bluuebunny/crossref_metadata_2025 huggingface dataset:
Co-Occurrence of L. monocytogenes with Other Bacterial Genera and Bacterial Diversity on Cleaned Conveyor Surfaces in a Swine Slaughterhouse
Authors: Cherifi Tamazight , Arsenault Julie , Quessy Sylvain , Fravalo Philippe
Published: March 2022

Bacterial pathogens, such as Listeria monocytogenes, can show resistance to disinfection and persistence on working surfaces, permitting them to survive and contaminate food products. Persistence—a complex phenomenon involving interactions between many bacteria within a biofilm—is modulated by in situ characteristics. This study aimed to describe, in silico, the microbiota identified in a swine slaughterhouse after sanitation procedures to better understand the presence of L. monocytogenes on these surfaces. Molecular tools for characterization of microbial communities were used to assess the relative contribution of different bacteria resulting from this phenomenon, and the 16S rRNA sequencing method was used on samples from meat conveyor belt surfaces collected on four sampling visits to study the co-occurrence between L. monocytogenes and other bacteria. From the background microbiota, a total of six genera were found to be negatively correlated with Listeria spp., suggesting Listeria growth inhibition, competition, or at least an absence of shared habitats. Based on these results, a complete scenario of interactions of Listeria with components of background microbiota was established. This work contributes to identifying avenues that could prevent the growth and persistence of L. monocytogenes on food-processing surfaces.
CRISPR-Cas9 Mediated Knockout of SagD Gene for Overexpression of Streptokinase in Streptococcus equisimilis
Authors: Chaudhari Armi , Vyas Sachin , Singh Vijai , Patel Amrutlal , Joshi Chaitanya , Joshi Madhvi
Published: March 2022

Streptokinase is an enzyme that can break down the blood clots in some cases of myocardial infarction (heart attack), pulmonary embolism, and arterial thromboembolism. Demand for streptokinase is higher globally than production due to increased incidences of various heart conditions. The main source of streptokinase is various strains of Streptococci. Expression of streptokinase in native strain Streptococcus equisimilis is limited due to the SagD gene-mediated post-translational modification of streptolysin, an inhibitor of streptokinase expression through the degradation of FasX small RNA (through CoV/RS), which stabilizes streptokinase mRNA. In order to improve the stability of mRNA and increase the expression of streptokinase, which is inhibited by SagA, we used CRISPR-Cas9 to successfully knockout the SagD gene and observed a 13.58-fold increased expression of streptokinase at the transcript level and 1.48-fold higher expression at the protein level in the mutant strain compared to wild type. We have demonstrated the successful gene knockout of SagD using CRISPR-Cas9 in S. equisimilis, where an engineered strain can be further used for overexpression of streptokinase for therapeutic applications.
Characterization of Some Salt-Tolerant Bacterial Hydrolases with Potential Utility in Cultural Heritage Bio-Cleaning
Authors: Ruginescu Robert , Enache Madalin , Popescu Octavian , Gomoiu Ioana , Cojoc Roxana , Batrinescu-Moteau Costin , Maria Gabriel , Dumbravician Maria , Neagu Simona
Published: March 2022

Salt-tolerant enzymes produced by halophilic and halotolerant microorganisms have been proposed to be used in various applications that involve high saline conditions. Considering their biotechnological significance and the current need for more efficient producers of such catalysts, the present study aimed to evaluate the extracellular proteolytic, esterolytic, cellulolytic and xylanolytic activities of some halotolerant strains, and to characterize their functional parameters. A total of 21 bacterial and fungal strains belonging to the genera Bacillus, Virgibacillus, Salinivibrio, Salinicoccus, Psychrobacter, Nocardiopsis, Penicillium, Aspergillus, and Emericellopsis were assayed by quantitative methods. Among them, the members of the Bacillus genus exhibited the highest catalytic activities. The exoenzymes produced by three selected Bacillus strains were active over wide ranges of salinity, temperature and pH. Proteases were active at 20–80 °C, pH 6–10, and 0–1 M NaCl, while esterases showed good catalytic activities at 20–80 °C, pH 7.5–10, and 0–4 M NaCl. Cellulases and xylanases were active at 20–80 °C, pH 5–10, and 0–5 M NaCl. Due to such properties, these hydrolases could be used in a newly proposed application, namely to clean aged consolidants and organic deposits accumulated over time from the surfaces of salt-loaded wall paintings.
Biological Control of Diamondback Moth—Increased Efficacy with Mixtures of Beauveria Fungi
Authors: Soth Sereyboth , Glare Travis R. , Hampton John G. , Card Stuart D. , Brookes Jenny J.
Published: March 2022

Diamondback moth (DBM) is an important horticultural pest worldwide as the larvae of these moths feed on the leaves of cruciferous vegetables. As DBM has developed resistance to more than 100 classes of synthetic insecticides, new biological control options are urgently required. Beauveria species are entomopathogenic fungi recognized as the most important fungal genus for controlling a wide range of agricultural, forestry, and veterinary arthropod pests. Previous research, aimed at developing new Beauveria-based biopesticides for DBM, has focused on screening single isolates of Beauveria bassiana. However, these fungal isolates have individual requirements, which may limit their effectiveness in some environments. This current study separately assessed 14 Beauveria isolates, from a range of habitats and aligned to four different species (Beauveria bassiana, B. caledonica, B. malawiensis, and B. pseudobassiana), to determine the most effective isolate for the control of DBM. Further assays then assessed whether selected combinations of these fungal isolates could increase the overall efficacy against DBM. Six Beauveria isolates (three B. bassiana and three B. pseudobassiana) achieved high DBM mortality at a low application rate with the first documented report of B. pseudobassiana able to kill 100% of DBM larvae. Further research determined that applications of low-virulent Beauveria isolates improved the control of DBM compared to mixtures containing high-virulent isolates. This novel approach increased the DBM pest mortality and shortened the time to kill.
The Escherichia coli Amino Acid Uptake Protein CycA: Regulation of Its Synthesis and Practical Application in l-Isoleucine Production
Authors: Hook Christine , Eremina Natalya , Zaytsev Petr , Varlamova Daria , Stoynova Nataliya
Published: March 2022

Amino acid transport systems perform important physiological functions; their role should certainly be considered in microbial production of amino acids. Typically, in the context of metabolic engineering, efforts are focused on the search for and application of specific amino acid efflux pumps. However, in addition, importers can also be used to improve the industrial process as a whole. In this study, the protein CycA, which is known for uptake of nonpolar amino acids, was characterized from the viewpoint of regulating its expression and range of substrates. We prepared a cycA-overexpressing strain and found that it exhibited high sensitivity to branched-chain amino acids and their structural analogues, with relatively increased consumption of these amino acids, suggesting that they are imported by CycA. The expression of cycA was found to be dependent on the extracellular concentrations of substrate amino acids. The role of some transcription factors in cycA expression, including of Lrp and Crp, was studied using a reporter gene construct. Evidence for the direct binding of Crp to the cycA regulatory region was obtained using a gel-retardation assay. The enhanced import of named amino acids due to cycA overexpression in the l-isoleucine-producing strain resulted in a significant reduction in the generation of undesirable impurities. This work demonstrates the importance of uptake systems with respect to their application in metabolic engineering.
Comparative Genome Analysis of the Photosynthetic Betaproteobacteria of the Genus Rhodocyclus: Heterogeneity within Strains Assigned to Rhodocyclus tenuis and Description of Rhodocyclus gracilis sp. nov. as a New Species
Authors: Kyndt John A. , Aviles Fabiola A. , Imhoff Johannes F. , Künzel Sven , Neulinger Sven C. , Meyer Terrance E.
Published: March 2022

The genome sequences for Rhodocyclus purpureus DSM 168T and four strains assigned to Rhodocyclus tenuis (DSM 110, DSM 111, DSM 112, and IM 230) have been determined. One of the strains studied (IM 230) has an average nucleotide identity (ANI) of 97% to the recently reported genome of the type strain DSM 109 of Rcy. tenuis and is regarded as virtually identical at the species level. The ANI of 80% for three other strains (DSM 110, DSM 111, DSM 112) to the type strain of Rcy. tenuis points to a differentiation of these at the species level. Rcy. purpureus is equidistant from Rcy. tenuis and the new species, based on both ANI (78–80%) and complete proteome comparisons (70% AAI). Strains DSM 110, DSM 111, and DSM 112 are very closely related to each other based on ANI, whole genome, and proteome comparisons but clearly distinct from the Rcy. tenuis type strain DSM 109. In addition to the whole genome differentiation, these three strains also contain unique genetic differences in cytochrome genes and contain genes for an anaerobic cobalamin synthesis pathway that is lacking from both Rcy. tenuis and Rcy. purpureus. Based on genomic and genetic differences, these three strains should be considered to represent a new species, which is distinctly different from both Rcy. purpureus and Rcy. tenuis, for which the new name Rhodocyclus gracilis sp. nov. is proposed.
Virological Characterization of Pigs with Erythema Multiforme
Authors: Halecker Sabrina , Papatsiros Vasileios , Psalla Dimitra , Krabben Ludwig , Kaufer Benedikt , Denner Joachim
Published: March 2022

Erythema multiforme in pigs is an acute, self-limiting disease characterized by red skin areas and often associated with anorexia, fever and respiratory problems. The cause of the disease remains unknown. In a recent study, animals of a commercial breeding herd in Greece were examined, and all animals were found seropositive for porcine reproductive and respiratory syndrome virus (PRRSV). However, neither PRRSV and porcine circovirus type 2 (PCV2) viremia nor antibodies against Aujeszky’s disease virus, African swine fever virus and classical swine fever virus were detected. Here, an extended examination of these pigs was performed on a wide range of porcine viruses using highly sensitive polymerase chain reaction (PCR)-based methods. Affected skin of five animals revealed the presence of porcine lymphotropic herpesvirus-1 (PLHV-1) in all cases, PLHV-2 in one animal and PLHV-3 in four animals. However, neither porcine cytomegalovirus (PCMV) nor porcine circoviruses (PCV1, PCV2, PCV3 and PCV4) were detected. In blood samples, PLHV-1 was present in two animals and PLHV-2, PCV2 and PCV3 in one individual, with PCMV, PCV1 and PCV4 in none of the animals. In one animal, four viruses were found in the blood (PLHV-1, PLHV-2, PCV2 and PCV3). A PRRSV viremia was also not detected. All animals carried porcine endogenous retrovirus C (PERV-C) in their genome, but recombinant PERV-A/C was not detected. The results suggest that porcine viruses may be involved in erythema multiforme in these animals and that further studies are needed to assess the role of these pathogens in the disease.
YfiB: An Outer Membrane Protein Involved in the Virulence of Shigella flexneri
Authors: Sen Tanuka , Verma Naresh K.
Published: March 2022

The intracellular pathogen Shigella flexneri, which is the causative agent of bacillary dysentery, significantly influences the worldwide implication of diarrheal infections, consequentially causing about 1.1 million deaths each year. Due to a nonavailability of an authorized vaccine and the upsurge of multidrug resistance amongst Shigella strains, there has been a huge demand for further genetic analyses which could help in the advancement of new/improved drugs, and finding vaccine candidates against the pathogen. The present study aims to illustrate the role of the yfiB gene in Shigella virulence, part of the periplasmic YfiBNR tripartite signalling system. This system is involved in the regulation of cyclic-di-GMP levels inside the bacterial cells, a vital messenger molecule impacting varied cellular processes such as biofilm formation, cytotoxicity, motility, synthesis of exopolysaccharide, and other virulence mechanisms such as adhesion and invasion of the bacteria. Through a combination of genetic, biochemical, and virulence assays, we show how knocking out the yfiB gene can disrupt the entire YfiBNR system and affect the native c-di-GMP levels. We found that this subsequently causes a negative effect on the biofilm formation, bacterial invasion, host–surface attachment, and the overall virulence of Shigella. This study also carried out a structural and functional assessment of the YfiB protein and determined critical amino acid residues, essential for proper functioning of this signalling system. The present work improves our understanding of the in vivo persistence and survival of Shigella, brings light to the c-di-GMP led regulation of Shigella virulence, and provides a prospective new target to design anti-infection drugs and vaccines against S. flexneri and other bacterial pathogens.
Isolation and Characterization of Commensal Bifidobacteria Strains in Gut Microbiota of Neonates Born Preterm: A Prospective Longitudinal Study
Authors: Wydau-Dematteis Sandra , Delannoy Johanne , Téolis Anne-Claire , Giuseppi Agnès , Campeotto Florence , Lapillonne Alexandre , Butel Marie-José , Aires Julio
Published: March 2022

Bifidobacterial population dynamics were investigated using a longitudinal analysis of dominant species isolated from feces of neonates born preterm (singletons (n = 10), pairs of twins (n = 11)) from birth up to 16 months of age. We performed quantification, isolation, and identification of the dominant bifidobacteria strains. The genetic relationship of the isolates was investigated via pulsed field gel electrophoresis (PFGE) genotyping, and PCR was used to screen the specific genetic marker tet genes. Additionally, all of the isolated strains were phenotypically characterized by their response to gastro-intestinal stresses and the MIC determination of tetracycline. In the same individual, our results showed a turnover of the bifidobacteria dominant population not only at species but also at strain levels. In addition, we found clonally related strains between twins. A minority of strains were tolerant to gastric (6%) and intestinal (16%) stresses. Thirteen percent of the strains were resistant to tetracycline. This work is original as it provides insights at the strain level of the early life in vivo dynamics of gut microbiota bifidobacteria in preterm neonates. It highlights the need to take into consideration the fluctuation of bifidobacteria populations that may occur for one individual.
Setting a Plausible Route for Saline Soil-Based Crop Cultivations by Application of Beneficial Halophyte-Associated Bacteria: A Review
Authors: Teo Han Meng , A. Aziz , A. Wahizatul A. , Bhubalan Kesaven , S. Siti Nordahliawate M. , C. I. Muhamad Syazlie , Ng Lee Chuen
Published: March 2022

The global scale of land salinization has always been a considerable concern for human livelihoods, mainly regarding the food-producing agricultural industries. The latest update suggested that the perpetual salinity problem claimed up to 900 million hectares of agricultural land worldwide, inducing salinity stress among salt-sensitive crops and ultimately reducing productivity and yield. Moreover, with the constant growth of the human population, sustainable solutions are vital to ensure food security and social welfare. Despite that, the current method of crop augmentations via selective breeding and genetic engineering only resulted in mild success. Therefore, using the biological approach of halotolerant plant growth-promoting bacteria (HT-PGPB) as bio-inoculants provides a promising crop enhancement strategy. HT-PGPB has been proven capable of forming a symbiotic relationship with the host plant by instilling induced salinity tolerance (IST) and multiple plant growth-promoting traits (PGP). Nevertheless, the mechanisms and prospects of HT-PGPB application of glycophytic rice crops remains incomprehensively reported. Thus, this review describes a plausible strategy of halophyte-associated HT-PGPB as the future catalyst for rice crop production in salt-dominated land and aims to meet the global Sustainable Development Goals (SDGs) of zero hunger.
Only Low Effects of Water Filters on the Enteric Carriage of Gastrointestinal Pathogen DNA in Colombian Indigenous People
Authors: Kann Simone , Concha Gustavo , Hartmann Maria , Köller Thomas , Alker Juliane , Schotte Ulrich , Kreienbrock Lothar , Frickmann Hagen , Warnke Philipp
Published: March 2022

Water filtration is a common strategy of water sanitation in resource-poor tropical settings. Here, we assessed the intermediate term effect of this preventive procedure including specific filter-related as well as general hygiene training on the molecular detection of enteric pathogens in stool samples from Colombian Indigenous people. From a total of 89 individuals from an Indigenous tribe called Wiwa, stool samples were assessed by real-time PCR for enteropathogenic microorganisms prior to the implementation of water filtration-based infection prevention. Three years after the onset of the preventive strategy, a follow-up assessment was performed. A significantly beneficial effect of water filtration could be shown for Ascaris spp. only (p = 0.035) and a tendency (p = 0.059) for Hymenolepis nana. No hints for effects on the gastrointestinal shedding of Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Campylobacter spp., Shigella spp./enteroinvasive Escherichia coli, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, and Taenia spp. were seen. In conclusion, the study indicates that water filtration can only be an element of a multi-modal hygiene concept to reduce enteric pathogen carriage in inhabitants of resource-poor tropical settings in spite of tendencies of beneficial effects.
Photoinactivation of Phage Phi6 as a SARS-CoV-2 Model in Wastewater: Evidence of Efficacy and Safety
Authors: Gomes Marta , Bartolomeu Maria , Vieira Cátia , Gomes Ana T. P. C. , Faustino Maria Amparo F. , Neves Maria Graça P. M. S. , Almeida Adelaide
Published: March 2022

The last two years have been marked by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. This virus is found in the intestinal tract; it reaches wastewater systems and, consequently, the natural receiving water bodies. As such, inefficiently treated wastewater (WW) can be a means of contamination. The currently used methods for the disinfection of WW can lead to the formation of toxic compounds and can be expensive or inefficient. As such, new and alternative approaches must be considered, namely, photodynamic inactivation (PDI). In this work, the bacteriophage φ6 (or, simply, phage φ6), which has been used as a suitable model for enveloped RNA viruses, such as coronaviruses (CoVs), was used as a model of SARS-CoV-2. Firstly, to understand the virus’s survival in the environment, phage φ6 was subjected to different laboratory-controlled environmental conditions (temperature, pH, salinity, and solar and UV-B irradiation), and its persistence over time was assessed. Second, to assess the efficiency of PDI towards the virus, assays were performed in both phosphate-buffered saline (PBS), a commonly used aqueous matrix, and a secondarily treated WW (a real WW matrix). Third, as WW is generally discharged into the marine environment after treatment, the safety of PDI-treated WW was assessed through the determination of the viability of native marine water microorganisms after their contact with the PDI-treated effluent. Overall, the results showed that, when used as a surrogate for SARS-CoV-2, phage φ6 remains viable in different environmental conditions for a considerable period. Moreover, PDI proved to be an efficient approach in the inactivation of the viruses, and the PDI-treated effluent showed no toxicity to native aquatic microorganisms under realistic dilution conditions, thus endorsing PDI as an efficient and safe tertiary WW disinfection method. Although all studies were performed with phage φ6, which is considered a suitable model of SARS-CoV-2, further studies using SARS-CoV-2 are necessary; nevertheless, the findings show the potential of PDI for controlling SARS-CoV-2 in WW.
Genomic Profiling of Antibiotic-Resistant Escherichia coli Isolates from Surface Water of Agricultural Drainage in North-Western Mexico: Detection of the International High-Risk Lineages ST410 and ST617
Authors: Magaña-Lizárraga José Antonio , Gómez-Gil Bruno , Rendón-Maldonado José Guadalupe , Delgado-Vargas Francisco , Vega-López Inés Fernando , Báez-Flores María Elena
Published: March 2022

Aquatic environments are recognized as one of the main reservoirs for the emergence and dissemination of high-risk lineages of multidrug-resistant (MDR) bacteria of public health concern. However, the genomic characteristics of antibiotic-resistant Escherichia coli isolates from aquatic origins remain limited. Herein, we examined the antibiotic resistance and virulence genomic profiles of three E. coli recovered from surface water in northwest Mexico. Antimicrobial susceptibility testing, whole-genome sequencing (WGS), and in-depth in silico analysis were performed. Two E. coli exhibited MDR phenotypes. WGS-based typing revealed genetic diversity, and phylogenetic analysis corroborated a notable divergent relationship among the studied E. coli. One E. coli strain, harboring enterotoxigenic and extraintestinal pathogenic-associated virulence genes, was assigned to the ST4 lineage. MDR E. coli, belonging to the international high-risk clones ST410 and ST617, carried genes and mutations conferring resistance to aminoglycosides, β-lactams, quinolones, sulfonamides, tetracyclines, and trimethoprim. This study describes, for the first time, the detection and genomic profiling of high-risk lineages of E. coli ST410 and ST617 from surface water in Mexico. Additionally, our results underscore the role of surface water as a reservoir for critical pathogenic and MDR E. coli clones and the need for the surveillance and monitoring of aquatic environments via WGS from the One Health perspective.
Effects of Perchlorate and Other Groundwater Inorganic Co-Contaminants on Aerobic RDX Degradation
Authors: Yadav Amit , Gupta Swati , Istvan Paula , Ronen Zeev
Published: March 2022

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) pollution is accompanied by other co-contaminants, such as perchlorate and chlorates, which can retard biodegradation. The effects of perchlorate and chlorate on aerobic RDX degradation remain unclear. We hypothesized that they have a negative or no impact on aerobic RDX-degrading bacteria. We used three aerobic RDX-degrading strains—Rhodococcus strains YH1 and T7 and Gordonia YY1—to examine this hypothesis. The strains were exposed to perchlorate, chlorate, and nitrate as single components or in a mixture. Their growth, degradation activity, and gene expression were monitored. Strain-specific responses to the co-contaminants were observed: enhanced growth of strain YH1 and inhibition of strain T7. Vmax and Km of cytochrome P450 (XplA) in the presence of the co-contaminants were not significantly different from the control, suggesting no direct influence on cytochrome P450. Surprisingly, xplA expression increased fourfold in cultures pre-grown on RDX and, after washing, transferred to a medium containing only perchlorate. This culture did not grow, but xplA was translated and active, albeit at lower levels than in the control. We explained this observation as being due to nitrogen limitation in the culture and not due to perchlorate induction. Our results suggest that the aerobic strain YH1 is effective for aerobic remediation of RDX in groundwater.
A Numerical Simulation of Electrical Resistivity of Fiber-Reinforced Composites, Part 2: Flexible Bituminous Asphalt
Authors: Ehsani Rojina , Miri Alireza , Tehrani Fariborz M.
Published: March 2022

Asphalt concrete pavements are vulnerable to freeze-thaw cycles. Consecutive cracking and penetration of corrosive agents can expedite the degradation of asphalt pavements and result in weight loss and reduced strength. Fiber reinforcement in flexible bituminous asphalt bridge cracks limits the crack width and enhances the toughness of the composite. Furthermore, steel fibers facilitate asphalt heating during maintenance and repair operations. Electrical resistivity is a vital parameter to measure the efficiency of these operations and to identify the state of degradation in fiber-reinforced asphalt concrete. The significant difference between conductivities of steel fibers and bituminous matrix warrants in-depth investigations of the influence of fiber reinforcement on the measured surface electrical resistivity of placed pavements. Numerical simulations endeavor to predict the resistivity and associated deviations due to randomly distributed fiber reinforcement. Results and discussions reveal the sources and magnitudes of fiber geometry and content adjustments. Outcomes investigate associated errors for practical applications.
Analysis of Conformational Preferences in Caffeine
Authors: Gómez Sara , Rojas-Valencia Natalia , Restrepo Albeiro
Published: March 2022

High level DLPNO–CCSD(T) electronic structure calculations with extended basis sets over B3LYP–D3 optimized geometries indicate that the three methyl groups in caffeine overcome steric hindrance to adopt uncommon conformations, each one placing a C–H bond on the same plane of the aromatic system, leading to the C–H bonds eclipsing one carbonyl group, one heavily delocalized C–N bond constituent of the fused double ring aromatic system, and one C–H bond from the imidazole ring. Deletion of indiscriminate and selective non-Lewis orbitals unequivocally show that hyperconjugation in the form of a bidirectional –CH3 ⇆ aromatic system charge transfer is responsible for these puzzling conformations. The structural preferences in caffeine are exclusively determined by orbital interactions, ruling out electrostatics, induction, bond critical points, and density redistribution because the steric effect, the allylic effect, the Quantum Theory of Atoms in Molecules (QTAIM), and the non-covalent interactions (NCI), all predict wrong energetic orderings. Tiny rotational barriers, not exceeding 1.3 kcal/mol suggest that at room conditions, each methyl group either acts as a free rotor or adopts fluxional behavior, thus preventing accurate determination of their conformations. In this context, our results supersede current experimental ambiguity in the assignation of methyl conformation in caffeine and, more generally, in methylated xanthines and their derivatives.
Evaluation of Vetiver Volatile Compound Production under Aeroponic-Grown Conditions for the Perfume Industry
Authors: Gavira Carole , Watteau Françoise , Lainé Jean-Marc , Bourgaud Frédéric , Legendre Laurent
Published: March 2022

Vetiver (Chrysopogon zizanioides (L.) Roberty) is a major tropical perfume crop. Access to its essential oil (EO)-filled roots is nevertheless cumbersome and land-damaging. This study, therefore, evaluated the potential of vetiver cultivation under soilless high-pressure aeroponics (HPA) for volatile organic compound (VOC) production. The VOC accumulation in the roots was investigated by transmission electron microscopy, and the composition of these VOCs was analyzed by gas chromatography coupled with mass spectrometry (GC/MS) after sampling by headspace solid-phase microextraction (HS-SPME). The HPA-grown plants were compared to plants that had been grown in potting soil and under axenic conditions. The HPA-grown plants were stunted, demonstrating less root biomass than the plants that had been grown in potting soil. The roots were slender, thinner, more tapered, and lacked the typical vetiver fragrance. HPA cultivation massively impaired the accumulation of the less-volatile hydrocarbon and oxygenated sesquiterpenes that normally form most of the VOCs. The axenic, tissue-cultured plants followed a similar and more exacerbated trend. Ultrastructural analyses revealed that the HPA conditions altered root ontogeny, whereby the roots contained fewer EO-accumulating cells and hosted fewer and more immature intracellular EO droplets. These preliminary results allowed to conclude that HPA-cultivated vetiver suffers from altered development and root ontology disorders that prevent EO accumulation.
Determination of Methyl Methanesulfonate and Ethyl Methylsulfonate in New Drug for the Treatment of Fatty Liver Using Derivatization Followed by High-Performance Liquid Chromatography with Ultraviolet Detection
Authors: Wang Yue , Feng Jing , Wu Song , Shao Huihui , Zhang Wenxuan , Zhang Kun , Zhang Hanyilan , Yang Qingyun
Published: March 2022

A new derivatization high-performance liquid chromatography method with ultraviolet detection was developed and validated for the quantitative analysis of methanesulfonate genotoxic impurities in an innovative drug for the treatment of non-alcoholic fatty liver disease. In this study, sodium dibenzyldithiocarbamate was used as a derivatization reagent for the first time to enhance the sensitivity of the analysis, and NaOH aqueous solution was chosen as a pH regulator to avoid the interference of the drug matrix. Several key experimental parameters of the derivatization reaction were investigated and optimized. In addition, specificity, linearity, precision, stability, and accuracy were validated. The determined results of the samples were consistent with those obtained from the derivatization gas chromatography–mass spectrometry analysis. Thus, the proposed method is a reliable and practical protocol for the determination of trace methanesulfonate genotoxic impurities in drugs containing mesylate groups.
Comparative Studies on Essential Oil and Phenolic Content with In Vitro Antioxidant, Anticholinesterase, Antimicrobial Activities of Achillea biebersteinii Afan. and A. millefolium subsp. millefolium Afan. L. Growing in Eastern Turkey
Authors: Toplan Gizem Gülsoy , Taşkın Turgut , İşcan Gökalp , Göger Fatih , Kürkçüoğlu Mine , Civaş Ayşe , Ecevit-Genç Gülay , Mat Afife , Başer Kemal Hüsnü Can
Published: March 2022

The potential applications of Achillea species in various industries have encouraged the examination of their phytochemical components along with their biological potential. In the present study, phenolic contents and essential oil compositions together with the in vitro biological activities of the aerial parts from Achillea biebersteinii Afan. and Achillea millefolium subsp. millefolium Afan. collected from Turkey were evaluated. Different solvent extracts (n-hexane, chloroform, methanol, water) were prepared and their antimicrobial, anticholinesterase, and antioxidant activities were studied. The LC-MS/MS results revealed the presence of 16 different phenolic compounds, including chlorogenic acid, rutin, quercetin, and luteolin glycosides, in methanolic extracts. According to GC-FID and GC/MS results, the primary components of the oils were identified as 1,8-cineole (32.5%), piperitone (14.4%), and camphor (13.7%) in A. biebersteinii and 1,8-cineole (12.3%) and β-eudesmol (8.9%) in A. millefolium subsp. millefolium. The infusion and methanolic extracts of both species were found to be rich in their total phenolic content as well as their antioxidant and anticholinesterase activity. In contrast, the n-hexane and chloroform extracts of both species showed strong antimicrobial activity with MIC values ranging from 15 to 2000 μg/mL. Our findings suggest that the investigated Achillea species could be evaluated as potent natural agents, and further studies into the promising extracts are needed.
Nobiletin Alleviates Astrocyte Activation and Oxidative Stress Induced by Hypoxia In Vitro
Authors: Wang Dandan , Gao Fengjuan , Hu Fangyuan , Wu Jihong
Published: March 2022

Increasing evidence indicates that nobiletin (NOB) is a promising neuroprotective agent. Astrocyte activation plays a key role in neurodegenerative disorders. Thus, this study aims to investigate the effects of NOB on astrocyte activation and the potential mechanisms. In this study, astrocytes were exposed to hypoxia injury for 24 h to induce activation in vitro. Glial fibrillary acidic protein (GFAP) was chosen as a marker of astrocyte activation. To evaluate the effects of NOB on the migration of activated astrocytes, we used a scratch wound healing assay and Transwell migration assay. In addition, the levels of reactive oxygen species (ROS), malondialdehyde (MDA), mitochondrial membrane potential, Nrf2 and HO-1 were measured to investigate the mechanisms of NOB in the activation of astrocytes. We found that NOB alleviated astrocyte activation and decreased GFAP expression during hypoxia. Simultaneously, NOB alleviated the migration of astrocytes induced by hypoxia. With NOB treatment, hypoxia-induced oxidative stress was partially reversed, including reducing the production of ROS and MDA. Furthermore, NOB significantly improved the mitochondrial dysfunction in activated astrocytes. Finally, NOB promoted Nrf2 nuclear translocation and HO-1 expression in response to continuous oxidative damage. Our study indicates, for the first time, that NOB alleviates the activation of astrocytes induced by hypoxia in vitro, in part by ameliorating oxidative stress and mitochondrial dysfunction. This provides new insights into the neuroprotective effects of NOB.
Anti-Neuroinflammatory Components from Clausena lenis Drake
Authors: Zhu Si-Si , Zhang Yi-Fan , Ding Meng , Zeng Ke-Wu , Tu Peng-Fei , Jiang Yong
Published: March 2022

Clausena lenis Drake (C. lenis) is a folk medicinal herb to treat influenza, colds, bronchitis, and malaria. The 95% and 50% ethanol extract of C. lenis showed significant nitric oxide (NO) inhibition activity in BV-2 microglial cells stimulated by lipopolysaccharide (LPS). Bio-guided isolation of the active extract afforded five new compounds, including a chlorine-containing furoquinoline racemate, (±)-claulenine A (1), an amide alkaloid, claulenine B (2), a prenylated coumarin, claulenin A (3), a furocoumarin glucoside, clauleside A (4), and a multi-prenylated p-hydroxybenzaldehyde, claulenin B (5), along with 33 known ones. Their structures were determined via spectroscopic methods, and the absolute configurations of new compounds were assigned via the electronic circular dichroism (ECD) calculations and single-crystal X-ray diffraction analysis. Compounds 2, 23, 27, 28, 33, and 34 showed potent anti-neuroinflammatory effects on LPS-induced NO production in BV-2 microglial cells, with IC50 values in the range of 17.6–40.9 μM. The possible mechanism was deduced to interact with iNOS through molecular docking.
Antimicrobial and Cytotoxic Activity of Novel Imidazolium-Based Ionic Liquids
Authors: Pałkowski Łukasz , Karolak Maciej , Skrzypczak Andrzej , Wojcieszak Marta , Walkiewicz Filip , Podemski Jonasz , Jaroch Karol , Bojko Barbara , Materna Katarzyna , Krysiński Jerzy
Published: March 2022

In this study, a series of 10 novel 1-methyl-3-octyloxymethylimidazolium derivatives carrying various anionic moieties (4-hydroxybenzenesulfonate, benzenesulfonate, carvacroloxyacetate, chloride, formate, propionate, thymoloxyacetate, vanillinoxyacetate, eugenoloxyacetate and trimethylacetate) were synthesized. Compounds were tested for their antimicrobial activity against six microbe strains (Staph-ylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, and Candida albicans), cytotoxic activity against the mouse melanoma cell line (B16 F10), and surface active properties. All synthesized compounds exhibited antimicrobial activity (expressed as minimum inhibitory concentration; in range of 0.10–27.82 mM/L), especially against Gram-positive bacteria and fungi. In addition, all compounds demonstrated cytotoxicity on B16 F10 cells (IC50 values 0.0101–0.0197 mM/L). Surface properties defined as CMC values, ranged from 0.72 to 32.35 mmol L-1. The obtained results provide an insight into the promising activity of a novel group of quaternary imidazolium derivatives having ionic liquid properties. The most potent compounds, containing a thymoloxyacetate and eugenoloxyacetate moiety, could be candidates for new antimicrobial agents or surfactants.
Effect of Ozone-Treated or Untreated Saskatoon Fruits (Amelanchier alnifolia Nutt.) Applied as an Additive on the Quality and Antioxidant Activity of Fruit Beers
Authors: Gorzelany Józef , Michałowska Dorota , Pluta Stanisław , Kapusta Ireneusz , Belcar Justyna
Published: March 2022

Fruit of Saskatoon (Amelanchier alnifolia Nutt.) are a good source of bioactive compounds, such as polyphenols, including anthocyanins, as well as vitamins, macro- and microelements and fibre. By treating Saskatoon fruits with gaseous ozone, and adding the material as an enhancer to barley beers, it is possible to impact the contents of bioactive compounds in the produced fruit beers. Sensory tests showed that beers made from barley with addition of Saskatoon fruit of the ‘Smoky’ cultivar were characterised by the most balanced taste and aroma. Physicochemical analyses of fruit beers, produced with Saskatoon fruit pulp added on the seventh day of fermentation, showed that the beers enhanced with ozone-treated and untreated ‘Smoky’ Saskatoon fruits had the highest contents of alcohol, 5.51% v/v and 5.66% v/v, respectively, as well as total polyphenol contents of 395 mg GAE/L and 401 mg GAE/L, respectively, and higher antioxidant activity (assessed using DPPH•, FRAP and ABTS+• assays). It was demonstrated that the ozonation process led to a decrease in the contents of neochlorogenic acid, on average by 91.00%, and of caffeic acid by 20.62%, relative to the beers enhanced with ‘Smoky’ Saskatoon fruits not subjected to ozone treatment. The present study shows that Saskatoon fruits can be used in the production of beer, and the Canadian cultivar ‘Smoky’ is recommended for this purpose.
Garcinia Biflavonoid 1 Improves Lipid Metabolism in HepG2 Cells via Regulating PPARα
Authors: Chen Hai-Xin , Yang Fan , He Xin-Qian , Li Ting , Sun Yong-Zhi , Song Jian-Ping , Huang Xin-An , Guo Wen-Feng
Published: March 2022

Garcinia biflavonoid 1 (GB1) is one of the active chemical components of Garcinia kola and is reported to be capable of reducing the intracellular lipid deposition, which is the most significant characteristic of non-alcoholic fatty liver disease. However, its bioactive mechanism remains elusive. In the current study, the lipid deposition was induced in HepG2 cells by exposure to oleic acid and palmitic acid (OA&PA), then the effect of GB1 on lipid metabolism and oxidative stress and the role of regulating PPARα in these cells was investigated. We found that GB1 could ameliorate the lipid deposition by reducing triglycerides (TGs) and upregulate the expression of PPARα and SIRT6, suppressing the cell apoptosis by reducing the oxidative stress and the inflammatory factors of ROS, IL10, and TNFα. The mechanism study showed that GB1 had bioactivity in a PPARα-dependent manner based on its failing to improve the lipid deposition and oxidative stress in PPARα-deficient cells. The result revealed that GB1 had significant bioactivity on improving the lipid metabolism, and its potential primary action mechanism suggested that GB1 could be a potential candidate for management of non-alcoholic fatty liver disease.
The Relationship between Dietary Habits and Work Engagement among Female Finnish Municipal Employees
Authors: Virtanen Jenni , Penttinen Markus A. , Laaksonen Marika , Erkkola Maijaliisa , Vepsäläinen Henna , Kautiainen Hannu , Korhonen Päivi
Published: March 2022

Background: Work engagement reflects work-related well-being. It is positively associated with health, life satisfaction, work efficiency, income level, and occupational prospects. However, little is known about the relationship between work engagement and diet. Methods: A cross-sectional study was conducted among female Finnish municipal employees (n = 630) in 2015. Work engagement was assessed using the Utrecht Work Engagement Index. The consumption of healthy and unhealthy food items was determined using a food frequency questionnaire. Sociodemographic factors, health behaviors, depressive and anxiety symptoms were assessed with self-administrated questionnaires. Results: Work engagement had a positive relationship with the daily consumption of healthy food items. This association remained significant even after adjusting for age, education years, financial situation, and physical activity. The frequency of consuming unhealthy food items showed no relationship with work engagement. Anxiety and depressive symptoms decreased linearly with the greater consumption of healthy foods. Conclusion: Frequent consumption of healthy food items is associated with higher work engagement, irrespectively of the consumption of unhealthy nutrients. These results encourage health care professionals to recommend healthy food items instead of forbidding unhealthy food, as well as employers to support healthy dietary habits among employees.
Efficacy of the Nutritional Risk Index, Geriatric Nutritional Risk Index, BMI, and GLIM-Defined Malnutrition in Predicting Survival of Patients with Head and Neck Cancer Patients Qualified for Home Enteral Nutrition
Authors: Przekop Zuzanna , Szostak-Węgierek Dorota , Milewska Magdalena , Panczyk Mariusz , Zaczek Zuzanna , Sobocki Jacek
Published: March 2022

Malnutrition is a poor prognostic factor in cancer disease. In case of head and neck cancer, there are multiple disease symptoms and side effects of treatment that increase the risk of malnutrition. The aim of the study is to assess the association between nutritional status at the time of qualification for home enteral nutrition (HEN) and overall survival of patients with head and neck cancer (HNC), and assessment usefulness of selected nutritional indices as prognostic factors. The retrospective survival analysis involved 157 patients with HNC referred to HEN between January 2018 and October 2021. The nutritional status assessment was performed at the qualification for HEN visit. We have analyzed results of body mass index (BMI), Nutritional Risk Index (NRI) for patients <65 years, Geriatric Nutritional Risk Index (GNRI) for patients ≥65 years and malnutrition defined by Global Leadership Initiative on Malnutrition (GLIM). The mean patient survival was 44.7 weeks and the median was 23.9 weeks. Patients with low NRI and GNRI score had a higher risk of death (NRI: p = 0.0229; GNRI: p = 0.371). NRI, GNRI, and malnutrition defined by GLIM were superior to BMI as prognostic markers for survival. Results suggest that the use of NRI, GNRI, and GLIM criteria could provide useful prognostic information. The longer survival since the qualifying visit for home enteral nutrition suggests that nutritional management could be initiated earlier.
The Association between Peptide Hormones with Obesity and Insulin Resistance Markers in Lean and Obese Individuals in the United Arab Emirates
Authors: Ali Ahmad Manal , Karavetian Mirey , Moubareck Carole Ayoub , Wazz Gabi , Mahdy Tarek , Venema Koen
Published: March 2022

Peptide hormones play a crucial role in body weight and glucose homeostasis. In this study, we aimed to explore this association and recruited 43 obese and 31 age- and sex-matched lean participants. We assessed their body mass index (BMI), waist circumference (WC), waist-to-height ratio (WtHR), percentage body fat (PBF), fasting blood levels of peptide hormones (GLP-1, GLP-2, insulin, leptin, ghrelin, CCK, and PYY), fasting blood sugar (FBS), and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR). We tested the associations between peptide hormones and markers of obesity and insulin resistance (IR) by using the Independent-Samples t-test and Mann-Whitney U test, partial correlation, and logistic regression. FBS, insulin, HOMA-IR, GLP-1, GLP-2, and leptin were significantly higher in the obese group; ghrelin and CCK were significantly higher in lean participants, and no difference was seen for PYY. Controlling for BMI, GLP-1 was positively correlated with WtHR, while ghrelin was inversely correlated with WtHR. GLP-1 was correlated with HOMA-IR. GLP-1 was associated with obesity and IR markers in the regression model. Our results show that obese and lean adults display significant differences in plasma peptide hormone levels. GLP-1 levels were independently associated with markers of obesity and IR. Restoring the appetite hormone balance in obesity may represent a potential therapeutic target.
Lactobacillus reuteri CCFM8631 Alleviates Hypercholesterolaemia Caused by the Paigen Atherogenic Diet by Regulating the Gut Microbiota
Authors: Wang Qianqian , He Yufeng , Li Xiu , Zhang Ting , Liang Ming , Wang Gang , Zhao Jianxin , Zhang Hao , Chen Wei
Published: March 2022

Cardiovascular disease has one of the highest global incidences and mortality rates. Atherosclerosis is the main cause of cardiovascular disease, and hypercholesterolaemia and hyperlipidaemia are the main risk factors for the development of atherosclerosis. Decreasing serum cholesterol and triglyceride concentrations is considered to be an effective strategy to prevent atherosclerotic cardiovascular disease. Previous studies have shown that many diseases are related to gut microbiota dysbiosis. The positive regulation of the gut microbiota by probiotics may prevent or treat certain diseases. In this study, Lactobacillus reuteri CCFM8631 treatment was shown to decrease plasma total cholesterol (TC), low-density lipoprotein–cholesterol, aspartate transaminase, alanine transaminase and trimethylamine N-oxide concentrations, decrease liver TC and malondialdehyde concentrations and recover liver superoxide dismutase concentrations in mice fed a Paigen atherogenic diet. In addition, L. reuteri increased the faecal short-chain fatty acid content (acetate, propionate and butyrate), which was accompanied by an increase in the relative abundance of faecal Deferribacteres, Lachnospiraceae NK4A136 group, Lactobacillus and Dubosiella; a decrease in the relative abundance of Erysipelatoclostridium and Romboutsia and the activation of butanoate and vitamin B6 metabolism, leading to the alleviation of hypercholesterolaemia.
Hydrolyzed Bound Phenolics from Rice Bran Alleviate Hyperlipidemia and Improve Gut Microbiota Dysbiosis in High-Fat-Diet Fed Mice
Authors: Zhao Guanghe , Zhang Ruifen , Huang Fei , Dong Lihong , Liu Lei , Jia Xuchao , Chi Jianwei , Ma Yongxuan , Deng Mei , Chen Yanxia , Ma Qin , Zhang Mingwei
Published: March 2022

It has been confirmed the lipid-lowering effect of rice bran free phenolics, but it is unknown whether rice bran bound phenolics, the phenolic profile of which differs from the free ones, have a similar effect. Thus, the hypolipidemic effect and potential mechanism of hydrolyzed bound phenolics (HBP) from rice bran was investigated in this study. The results showed that HBP supplementation significantly improved serum lipid profiles of high-fat-diet fed mice. HBP inhibited the activation of nuclear receptors liver X receptor-α (LXRα), sterol regulatory element binding protein 1c (SREBP-1c), and peroxisome proliferators-activated receptors-γ (PPARγ), and, therefore, changed the expressions of their downstream genes, including LDLR, CD36, ACC1, FAS, and DGAT2 in the liver. Moreover, HBP supplementation reversed the high-fat-diet induced gut microbiota dysbiosis. These findings suggest that HBP might alleviate the hyperlipidemia via inhibiting the hepatic de novolipogenesis, regulating the uptake of cholesterol and fatty acid in the liver and their absorption in the gut. The attenuation of microbiota dysbiosis might contribute to the above effects.