Equal Concentrations Research Articles

Quantitative Kinetics of the Rapid Bromination of Isoxazole in Bromide-Free Aqueous Medium using Hydrodynamic Voltammetry

The kinetics of rapid bromination of isoxazole by molecular bromine at pH 4.7 have been studied quantitatively. Bromination of isoxazole is an electrophilic substitution reaction where the inductive effect and resonance effect govern reactivity and orientation respectively. Isoxazoles have a broad range of applications in pharmaceuticals and therapeutics. Hence the quantitative kinetic study will help in these applications. The bromination reactions are rapid electrophilic substitution reactions with a half-life of a few seconds, therefore the kinetic data regarding the bromination was lacking thus far. A distinct technique known as hydrodynamic voltammetry was used to perform the kinetics of the reaction. Only molecular bromine gets reduced on the working electrode during the reactions. As the solution lacked bromide ions, a decrease in the concentration of molecular Br2 was measured with reference to diffusion current using a rotating platinum electrode (RPE). During the work, a saturated calomel electrode (SCE) was used as a reference electrode. Bromination of isoxazole followed second-order kinetics with an equal initial concentration of reactants. Thermodynamic parameters were obtained based on rate constants at five different temperatures. The obtained rate constants and activation energy were used to comment on the mechanism of the electrophilic substitution reaction. Thus, the present work quantitatively verifies the reactivity of the isoxazole in the bromination reaction.

Influence of host-specific and locally isolated multi-strain probiotics on piglet performance, mortality, inflammatory response, and gut microbiome.

This study aimed to assess the impact of host-specific and locally isolated multi-strain probiotics on piglet performance, mortality, inflammatory responses, and gut microbiome. A total of 52 piglet litters-34 from Landrace sows and 18 from Large White sows-were allocated to two groups: a control group and a multi-strain probiotic group. The probiotic group comprised seven strains of lactic acid bacteria (MLAB): Lactobacillus brevis, Lactobacillus reuteri, Lactobacillus paraplantarum, Lactococcus lactis, Lactobacillus pentosus, Weissella cibaria, and Pediococcus pentosaceus. Each strain was included in equal concentrations, resulting in a final liquid mixture containing 109 CFU/mL. The MLAB group received the probiotics orally starting from 7 days of age until weaning at four weeks. Following weaning, supplementation continued via feed spraying for an additional four weeks. MLAB supplementation did not significantly affect piglet performance but showed a trend towards reducing the mortality rate (p = 0.06). It influenced the inflammatory response by upregulating the expression of anti-inflammatory cytokines interleukin (IL)-4 and IL-10 (p<0.05). Microbial community analysis indicated that MLAB supplementation increased both microbial diversity (Simpson index: p = 0.06) and species richness (Chao1 index: p = 0.02). Piglets receiving MLAB had a significantly higher abundance of the phylum Firmicutes (p<0.01) compared to the control group, while the abundance of the phylum Bacteroidota was markedly reduced (p<0.01). In addition, the relative abundance of the bacterial genera Prevotellaceae_NK3B31 (p<0.01) and Chlamydia (p = 0.03) was lower in the MLAB group. Overall, these results suggest that while MLAB supplementation does not directly improve piglet growth performance, it has the potential to improve immune function and promote a healthier gut microbiota in weaning piglets, which could ultimately reduce mortality rates.

Cupric citrate supplementation improves growth performance, nutrient utilization, antioxidant capacity, and intestinal microbiota of broilers.

This study aimed to examine the impact of cupric citrate on broilers and compare it with the copper sulfate groups and a control group. A total of 360 1-day-old Ross 308 broilers were randomly assigned into 5 groups, each with 6 replicates of 12 broilers per treatment. The control group was fed a basal diet without any copper supplementation. In contrast, the other groups received basal diets supplemented with either 50 mg/kg (CS-50) or 100 mg/kg (CS-100) of copper in the form of copper sulfate, or 50 mg/kg (CC-50) or 100 mg/kg (CC-100) of copper in the form of cupric citrate, for a period of 42 days. The results showed that copper supplementation affected the average daily gain from day 1 to 21 (p = 0.026) and day 1 to 42 (p = 0.025) in a source-dependent manner. Copper source also influenced the energy digestibility (p = 0.004), with the CC-100 being the most effective treatment. Notably, birds in the CC-100 groups had significantly reduced concentrations of Escherichia coli (p < 0.05) in the cecum, and the Lactobacillus in the ileum, compared to the control group. Dietary copper supplementation also increased the pH in the duodenum (p < 0.05) irrespective of the sources and levels. In addition, the source of copper affected the activities of ceruloplasmin (p = 0.014) and Cu-Zn superoxide dismutase (p = 0.025) in the serum, with the CC-100 group showing the highest levels of both enzymes. Copper supplementation generally improves the growth, nutrient utilization, intestinal microflora, gastrointestinal pH, and antioxidant defences of broilers. Moreover, cupric citrate is as effective as copper sulfate even at equal or lower concentrations.

Optimization of 18 S rRNA metabarcoding for the simultaneous diagnosis of intestinal parasites

Recent advancements in next-generation sequencing (NGS) technologies have created new opportunities for comprehensive screening of multiple parasite species. In this study, we cloned the 18 S rDNA V9 region of 11 species of intestinal parasites into plasmids. Equal amounts and concentrations of these 11 plasmids were pooled, and amplicon NGS targeting the 18 S rDNA V9 region was performed using the Illumina iSeq 100 platform. A total of 434,849 reads were identified, and all 11 parasite species were detected, although the number of output reads for each parasite varied. The read count ratio, in descending order, was as follows: Clonorchis sinensis, 17.2%; Entamoeba histolytica, 16.7%; Dibothriocephalus latus, 14.4%; Trichuris trichiura, 10.8%; Fasciola hepatica, 8.7%; Necator americanus, 8.5%; Paragonimus westermani, 8.5%; Taenia saginata, 7.1%; Giardia intestinalis, 5.0%; Ascaris lumbricoides, 1.7%; and Enterobius vermicularis, 0.9%. We found that the DNA secondary structures showed a negative association with the number of output reads. Additionally, variations in the amplicon PCR annealing temperature affected the relative abundance of output reads for each parasite. These findings can be applied to improve parasite detection methodologies and ultimately enhance efforts to control and prevent intestinal parasitic infections.

Antimicrobial Activity of Commercial Essential Oils Available in the Market of Kathmandu

Given the increasing global apprehension surrounding antibiotic resistance, there has been growing interest in exploring natural compounds, specifically essential oils, as potential substitutes for combating microbial pathogens. This research work aimed to evaluate the antimicrobial activity exhibited by commercial essential oils and by diluting them with the carrier oils at equal concentration. The screening of antimicrobial activity was conducted using the agar well diffusion method with eight essential oils. Using the broth dilution method, the antibacterial activity of eight commonly used essential oils: lavender, tea tree, eucalyptus, peppermint, thyme, citrus, rosemary, and cinnamon were assessed. Both Gram-positive and Gram-negative species were selected for antimicrobial activity. The minimum inhibitory concentration (MIC) was determined by the broth dilution method. Following MIC determination, a sub-culture was performed on nutrient agar plates to ascertain the minimum bactericidal concentration (MBC). All the antibiotics used in the study demonstrated sensitivity when subjected to the antibiotic susceptibility test. Thyme, cinnamon and peppermint strongly inhibited the growth of S. epidermidis 46 mm, 30mm, and 34 mm respectively. Findings revealed distinct variations in the MIC values among the different essential oils and bacteria tested. Thyme and tea tree oils exhibited the broadest antibacterial spectrum, inhibiting both Gram-positive and Gram-negative bacteria at relatively low concentrations. Thyme (15.625 μl/ml) and tea tree (31.25 μl/ml) oil demonstrated promising activity against Gram-positive pathogen, while tea tree oil (15.625 μl/ml) displayed better efficacy against Gram-negative bacteria. A significant outcome was observed when the essential oils were diluted with carrier oils, indicating promising results in terms of antibacterial activity. These findings highlight the potential of essential oils, particularly thyme and tea tree oils, as effective natural antimicrobial agents.

Depletion forces beyond the diluted limit

The determination of the effective interactions in many-body systems still possesses a tremendous challenge in Condensed Matter Physics. The problem mainly resides in the fact that, on the one hand, there is not a unique route to obtain the effective forces between the ”observable” constituents and, on the other hand, one typically considers that the diluted limit of those components always corresponds to the effective forces at all particle concentrations. In the particular case of colloidal dispersions, it is very important to have experimental, theoretical, and computational tools that allow us to determine, accurately and without using significant approximations, the effective interactions between colloids. In this contribution, we report on a detailed study of depletion potentials in colloidal mixtures at finite concentration, namely, binary and ternary mixtures of disks (in two dimensions or 2D) and spheres (in three dimensions or 3D). To this end, the depletion forces between the large colloidal species are determined through a recently developed scheme based on the so-called contraction of the description, which has been extended and built at the level of the bare forces, i.e., we basically consider that all particles interact through the bare potentials and the net force acting on a given particle is determined by the second’s Newton law. This scheme can be easily adapted to Molecular Dynamics simulations. To verify the physical consistency of the formalism, we explicitly show that in the diluted limit of large particles, the resulting depletion potential reproduces correctly the AO-Vrij limit. As further proof of the accuracy of the results, a comparison of the structure of the large colloids in the whole mixture with the one that results using exclusively the depletion potential is carried out. In 3D, the results are explicitly compared with the potential of mean force and those obtained with the integral equations formalism. We also report a new colloidal stability mechanism based on the use of two different species of depletant agents at low and equal concentrations. Although we highlight the fact that the depletion potential depends on the concentration of the large species, we show that this dependence can be eliminated when the colloidal dispersion is open to a reservoir of small particles, i.e., when the chemical potential of small particles is fixed. Finally, we report the effects of polydispersity on the depletion interaction between large colloids.

Optical and gamma-ray shielding properties of calcium sodium borate glasses with varied equal concentrations of ZnO and BaO

This study presents the optical and radiation shielding characteristics of a newly developed borate glass, doped with equal quantities of ZnO and BaO, and modified with calcium and sodium. The glass samples were prepared using the melt quenching technique, with the composition formula (80-x-y)B2O3-10CaO-10Na2O-xZnO-yBaO, where x and y were varied at 5, 10, 15, and 20 mol%. Comprehensive analyses of the optical and gamma shielding properties were carried out using UV–visible spectrophotometry and Phy-X software, respectively. The UV–visible spectroscopic data allowed for the calculation of various parameters including the direct and indirect optical energy band gaps, refractive index, dielectric constants, and polarizability. It was observed that the direct optical energy band gap decreased from 3.91 to 3.10 eV, while the indirect band gap fell from 3.41 to 2.91 eV with increasing ZnO and BaO content. Conversely, the refractive index rose from 2.29 to 2.42 with higher concentrations of ZnO and BaO. The linear attenuation coefficients (LACs) across the range of 0.015–15 MeV exhibited an inverse relationship. Among the glass samples, B40Zn20Ba20 exhibited the lowest tenth value layer (TVL) and the highest effective atomic number (Zeff), indicating its superior performance as a radiation shielding material. Overall, the produced glass samples demonstrate commendable properties for both optical applications and radiation shielding.

Exploring the reactivity and interactions of a poly(fluorene‐co‐tetrazine)‐conjugated polymer with Single‐walled Carbon Nanotubes

AbstractConjugated tetrazine‐containing polymers that undergo inverse electron demand Diels‐Alder (IEDDA) reactions with trans‐cyclooctenes are interesting not only for their intrinsic optoelectronic properties, but also their interactions with π‐conjugated surfaces. Here, we prepared a series of poly(fluorene‐co‐tetrazine) polymers and carried out IEDDA reactions to decorate them with hydroxyl, hexadecyl, or triethylene glycol side chains. The polymers were investigated pre‐ and post‐IEDDA coupling in terms of their ability to disperse single‐walled carbon nanotubes (SWNTs) in organic solvent. It was found that polymer molecular weight, side chain structure, and degree of conjugation all impacted the quality of SWNT dispersions. While the starting poly(fluorene‐co‐tetrazine) polymer produced concentrated dispersions, the post‐IEDDA polymer containing dihydropyridazine groups did not produce dispersions of equal concentration. However, upon oxidation to the fully aromatic pyridazines, the polymers regained their ability to form concentrated dispersions. Furthermore, the post‐IEDDA polymers exhibited increased selectivity toward metallic SWNTs relative to the starting polymer. Due to the efficiency of the IEDDA reaction, it was also possible to use this chemistry to derivatize the nanotube complex with poly(fluorene‐co‐tetrazine) post‐dispersion. Overall, this work demonstrates the first use of reactive polytetrazines to disperse SWNTs, allowing rapid modification of polymer‐nanotube complexes.

Controlling the thiourea for optimized growth of CdS nanorod arrays for improved photoelectrochemical water splitting

Energy and the environment are very important issues to secure, preserve and improve our modern lifestyle. The conversion of sunlight into hydrogen and oxygen via photoelectrochemical (PEC) water splitting is one of the most potential routes for clean energy. Cadmium sulfide (CdS) is a promising semiconductor for utilization as a photoanode. In this work, CdS has been grown via the hydrothermal method by optimizing the thiourea concentration. The growth of CdS with equal concentration of Cd2+ and S2-demonstrates the crowded hexagonal-shaped nanorod arrays with small diameter and relatively longer length, and it exhibits the highest photocurrent density due to some factors, such as high length-to-diameter ratio, large reaction area, suitable flat band potential, slow charge recombination rate, fast charge transfer, suitable conduction and valence band edges, and surface reaction kinetics. This work will be of potential to further develop improved nanocomposites of CdS nanorods for hydrogen production and research in related fields.

90 Awardee Talk: Shedding light into the black box: The dual-flow continuous culture system and ruminal fermentation

Abstract Nutrients derived from ruminal fermentation supply a substantial amount of metabolizable nutrients for ruminants. These include amino acids, carbohydrates, fatty acids, vitamins, and fermentation end products, such as ammonia and volatile fatty acids. Furthermore, ruminal fermentation modifies dietary nutrients in such a way that nutrients reaching the small intestine for absorption may substantially differ from those being consumed in the diet. Therefore, understanding ruminal fermentation kinetics and quantification of nutrients flowing out of the rumen is an important aspect of ruminant nutrition. Accurate quantification of these nutrients, using in vivo experiments, is laborious, costly, and invasive, requiring surgical procedures such as ruminal, abomasal, and intestinal cannulas. To decrease the time and cost of experiments as well as the use of animals, in vitro methodologies are important tools in ruminant nutrition. Therefore, the objectives of this presentation are i) describe different in vitro methodologies, ii) discuss the advantages of in vitro methodologies, iii) discuss shortcomings of in vitro methodologies, and iv) describe potential developments that may improve in vitro methodologies. Having been used for decades, in vitro methodologies such as pure, batch, and continuous cultures have been well documented to investigate a wide array of aspects of nutrition, including the effects of different dietary compositions, individual fermentation end products, and impacts on the microbiome. However, both batch and pure cultures can result in a build-up of end products that may inhibit fermentation. Continuous culture, however, allows for the removal of end products but, similar to pure and batch cultures, is applicable only to ruminal fermentation and cannot provide information regarding intestinal digestion. The dual-flow continuous culture system (DFCC), developed in the 1970s and used by various laboratories in the USA and Europe consists of fermentation vessels of approximately 1.5 L that are capable of simulating ruminal fermentation for periods longer than 10 d. It has been extensively used and evaluated by meta-analyses. When compared with other in vitro methodologies, the DFCC has longer fermentation time, larger volume, and the possibility of sampling from the liquid and the solid flows. When compared with in vivo experiments the DFCC is faster, less costly, less invasive, allows for greater range of treatments, allows for more controlled condition, and isolates ruminal function. Limitations of the DFCC include it does not predict in vivo response, it assumes equal saliva flow and concentration, no effects of intake, no ruminal absorption, less retention of protozoa, possible differences in microbiome, and limited gas emission data. Therefore, while in vitro methodologies provide useful data into the field of ruminant nutrition, some in vitro methodologies are more robust than others. Furthermore, in vivo experiments are still the ideal approach for assessing nutrient utilization and necessary to evaluate animal responses.

500 Effect of different iron sources on growth performance, gut health, and microbiota in nursery pigs

Abstract This study evaluated the effect of different iron sources, including iron polysaccharide complexes (SQM iron) and ferrous sulfate (FS) on growth performance, gut health, pathogenic bacteria load, colon fecal microbiota, and fecal iron concentration in nursery pigs. Nursery pigs (n = 320, TN70 cross, Topigs Norsvin Canada) weaned at 21 ± 2 d were randomly assigned to four treatments arranged in 2 x 2 factorial, with the factors being sanitation conditions (clean vs. dirty) and iron sources (SQM iron vs. FS, each at 100 mg iron/kg) to give 8 pens (10 piglets per pen) per treatment. Pigs were fed their respective diets in two phases over a 28-d period. Fecal scores, body weight (BW) and feed intake were recorded weekly. Iron content, gene expression of butyryl-CoA:acetate CoA-transferase and bisulfite reductase, and pathogenic bacteria counts were determined in fresh fecal samples collected on d 0, d 14, and d 21. On d 28, one pig randomly selected from each pen was euthanized to sample colon digesta for microbiota analysis and jejunal tissue and serum for gut integrity and permeability analysis. Data were analyzed using PROC MIXED of SAS followed by Tukey’s multiple comparison test except for raw 16s sRNA sequences, which were processed using QIIME2 and analyzed with Kruskal-Wallis test. Overall, from d 0 to14 SQM iron resulted in higher average daily gain (ADG) and average daily feed intake (ADFI; P &amp;lt; 0.05). SQM iron resulted in higher fecal iron at d 21 and improved fecal scores at d 27 (P &amp;lt; 0.05). SQM iron increased the relative abundance of fecal Campylobacter jejuni when comparing d 21 to d 0, and O157 E. Coli when comparing d 14 to d 0 (P &amp;lt; 0.05). Ferrous sulfate resulted in greater changes in relative abundance of butyryl-CoA:acetate CoA-transferase and bisulfate reductase than the SQM iron treatment under clean conditions when comparing d 21 to d 0 (P &amp;lt; 0.05). Dietary iron treatments did not significantly affect gastrointestinal integrity and permeability in jejunum and did not alter fecal microbiota composition on d 28. In conclusion, substituting 100 mg/kg added iron as FS with equal concentration of SQM iron improved ADG in the initial 2 wk and overall ADFI and promoted gut health indicators.

Analytical evaluation of oil and water obtained from demulsification of crude oil using cashew (&lt;i&gt;Anacardium occidentale&lt;/i&gt;) nutshell liquid

The physiochemical characteristics of oil and water obtained from the demusifier treatment of crude oil using equal concentrations of Cashew nutshell liquid (CSNL) and a commercial grade demulsifier (DX077) were evaluated. Results obtained shows that basic sediment and water (BSW) of the crude after treatment with CSNL and DXO77 were 17.5 and 17.9 % respectively which was 0.4% apart. The total oil and grease (TOG) and total petroleum hydrocarbon (TPH) of the produced water after treatment with CSNL were 24.4 and 21.7 mg/l respectively while results after treatment with DXO77 were 23.7 and 20.5 mg/l respectively. The TOG and TPH of the produced water after demulsifier treatment with CSNL and DX077 were within acceptable limit of &lt; 30.0 mg/l as specified by Department of Petroleum Resources (DPR). Analyses of the same crude oil before demulsifier treatment gave a lower BSW of 9.50 % as well as an emulsion band of 16.2 % indicating improper oil/water separation. The TOG and TPH of the produced water before demulsifier treatment were 42.5 and 38.7 mg/l respectively which were above DPR specification. Disposal of produced water with high TOG and TPH into the environment can lead to the release of volatile organic compounds (VOCs) into the ecosystem resulting in environmental pollution as well as other health hazards. Results obtained also show that the water density, pH, electrical conductivity, salinity and chloride of the produced water after demulsifier treatment with CSNL and DX077 were within DPR specification. Reinjecting produced water with high salinity, chloride and conductivity into the reservoir can reduce oil recovery especially from sandstone and carbonate reservoirs. CSNL proved to be an effective demulsifier and is preferrable to the synthetic demulsifier (DX077) because it is more environmentally friendly owing to its green nature.

Mixed former effect in barium borophosphate glasses on the red (Eu3+)-blue (Eu2+) emission for LEDs

The synergistic mixing of P2O5 and B2O3 glass formers at a constant concentration of BaO as a glass modifier plays a vital role in generating a single-phase phosphor doped with Eu2O3 for white light-emitting diodes (WLEDs). Herein, a glass system of nominal composition xP2O5∙(50-x)B2O3∙49.5BaO∙0.5Eu2O3 (0 ≤ x ≤ 50 mol%) was prepared using a melt-quenching technique under normal atmospheric conditions. While the XRD patterns demonstrated the amorphous nature, the SEM micrographs proved the formation of cluster structures. Structural characterization using ATR-FTIR spectroscopy revealed the presence of main vibrational structural units associated with borate and phosphate glass formers, including BO3, BO4−, P=O, PO2−, and PO32−. The IR spectra at x = 0 and 50 mol% exhibit characteristic metaborate and metaphosphate absorption peaks, respectively. In contrast, the mixing of both B2O3 and P2O5 at equal concentrations (x = 25 mol%) has resulted in the absence of absorption peaks assigned to the BO3 and P=O units, thus indicating the mixed structural effect. This is assigned to the formation of B-O-P bonds, with the BO4−, PO2−, and PO32− as the main structural units. These structural changes affect the reduction process of Eu3+ to Eu2+, as demonstrated by the corresponding photoluminescence spectra and observed by the cathodoluminescence micrographs. Photoluminescence studies showed tunable emission from red (Eu3+) to white light (Eu2+ and Eu3+) under UV excitation, with the P2O5 content rising from 0 to 50 mol% due to the growing Eu2+ population formed by the reduction of Eu3+. The optical basicity was observed to decrease with the P2O5 content, favouring the reduction process. The demonstrated tunable single-host white phosphor makes this an attractive system for energy-efficient WLED applications.

Heavy metals altered the xenobiotic metabolism of rats by targeting the GST enzyme: An in vitro and in silico study

Among all the heavy metals, Pb, Cd, and As are the most harmful pollutants in the environment. They reach into the organisms via various levels of food chains i.e. air and water. Glutathione-s-transferase (GST, E.C. 2.5.1.18), a key enzyme of xenobiotics metabolism, plays an important role in the removal of several toxicants. The present study aimed to evaluate any inhibitory action of these heavy metals on the GST enzyme isolated from the hepatic tissues of rats. A 10 % (w/v) homogenate of rat liver was prepared in cold and centrifuged at 4 °C at 9000xg for 30 min. The supernatant was collected and kept frozen at −20 °C or used fresh for carrying out different experiments. The activity of GST was monitored spectrophotometrically at 340 nm using 220 μg of soluble protein with varying equal substrate concentrations (0.125–2 mM) in phosphate buffer (50 mM, pH 6.5). To assess the impact of heavy metals on the enzyme activity, different concentrations of Cd (0–0.6 mM) and Pb (0–2 mM) were added to the reaction mixture followed by monitoring the residual activity. The optimum temperature and pH of rat liver GST were found to be 37 °C and 6.5, respectively. The Km value for GST was 0.69 mM and the Vmax was found to be 78.67 U/mg. The Cd and Pb significantly altered the kinetic behaviour of the enzyme. The Vmax and Kcat/Km parameters of GST were recorded to be decreased after interaction with Cd and Pb individually and showed a mixed type of inhibition pattern suggesting that these inhibitors may have a greater binding affinity either for the free enzyme or the substrate-enzyme complex. These metals showed a time-dependent enzyme inhibition profile. Cd was found to be the most potent inhibitor when compared to other treated metals; the order of inhibitory effect of metal ions was Cd>Pb>As. The in silico ion docking analysis for determining the probable interactions of Cd and Pb with fragmented GST validated that Cd exhibited higher inhibition potential for the enzyme as compared to Pb. The results of the present study indicated that exposure of both the Cd and Pb may cause significant inhibition of hepatic GST; the former with higher inhibitory potential than the later. However, As proved to be least effective against the enzyme under the aforesaid experimental conditions.

Application of the Agilent 2100 Bioanalyzer instrument as quality control for next-generation sequencing.

Next-generation sequencing (NGS) technologies have expanded the spectrum of forensic DNA analysis by facilitating efficient and precise genotyping of a large number of genetic markers. Yet, challenges persist regarding complex sample processing and assurance of equal molar concentrations across pooled samples. Since optimal cluster density is crucial for sequencing performance, the determination of both quantity and quality is indispensable for library preparation. In this study, we investigated the application of the Agilent 2100 Bioanalyzer for library quality control, as studies for forensic approaches, particularly for highly degraded postmortem samples, are rare. Our analysis encompassed assessing total DNA concentrations, fluorescence unit (FU) values, and adapter dimer concentrations in purified DNA libraries derived from buccal swabs and tissue samples of decomposed corpses. The sensitivity study tested a serial dilution derived from buccal swabs and revealed a decrease in FU values and an increase in adapter dimers with declining DNA input concentrations. Deviations in total DNA concentrations and average peak heights between the Agilent 2100 Bioanalyzer runs indicated a lack of repeatability in data and presented challenges in accurate quantification, which was also observed in previous studies. Yet, the analysis of degraded samples from decomposed human remains has shown the ability to detect adapter dimer concentrations, which can be crucial for the quality of subsequent NGS library preparation and sequencing success. Therefore, the Agilent 2100 Bioanalyzer proves to be a valuable tool for NGS quality control.

Electrospray ionisation suppression in aquatic dissolved organic matter studies – Investigation via liquid chromatography–mass spectrometry

Ionisation suppression is a persistent issue in electrospray ionisation mass spectrometry, which decreases the signal of co-eluting analytes. In non-targeted analysis, where analyte and organic matrix identity is unknown, this poses a very challenging analytical problem when it comes to quantitatively assessing differences between samples, including in a compositional sense. In this study, I demonstrate the problems that arise due to ionisation suppression using a very simple sample mixing scheme between a fresh, metabolite rich sample (a leaf leachate) and a forest pond water. Samples were analysed after solid phase extraction on Agilent PPL and using high performance liquid chromatography coupled to electrospray ionisation – Orbitrap mass spectrometry, charged aerosol detector and diode array detector, the latter two allowing quantification of eluting material. I found that more than half of the well-resolved analytes expected to be present (at equal concentration) were completely lost from detection after mixing with pond water DOM. The average recovery of analytical signal (i.e., the signal weighted average), was about 50%, and was highly variable between analytes. Ionisation suppression also affected the signal obtained from the geochemical background DOM, and material recovery decreased slightly when mixing samples and extracting at a higher volume on PPL. Overall, the results showed that ionisation suppression is an extremely important problem for comparison of biogeochemical samples, even when only considering presence and absence of detected features. A multi detector approach and liquid chromatographic separation adds great value in comparison to use of only high resolution mass spectrometry (in direct infusion mode).

Physicochemical, techno-functional, and proteolytic effects of various hydrocolloids as fat replacers in low-fat cheddar cheese

This study aimed to investigate the physiochemical properties, techno-functional attributes, and proteolytic impacts of incorporating different hydrocolloids as fat substitutes in low-fat cheddar cheese. The effects of hydrocolloids (gums) and dietary fiber, both individually and in combination, were studied for the production of low-fat cheddar cheese with the same textural and functional attributes as full-fat cheese. Physicochemical, techno-functional attributes, organic acid, and proteolysis analyses were performed at 0 and 90-day storage intervals. The results of different combinations were compared with positive (4% fat) and negative (2% fat) controls, and it was observed that using fat replacers improved the textural and functional properties of low-fat cheddar cheese. A significant difference was observed in the meltability and flowability of low-fat cheese samples with different analytes. The proteolysis pattern in low-fat cheese also differed from that of its full-fat counterpart. Three treatments were considered the best: T2 (guar gum at 0.45%), T6 (guar gum, inulin, and resistant starch at 33%), and T7 (all analytes at equal concentrations). The findings revealed that hydrocolloids notably improved the texture, meltability, and sensory characteristics of low-fat cheddar cheese. Additionally, SEM images revealed that the microstructure variations can be attributed to changes in the hydrocolloids water retention and fat content. The Pearson correlation coefficient revealed relationships between different functional properties, contributing to understanding how hydrocolloids as fat replacers impact the overall quality of low-fat cheddar cheese. This study offers useful information for the dairy sector to manufacture low-fat cheeses that have enhanced customer acceptance and provide nutritional advantages.

Hierarchical HZSM-5 catalysts enhancing monocyclic aromatics selectivity in co-pyrolysis of wheat straw and polyethylene mixture

The limitation in bio-oil quality hampers its further utilization in the fuel or chemical industry. This study employed a combination of tetra propylammonium hydroxide (TPAOH) and NaOH solutions for HZSM-5 desilication to enhance the yield of monocyclic aromatic hydrocarbons in the catalytic co-pyrolysis of wheat straw with polyethylene. Results revealed hierarchical HZSM-5, prepared with equal mole concentrations of NaOH and TPAOH, exhibited optimal catalytic performance. The bio-oil from this process had an aromatic content of 72 %, with monocyclic aromatic hydrocarbons (MAHs) constituting 56.26 %. Further optimization was achieved through iron modification of hierarchical HZSM-5. The addition of 0.5 % iron-modified hierarchical HZSM-5 increased the aromatic hydrocarbons to 80.57 %, with BTX compounds (benzene, toluene, and xylene) reaching a selectivity of 61.9 %. This approach reduced polycyclic aromatic hydrocarbons (PAHs), contributing to less coke formation, presenting a promising method for high-quality bio-oil.

Antifungal Effect of Poly(methyl methacrylate) with Farnesol and Undecylenic Acid against Candida albicans Biofilm Formation.

The control of Candida albicans biofilm formation on dentures made of poly(methyl methacrylate) (PMMA) is an important challenge due to the high resistance to antifungal drugs. Interestingly, the natural compounds undecylenic acid (UDA) and farnesol (FAR) both prevent C. albicans biofilm formation and could have a synergetic effect. We therefore modified PMMA with a combination of UDA and FAR (UDA+FAR), aiming to obtain the antifungal PMMA_UDA+FAR composites. Equal concentrations of FAR and UDA were added to PMMA to reach 3%, 6%, and 9% in total of both compounds in composites. The physico-chemical properties of the composites were characterized by Fourier-transform infrared spectroscopy and water contact angle measurement. The antifungal activity of the composites was tested on both biofilm and planktonic cells with an XTT test 0 and 6 days after the composites' preparation. The effect of the UDA+FAR combination on C. albicans filamentation was studied in agar containing 0.0125% and 0.4% UDA+FAR after 24 h and 48 h of incubation. The results showed the presence of UDA and FAR on the composite and decreases in the water contact angle and metabolic activity of both the biofilm and planktonic cells at both time points at non-toxic UDA+FAR concentrations. Thus, the modification of PMMA with a combination of UDA+FAR reduces C. albicans biofilm formation on dentures and could be a promising anti-Candida strategy.

Biomodulatory Effects of Molecular Delivery in Human T Cells Using 3D-Printed Acoustofluidic Devices

ObjectiveCell-based therapies have shown significant promise for treating many diseases, including cancer. Current cell therapy manufacturing processes primarily utilize viral transduction to insert genomic material into cells, which has limitations, including variable transduction efficiency and extended processing times. Non-viral transfection techniques are also limited by high variability or reduced molecular delivery efficiency. Novel 3D-printed acoustofluidic devices are in development to address these challenges by delivering biomolecules into cells within seconds via sonoporation. MethodsIn this study, we assessed biological parameters that influence the ultrasound-mediated delivery of fluorescent molecules (i.e., calcein and 150 kDa FITC-Dextran) to human T cells using flow cytometry and confocal imaging. ResultsLow cell plating densities (100,000 cells/mL) enhanced molecular delivery compared to higher cell plating densities (p < 0.001), even though cells were resuspended at equal concentrations for acoustofluidic processing. Additionally, cells in the S phase of the cell cycle had enhanced intracellular delivery compared to cells in the G2/M phase (p < 0.001) and G0/G1 phase (p < 0.01), while also maintaining higher viability compared to G0/G1 phase (p < 0.001). Furthermore, the calcium chelator (EGTA) decreased overall molecular delivery levels. Confocal imaging indicated that the actin cytoskeleton had important implications on plasma membrane recovery dynamics after sonoporation. In addition, confocal imaging indicates that acoustofluidic treatment can permeabilize the nuclear membrane, which could enable rapid intranuclear delivery of nucleic acids. ConclusionsThe results of this study demonstrate that a 3D-printed acoustofluidic device can enhance molecular delivery to human T cells, which may enable improved techniques for non-viral processing of cell therapies.