial activity was evaluated by determining minimum inhibition (by broth microdilution) and minimum bactericidal (by growth on agar) concentrations. The assessment revealed that MIC values for L. monocytogenes varied between 3.9 and 62.5 μg/mL as well as for S. aureus ranged between 7.8 and 250 μg/mL, with the exception of one compound with much weaker MIC of 500 μg/mL. The MBC values for L. monocytogenes have been found to be of 7.8–250 μg/mL, while S. aureus demonstrated the higher resistance and MBCs varied in the range of 7.8–500 μg/mL. The determined MBC/MIC ratios showed that eleven compounds were classified bactericidal agents for all tested bacteria. [ABSTRACT FROM AUTHOR]

This research aims to produce an antioxidant and biodegradable polysaccharide film by using macroalgae agar and sulfated polysaccharide. Agar and sulfated polysaccharide (fucoidan) were extracted from Gracilaria corticata and Sargassum angustifolium macroalgae. Five treatments were conducted: (A) agar film (1%, W:V), (C) chitosan film (1%, W:V + 1% acetic acid), (AC) agar:chitosan composite (50:50, V:V), (ACF) AC film with fucoidan (0.5%, W:V), and (ACFA) ACF film with citric acid (30% of the dry weight of film) as a cross‐linking agent. Then, 0.75% (V:V) of glycerol was added to all films. The physical, mechanical, antioxidant, color variations, microstructure (SEM), and Fourier transform infrared (FT‐IR) spectroscopy were investigated. Based on the results, modifying the agar film with chitosan improved the mechanical strength, humidity, and solubility in the AC composite film (p <.05). Further, adding sulfated polysaccharide and citric acid cross‐linking agent to the agar–chitosan composite led to a significant decrease in solubility, humidity, and permeability to water vapor in ACFA films (p <.05), indicating strong cross‐linking and reduction in film pores based on the SEM pictures and FTIR results. However, the physical and mechanical properties of the agar‐based film obtained from Gracilaria algae can be improved by adding chitosan and citric acid cross‐linking agent, and the addition of fucoidan obtained from Sargassum algae has improved its antioxidant properties. This biodegradable film can be a good candidate for preserving perishable products. [ABSTRACT FROM AUTHOR]

The present study was aimed at investigating the optimization of extraction variables for food grade quality agar from Gracilaria tenuistipitata, so far, the first study on Bangladeshi seaweed. Water (native)‐ and NaOH (alkali)‐pretreated agars were comparatively analyzed by several physicochemical parameters. All extraction variables significantly affected the agar yield in both extraction conditions. Alkali‐pretreated agar provided a better yield (12–13% w/w) and gel strength (201 g/cm2) in extraction conditions as followed by 2% NaOH pretreatment at 30°C for 3 h, seaweed to water ratio at 1:150, and extraction temperature at 100°C for 2 h. Gelling and melting temperatures, color, and pH values of both agars were found to be comparable with commercial agar. Significantly higher sulfate contents including organic and inorganic and total carotenoids were reported in native (3.14% and 1.29 μg/mL) than that in alkali‐pretreated agar (1.27% and 0.62 μg/mL). FTIR spectrum demonstrated the purity of the agar as characterized by the stronger relative intensity with higher degree of conversion of L‐galactose 6‐sulfate to 3,6‐anhydrogalactose in alkali pretreatment group than that of native ones. Moreover, antioxidant activity (% DPPH scavenging) was observed and confirmed by IC50 values of 5.42 and 9.02 mg/mL in water‐ and alkali‐pretreated agars, respectively. The results suggested that agar from G. tenuistipitata with optimized alkali extraction conditions could promote cost‐effective yield with improved physicochemical characteristics and biofunctional values upon consumption by the consumers as food materials. [ABSTRACT FROM AUTHOR]

Nanocomposite biohydrogels comprised of Xanthan gum, Agar, and Gelatin including different weight ratios of Zinc oxide nanoparticles (ZnO−NPs) have been prepared via gamma irradiation technique in presence of methylene bis acrylamide (MBAm) as a crosslinking agent to be employed as antibacterial environmentally benign formulations. The prepared hydrogels were characterized by several techniques: Fourier‐transform infrared spectroscopy for confirming the chemical structure, X‐ray diffraction for crystallographic features and thermal gravimetric analysis for thermal properties. The AFM and SEM were used for investigating the surface features and morphology of the biohydrogels. The results showed that both Gamma irradiation and incorporation of ZnO−NPs improve the gel (wt %) of the prepared biohydrogel nanocomposite from 42.8±1.5 wt% for the unirradiated composite to 81.2±0.9 wt% after exposed to 20 kGy and reaches to 92±0.68 wt% in addition of 0.5×10−2 g of ZnO−NPs. Also, the effect of ZnO−NPs on the antimicrobial activity of the biohydrogel was verified. It was found that the prepared nanocomposite biohydrogels possess good thermal stability and remarkable antimicrobial activity towards different pathogens, thus they can be used in possible biological applications. [ABSTRACT FROM AUTHOR]

The green route for the synthesis of metal nanoparticles has emerged as a promising pathway due to its simple, non‐toxic, low‐cost, and environment‐friendly nature. In this study, eco‐friendly iron nanoparticles were successfully synthesized from the extract of green tea leaves. The synthesized nanoparticles were characterized by UV‐Visible, FT‐IR, SEM, EDX, TEM, and XRD techniques. The morphological analysis of the nanoparticles by SEM and TEM studies revealed that they were spherical in shape with particle size in the 30–70 nm range. The XRD investigation established the amorphous nature of the nanoparticles. The role of the phytochemicals of the tea extract as the reducing and stabilizing agents for the nanoparticles was confirmed by the FT‐IR analysis. The iron nanoparticles exhibited significant catalytic potential in the degradation of organic dyes‐ malachite green and methylene blue. The reactions followed pseudo‐first‐order kinetics and the percent degradation of 85.20 % for malachite green and 89.42 % for methylene blue was achieved in 20 min. and 10 min. respectively. The nano‐catalyst was reusable for three consecutive cycles of degradation reaction without any significant loss in its activity. The anti‐oxidant activity of the nanoparticles was studied by the DPPH assay wherein the nanoparticles exhibited a promising free radical scavenging ability of about 77 % with the IC50 value of 589.70 μg/mL. The anti‐bacterial activity of the nanoparticles was analyzed by the agar well diffusion method against four bacterial strains: gram‐positive Staphylococcus aureus, Bacillus subtilis, and gram‐negative Escherichia coli, Pseudomonas aeruginosa. The zones of inhibition and the Minimum Inhibitory Concentration (MIC) values of the nanoparticles were determined. The nanoparticles showed moderate toxicity against all the tested bacterial strains. The synthesis of the iron nanoparticles was rapid and simple and the results of the investigated applications were significant and promising. [ABSTRACT FROM AUTHOR]

PVA‐BB/GO‐Fe3+/agar hydrogels (BB is configured by 1:1 configuration of 0.04 mol L−1 boric acid solution and 0.04 mol L−1 borax solution) are designed with excellent mechanical and electrical properties due to their triple cross‐linked network, and they maintain stable performance in various tests, which is sufficient to support the use of hydrogels in multiple scenarios. Meanwhile, the tension responsiveness of the PVA‐BB/GO‐Fe3+/agar hydrogel allows it to rapidly transmit signal changes upon tension changes. Finally, maintaining increased energy dissipation and strength enhancement during the loading–unloading cycle provides the possibility of long, stable operation. In summary, it is believed that PVA‐BB/GO‐Fe3+/agar hydrogel has significant potential in the field of flexible sensors and is expected to be fabricated as a sensor for motion monitoring. [ABSTRACT FROM AUTHOR]

In the present study, a series of novel 1‐Methyl‐3‐(3‐oxo‐3‐phenylprop‐1‐enyl) quinoline‐2‐(1H)‐ones derivatives (3 a–3 i) were synthesized via condensation, followed by the ring closure to produce pyrazoline derivatives (4 a–4 i). All the newly synthesized pyrazolines derivatives were determined by spectroscopic techniques and were tested for antimicrobial activity by agar well diffusion assay and MIC, from which it was observed that the compounds 4 g and 4 i showed substantial antimicrobial activity against bacterial strains. Whereas compound 4 i exhibited remarkable antifungal activity. Post this, computational methodologies were employed for studying the pharmacokinetic profiles and assessing the drug likeness of the title compounds. They were subjected to molecular docking analysis and it was observed that the proposed antibacterial compounds 4 g and 4 i showed significant binding to the active site of NAD+ synthetase enzyme of the bacteria and the proposed antifungal compound 4 i showed significant binding towards the active site of Cytochrome P450 51 in fungi. [ABSTRACT FROM AUTHOR]

Fungi of genera Phlyctema and Neofabraea are the causal agents of bull's eye rot, a major postharvest disease of pome fruits. To investigate their morphological and genetic diversity, isolates obtained in Italy and Chile from decayed fruit and rainwater between 2014 and 2019 were grown on two agar media, inoculated onto four fruit cultivars and compared using four marker genes. Consistent intra‐ and interspecies phenotypic differences were recorded among isolates identified as P. vagabunda (two main morphotypes, PvM‐I and PvM‐II, were distinguished) and N. kienholzii. In particular, the Chilean isolates belonging to PvM‐I showed low sporulation in vitro, while isolates belonging to PvM‐II showed the most abundant sporulation and also formed conidiomata deep within fruit tissue. Host cultivar influenced the disease incidence in unwounded, inoculated fruit. Cripps Pink and Golden Delicious apples favoured the formation of P. vagabunda conidiomata and macroconidia, while Granny Smith apples and/or Kaiser pears restricted sporulation of some isolates of PvM‐I. Mycelial cords of P. vagabunda and N. kienholzii were consistently recorded in inoculated fruit, suggesting their possible involvement as a source of inoculum. Propagules of P. vagabunda were present in rainwater collected from apple plants from September to October in Italy. According to sequence analysis of ITS, EF‐1α, TUB2 and ACT1 regions of the fungi, 12 distinct sequence types were identified, three of which were characteristic of isolates from the Southern Hemisphere. The condensed maximum‐likelihood phylogenetic tree separated the 50 P. vagabunda isolates into six phylogroups, suggesting a correlation with their geographical distribution. [ABSTRACT FROM AUTHOR]

The aim of this in vitro study was to evaluate the interaction between ciprofloxacin and amoxicillin against beta‐lactamase‐producing Staphylococcus aureus. Concentration‐dependent curves for each individual drug were carried out to obtain the mean inhibitory concentration in the agar well diffusion assay. Then, different ratios of the ciprofloxacin–amoxicillin combination (0.5:0.5, 0.8:0.2, 0.2:0.8, 0.9:0.1, 0.1:0.9, 0.95:0.05, and 0.05:0.95) were assessed. Data were analyzed using the isobolographic analysis and interaction index. The isobolographic evaluation shows that the 0.9:0.1 and 0.95:0.05 ratios of the ciprofloxacin–amoxicillin combination produced a synergistic antimicrobial interaction, the 0.8:0.2, 0.2:0.8, 0.1:0.9, and 0.05:0.95 proportions showed an additive antibacterial effect, and the 0.5:0.5 proportion induced antagonistic antimicrobial effects. The interaction index showed similar outcomes to the isobolographic analysis. In conclusion, the data of this study mainly show antimicrobial additive results of the ciprofloxacin–amoxicillin combination against beta‐lactamase‐producing S. aureus. [ABSTRACT FROM AUTHOR]

Christ's thorn (Paliurus spina‐christi Mill.) tree is an important and valuable forestry species and has significant potential in afforestation and reforestation systems. In recent years, dieback symptoms were observed on Paliurus spina‐christi trees in Gilan‐e Gharb region of Kermanshah province, west Iran. During 2020 to 2021, Christ's thorn trees that exhibit branch dieback symptoms were sampled for fungal isolations. The pathogenicity tests, biochemical assay of growth, acid production on creatine sucrose agar (CREA) medium and morphological and molecular investigations (ITS and β‐tubulin regions) identified Paecilomyces formosus as a dieback causal agent. This is the first report of P. formosus pathogenicity on Paliurus spina‐christi trees. Furthermore, disease severity was assayed on 11 tree species under three different temperatures 15, 25 and 35°C. Disease severity caused by all isolates on detached branches was greater at 35°C than at 15 and 25°C. This study presents the host range of this pathogen and showed that these potential hosts are prone to this pathogen under high temperatures, which forest trees experienced in recent decades. [ABSTRACT FROM AUTHOR]

Summary: There is little information on the survival of probiotics in plant‐based foods after simulated gastric and intestinal conditions, likewise the microstructure arrangement in the no‐dairy fermented food. This work aimed to study if the agar–agar in a fermented coconut jelly confers protection to probiotics, phenolic and antioxidant compounds during in vitro digestion. Samples containing higher agar–agar amounts tend to retain (P < 0.05) antioxidant and phenolic compounds in their network better, even after the in vitro digestion. Also, a compact and homogeneous microstructure was observed by the Confocal Laser Scanning Microscopy. The texture profile analysis shows that 1% of agar samples presented the maximum hardness (P < 0.05) due to more bonding points and intermolecular interactions. Finally, the survival of probiotics remained above the recommended values (106–107 CFU g−1) after the in vitro digestion of a product with probiotic potential. [ABSTRACT FROM AUTHOR]

Hull‐less barley grass polysaccharide extracts (HBP) are prepared, and the physicochemical and antioxidant properties of agar films with different HBP contents (0, 0.5, 1, 1.5, and 2% w/w of agar mass) are investigated. Infrared spectroscopy shows that HBP interacts with the agar matrix via intermolecular hydrogen bonds. The incorporation of HBP causes rougher surface morphology with increased film thickness (p < 0.05). The films with HBP exhibit decreased brightness with a greenish and yellowish hue. The flexibility of the films is improved gradually as HBP contents increase from 0 to 1.5% w/w (p < 0.05), however, a slight reduction is observed with 2% w/w HBP addition (p > 0.05). The water resistance and surface hydrophobicity of the films are enhanced by adding HBP (p < 0.05). The antioxidant test reveals the strong antioxidant capacity of the HBP‐loaded films, which is positively correlated with the HBP contents (p < 0.05), with 2% w/w HBP leading to 59.28% DPPH radical scavenging and 69.00% ABTS radical scavenging. Results suggest that HBP is a promising functional agent for developing antioxidant biopolymer films, and the agar/HBP edible films prepared in this study have potential in food packaging fields. [ABSTRACT FROM AUTHOR]

To determine if additional agar plates could allow earlier detection of anaerobes in spinal surgical site infections (SSIs), we performed a prospective study (November 2017‐January 2019) of patients with early spinal SSIs. In addition to routine 14‐day cultures, surgical samples were inoculated onto three additional plates (CDC anaerobe agar with 5% sheep blood [CDC], CDC anaerobe laked sheep blood agar with kanamycin/vancomycin [BBL], and Bacteroides bile esculin [BBE] agar with amikacin (BD, USA)) incubated under anaerobic conditions (72 h, 37°C). The primary endpoint was detection of anaerobes by these methods, as compared to routine culture. Anaerobes were identified in 7/61 patients (11%) using the routine procedure and in one extra case with additional plates (overall detection rate 8/61, 13%). Sensitivity was greater for the CDC plate than for the BBL and BBE plates. When routine culture was positive, the CDC plate was always positive, and in three cases showed at least one additional anaerobe. Using additional agar plates, anaerobes were identified in early spinal SSI in 13% of patients. Within 3 days, CDC agar plate enabled detection of anaerobes in one extra case and at least one additional anaerobe in three other cases, compared to routine 14‐day culture. [ABSTRACT FROM AUTHOR]

Summary: Chitosan and phenolic acid were catalysed by 1‐(3‐dimethylaminopropyl)‐3‐ethylcarbodiimide hydrochloride (EDC)/1‐hydroxybenzotriazole (HOBt) to produce graft copolymer with high performance. The grafting ratios of gallic acid‐grafted chitosan copolymer (GA‐CS, 16.58%) and protocatechuic acid‐grafted chitosan copolymer (PCA‐CS, 14.36%) were determined by ultraviolet–visible spectrometer. The characterisation of products was analysed by Fourier transform infrared spectrometer, proton nuclear magnetic resonance and X‐ray photoelectron spectroscopy. The water solubility of natural chitosan (NA‐CS, 5.63%), GA‐CS (45.05%) and PCA‐CS (85.96%) confirmed that the water solubility of the modified chitosan enhanced after the existence of phenolic acid. 1,1‐diphenyl‐2‐picryl hydrazine (DPPH) scavenging test, β‐carotene rinsing experiment, total reducing power determination and standard agar well diffusion assay method were used to evaluate the antioxidant capacity and antibacterial ability of copolymer. Notably, the antibacterial ability of PCA‐CS and GA‐CS remained relatively consistent, but GA‐CS has the highest antioxidant capacity. This study shows that the EDC/HOBt reaction system can successfully graft phenolic acids onto chitosan without affecting the bioactivity of phenolic acids. In addition, the preparation of food preservation coatings by phenolic acid–chitosan copolymers with high antibacterial and antioxidant properties will have great development potential in the field of food technology. [ABSTRACT FROM AUTHOR]

Both in vitro and in vivo studies have shown that the probiotic Limosilactobacillus reuteri can improve oral health. Limosilactobacillus reuteri species are known to produce the antimicrobial "reuterin" from glycerol. In order to further increase its antimicrobial activity, this study evaluated the effect of the combined use of glycerol and Limosilactobacillus reuteri (ATCC PTA 5289) in view of using a synergistic synbiotic over a probiotic. An antagonistic agar growth and a multispecies biofilm model showed that the antimicrobial potential of the probiotic was significantly enhanced against periodontal pathobionts and anaerobic commensals when supplemented with glycerol. Synbiotic biofilms also showed a significant reduction in inflammatory expression of human oral keratinocytes (HOK‐18A), but only when the keratinocytes were preincubated with the probiotic. Probiotic preincubation of keratinocytes or probiotic and synbiotic treatment of biofilms alone was insufficient to significantly reduce inflammatory expression. Overall, this study shows that combining glycerol with the probiotic L. reuteri into a synergistic synbiotic can greatly improve the effectiveness of the latter. [ABSTRACT FROM AUTHOR]

Background: Malassezia (M.) pachydermatis as a frequent reason for dermatological consultation in dogs and cats was recently shown to be lipid‐dependent, too. Lipolytic activity is a prerequisite for activating antimicrobial effectivity of fatty acid esters. Objectives: It was therefore of interest whether it is possible to induce this mechanism in M. pachydermatis and to identify possible differences between minimal and strong lipid‐dependent strains. Methods: In an agar dilution test, the minimal inhibitory concentrations of six fatty acid esters were determined for seventeen M. pachydermatis strains. GC analysis of parent compounds and liberated fatty acids was used to quantify ester cleavage. Results: Hydrolysis was observed in all test strains in a homogenous manner but was dependent on the chemical structure. Lowest MICs (500 ppm after 14 days of incubation) were obtained applying glyceryl monocaprylate and 3‐hydroxylpropyl caprylate, while the corresponding esters of undecylenic acid showed nearly twice the value. As shown by GC analysis with the reference strains CBS 1879 and CBS 1892 and 3‐hydroxypropyl caprylate, hydrolysis and caprylic acid formation starts immediately and was dependent on yeast density. Furthermore, nine azole‐resistant strains isolated from dogs with treatment failures showed MIC values comparable to the other strains and no resistance to monohydric fatty acid esters. Conclusions: Medium‐chain fatty acid esters may represent a new therapeutic option for veterinary use even in azole‐resistant strains. The in vivo verification in M. pachydermatis‐associated dermatitis in dogs and cats will be the next step for the successful development of new therapeutics. [ABSTRACT FROM AUTHOR]

Phosphate is widely used in surimi products to improve the gel properties. However, excess addition of phosphate occurs, which can harm the consumer's health. This study aimed to evaluate the effects of agar gum and fucoidan on maintaining the gel properties of surimi products instead of phosphate. Interestingly, our results showed that 0.125% of agar gum and fucoidan to replace phosphate could enhance water‐holding capacity and maintain gel strength and textual properties of surimi products well. Especially at frozen storage for 1 year, 0.125% of agar gum reduced the expressible moisture content of surimi products by around 10% (p <.05). Sensory evaluation showed that 0.125% of agar gum and fucoidan instead of phosphate can improve tissue and fondness of surimi products in refrigerated storage for 24 h but not in frozen storage for 1 year. The addition of agar gum and fucoidan at a high concentration >0.50% increased the WHC, but significantly decreased gel strength and springiness of surimi products (p <.05). Particularly, 1.00% of agar gum and fucoidan reduced gel strength by around 20% (p <.05). It might be due to the destruction of the gel network structure of surimi protein following the excess addition of these polysaccharides. It can be concluded that 0.125% of agar gum and fucoidan can replace phosphate to develop high‐quality surimi products, and excessive addition of them have negative effects. [ABSTRACT FROM AUTHOR]

Biocatalytic oxidation reactions are in high demand. Among the applied enzymes, the heme‐thiolate enzyme subfamily of unspecific peroxygenases (UPOs), which relies on hydrogen peroxide as cosubstrate and oxidant, has generated great interest. Almost two decades after their first description, databases provide thousands of putative UPO sequences, but only a few enzymes have been characterized. To address this gap, efficient screening methods for the identification of novel UPOs are necessary. Here, a new screening strategy is presented based on solid cultivation of wild‐type fungi. The identification of a promising candidate strain highlights the applicability of this approach. [ABSTRACT FROM AUTHOR]

Background: Among bacterial pathogens, Staphylococcus aureus which lives in the mammary gland is the leading cause of bovine mastitis worldwide, which causes enormous economic losses to the dairy industry. Objectives and methods: The study was carried out cross‐sectionally to determine the occurrence of methicillin‐resistant S. aureus (MRSA) and the risk factors for mastitis infection in dairy cows in and around Adama from October 2014 to June 2015. This particular study included 384 animals. Milk samples were collected and screened with California mastitis test. Then bacteria were cultured and identified using biochemical tests and disc diffusion test was used to determine the antimicrobial sensitivity for MRSA. Results: The prevalence of mastitis was 121 (31.5%). Among them, 37 cases (30.6%) were clinical mastitis and 84 cases (69.4%) were subclinical mastitis. Among these positive cases, 37 cases (30.6%) of S. aureus were isolated. The prevalence of mastitis was significantly related to the breed, age, floor type and sanitation status of the milking (p <0.05). The Kirby–Bauer disc diffusion method was performed on Mueller Hinton agar medium according to NCCLS guidelines to test antibiotic sensitivity. The 32.4% of S. aureus isolates were resistant to oxacillin. Isolates of MRSA are more resistant to amoxicillin (75%), oxytetracycline (66.7%) and sulfa (50%). However, they were sensitive to kanamycin (75%), streptomycin (58.3%) and nalidixic acid (50%). Insufficient dosage, short treatment time and chronic infection in dairy cows in the herd are the main reasons for the large number of resistant strains. Conclusion and recommendation: Generally, mastitis was prevalent in the area, and it was resistant to commonly used antibiotics. Therefore, hygienic, prevention and alternative treatment methods should be implemented. [ABSTRACT FROM AUTHOR]

Background: Azole resistance screening in Aspergillus fumigatus isolates can be routinely carried out by using azole‐containing plates (E.Def 10.2 method), that requires filtering conidial suspensions prior inoculum adjustment. Objectives: We evaluated whether skipping the filtration step of conidial suspensions negatively influences the performance of the E.Def 10.2. Patients/Methods: A. fumigatus sensu stricto isolates (n = 92), classified as azole‐susceptible or azole‐resistant according to the EUCAST microdilution E.Def 9.4 method, were studied. Azole‐resistant isolates had either wild type cyp51A gene sequence (n = 3) or the TR34‐L98H (n = 26), G54R (n = 5), TR46‐Y121F‐T289A (n = 1), F46Y‐M172V‐N248T‐D255E‐E427K (n = 1), F165L (n = 1) or G448S (n = 1) cyp51A gene substitutions. In‐house azole‐containing agar plates were prepared according to the EUCAST E.Def 10.2 procedure. Conidial suspensions were obtained by adding distilled water (Tween 20 0.1%). Subsequently, the suspensions were either filtered or left unfiltered prior to inoculum adjustment to 0.5 McFarland. Using microdilution as the gold standard, agreement, sensitivity and specificity of the agar plates inoculated with two inoculums were assessed. Results: Agreements for the agar screening method with either unfiltered or filtered conidial suspensions were high for itraconazole (100%), voriconazole (100%) and posaconazole (97.8%). Sensitivity (100%) and specificity (98.2%) of the procedure to rule in or out resistance when unfiltered suspensions were used were also high. Isolates harbouring the TR34‐L98H, G54R and TR46‐Y121F‐T289A substitutions were detected with the modified method. Conclusions: Unfiltered conidial suspensions does not negatively influence the performance of the E.Def 10.2 method when screening for A. fumigatus sensu stricto. [ABSTRACT FROM AUTHOR]

Current strategies to improve the throughput of continuous directed evolution technologies often involve complex mechanical fluid‐controlling system or robotic platforms, which limits their popularization and application in general laboratories. Inspired by our previous study on bacterial range expansion, in this study, we report a system termed SPACE for rapid and extensively parallelizable evolution of biomolecules by introducing spatial dimensions into the landmark phage‐assisted continuous evolution system. Specifically, M13 phages and chemotactic Escherichia coli cells were closely inoculated onto a semisolid agar. The phages came into contact with the expanding front of the bacterial range, and then comigrated with the bacteria. This system leverages competition over space, wherein evolutionary progress is closely associated with the production of spatial patterns, allowing the emergence of improved or new protein functions. In a prototypical problem, SPACE remarkably simplified the process and evolved the promoter recognition of T7 RNA polymerase (RNAP) to a library of 96 random sequences in parallel. These results establish SPACE as a simple, easy to implement, and massively parallelizable platform for continuous directed evolution in general laboratories. Synopsis: The spatial competition among phages that infect range‐expanding host bacteria is exploited to develop a spatial continuous directed evolution system that is simple, easy to implement, and massively parallelizable. SPACE is a system for rapid and parallelizable evolution of biomolecules, which introduces spatial dimensions into the phage‐assisted continuous evolution system.The system leverages competition over space, wherein evolutionary progress is closely associated with the production of spatial patterns, allowing the emergence of improved or new protein functions.SPACE is applied to evolve the promoter recognition of T7 RNA polymerase to a library of 96 random sequences in parallel. [ABSTRACT FROM AUTHOR]

Fungi offer a deep source of natural products but remain underutilized. Most biosynthetic gene clusters (BGCs) that can be detected are silent or "cryptic" in standard lab cultures and their products are thus not interrogated in routine screens. As genetic alterations are difficult and some strains can only be grown on agar, we have herein applied an agar‐based high‐throughput chemical genetic screen to identify inducers of fungal BGCs. Using R. solani and S. sclerotiorum as test cases, we report 13 cryptic metabolites in four compound groups, including sclerocyclane, a natural product with a novel scaffold. Steroids were the best elicitors and follow‐up studies showed that plant‐steroids trigger sclerocyclane synthesis, which shows antibiotic activity against B. plantarii, an ecological competitor of S. sclerotiorum. Our results open new paths to exploring the chemical ecology of fungal‐plant interactions and provide a genetics‐free approach for uncovering cryptic fungal metabolites. [ABSTRACT FROM AUTHOR]

Fungi offer a deep source of natural products but remain underutilized. Most biosynthetic gene clusters (BGCs) that can be detected are silent or "cryptic" in standard lab cultures and their products are thus not interrogated in routine screens. As genetic alterations are difficult and some strains can only be grown on agar, we have herein applied an agar‐based high‐throughput chemical genetic screen to identify inducers of fungal BGCs. Using R. solani and S. sclerotiorum as test cases, we report 13 cryptic metabolites in four compound groups, including sclerocyclane, a natural product with a novel scaffold. Steroids were the best elicitors and follow‐up studies showed that plant‐steroids trigger sclerocyclane synthesis, which shows antibiotic activity against B. plantarii, an ecological competitor of S. sclerotiorum. Our results open new paths to exploring the chemical ecology of fungal‐plant interactions and provide a genetics‐free approach for uncovering cryptic fungal metabolites. [ABSTRACT FROM AUTHOR]

Background: Lipophilic basidiomycetous yeasts of the Malassezia genus can cause various skin diseases, such as seborrheic dermatitis, pityriasis versicolor, folliculitis and atopic dermatitis, and even life‐threatening fungemia in newborns and immunocompromised individuals. Routine mycological media used in clinical practice do not contain sufficient lipid ingredients required for the growth of Malassezia species. A recently developed medium, FastFung agar, is promising for culturing fastidious fungal species. Methods: In this study, we compared FastFung agar and mDixon agar for culturing Malassezia species from nasolabial fold and retroauricular specimens of 83 healthy individuals and 187 and 57 patients with acne vulgaris and seborrheic dermatitis, respectively. Results: Malassezia species were identified using conventional tests and matrix‐assisted laser desorption/ionisation mass spectrometry. In total, 96 of 654 samples (14.6%) contained Malassezia species. The total isolation rate was significantly higher in patients with seborrheic dermatitis (40.4%) than in healthy volunteers (21.7%; p <.05), and the rate of M. furfur isolation was significantly higher for patients with acne vulgaris (13.9%) and seborrheic dermatitis (24.6%) than for healthy individuals (1.5%; p <.05). FastFung agar was superior to mDixon agar in M. furfur isolation (p =.004) but showed similar performance in the case of non‐M. furfur species (p >.05). Among cultured Malassezia species, perfect agreement between mDixon agar and FastFung agar was found only for M. globosa (κ = 0.90). Conclusion: Our results indicate that FastFung agar favours the growth of Malassezia species and should be useful in clinical mycology laboratories. [ABSTRACT FROM AUTHOR]

Gracilariales is a clade of florideophycean red macroalgae known for being the main source of agar. We present a de novo genome assembly and annotation of Gracilaria domingensis, an agarophyte alga with flattened thallus widely distributed along Central and South American Atlantic intertidal zones. In addition to structural analysis, an organizational comparison was done with other Rhodophyta genomes. The nuclear genome has 78 Mbp, with 11,437 predicted coding genes, 4,075 of which did not have hits in sequence databases. We also predicted 1,567 noncoding RNAs, distributed in 14 classes. The plastid and mitochondrion genome structures were also obtained. Genes related to agar synthesis were identified. Genes for type II galactose sulfurylases could not be found. Genes related to ascorbate synthesis were found. These results suggest an intricate connection of cell wall polysaccharide synthesis and the redox systems through the use of L‐galactose in Rhodophyta. The genome of G. domingensis should be valuable to phycological and aquacultural research, as it is the first tropical and Western Atlantic red macroalgal genome to be sequenced. [ABSTRACT FROM AUTHOR]

Introduction: Calreticulin (CALR) mutations in myeloproliferative neoplasms (MPN) have been reported to be key markers in the molecular diagnosis, particularly in patients lacking JAK2 V617F mutation. In most current reports, CALR mutations were analysed by either allele‐specific PCR (AS‐PCR), or the more expensive quantitative real‐time PCR, pyrosequencing and next‐generation sequencing. Hence, we report the use of an alternative method, the conformation sensitive gel electrophoresis (CSGE) for the detection of CALR mutations in BCR‐ABL1‐negative MPN patients. Methods: Forty BCR‐ABL1‐negative MPN patients' DNA: 19 polycythemia vera (PV), 7 essential thrombocytosis (ET) and 14 primary myelofibrosis (PMF), were screened for CALR mutations by CSGE. PCR primers were designed to amplify sequences spanning between exons 8 and 9 to target the mutation hotspots in CALR. Amplicons displaying abnormal CSGE profiles by electrophoresis were directly sequenced, and results were analysed by BioEdit Sequence Alignment Editor v7.2.6. CSGE results were compared with AS‐PCR and confirmed by Sanger sequencing. Results: CSGE identified 4 types of mutations; 2 PMF patients with either CALR type 1 (c.1099_1150del52) or type 2 (c.1155_1156insTTGTC), 1 ET patient with nucleotide deletion (c.1121delA) and insertion (c.1190insA) and 1 PV patient with p.K368del (c.1102_1104delAAG) and insertion (c.1135insA) inframe mutations. Three patients have an altered KDEL motif at the C‐terminal of CALR protein. In comparison, AS‐PCR only able to detect two PMF patients with mutations, either type 1 and type 2. Conclusion: CSGE is inexpensive, sensitive and reliable alternative method for the detection of CALR mutations in BCR‐ABL1‐negative MPN patients. [ABSTRACT FROM AUTHOR]

Culture-based Salmonella isolation consists of nonselective pre-enrichment, followed by selective enrichment in Rappaport-Vassiliadis (RV) or tetrathionate (TT) broths, and subsequent plating on selective indicator agar. This study aimed to assess the recovery of two strains belonging to serovars Montevideo (strain ATCC-8387) and Typhimurium (strain ATCC-14028) when grown together in different media. The two strains were co-inoculated 1:1, 10:1, 100:1, and the reciprocal, and CRISPR-SeroSeq was used to assess the relative frequency of both strains after each culture step. Individually, there was no growth difference between both strains in universal preenrichment (UP) or RV broths, though both strains had a higher growth in the former. In TT, both strains had reduced growth, especially ser. Montevideo-8387. When the strains were combined, the growth of ser. Typhimurium-14028 strain was higher in UP and TT broths, while the ser. Montevideo strain was higher in RV broth. The ser. Typhimurium strain also grew better on xylose lysine tergitol-4 (XLT-4) agar. Using media that are commonly used for Salmonella isolation, this work reveals strain-tostrain differences in growth and that in three conditions (UP, RV, XLT-4), these differences were only manifested when the strains were in competition with each other. These findings illustrate the importance of dual selective enrichments to be able to capture all Salmonella present in a sample. [ABSTRACT FROM AUTHOR]

To develop a sterilization method for controlling post-harvest stem-end rot of mango, we examined to sterilize fungi on peduncle by using the dielectric barrier discharge (DBD). Additionally, to investigate the sterilization factors, we performed sterilization against stem-end rot on agar medium by using the DBD generated in air, argon gas, or oxygen gas. It was found that the DBD can sterilize fungi with high probability. Ultra-violet ray and O3 do not contribute to the sterilization. High energy electrons, and radicals such as O and OH play an essential role for the sterilization. [ABSTRACT FROM AUTHOR]

Summary: Phage contamination is a common complication for the fermentation and pharmaceutical industries. The risk of bacteriophage contamination in laboratory processes increases with multiple rounds of genetic manipulation such as deletion and complementation. The contamination of temperate phages does not lead to immediate host cell lysis but could become a serious issue when the lytic cycle is activated under specific conditions. Our objective was to develop a quick and reliable detection method for checking possible temperate phage contamination. Here, using motility plates, we found that when the strain carries a newly acquired temperate phage, its presence can be easily detected by the formation of a clear 'lysis zone' when swimming against the original strain on the same swimming plates. Compared to the traditional double agar layer method and genomic sequencing‐based methods, the duration of the motility‐based assay is shorter and the procedure is simplified. More importantly, for the bacterial strains that already contain active prophages, this method can still easily detect the newly acquired phages without tedious phage identification procedure. These features make this method highly applicable to laboratory and industrial processes. [ABSTRACT FROM AUTHOR]

The presence of sulfate groups in agar compromises the agar quality by affecting the crosslinking during gelling process. Some arylsulfatases can catalyze the hydrolysis of sulfate bonds in agar to improve the agar quality. Immobilized arylsulfatases prove beneficial advantages for their industrial applications. Here, a previously characterized mutant arylsulfatase K253H/H260L was immobilized on the synthesized magnetic Fe3O4 nanoparticles after functionalization by tannic acid (MNPs@TA). The surface properties and molecular structures of the immobilized arylsulfatase (MNPs@TA@ARS) were examined by scanning electron microscopy and Fourier transform infrared spectroscopy. Enzymatic characterization showed that MNPs@TA@ARS exhibited shifted optimal temperature and pH with deviated apparent Km and Vmax compared to its free counterpart. The immobilized arylsulfatase demonstrated improved thermal and pH stability and enhanced storage stability with modest reusability. In addition, MNPs@TA@ARS displayed enhanced tolerance to various inhibitors and detergents. The utilization of the immobilized arylsulfatase for agar desulfation brought the treated agar with improved quality. [ABSTRACT FROM AUTHOR]

Summary: Neoagarobiose (NA2) derived from agar marine biomass is a rare reagent that acts as an anti‐melanogenesis reagent and moisturizer. Here, for the economical manufacturing of NA2, we developed the co‐secretory production system of endo‐type β‐agarases (DagA) and exo‐type β‐agarases (EXB3) in Corynebacterium glutamicum. For this purpose, we first developed a secretory system of DagA via Tat pathway. To improve the secretion efficiency, we coexpressed two Tat pathway components (TatA and TatC), and to improve the purity of secreted DagA in the culture supernatant, two endogenous protein genes (Cg2052 and Cg1514) were removed. Using the engineered strain (C. glutamicum SP002), we confirmed that DagA as high as 1.53 g l‐1 was successfully produced in the culture media with high purity (72.7% in the supernatant protein fraction). Next, we constructed the expression system (pHCP‐CgR‐DagA‐EXB3) for the simultaneous secretion of EXB3 via Sec‐pathway together with DagA, and it was clearly confirmed that DagA and EXB3 were successfully secreted as high as 54% and 24.5%, respectively. Finally, using culture medium containing DagA and EXB3, we successfully demonstrated the conversion of high‐concentration agar (40 g l‐1) into NA2 via a two‐stage hydrolysis process. Agar is a potential and renewable raw macromolecule in macroalgae, and neoagarobiose (NA2) generated by hydrolysis of agar, is a rare reagent acting as a moisturizing and anti‐melanogenesis. Here, we report the development of two types of β‐agarases (endo‐ and exo‐type) secretory system in C. glutamicum for economical production of NA2. [ABSTRACT FROM AUTHOR]

Purpose: Chemical shift‐encoded magnetic resonance imaging enables accurate quantification of liver fat content though estimation of proton density fat‐fraction (PDFF). Computed tomography (CT) is capable of quantifying fat, based on decreased attenuation with increased fat concentration. Current quantitative fat phantoms do not accurately mimic the CT number of human liver. The purpose of this work was to develop and validate an optimized phantom that simultaneously mimics the MRI and CT signals of fatty liver. Methods: An agar‐based phantom containing 12 vials doped with iodinated contrast, and with a granular range of fat fractions was designed and constructed within a novel CT and MR compatible spherical housing design. A four‐site, three‐vendor validation study was performed. MRI (1.5T and 3T) and CT images were obtained using each vendor's PDFF and CT reconstruction, respectively. An ROI centered in each vial was placed to measure MRI‐PDFF (%) and CT number (HU). Mixed‐effects model, linear regression, and Bland‐Altman analysis were used for statistical analysis. Results: MRI‐PDFF agreed closely with nominal PDFF values across both field strengths and all MRI vendors. A linear relationship (slope = −0.54 ± 0.01%/HU, intercept = 37.15 ± 0.03%) with an R2 of 0.999 was observed between MRI‐PDFF and CT number, replicating established in vivo signal behavior. Excellent test‐retest repeatability across vendors (MRI: mean = −0.04%, 95% limits of agreement = [−0.24%, 0.16%]; CT: mean = 0.16 HU, 95% limits of agreement = [−0.15HU, 0.47HU]) and good reproducibility using GE scanners (MRI: mean = −0.21%, 95% limits of agreement = [−1.47%, 1.06%]; CT: mean = −0.18HU, 95% limits of agreement = [−1.96HU, 1.6HU]) were demonstrated. Conclusions: The proposed fat phantom successfully mimicked quantitative liver signal for both MRI and CT. The proposed fat phantom in this study may facilitate broader application and harmonization of liver fat quantification techniques using MRI and CT across institutions, vendors and imaging platforms. [ABSTRACT FROM AUTHOR]

In this study, the antimicrobial effect and DNA gyrase inhibitor potential of vanillin‐based pyridyl–substituted fluoro‐indolines were evaluated. These compounds are synthesized and established through green‐chemistry approaches. The inhibition effect on both DNA gyrase A and B was evaluated in silico and in vitro. Agar well diffusion method–based antimicrobial activity against Gram‐ve Pseudomonas aeruginosa (MTCC 424) and Escherichia coli (MTCC 443), Gram+ve Streptococcus pyogenes (MTCC 442) and Staphylococcus aureus (MTCC 96), and a clinical isolate of Candida albicans (Fungi) was evaluated. The cytotoxicity of the compounds was assessed over macrophages using the MTT assay. In the results, the target compounds exhibited a broad‐spectrum antimicrobial activity against both bacterial types and fungal. [ABSTRACT FROM AUTHOR]

Summary: Few microbes can grow under laboratory conditions, highlighting the fact that the majority of microbes in environment are still uncultured and untapped resources. This study used alternative cultivation methods, diffusion chambers (DC), dilution‐to‐extinction culture (DTE) and modified agar preparation step (PS media) to cultivate previously uncultured marine bacterial species. These methods were applied to samples from a coastal intertidal zone, and the results were compared with those from standard direct plating (SDP) cultivation. Among the strains isolated with DC, DTE and PS media methods, 28%, 48% and 33% were novel species, respectively, while the SDP method resulted in the isolation of only 9% of novel species. Most isolates were unique to the method used for their cultivation. This implies that each method is selective in its own way, which is different from SDP, thus able to access species that are difficult to obtain using conventional approaches. Comparing the diversity showed that 75 genera were recovered by the alternative methods, 2.7 times higher than that of the SDP cultivation, which constituted 45% of total diversity from culture‐independent sequencing. We conclude that combining alternative cultivation methods represents a highly promising key for accessing 'microbial dark matter'. [ABSTRACT FROM AUTHOR]