Commercial Extraction Research Articles

Studying Cationic Liposomes for Quick, Simple, and Effective Nucleic Acid Preconcentration and Isolation.

To ensure high quality of food and water, the identification of traces of pathogens is mandatory. Rapid nucleic acid-based tests shorten traditional detection times while maintaining low detection limits. Challenging is the loss of nucleic acids during necessary purification processes, since elution off solid surfaces is not efficient. We therefore propose the development of a vanishing surface strategy in which cationic liposomes efficiently capture nucleic acids. A lipoplex is formed that can be easily centrifuged down and washed if needed. Adding the lipoplex to detergent solutions or nucleic acid amplification reactions dissolves the liposomes, releasing 100% of the nucleic acid into the reaction. After initial protocol optimization, it was applied to isolate DNA from Escherichia coli, Staphylococcus aureus, and adenovirus in buffer followed by qPCR detection. This enabled the detection of these pathogens down to concentrations of 1 CFU or 1 PFU/mL, respectively. Comparing it to a standard commercial DNA extraction kit, it was superior, as evidenced by lower Ct-values in the qPCR for all pathogen concentrations. Scaling up to larger volumes, samples containing bacteria were first concentrated through nitrocellulose filters (pore size = 0.45 μm). Tap water, lake water, and rinse water of fresh produce were investigated, leading to relevant limits of detection of 100 CFU in 100 mL of tap water, 1000 CFU in 100 mL of lake water, and 100 CFU in 10 g of iceberg lettuce, respectively. Since the liposome protocol is a homogeneous, simple incubation step, it is a valuable alternative to standard commercial nucleic acid extraction kits.

Impacts of different intensities of commercial Sphagnum moss extraction on CO2 fluxes in a northern Patagonia peatland.

Peatlands are key ecosystems for global climate regulation because they provide the most efficient carbon sink on the planet. Despite this, they have been widely degraded by various anthropogenic disturbances, causing imbalances in their ecological functioning. A more recent type of disturbance corresponds to the commercial extraction of Sphagnum mosses, which has been carried out in temperate peatlands distributed in Australasia and Patagonia. In the extreme north of Chilean Patagonia, many peatlands have been subject to intensive commercial extraction of the moss Sphagnum magellanicum, causing their degradation and impacts on CO2 fluxes, a situation that has been barely studied. We conducted a field study on CO2 exchange at three sites, two of them representing different intensities of commercial extraction of the moss S. magellanicum and an undisturbed site in an anthropogenic peatland located in the northern Patagonian region of Chile. CO2 fluxes were measured in situ using a transparent and opaque closed chamber. Gross primary productivity (GPP), ecosystem respiration (Reco), and net ecosystem exchange (NEE) were modeled at high temporal resolution by determining daily and cumulative CO2 flux rates during the vegetation growing season. The NEE records showed significant differences between the three sites studied, attributed to differences in the GPP. Sites with high (<30% residual moss cover) and low (∼50% residual moss cover) extraction intensity were a source of CO2, with cumulative NEE values of 416.6±13.1 and 248.2±9.9g CO2-C m-2 season-1 respectively, while the undisturbed area was a moderate CO2 sink with a cumulative NEE of -86.4±2.8g CO2-C m-2 season-1. Our results provide evidence that increasing levels of Sphagnum moss commercial extraction generate increases in CO2 emissions from anthropogenic peatlands into the atmosphere, and could be used to advance the improvement of current regulations that regulate moss harvesting in Chile.

In vivo Evaluation of Saccharomyces-Modified Tempeh as Potential Prebiotic and Probiotic Food using Mus musculus as an Animal Model

Saccharomyces-modified tempeh (SM tempeh), which is produced by adding Saccharomyces cerevisiae during soybean fermentation, is considered to have the potential as a source of prebiotics. The research aims to determine the prebiotic activity score (PAS) of SM tempeh extract against the probiotics S. cerevisiae and Lactobacillus casei, as well as to evaluate the resistance of S. cerevisiae and Escherichia coli in the intestines of mice fed tempeh. The PAS evaluation was carried out using a factorial complete randomized block design with three replications and one-way ANOVA for data analysis followed by the least significant difference test (5%). Meanwhile, microbial survivability was carried out in vivo using male Mus musculus strain mice fed standard feed, and standard feed with tempeh extract supplementation. The results showed that the supplemetation of either SM or commercial tempeh extract to the growth media significantly affected on the microbial load of S. cerevisiae, L. casei and E. coli, but the concentrations of tempeh extract had no significant effect. Apart from that, the concentrations of tempeh extract had no effect on the PAS of S. cerevisiae and L. casei, meaning that it was able to promote the growth of probiotics in the amount added to the media in the range of 2–10%. In addition, the feeding type had a significant effect on the survival of S. cerevisiae and E. coli in the intestines. S. cerevisiae carried on SM tempeh was detected surviving in the mice intestine at a rate of 6.12 log CFU/g, indicating that the tempeh was a probiotic food. Most likely SM tempeh is a synbiotic food.

Formulation and Evaluation of the Antioxidant Activity of an Emulsion Containing a Commercial Green Tea Extract.

The addition of an extract to an emulsion is intended to improve its fragrance and care qualities. Green tea is a beverage known all over the world. It is tasty and has beneficial effects on human health due to its high polyphenol content. The compounds present in this variety of tea have also made it an interesting cosmetic ingredient. The polyphenols contained in green tea have antioxidant properties and can delay the ageing process in human skin. Various preparations with this ingredient can be found on the market-from creams to hair care products. Making one's own cosmetics is also a trend. In the following study, three creams containing green tea extracts from three different manufacturers were prepared, and the total polyphenol (TP) contents, the phenolic profile of the extracts used and the antioxidant activity of these preparations were examined using two methods: DPPH• and ABTS•+ cationic radicals. The study showed that the antioxidant activity of the glycerin-water extracts measured by the selected methods was higher than that of the oil extract. Among the creams, the product with green tea extract from Firm 2 (glycerin-water extract) showed the best antioxidant properties.

Metabolic profiling of endophytic fungi acting as antagonists of the banana pathogen Colletotrichum musae.

Three endophytic strains, Phomopsis sp., Fusarium proliferatum, and Tinctoporellus epimiltinus, isolated from various plants in the rainforest of the Philippines, were investigated regarding their ability to repress growth of the pathogenic fungus Colletotrichum musae on banana fruits causing anthracnose disease. An in vitro plate-to-plate assay and an in vivo sealed box assay were conducted, using commercial versus natural potato dextrose medium (PDA). All tested endophytes were able to significantly reduce C. musae growth compared to the control. However, the type of medium had no significant effect on lesion size of C. musae on banana. An interaction effect between fungal strain and medium could be shown. On the commercial medium, no differences between the biocontrol ability of the fungi and control treatments could be found, while there were significant differences between the fungal strains on natural medium. Lesions on banana incubated with Phomopsis sp. on natural medium were significantly but only slightly larger than those on banana incubated with F. proliferatum. Volatiles released by these two strains and one pathogenic strain of F. graminearum were collected using polydimethylsiloxane tubes and analyzed via gas chromatography mass spectrometry (GC-MS). Twelve volatile metabolites were detected. Benzaldehyde was the most prominent volatile emitted from the commercial and plain medium. 2-Undecanone, 2-nonanone, and phenylethyl-alcohol were detected in individual samples in both media. 1-Decanol and acoradiene were exclusive to the commercial medium, with acoradiene also being unique to F. proliferatum. Five volatileorganic compounds (VOCs)were emitted from all tested fungal species: 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, and phenylethyl-alcohol. Beta-acorenol was detected in F. proliferatum grown on both media. To reveal whether the medium (commercial PDA versus potato extract) affected the metabolism of the fungi, metabolic footprints were assessed via high performance liquid chromatography with quadrupole time of flight mass spectrometry MS (HPLC-QTOF-MS). A total of 388 metabolic signals were recorded. The intensities of 80-90% of these signals differed significantly between the two types of media. Metabolic footprints varied in response to different potato dextrose medium preparations. The two promising fungal strains may be used to reduce postharvest decay and losses in fruits.

Detection of Porcine Norovirus GII.18 Strains in Pigs Using Broadly Reactive RT-qPCR Assay for Genogroup II Noroviruses

Abstract Noroviruses, belonging to the family Caliciviridae, are classified into at least ten genogroups (G) based on their major capsid protein (VP1). The common genogroup to be identified in both humans and pigs is GII, although porcine noroviruses (PoNoVs) belong to genotypes of their own (GII.11, GII.18, and GII.19). So far, PoNoVs have not been studied much in Finland, possibly due to their rather symptomless nature in pigs. In the present study, we enrolled a total of 189 fecal samples collected from pigs from Finnish farms. Samples were taken from 12 farms in 2010, 2019 and 2020. We analyzed feces from growing pigs ranging from 2.1 to 6 months of age. RNA was extracted from fecal suspensions using a commercial viral RNA extraction kit, followed by RT (reverse transcription)-qPCR. The genotypes were determined by Sanger sequencing of the PCR fragments amplified by conventional PCR. Of the 12 farms, 6 (50%) had at least one PoNoV-infected pig. Altogether 18 (9.5%) of the 189 pigs tested positive for PoNoVs. Pigs mostly aged over 4 months were infected with PoNoVs. Eventually, 12 positive samples were determined as genotype GII.18. We could demonstrate the presence of PoNoVs in Finnish pigs. In future, more studies in which longer sequences from PoNoV genome can be obtained, are required.

Comparison of Plant Genomic DNA Extraction Kits Using the Proofman-LMTIA Amplification Assay.

The extraction of DNA is the basis of molecular biology research. The quality of the extracted DNA is one of the key factors for the success of molecular biology experiments. To select a suitable DNA extraction method for Chinese medicinal herbs and seeds. In this experiment, four commercial DNA extraction kits were used to extract the genomic DNA (gDNA) from the Pinellia ternata (Thunb.) Breit. powder; Arisaema amurense Maxim. powder as well as the seeds of Glycine max (L.) Merr. On the one hand, the concentration and purity of DNA extracted by these four kits were compared. On the other hand, nucleic acid amplification experiments were performed on three samples extracted by each of the four kits by Proofman-LMTIA methods, which is a novel nucleic acid isothermal amplification technique. The concentration and purity of DNA extracted by different kits were used to determine which methods were suitable for the dry powder of Chinese herbal medicines and seeds. The efficiency of the amplification curve to show whether the extracted DNA can be used in nucleic acid amplification experiments. The results showed that the Proofman-LMTIA methods were of high specificity and the optimal reaction temperatures were 63, 59, and 59°C for P. ternata (Thunb.) Makino; A. amurense Maxim. and G. max (L.) Merr., respectively. The concentration and purity of the gDNAs extracted with all kits were within the acceptable ranges; meanwhile, the amplification of the gDNA extracted by Kit II was of the highest efficiency. In this experiment, the principle, concentration, purity, and time taken for extracting DNA with four kits were compared. The automated extraction kit based on the magnetic method is suitable for extracting DNA from Chinese medicinal herbs and seeds. The extracted DNA is suitable for nucleic acid amplification detection.

Quantifying the Pore Characteristics and Heterogeneity of the Lower Cambrian Black Shale in the Deep-Water Region, South China.

Recently, significant breakthroughs have been made in the exploration of shale gas in the Lower Cambrian black shale of the Sichuan Basin, indicating a promising commercial extraction potential. However, there remains considerable controversy regarding the pore structural characteristics for this shale formation, especially in the deep-water region. To address this, this paper focused on core samples from two shale gas wells (Xa1 and Xb1) located in the slope-basin facies zone during the Early Cambrian. The analysis involved X-ray diffraction (XRD) analysis, microscopic imaging, and N2-CO2 adsorption-desorption experiments to qualitatively and quantitatively characterize the pore structural characteristics, pore size distribution, and pore types in the deep-water region. The results show that Niutitang shales can be classified into four lithofacies based on mineral composition: siliceous shale, muddy siliceous shale, mixed siliceous shale, and siliceous calcareous shale. Meanwhile, the black shale exhibits diverse pore types, predominantly organic pores, along with significant development of interparticle pores, intraparticle pores, intercrystalline pores, and a few microfractures. In the Niutitang Formation shale of deep-water regions, mesopores contribute the most to the pore volume, followed by macropores, exceeding 89%. Micropores contribute the most to the specific surface area, followed by mesopores, accounting for more than 98%. Each lithofacies shale displays a similar multipeaked distribution pattern, with siliceous shale generally having the most significant pore volume and specific surface area and the highest heterogeneity. Notably, the total organic carbon (TOC) content is the primary factor controlling the micropore structure of the Niutitang Formation shale in the study area, with a less significant impact on mesopores and macropores. Siliceous shale, which has a high average TOC content of 9.58 wt %, exhibits the greatest pore volume and specific surface area. Quartz contributes to the development of micropores in the shale of the study area, whereas clay minerals somewhat inhibit pore development. Overall, the mineral composition has a minor impact on pore development. These findings provide theoretical support for the evaluation of deep shale gas reservoirs and the prediction of favorable areas in the Lower Cambrian Niutitang Formation in Western Hunan.

Bronchial Challenge in the Diagnosis of Cannabis Allergy.

We have read with great interest the recent review published by Dr. Ebo and collaborators on cannabis allergy. It highlights the difficulties in getting a valid diagnosis because some patients do not admit its consumption, which may not be legalized, there are no commercial extracts and there are problems in cutaneous tests interpretation due to cross-reactivity with other related allergens. In addition to this, bronchial challenge is considered the confirmatory diagnosis, but it is complex to perform. This also occurs in diagnosis of occupational asthma, such as in baker's asthma, in which we have investigated a reliable marker to avoid the bronchial challenge. However, we think it is necessary to do this test to ensure a correct etiological diagnosis of cannabis asthma, and we have carried out bronchial challenge with cannabis on more than 600 patients since 2009, all them active consumers. In our country cannabis consumption has not been legalized, but to do this challenges we asked the National Court for permission, which we obtained easily. Although molecular diagnosis has helped in revealing the problems caused by cross-reactivity, the gold standard continues to be the provocation. In our articles we detail how to do it safely, and we encourage our colleagues to reach a definitive diagnosis that cannabis is the cause of their bronchial symptoms.

Comparative evaluation of PCR and loop-mediated isothermal amplification (LAMP) assays for detecting Pasteurella multocida in poultry

ABSTRACT Aims To develop a colourimetric loop-mediated isothermal amplification (LAMP) assay for the detection of Pasteurella multocida in clinical poultry samples and compare the performance of this assay with PCR. A secondary aim was to evaluate a simple DNA extraction method that could enable LAMP-based testing in the field without the need for specialised laboratory equipment. Methods Primer sets for both LAMP and PCR were designed to amplify the KMT1 gene of P. multocida. DNA was extracted from 12 P. multocida isolates using a commercial extraction kit, and subjected to analysis using both LAMP and PCR. The analytical specificity of the LAMP assay was evaluated by testing it against a panel of 12 unrelated bacterial species, and the analytical sensitivity (limit of detection) was determined through testing of serial dilutions of the target DNA and compared to that of PCR. Subsequently, cloacal swabs (n = 40) from a commercial turkey flock were subjected to analysis using both LAMP and PCR assays, using a rapid DNA extraction method and a commercial extraction kit. Clinical sensitivity and specificity of the LAMP assay were calculated in comparison to PCR. Results A single DNA fragment of the expected size (∼ 200 base pairs), was amplified by PCR from 12 P. multocida isolates, which were also all positive by the LAMP assay. The identity of all PCR amplicons was confirmed by sequencing. Both PCR and LAMP showed similar analytical sensitivity, with a LOD of 20 pg of target DNA. As neither PCR nor LAMP assays produced positive results with 12 non-related bacterial species, the analytical specificity was assessed as 100%. However, LAMP demonstrated lower clinical specificity (94.74%) compared to PCR (100%) when 40 clinical samples were tested. None of the DNA samples extracted using the simplified DNA extraction method were amplified by either LAMP or PCR. Conclusion The LAMP assay developed in this study exhibits comparable performance to PCR in detecting P. multocida. Clinical relevance The use of a rapid and portable DNA extraction method, in conjunction with LAMP assays, could create opportunities for point-of-care testing for fowl cholera in field settings.

Enhancing wine malolactic fermentation: Variable effect of yeast mannoproteins on Oenococcus oeni strains

Lactic acid bacteria (LAB), principally Oenococcus oeni, play crucial roles in wine production, contributing to the transformation of L-malic acid into L-lactic acid during malolactic fermentation (MLF). This fermentation is influenced by different factors, including the initial LAB population and wine stress factors, such as nutrient availability. Yeast mannoproteins can enhance LAB survival in wine. This study explored in model conditions the impact of a commercial mannoprotein extract on MLF dynamics in ten O. oeni strains. The results revealed strain-specific responses in fermentation kinetics and mannoprotein utilization. Mannoprotein addition influenced MLF outcomes, depending on the strain and concentration. The variability in MLF confirmed different technological aptitude of the strains used. The α-mannosidase enzymatic activity was determined and showed higher values in the supernatant than in whole cells. Moreover, α-mannosidase activity varied among strains, suggesting differential regulation in response to fermentation conditions. These findings highlight the importance of understanding mannoprotein interactions with O. oeni for optimizing MLF efficiency and enhancing wine quality. Further research under cellar conditions is needed to evaluate the potential of yeast mannoproteins to promote MLF.

Propolis Extract: Weaving Antioxidant Power into Polymeric Composites Through Electrospinning.

The manufacture of composites with bioactive compounds represents a promising strategy for developing advanced materials in biomedical, food, and industrial applications. However, challenges such as stability, bioactivity retention, and controlled release hinder their effectiveness. Electrospinning emerges as a viable technique for encapsulating bioactive compounds, offering advantages such as high surface area, porosity, and gradual release, which are critical for maintaining the bioactivity of embedded compounds. Regarding bioactive composition, propolis has been highlighted as a potential source and has great potential as a biopolymer ingredient due to its antioxidant and antimicrobial properties. This study analyzed the composition and antioxidant activity of three commercial propolis extracts to select the most suitable extract for fiber composite production using zein and polyethylene oxide (PEO), both recognized as safe. The characterization of the electrospun fibers, including morphology, thermal properties, and antioxidant release, was conducted through various analytical techniques. The findings highlight the effectiveness of electrospinning for developing composite materials with bioactive compounds, paving the way for innovations in antioxidant technologies across multiple sectors.

Effectiveness of Commercial Plant Extracts in Management of Plutella xylostella (Lepidoptera: Plutellidae) on Broccoli in Mexico1

Plutella xylostella (L.) (Lepidoptera: Plutellidae) is the primary pest of broccoli (Brassica oleracea var. italica von Plenck), and it holds the distinction of having the highest number of documented cases of insecticide resistance among insect pests, which complicates its management. The utilization of plant extracts is considered an alternative approach. Therefore, this study aimed to determine the efficacy of commercial plant extracts for the control of P. xylostella. Nine commercial extract products were evaluated, and their effects were compared with Inex-A® + distilled water (control group) and Exalt® (commercial control). Toxicity assays were conducted using third-instar larvae by topical application bioassay. Oviposition inhibition was assessed along with adult female repellency by exposing females to treated surfaces. To determine the residual effect on feeding and oviposition inhibition, third-instar larvae and 3- to 6-d-old adult females were used, respectively, with residual effects measured at 2, 4, 6, 8, and 10 d postapplication. Nimicide 80 (Ultraquimia, Morelos, Mexico) exhibited a mortality rate of 35% of third-instar larvae. Garlimax (Plant Health Care, Azcapotzalco, Mexico) inhibited oviposition by 80%, and demonstrated 85% repellency. The residual effect of Garlimax persisted for 4 d, resulting in a reduced percentage of the consumed area and egg oviposition. The findings of this research highlight the effectiveness of these two products, Nimicide 80 and Garlimax; however, field evaluations are recommended to verify the consistency of their biological effects.

Functional Olive Oil Production via Emulsions: Evaluation of Phenolic Encapsulation Efficiency, Storage Stability, and Bioavailability.

Olive oil is valued for its health benefits, largely due to its bioactive compounds, including hydroxytyrosol (HTyr) and oleuropein (OLE), which have antioxidant, anti-inflammatory, and cardioprotective properties. However, many of these compounds are lost during the production process. This study developed a functional olive oil-derived product using water-in-oil emulsions (W/O) to incorporate commercial extracts rich in HTyr and OLE. HTyr and OLE were encapsulated in a W/O emulsion to preserve their bioactivity. The encapsulation efficiency (EE) was evaluated, and the performance of the emulsion was tested using an in vitro gastrointestinal digestion model. Bioaccessibility was measured by calculating the recovery percentage of HTyr and OLE during the digestion stages. The results showed that OLE exhibited higher EE (88%) than HTyr (65%). During digestion, HTyr exhibited a gradual and controlled release, with bioaccessibility exceeding 80% in the gastric phase and a maintained stability throughout the intestinal phase. In contrast, OLE displayed high bioaccessibility in the gastric phase but experienced a notable decrease during the intestinal phase. Overall, the W/O emulsion provided superior protection and stability for both compounds, particularly for the secoiridoids, compared to the non-emulsified oil. The W/O emulsion improved the encapsulation and bioaccessibility of HTyr and OLE, constituting a promising method for enriching olive oil with bioactive phenolic compounds. Therefore, this method could enhance olive oil's health benefits by increasing the availability of these bioactive compounds during digestion, offering the potential for the development of fortified foods.

Inside the Atacama Desert: uncovering the living microbiome of an extreme environment.

The Atacama Desert in Chile is one of the driest and most inhospitable places on Earth. To analyze the diversity and distribution of microbial communities in such an environment, one of the most important and challenging steps is DNA extraction. Using commercial environmental DNA extraction protocols, a mixture of living, dormant, and dead cells of microorganisms is extracted, but separation of the different DNA pools is almost impossible. To overcome this problem, we applied a novel method on soils across a west-east moisture transect in the Atacama Desert to distinguish between extracellular DNA (eDNA) and intracellular DNA (iDNA) at the cell extraction level. Here, we show that a large number of living and potentially active microorganisms, such as Acidimicrobiia, Geodermatophilaceae, Frankiales, and Burkholderiaceae, occur in the hyperarid areas. We observed viable microorganisms involved as pioneers in initial soil formation processes, such as carbon and nitrogen fixation, as well as mineral-weathering processes. In response to various environmental stressors, microbes coexist as generalists or specialists in the desert soil environment. Our results show that specialists compete in a limited range of niches, while generalists tolerate a wider range of environmental conditions. Use of the DNA separation approach can provide new insights into different roles within viable microbial communities, especially in low-biomass environments where RNA-based analyses often fail.IMPORTANCEThe novel e- and iDNA separation technique offers insights into the living community at the cell extraction level in the hyperarid Atacama Desert. This approach provides a new framework for analyzing the composition and structure of the potentially active part of the microbial communities as well as their specialization, ecological network and community assembly process. Our findings underscore the significance of utilizing alternative genomic techniques in low-biomass environments where traditional DNA- and RNA-based analyses may not be feasible. The results demonstrate the viability of the proposed study framework and show that specialized microorganisms are important in initial soil formation processes, including microbial-driven mineral weathering, as well as the fixation of carbon and nitrogen.

Comparative evaluation of commercial DNA isolation approaches for nanopore-only bacterial genome assembly and plasmid recovery

The advent of Oxford Nanopore Technologies has undergone significant improvements in terms of sequencing costs, accuracy, and sequencing read lengths, making it a cost-effective, and readily accessible approach for analyzing microbial genomes. A major challenge for bacterial whole genome sequencing by Nanopore technology is the requirement for a higher quality and quantity of high molecular weight DNA compared to short-read sequencing platforms. In this study, using eight pathogenic bacteria, we evaluated the quality, quantity, and fragmented size distribution of extracted DNA obtained from three different commercial DNA extraction kits, and one automated robotic platform. Our results demonstrated significant variation in DNA yield and purity among the extraction kits. The ZymoBIOMICS DNA Miniprep Kit (ZM) provided a higher purity of DNA compared to other kit-based extractions. All kit-based DNA extractions were successfully performed on all twenty-four samples using a single MinION flow cell, with the Nanobind CBB Big DNA kit (NB) yielding the longest raw reads. The Fire Monkey HMW-DNA Extraction Kit (FM) and the automated Roche MagNaPure 96 platform (RO) outperformed in genome assembly, particularly in gram-negative bacteria. Based on our finding, we recommend a minimum read coverage and raw read N50, obtained from the appropriate DNA extraction kit for each bacterial species, to optimize genome assembly and plasmid recovery. This approach will assist end-users in selecting the most effective kit-based extraction method for bacterial whole-genome assembly using only long-read nanopore sequences.

Alginate Formulation for Wound Healing Applications.

Significance: Alginate, sourced from seaweed, holds significant importance in industrial and biomedical domains due to its versatile properties. Its chemical composition, primarily comprising β-D-mannuronic acid and α-L-guluronic acid, governs its physical and biological attributes. This polysaccharide, extracted from brown algae and bacteria, offers diverse compositions impacting key factors such as molecular weight, flexibility, solubility, and stability. Recent Advances: Commercial extraction methods yield soluble sodium alginate essential for various biomedical applications. Extraction processes involve chemical treatments converting insoluble alginic acid salts into soluble forms. While biosynthesis pathways in bacteria and algae share similarities, differences in enzyme utilization and product characteristics are noted. Critical Issues: Despite its widespread applicability, challenges persist regarding alginate's stability, biodegradability, and bioactivity. Further understanding of its interactions in complex biological environments and the optimization of extraction and synthesis processes are imperative. Additionally, concerns regarding immune responses to alginate-based implants necessitate thorough investigation. Future Directions: Future research endeavors aim to enhance alginate's stability and bioactivity, facilitating its broader utilization in regenerative medicine and therapeutic interventions. Novel approaches focusing on tailored hydrogel formations, advanced drug delivery systems, and optimized cellular encapsulation techniques hold promise. Continued exploration of alginate's potential in tissue engineering and wound healing, alongside efforts to address critical issues, will drive advancements in biomedical applications.

A novel ionic liquid-based approach for DNA and RNA extraction simplifies sample preparation for bacterial diagnostics

DNA- and RNA-based diagnostics play a pivotal role in accurately detecting and characterizing health-relevant bacteria, offering insights into bacterial presence, viability and treatment efficacy. Herein, we present the development of a novel extraction protocol for both DNA and RNA, designed to enable simple and rapid molecular diagnostics. The extraction method is based on the hydrophilic ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate and silica-coated magnetic beads. First, we developed an IL-based cell lysis protocol for bacteria that operates at room temperature. Subsequently, we established a magnetic bead purification procedure to efficiently and reproducibly extract DNA and RNA from the IL-lysates. The IL not only lyses the cells, but also facilitates the adsorption of nucleic acids (NAs) onto the surface of the magnetic beads, eliminating the need for a chaotropic binding buffer and allowing for purification of NAs without significant effort and materials required. Lastly, we combined the cell lysis step and the purification step and evaluated the novel IL-based extraction method on periopathogenic bacterial cultures, comparing it to commercial DNA and RNA extraction kits via (RT)-qPCR. In comparison to the reference methods, the IL-based extraction protocol yielded similar or superior results. Furthermore, costs are lower, required materials and equipment are minimal and the process is fast (30 min), simple and automatable. These characteristics favour the developed method for use in routine and high-throughput testing as well as in point-of-care, on-site and low-resource settings, thereby advancing the field of molecular diagnostics.Graphical

Do commercial dog extracts cross-react with Felis domesticus allergen 1.

Background: Half of U.S. households own a dog despite 10% of individuals being sensitized to dog. Assessment and treatment options for dog allergy include the use of commercial dog extracts which have inconsistent performance, making diagnosing and managing dog allergy challenging. Contamination of dog extracts with other allergens has previously been reported. Objective: We sought to determine whether commercial dog extracts contain other aeroallergens. Methods: An extract purity and quantification study on acetone precipitated dog hair and dander extract (AP dog) was performed, 6 aeroallergens; Alternaria (Alt a 1), Ragweed (Amb a 1), German Cockroach (Bla g 2), Dust Mite (Der p t), Cat (Fel d 1), and Rye Grass (Lol p 1). Following, conventional dog hair and dander extract (CV dog) and the new ultrafiltered dog hair and dander extract (UF dog) were also assessed based on the initial results of AP dog. SDS-PAGE was performed on all three dog extracts to compare allergen content. Lastly, serology results and aeroallergen immunotherapy prescriptions were compared. Results: The ELISA trays with Alt a 1, Amb a 1, Bla g 2, Der p 1, and Lol p 1 antibodies did not capture AP dog, while the ELISA tray with Fel d 1 antibody captured AP dog, CV dog, and UF dog. SDS-PAGE results of the 3 dog extracts did not reveal a band at the molecular weight for Fel d 1. Conculsion: Contamination of commercial dog extracts was not found. However, our findings are supportive of commercial dog extracts containing a Fel d 1-like dog allergen that is cross-reactive to Fel d 1. Cross-reactivity between commercial dog extracts and Fel d 1 could be responsible for double positivity to cat and dog in serology. Additional studies are needed to better illustrate this Fel d 1-like dog allergen.

Comparative Analysis of Pharmaceutical Persistence in Canadian Wastewater Treatment Plants Utilizing Molecularly Imprinted Polymers and Commercial Solid Phase Extraction Sorbents

In this study, the performances of a synthesized Molecularly Imprinted Polymer (MIP) and a commercial Solid Phase Extraction (SPE) cartridge were compared in terms of the removal efficiency and quantification of pharmaceutical compounds in wastewater samples. Concentration analysis of Emtricitabine, Tenofovir, Naproxen, Diclofenac, Ibuprofen, and Efavirenz was conducted in influent, primary effluent, secondary effluent, and post-chlorination water samples using liquid chromatography-UV detection. Regressions analysis revealed high linearity (R2 values from 0.9980 to 0.9999) for the compounds, alongside recoveries ranging from 90.9% to 100%. The method detection limits (MDLs) were also determined. Removal efficiencies were computed, with Naproxen and Ibuprofen demonstrating complete removal (100%), while other compounds displayed different of removal efficiency levels. The MIP was found to perform slightly better than traditional SPE cartridges in terms of selectivity, efficiency, and overall performance. This study provides valuable insights into the concentrations of target compounds at various treatment stages, emphasizing the importance of choosing extraction cartridges in wastewater treatment processes.