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20 pages, 5436 KiB

Open AccessArticle

Phytochemical Profiling and Bioactive Potential of Grape Seed Extract in Enhancing Salinity Tolerance of Vicia faba

by Doaa E. Elsherif, Fatmah A. Safhi, Prasanta K. Subudhi, Abdelghany S. Shaban, Mai A. El-Esawy and Asmaa M. Khalifa

Plants 2024, 13(12), 1596; https://doi.org/10.3390/plants13121596 (registering DOI) - 8 Jun 2024

Abstract

Salinity stress poses a significant threat to crop productivity worldwide, necessitating effective mitigation strategies. This study investigated the phytochemical composition and potential of grape seed extract (GSE) to mitigate salinity stress effects on faba bean plants. GC–MS analysis revealed several bioactive components in [...] Read more.

Salinity stress poses a significant threat to crop productivity worldwide, necessitating effective mitigation strategies. This study investigated the phytochemical composition and potential of grape seed extract (GSE) to mitigate salinity stress effects on faba bean plants. GC–MS analysis revealed several bioactive components in GSE, predominantly fatty acids. GSE was rich in essential nutrients and possessed a high antioxidant capacity. After 14 days of germination, GSE was applied as a foliar spray at different concentrations (0, 2, 4, 6, and 8 g/L) to mitigate the negative effects of salt stress (150 mM NaCl) on faba bean plants. Foliar application of 2–8 g/L GSE significantly enhanced growth parameters such as shoot length, root length, fresh weight, and dry weight of salt-stressed bean plants compared to the control. The Fv/Fm ratio, indicating photosynthetic activity, also improved with GSE treatment under salinity stress compared to the control. GSE effectively alleviated the oxidative stress induced by salinity, reducing malondialdehyde, hydrogen peroxide, praline, and glycine betaine levels. Total soluble proteins, amino acids, and sugars were enhanced in GSE-treated, salt-stressed plants. GSE treatment under salinity stress modulated the total antioxidant capacity, antioxidant responses, and enzyme activities such as peroxidase, ascorbate peroxidase, and polyphenol oxidase compared to salt-stressed plants. Gene expression analysis revealed GSE (6 g/L) upregulated photosynthesis (chlorophyll a/b-binding protein of LHCII type 1-like (Lhcb1) and ribulose bisphosphate carboxylase large chain-like (RbcL)) and carbohydrate metabolism (cell wall invertase I (CWINV1) genes) while downregulating stress response genes (ornithine aminotransferase (OAT) and ethylene-responsive transcription factor 1 (ERF1)) in salt-stressed bean plants. The study demonstrates GSE’s usefulness in mitigating salinity stress effects on bean plants by modulating growth, physiology, and gene expression patterns, highlighting its potential as a natural approach to enhance salt tolerance. Full article

(This article belongs to the Special Issue Mechanism of Drought and Salinity Tolerance in Crops)

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14 pages, 4084 KiB

Open AccessArticle

Regional Hydro-Chemistry of Hydrothermal Springs in Northeastern Algeria, Case of Guelma, Souk Ahras, Tebessa and Khenchela Regions

by Ibtissem Djaafri, Karima Seghir, Vincent Valles and Laurent Barbiero

Earth 2024, 5(2), 214-227; https://doi.org/10.3390/earth5020011 (registering DOI) - 8 Jun 2024

Abstract

Hydrothermal units are characterized by the emergence of several large-flow thermo-mineral springs (griffons), each with varying temperature and physico-chemical characteristics depending on the point of emergence. It seems, however, that there is variability between the different systems, although it is not easy to [...] Read more.

Hydrothermal units are characterized by the emergence of several large-flow thermo-mineral springs (griffons), each with varying temperature and physico-chemical characteristics depending on the point of emergence. It seems, however, that there is variability between the different systems, although it is not easy to characterize it because the variability within each system is high. The regional dimension of the chemical composition of thermal waters is, therefore, an aspect that has received very little attention in the literature due to the lack of access to the deep reservoir. In this study, we investigated the spatial variability, on a regional scale, in the characteristics of thermal waters in northeastern Algeria, and more specifically the hydrothermal systems of Guelma, Souk Ahras, Khenchela and Tébessa. Thirty-two hot water samples were taken between December 2018 and October 2019, including five samples of low-temperature mineral spring water. Standard physico-chemical parameters, major anions and cations and lithium were analyzed. The data were log-transformed data and processed via principal component analysis, discriminant analysis and unsupervised classification. The results show that thermal waters are the result of a mixture of hot waters, whose chemical profile has a certain local character, and contaminated by cold surface waters. These surface waters may also have several chemical profiles depending on the location. In addition to the internal variability in each resource, there are differences in water quality between these different hydrothermal systems. The Guelma region differs the most from the other thermal regions studied, with a specific calcic sulfate chemical profile. This question is essential for the rational development of these regional resources in any field whatsoever. Full article

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16 pages, 3044 KiB

Open AccessArticle

Newly Woody Artificial Diet Reveals Antibacterial Activity of Hemolymph in Larvae of Zophobas atratus (Fabricius, 1775) (Coleoptera: Tenebrionidae)

by Alexander Kuprin, Vladislava Baklanova, Maria Khandy, Andrei Grinchenko and Vadim Kumeiko

Insects 2024, 15(6), 435; https://doi.org/10.3390/insects15060435 (registering DOI) - 8 Jun 2024

Abstract

The rearing of saproxylic insects in laboratory conditions is an important task for studying the biology of insects. Through understanding nutritional needs, it is possible to optimize beetle rearing in laboratory conditions. In this study, an artificial fungi-based diet (FD) was developed for [...] Read more.

The rearing of saproxylic insects in laboratory conditions is an important task for studying the biology of insects. Through understanding nutritional needs, it is possible to optimize beetle rearing in laboratory conditions. In this study, an artificial fungi-based diet (FD) was developed for the cultivation of the darkling beetle Zophobas atratus (Fabricius, 1775) (Coleoptera: Tenebrionidae) in laboratory conditions as a model object for studying the biology of saproxylophagous beetles. To assess the influence of the diet, a number of physiological parameters were measured, including development time, body size, and weight of all stages of the beetle’s life cycle, as well as its immune status. The immune status of Z. atratus was assessed on the basis of larval hemolymph antibacterial activity against six different bacterial strains assessed using disk-diffusion and photometric tests. Our findings show that the FD reduces development time and boosts the immune status as compared to beetles reared on a standard diet (SD). Samples from FD-reared larvae had pronounced antibacterial activity as compared to samples from SD-reared larvae. This work is of fundamental importance for understanding the correlations between nutrition and development of saproxylic Coleoptera and is the first report on immune status regulation in this group of insects. Full article

(This article belongs to the Special Issue Insect Rearing: Reserve Forces with Commercial and Ecological Values)

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24 pages, 6241 KiB

Open AccessArticle

Discovery and Characterization of the ddx41 Gene in Atlantic Salmon: Evolutionary Implications, Structural Functions, and Innate Immune Responses to Piscirickettsia salmonis and Renibacterium salmoninarum Infections

by Alejandro J. Yañez, Claudia A. Barrientos, Adolfo Isla, Marcelo Aguilar, Sandra N. Flores-Martin, Yassef Yuivar, Adriana Ojeda, Pablo Ibieta, Mauricio Hernández, Jaime Figueroa, Rubén Avendaño-Herrera and Marcos Mancilla

Int. J. Mol. Sci. 2024, 25(12), 6346; https://doi.org/10.3390/ijms25126346 (registering DOI) - 8 Jun 2024

Abstract

The innate immune response in Salmo salar, mediated by pattern recognition receptors (PRRs), is crucial for defending against pathogens. This study examined DDX41 protein functions as a cytosolic/nuclear sensor for cyclic dinucleotides, RNA, and DNA from invasive intracellular bacteria. The investigation determined [...] Read more.

The innate immune response in Salmo salar, mediated by pattern recognition receptors (PRRs), is crucial for defending against pathogens. This study examined DDX41 protein functions as a cytosolic/nuclear sensor for cyclic dinucleotides, RNA, and DNA from invasive intracellular bacteria. The investigation determined the existence, conservation, and functional expression of the ddx41 gene in S. salar. In silico predictions and experimental validations identified a single ddx41 gene on chromosome 5 in S. salar, showing 83.92% homology with its human counterpart. Transcriptomic analysis in salmon head kidney confirmed gene transcriptional integrity. Proteomic identification through mass spectrometry characterized three unique peptides with 99.99% statistical confidence. Phylogenetic analysis demonstrated significant evolutionary conservation across species. Functional gene expression analysis in SHK-1 cells infected by Piscirickettsia salmonis and Renibacterium salmoninarum indicated significant upregulation of DDX41, correlated with increased proinflammatory cytokine levels and activation of irf3 and interferon signaling pathways. In vivo studies corroborated DDX41 activation in immune responses, particularly when S. salar was challenged with P. salmonis, underscoring its potential in enhancing disease resistance. This is the first study to identify the DDX41 pathway as a key component in S. salar innate immune response to invading pathogens, establishing a basis for future research in salmonid disease resistance. Full article

(This article belongs to the Special Issue Immune System and Immune Responses in Fish and Their Role in Comparative Immunity Study)

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12 pages, 1668 KiB

Open AccessArticle

Effect of ADORA2A Gene Polymorphism and Acute Caffeine Supplementation on Hormonal Response to Resistance Exercise: A Double-Blind, Crossover, Placebo-Controlled Study

by Mohammad Rahman Rahimi, Ekaterina A. Semenova, George John, Fateme Fallah, Andrey K. Larin, Edward V. Generozov and Ildus I. Ahmetov

Nutrients 2024, 16(12), 1803; https://doi.org/10.3390/nu16121803 (registering DOI) - 8 Jun 2024

Abstract

Previous studies have reported that TT genotype carriers of the adenosine A2a receptor (ADORA2A) gene rs5751876 polymorphism have better ergogenic and anti-inflammatory responses to caffeine intake compared to C allele carriers. The aim of the present study was twofold: (1) to [...] Read more.

Previous studies have reported that TT genotype carriers of the adenosine A2a receptor (ADORA2A) gene rs5751876 polymorphism have better ergogenic and anti-inflammatory responses to caffeine intake compared to C allele carriers. The aim of the present study was twofold: (1) to investigate the association of the ADORA2A rs5751876 polymorphism with acute caffeine supplementation on hormonal (growth hormone and testosterone) response to resistance exercise (RE); (2) to examine the relationship between the rs5751876 polymorphism and the resting levels of growth hormone and testosterone in athletes who are light caffeine consumers. A double-blind, crossover, placebo-controlled study involving 30 resistance-trained men (age 21.7 ± 4.1) was conducted to assess the impact of caffeine supplementation on serum growth hormone (GH) and testosterone (TS) levels before, immediately after, and 15 min post-RE. One hour before engaging in resistance exercise, subjects were randomly administered 6 mg of caffeine per kg of body mass or a placebo (maltodextrin). After a 7-day washout period, the same protocol was repeated. Resting testosterone and growth hormone levels were examined in the sera of 94 elite athletes (31 females, age 21.4 ± 2.8; 63 males, age 22.9 ± 3.8). Caffeine consumption led to significantly greater increases in GH and TS in men with the TT genotype compared to C allele carriers. Furthermore, in the group of athletes, carriers of the TT genotype had significantly higher testosterone (p = 0.0125) and growth hormone (p = 0.0365) levels compared to C allele carriers. In conclusion, the ADORA2A gene rs5751876 polymorphism may modify the effect of caffeine intake on the hormonal response to exercise. Full article

(This article belongs to the Special Issue Nutrition and Supplementation Strategies to Enhance Resistance Training Adaptations)

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13 pages, 2324 KiB

Open AccessArticle

Effect of Thermal Treatment on Gelling and Emulsifying Properties of Soy β-Conglycinin and Glycinin

by Wei Zhang, Mengru Jin, Hong Wang, Siqi Cheng, Jialu Cao, Dingrong Kang, Jingnan Zhang, Wei Zhou, Longteng Zhang, Rugang Zhu, Donghong Liu and Guanchen Liu

Foods 2024, 13(12), 1804; https://doi.org/10.3390/foods13121804 (registering DOI) - 8 Jun 2024

Abstract

This study investigated the impact of different preheat treatments on the emulsifying and gel textural properties of soy protein with varying 11S/7S ratios. A mixture of 7S and 11S globulins, obtained from defatted soybean meal, was prepared at different ratios. The mixed proteins [...] Read more.

This study investigated the impact of different preheat treatments on the emulsifying and gel textural properties of soy protein with varying 11S/7S ratios. A mixture of 7S and 11S globulins, obtained from defatted soybean meal, was prepared at different ratios. The mixed proteins were subjected to preheating (75 °C, 85 °C, and 95 °C for 5 min) or non-preheating, followed by spray drying or non-spray drying. The solubility of protein mixtures rich in the 7S fraction tended to decrease significantly after heating at 85 °C, while protein mixtures rich in the 11S fraction showed a significant decrease after heating at 95 °C. Surprisingly, the emulsion stability index (ESI) of protein mixtures rich in the 7S fraction significantly improved twofold during processing at 75 °C. This study revealed a negative correlation between the emulsifying ability of soy protein and the 11S/7S ratio. For protein mixtures rich in either the 7S or the 11S fractions, gelling proprieties as well as emulsion activity index (EAI) and ESI showed no significant changes after spray drying; however, surface hydrophobicity was significantly enhanced following heating at 85 °C post-spray drying treatment. These findings provide insights into the alterations in gelling and emulsifying properties during various heating processes, offering great potential for producing soy protein ingredients with enhanced emulsifying ability and gelling property. They also contribute to establishing a theoretical basis for the standardized production of soy protein isolate with specific functional characteristics. Full article

(This article belongs to the Special Issue Functionality and Food Applications of Plant Proteins (Volume II))

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19 pages, 1478 KiB

Open AccessArticle

Clustering Mixed-Type Data via Dirichlet Process Mixture Model with Cluster-Specific Covariance Matrices

by Nurul Afiqah Burhanuddin, Kamarulzaman Ibrahim, Hani Syahida Zulkafli and Norwati Mustapha

Symmetry 2024, 16(6), 712; https://doi.org/10.3390/sym16060712 (registering DOI) - 8 Jun 2024

Abstract

Many studies have shown successful applications of the Dirichlet process mixture model (DPMM) for clustering continuous data. Beyond continuous data, in practice, one can expect to see different data types, including ordinal and nominal data. Existing DPMMs for clustering mixed-type data assume a [...] Read more.

Many studies have shown successful applications of the Dirichlet process mixture model (DPMM) for clustering continuous data. Beyond continuous data, in practice, one can expect to see different data types, including ordinal and nominal data. Existing DPMMs for clustering mixed-type data assume a strict covariance matrix structure, resulting in an overfit model. This article explores a DPMM for mixed-type data that allows the covariance matrix to differ from one cluster to another. We assume an underlying latent variable framework for ordinal and nominal data, which is then modeled jointly with the continuous data. The identifiability issue on the covariance matrix poses computational challenges, thus requiring a nonstandard inferential algorithm. The applicability and flexibility of the proposed model are illustrated through simulation examples and real data applications. Full article

(This article belongs to the Section Mathematics)

11 pages, 1898 KiB

Open AccessArticle

The Clinical Role of Adjuvant Chemotherapy after Sublobar Resection for Non-Small-Cell Lung Cancer ≤ 20 mm with Lymph Node Metastases: A Propensity-Matched Analysis of the National Cancer Database

by Shinkichi Takamori, Junjia Zhu, Asato Hashinokuchi and Takefumi Komiya

Cancers 2024, 16(12), 2176; https://doi.org/10.3390/cancers16122176 (registering DOI) - 8 Jun 2024

Abstract

Sublobar resection is a standard surgical procedure for small-sized non-small-cell lung cancer (NSCLC). However, the clinical role of adjuvant chemotherapy for small-sized NSCLC with pathological lymph node (LN) metastasis after sublobar resection is unknown. The National Cancer Database was queried for NSCLC patients [...] Read more.

Sublobar resection is a standard surgical procedure for small-sized non-small-cell lung cancer (NSCLC). However, the clinical role of adjuvant chemotherapy for small-sized NSCLC with pathological lymph node (LN) metastasis after sublobar resection is unknown. The National Cancer Database was queried for NSCLC patients between 2004 and 2018. Eligibility included sublobar resection with pathological LN metastasis, R0 resection, Charlson comorbidity score = 0, clinical stage T1a-b, and tumor size ≤ 20 mm. The Kaplan–Meier method with a log-rank test and multivariable Cox proportional hazards analyses were used for assessing survival. The samples were evaluated before and after propensity score matching (PSM) with respect to age, sex, histologic type, and pathological LN status. Of 810 patients who met the eligibility criteria, 567 (70.0%) underwent adjuvant chemotherapy. After PSM, patients with adjuvant chemotherapy had a significantly longer survival than those without (median survival: 64.3 vs. 34.0 months, hazard ratio for death: 0.61, p < 0.0001). Multivariate analyses after PSM showed that younger age (p = 0.0206), female (p = 0.0005), and adjuvant chemotherapy (p < 0.0001) were independent prognostic factors for longer survival. Adjuvant chemotherapy has a prognostic impact in patients with small-sized NSCLC and pathological lymph node metastasis who undergo sublobar resection. Full article

(This article belongs to the Special Issue A New Era in Treatment of Early-Stage Non-small Cell Lung Cancer)

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21 pages, 2664 KiB

Open AccessArticle

Early Inhibition of Phosphodiesterase 4B (PDE4B) Instills Cognitive Resilience in APPswe/PS1dE9 Mice

by Ben Rombaut, Melissa Schepers, Assia Tiane, Femke Mussen, Lisa Koole, Sofie Kessels, Chloë Trippaers, Ruben Jacobs, Kristiaan Wouters, Emily Willems, Lieve van Veggel, Philippos Koulousakis, Dorien Deluyker, Virginie Bito, Jos Prickaerts, Inez Wens, Bert Brône, Daniel L. A. van den Hove and Tim Vanmierlo

Cells 2024, 13(12), 1000; https://doi.org/10.3390/cells13121000 (registering DOI) - 8 Jun 2024

Abstract

Microglia activity can drive excessive synaptic loss during the prodromal phase of Alzheimer’s disease (AD) and is associated with lowered cyclic adenosine monophosphate (cAMP) due to cAMP phosphodiesterase 4B (PDE4B). This study aimed to investigate whether long-term inhibition of PDE4B by A33 (3 [...] Read more.

Microglia activity can drive excessive synaptic loss during the prodromal phase of Alzheimer’s disease (AD) and is associated with lowered cyclic adenosine monophosphate (cAMP) due to cAMP phosphodiesterase 4B (PDE4B). This study aimed to investigate whether long-term inhibition of PDE4B by A33 (3 mg/kg/day) can prevent synapse loss and its associated cognitive decline in APPswe/PS1dE9 mice. This model is characterized by a chimeric mouse/human APP with the Swedish mutation and human PSEN1 lacking exon 9 (dE9), both under the control of the mouse prion protein promoter. The effects on cognitive function of prolonged A33 treatment from 20 days to 4 months of age, was assessed at 7–8 months. PDE4B inhibition significantly improved both the working and spatial memory of APPswe/PSdE9 mice after treatment ended. At the cellular level, in vitro inhibition of PDE4B induced microglial filopodia formation, suggesting that regulation of PDE4B activity can counteract microglia activation. Further research is needed to investigate if this could prevent microglia from adopting their ‘disease-associated microglia (DAM)’ phenotype in vivo. These findings support the possibility that PDE4B is a potential target in combating AD pathology and that early intervention using A33 may be a promising treatment strategy for AD. Full article

(This article belongs to the Special Issue Phosphodiesterases (PDEs): Therapeutic Targets in Human Health and Disease)

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20 pages, 928 KiB

Open AccessArticle

Gestational Age, Infection, and Suboptimal Maternal Prepregnancy BMI Independently Associate with Placental Histopathology in a Cohort of Pregnancies without Major Maternal Comorbidities

by Eleanor Duffley, David Grynspan, Hailey Scott, Anthea Lafrenière, Cherley Borba Vieira de Andrade, Enrrico Bloise and Kristin L. Connor

J. Clin. Med. 2024, 13(12), 3378; https://doi.org/10.3390/jcm13123378 (registering DOI) - 8 Jun 2024

Abstract

Background: The placenta undergoes morphological and functional adaptations to adverse exposures during pregnancy. The effects ofsuboptimal maternal body mass index (BMI), preterm birth, and infection on placental histopathological phenotypes are not yet well understood, despite the association between these conditions and poor offspring [...] Read more.

Background: The placenta undergoes morphological and functional adaptations to adverse exposures during pregnancy. The effects ofsuboptimal maternal body mass index (BMI), preterm birth, and infection on placental histopathological phenotypes are not yet well understood, despite the association between these conditions and poor offspring outcomes. We hypothesized that suboptimal maternal prepregnancy BMI and preterm birth (with and without infection) would associate with altered placental maturity and morphometry, and that altered placental maturity would associate with poor birth outcomes. Methods: Clinical data and human placentae were collected from 96 pregnancies where mothers were underweight, normal weight, overweight, or obese, without other major complications. Placental histopathological characteristics were scored by an anatomical pathologist. Associations between maternal BMI, placental pathology (immaturity and hypermaturity), placental morphometry, and infant outcomes were investigated for term and preterm births with and without infection. Results: Fetal capillary volumetric proportion was decreased, whereas the villous stromal volumetric proportion was increased in placentae from preterm pregnancies with chorioamnionitis compared to preterm placentae without chorioamnionitis. At term and preterm, pregnancies with maternal overweight and obesity had a high percentage increase in proportion of immature placentae compared to normal weight. Placental maturity did not associate with infant birth outcomes. We observed placental hypermaturity and altered placental morphometry among preterm pregnancies with chorioamnionitis, suggestive of altered placental development, which may inform about pregnancies susceptible to preterm birth and infection. Conclusions: Our data increase our understanding of how common metabolic exposures and preterm birth, in the absence of other comorbidities or complications, potentially contribute to poor pregnancy outcomes and developmental programming. Full article

(This article belongs to the Special Issue New Insights into Pregnancy Complications)

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26 pages, 1655 KiB

Open AccessArticle

A Physiologically Based Pharmacokinetic (PBPK) Study to Assess the Adjuvanticity of Tree Peptides in an Oral Vaccine

by Leonor Saldanha, Ülo Langel and Nuno Vale

Pharmaceutics 2024, 16(6), 780; https://doi.org/10.3390/pharmaceutics16060780 (registering DOI) - 8 Jun 2024

Abstract

Following up on the first PBPK model for an oral vaccine built for alpha-tocopherol, three peptides are explored in this article to verify if they could support an oral vaccine formulation as adjuvants using the same PBPK modeling approach. A literature review was [...] Read more.

Following up on the first PBPK model for an oral vaccine built for alpha-tocopherol, three peptides are explored in this article to verify if they could support an oral vaccine formulation as adjuvants using the same PBPK modeling approach. A literature review was conducted to verify what peptides have been used as adjuvants in the last decades, and it was noticed that MDP derivatives have been used, with one of them even being commercially approved and used as an adjuvant when administered intravenously in oncology. The aim of this study was to build optimized models for three MDP peptides (MDP itself, MTP-PE, and murabutide) and to verify if they could act as adjuvants for an oral vaccine. Challenges faced by peptides in an oral delivery system are taken into consideration, and improvements to the formulations to achieve better results are described in a step-wise approach to reach the most-optimized model. Once simulations are performed, results are compared to determine what would be the best peptide to support as an oral adjuvant. According to our results, MTP-PE, the currently approved and commercialized peptide, could have potential to be incorporated into an oral formulation. It would be interesting to proceed with further in vivo experiments to determine the behavior of this peptide when administered orally with a proper formulation to overcome the challenges of oral delivery systems. Full article

(This article belongs to the Special Issue Approaches to Individualized Drug Therapy Based on Population Pharmacometrics)

13 pages, 5470 KiB

Open AccessArticle

A Simple and Rapid Microscale Method for Isolating Bacterial Lipopolysaccharides

by Daniil Grumov, Alexey Kostarnoy, Petya Gancheva and Alexey Kondratev

Int. J. Mol. Sci. 2024, 25(12), 6345; https://doi.org/10.3390/ijms25126345 (registering DOI) - 8 Jun 2024

Abstract

Bacterial endotoxins (lipopolysaccharides (LPSs)) are important mediators of inflammatory processes induced by Gram-negative microorganisms. LPSs are the key inducers of septic shock due to a Gram-negative bacterial infection; thus, the structure and functions of LPSs are of specific interest. Often, highly purified bacterial [...] Read more.

Bacterial endotoxins (lipopolysaccharides (LPSs)) are important mediators of inflammatory processes induced by Gram-negative microorganisms. LPSs are the key inducers of septic shock due to a Gram-negative bacterial infection; thus, the structure and functions of LPSs are of specific interest. Often, highly purified bacterial endotoxins must be isolated from small amounts of biological material. Each of the currently available methods for LPS extraction has certain limitations. Herein, we describe a rapid and simple microscale method for extracting LPSs. The method consists of the following steps: ultrasonic destruction of the bacterial material, LPS extraction via heating, LPS purification with organic solvents, and treatment with proteinase K. LPSs that were extracted by using this method contained less than 2–3% protein and 1% total nucleic acid. We also demonstrated the structural integrity of the O-antigen and lipid A via the sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS–PAGE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI–MS) methods, respectively. We demonstrated the ability of the extracted LPSs to induce typical secretion of cytokines and chemokines by primary macrophages. Overall, this method may be used to isolate purified LPSs with preserved structures of both the O-antigen and lipid A and unchanged functional activity from small amounts of bacterial biomass. Full article

(This article belongs to the Collection Feature Papers in Molecular Microbiology)

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16 pages, 664 KiB

Open AccessReview

Interferon-Stimulated Genes that Target Retrovirus Translation

by Niklas Jäger, Stefan Pöhlmann, Marina V. Rodnina and Shreya Ahana Ayyub

Viruses 2024, 16(6), 933; https://doi.org/10.3390/v16060933 (registering DOI) - 8 Jun 2024

Abstract

The innate immune system, particularly the interferon (IFN) system, constitutes the initial line of defense against viral infections. IFN signaling induces the expression of interferon-stimulated genes (ISGs), and their products frequently restrict viral infection. Retroviruses like the human immunodeficiency viruses and the human [...] Read more.

The innate immune system, particularly the interferon (IFN) system, constitutes the initial line of defense against viral infections. IFN signaling induces the expression of interferon-stimulated genes (ISGs), and their products frequently restrict viral infection. Retroviruses like the human immunodeficiency viruses and the human T-lymphotropic viruses cause severe human diseases and are targeted by ISG-encoded proteins. Here, we discuss ISGs that inhibit the translation of retroviral mRNAs and thereby retrovirus propagation. The Schlafen proteins degrade cellular tRNAs and rRNAs needed for translation. Zinc Finger Antiviral Protein and RNA-activated protein kinase inhibit translation initiation factors, and Shiftless suppresses translation recoding essential for the expression of retroviral enzymes. We outline common mechanisms that underlie the antiviral activity of multifunctional ISGs and discuss potential antiretroviral therapeutic approaches based on the mode of action of these ISGs. Full article

(This article belongs to the Special Issue Innate Sensing and Restriction of Retroviruses)

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27 pages, 3934 KiB

Open AccessArticle

Evidence That a Peptide-Drug/p53 Gene Complex Promotes Cognate Gene Expression and Inhibits the Viability of Glioblastoma Cells

by Ana Neves, Tânia Albuquerque, Rúben Faria, Cecília R. A. Santos, Eric Vivès, Prisca Boisguérin, Diana Carneiro, Daniel F. Bruno, Maria D. Pavlaki, Susana Loureiro, Ângela Sousa and Diana Costa

Pharmaceutics 2024, 16(6), 781; https://doi.org/10.3390/pharmaceutics16060781 (registering DOI) - 8 Jun 2024

Abstract

Glioblastoma multiform (GBM) is considered the deadliest brain cancer. Conventional therapies are followed by poor patient survival outcomes, so novel and more efficacious therapeutic strategies are imperative to tackle this scourge. Gene therapy has emerged as an exciting and innovative tool in cancer [...] Read more.

Glioblastoma multiform (GBM) is considered the deadliest brain cancer. Conventional therapies are followed by poor patient survival outcomes, so novel and more efficacious therapeutic strategies are imperative to tackle this scourge. Gene therapy has emerged as an exciting and innovative tool in cancer therapy. Its combination with chemotherapy has significantly improved therapeutic outcomes. In line with this, our team has developed temozolomide–transferrin (Tf) peptide (WRAP5)/p53 gene nanometric complexes that were revealed to be biocompatible with non-cancerous cells and in a zebrafish model and were able to efficiently target and internalize into SNB19 and U373 glioma cell lines. The transfection of these cells, mediated by the formulated peptide-drug/gene complexes, resulted in p53 expression. The combined action of the anticancer drug with p53 supplementation in cancer cells enhances cytotoxicity, which was correlated to apoptosis activation through quantification of caspase-3 activity. In addition, increased caspase-9 levels revealed that the intrinsic or mitochondrial pathway of apoptosis was implicated. This assumption was further evidenced by the presence, in glioma cells, of Bax protein overexpression—a core regulator of this apoptotic pathway. Our findings demonstrated the great potential of peptide TMZ/p53 co-delivery complexes for cellular transfection, p53 expression, and apoptosis induction, holding promising therapeutic value toward glioblastoma. Full article

(This article belongs to the Special Issue Recent Advances in Nanotechnology Therapeutics)

26 pages, 1639 KiB

Open AccessArticle

Towards the Improvement of Patient Experience Evaluation Items for Patient-Centered Care in Head and Neck Cancer: A Qualitative Comparative Study

by Eun-Jeong Kim, Yoo-Ri Koo and Inn-Chul Nam

Healthcare 2024, 12(12), 1164; https://doi.org/10.3390/healthcare12121164 (registering DOI) - 8 Jun 2024

Abstract

Owing to long-term treatment, frequent consultations, and complications, the evaluation of patients with head and neck cancer (HNC) must be improved. This study explored an opportunity for improving patient experience (PE) evaluation of patients with HNC to achieve a patient-centered, integrative evaluation model [...] Read more.

Owing to long-term treatment, frequent consultations, and complications, the evaluation of patients with head and neck cancer (HNC) must be improved. This study explored an opportunity for improving patient experience (PE) evaluation of patients with HNC to achieve a patient-centered, integrative evaluation model based on patient needs. The study comprised four phases: (1) a systematic literature review of PE factors for patient quality of life (QoL) and establishment of PE factor categories as a framework, (2) a review of current cancer or HNC PE evaluation tools, (3) identification of potential PE evaluation items based on patient needs by conducting user research, and (4) suggestion of integrative HNC PE evaluation items through expert validation. As a result, the 39 potential items were initially identified from the literature review and user research. After conducting two survey rounds with experts, 25 items were suggested as HNC PE evaluation items. These underscore the importance of highlighting the patient’s participation, the medical staff’s comprehensive information delivery, empathy, and collaborative communication, the hospital’s support of communication channels, the medical environment for patient emotional support, the education program, and systematic patient satisfaction data management. PE evaluation items that consider the diverse perspectives of stakeholders involved in HNC treatment and factors of comprehensive PE will contribute to improving HNC patient-centered care (PCC). Full article

(This article belongs to the Special Issue Quality of Life and Health Promotion in Chronic Illness or Critical Medical Conditions: Implications for Integrated Health Care Delivery)

15 pages, 1249 KiB

Open AccessArticle

Use of Deep-Amplicon Sequencing (DAS), Real-Time PCR and In Situ Hybridization to Detect H. pylori and Other Pathogenic Helicobacter Species in Feces from Children

by Yolanda Moreno Trigos, Miguel Tortajada-Girbés, Raquel Simó-Jordá, Manuel Hernández Pérez, Irene Hortelano, Miguel García-Ferrús and María Antonia Ferrús Pérez

Diagnostics 2024, 14(12), 1216; https://doi.org/10.3390/diagnostics14121216 (registering DOI) - 8 Jun 2024

Abstract

Background: Detecting Helicobacter pylori in fecal samples is easier and more comfortable than invasive techniques, especially in children. Thus, the objective of the present work was to detect H. pylori in feces from children by molecular methods as an alternative for diagnostic and [...] Read more.

Background: Detecting Helicobacter pylori in fecal samples is easier and more comfortable than invasive techniques, especially in children. Thus, the objective of the present work was to detect H. pylori in feces from children by molecular methods as an alternative for diagnostic and epidemiological studies. Methods: Forty-five fecal samples were taken from pediatric patients who presented symptoms compatible with H. pylori infection. HpSA test, culture, real-time quantitative PCR (qPCR), fluorescence in situ hybridization (FISH), direct viable count associated with FISH (DVC-FISH), and Illumina-based deep-amplicon sequencing (DAS) were applied. Results: No H. pylori colonies were isolated from the samples. qPCR analysis detected H. pylori in the feces of 24.4% of the patients. In comparison, DVC-FISH analysis showed the presence of viable H. pylori cells in 53.3% of the samples, 37% of which carried 23S rRNA mutations that confer resistance to clarithromycin. After DAS, H. pylori-specific 16S rDNA sequences were detected in 26 samples. In addition, DNA from H. hepaticus was identified in 10 samples, and H. pullorum DNA was detected in one sample. Conclusion: The results of this study show the presence of H. pylori, H. hepaticus, and H. pullorum in children’s stools, demonstrating the coexistence of more than one Helicobacter species in the same patient. The DVC-FISH method showed the presence of viable, potentially infective H. pylori cells in a high percentage of the children’s stools. These results support the idea that fecal–oral transmission is probably a common route for H. pylori and suggest possible fecal–oral transmission of other pathogenic Helicobacter species. Full article

(This article belongs to the Special Issue Diagnosis and Management of Gastrointestinal Infections)

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21 pages, 1228 KiB

Open AccessFeature PaperArticle

Valuable Nutrients, Aroma Profile, and Functional Bioactives Extracted by Eco-Friendly Extraction Techniques from Wild Olive Fruits (Olea europaea L. var. sylvestris)

by Maria Irakli, Theano Samara, Elisavet Bouloumpasi, Kalliopi Kadoglidou, Paschalina Chatzopoulou and Ioannis Spanos

Processes 2024, 12(6), 1181; https://doi.org/10.3390/pr12061181 (registering DOI) - 8 Jun 2024

Abstract

Wild olive tree, or oleaster (var. sylvestris), native to the Mediterranean region, is considered a traditional source of healthy food. Wild olive fruit (WOF) exhibits several biological properties associated with its chemical composition. Although Greece has important olive genetic resources, including oleaster [...] Read more.

Wild olive tree, or oleaster (var. sylvestris), native to the Mediterranean region, is considered a traditional source of healthy food. Wild olive fruit (WOF) exhibits several biological properties associated with its chemical composition. Although Greece has important olive genetic resources, including oleaster populations, limited information is available on the chemical characterization of WOF. Therefore, the present investigation was undertaken to study the nutritional, bioactive (phenolics, tocopherols, and pigments), volatile profile, and antioxidant properties of WOF collected from Greece. Moreover, eco-friendly processes, including ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), and accelerated solvent extraction (ASE), were applied to obtain oleuropein-rich extracts from WOF. Evaluation of phenolic composition in WOF extracts was carried out by LC-DAD-ESI-MS, while antioxidant activity was evaluated by applying DPPH, ABTS, and FRAP methods. Our outcomes show that Greek WOF is rich in oleic acid (71.55%), total phenolics (64.89 mg GAE/g dw), tocopherols (107.05 mg/kg dw), and carotenoids (85.90 mg/kg dw). Oleuropein (72.03 mg/g dw) was the main phenolic compound in the WOF extracts. Also, the WOF was characterized by an increased level of volatile compounds, mainly terpenoids (46.73%). UAE was more efficient than MAE and ASE for recovering oleuropein-rich extracts with high antioxidant activity. These results emphasized the high potential of WOF as an alternative bioactive ingredient for use in the food industry. Full article

(This article belongs to the Special Issue Technologies for Production, Processing, and Extractions of Nature Product Compounds, 2nd Volume)

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14 pages, 255 KiB

Open AccessArticle

Influenza Vaccine Hesitancy among Cancer Survivors in China: A Multicenter Survey

by Xin Guo, Qi Han, Yuqin Wang, Rui Zhang, Yuenan Huang and Botang Guo

Vaccines 2024, 12(6), 639; https://doi.org/10.3390/vaccines12060639 (registering DOI) - 8 Jun 2024

Abstract

Background: Cancer survivors are at higher risk of developing severe complications from influenza due to their compromised immune systems. Despite their increased vulnerability to influenza and the availability of vaccines, vaccine hesitancy among cancer survivors remains a significant public health concern in China. [...] Read more.

Background: Cancer survivors are at higher risk of developing severe complications from influenza due to their compromised immune systems. Despite their increased vulnerability to influenza and the availability of vaccines, vaccine hesitancy among cancer survivors remains a significant public health concern in China. Methods: A multicenter, cross-sectional study was conducted among cancer survivors in China from January to December 2023. A total of 500 participants were recruited from the oncology departments of five tertiary hospitals. A structured, self-administered questionnaire was used to collect data on socio-demographic characteristics, cancer-related information, medical history, lifestyle factors, and influenza vaccine hesitancy. Univariate and multivariate logistic regression analyses were performed to identify factors associated with influenza vaccine hesitancy. Results: The response rate was 97.0% (485/500). Among all participants, 204 (42.06%) reported vaccine hesitancy. The results of multiple logistic regression showed that the longer the end of anti-cancer treatment, without a history of adverse vaccine reactions, and the level of family support played a protective role in vaccine hesitancy. Current rehabilitation status, frequent colds, not being informed by doctors about vaccination, exercising, lack of community vaccination education programs, and concerns about vaccine safety were risk factors that increase vaccine hesitancy. Conclusions: A high proportion of cancer survivors in our study reported influenza vaccine hesitancy. Addressing concerns about vaccine safety, improving access to vaccination services, and enhancing doctor–patient communication are crucial for increasing influenza vaccine uptake in this vulnerable population. Full article

(This article belongs to the Special Issue Vaccination Uptake and Public Health)

16 pages, 2949 KiB

Open AccessArticle

Tomato SlWRKY3 Negatively Regulates Botrytis cinerea Resistance via TPK1b

by Dan Luo, Jun Cai, Wenhui Sun, Qihong Yang, Guoyu Hu and Taotao Wang

Plants 2024, 13(12), 1597; https://doi.org/10.3390/plants13121597 (registering DOI) - 8 Jun 2024

Abstract

Botrytis cinerea is considered the second most important fungal plant pathogen, and can cause serious disease, especially on tomato. The TPK1b gene encodes a receptor-like kinase that can positively regulate plant resistance to B. cinerea. Here, we identified a tomato WRKY transcription [...] Read more.

Botrytis cinerea is considered the second most important fungal plant pathogen, and can cause serious disease, especially on tomato. The TPK1b gene encodes a receptor-like kinase that can positively regulate plant resistance to B. cinerea. Here, we identified a tomato WRKY transcription factor SlWRKY3 that binds to the W-box on the TPK1b promoter. It can negatively regulate TPK1b transcription, then regulate downstream signaling pathways, and ultimately negatively regulate tomato resistance to B. cinerea. SlWRKY3 interference can enhance resistance to B. cinerea, and SlWRKY3 overexpression leads to susceptibility to B. cinerea. Additionally, we found that B. cinerea can significantly, and rapidly, induce the upregulation of SlWRKY3 expression. In SlWRKY3 transgenic plants, the TPK1b expression level was negatively correlated with SlWRKY3 expression. Compared with the control, the expression of the SA pathway marker gene PR1 was downregulated in W3-OE plants and upregulated in W3-Ri plants when inoculated with B. cinerea for 48 h. Moreover, SlWRKY3 positively regulated ROS production. Overall, SlWRKY3 can inhibit TPK1b transcription in tomato, and negatively regulate resistance to B. cinerea by modulating the downstream SA and ROS pathways. Full article

(This article belongs to the Section Horticultural Science and Ornamental Plants)

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29 pages, 12228 KiB

Open AccessArticle

RadWet-L: A Novel Approach for Mapping of Inundation Dynamics of Forested Wetlands Using ALOS-2 PALSAR-2 L-Band Radar Imagery

by Gregory Oakes, Andy Hardy, Pete Bunting and Ake Rosenqvist

Remote Sens. 2024, 16(12), 2078; https://doi.org/10.3390/rs16122078 (registering DOI) - 8 Jun 2024

Abstract

The ability to accurately map tropical wetland dynamics can significantly contribute to a number of areas, including food and water security, protection and enhancement of ecosystems, flood hazard management, and our understanding of natural greenhouse gas emissions. Yet currently, there is not a [...] Read more.

The ability to accurately map tropical wetland dynamics can significantly contribute to a number of areas, including food and water security, protection and enhancement of ecosystems, flood hazard management, and our understanding of natural greenhouse gas emissions. Yet currently, there is not a tractable solution for mapping tropical forested wetlands at high spatial and temporal resolutions at a regional scale. This means that we lack accurate and up-to-date information about some of the world’s most significant wetlands, including the Amazon Basin. RadWet-L is an automated machine-learning classification technique for the mapping of both inundated forests and open water using ALOS ScanSAR data. We applied and validated RadWet-L for the Amazon Basin. The proposed method is computationally light and transferable across the range of landscape types in the Amazon Basin allowing, for the first time, regional inundation maps to be produced every 42 days at 50 m resolution over the period 2019–2023. Time series estimates of inundation extent from RadWet-L were significantly correlated with NASA-GFZ GRACE-FO water thickness (Pearson’s r = 0.96, p < 0.01), USDA G-REALM lake hight (Pearson’s r between 0.63 and 0.91, p < 0.01), and in situ river stage measurements (Pearson’s r between 0.78 and 0.94, p < 0.01). Additionally, we conducted an evaluation of 11,162 points against the input ScanSAR data revealing spatial and temporal consistency in the approach (F1 score = 0.97). Serial classifications of ALOS-2 PALSAR-2 ScanSAR data by RadWet-L can provide unique insights into the spatio-temporal inundation dynamics within the Amazon Basin. Understanding these dynamics can inform policy in the sustainable use of these wetlands, as well as the impacts of inundation dynamics on biodiversity and greenhouse gas budgets. Full article

(This article belongs to the Special Issue Remote Sensing for Mapping and Monitoring Wetlands and Their Ecosystems)

13 pages, 4482 KiB

Open AccessArticle

Preparation and Characterization of Chitosan/Hydroxypropyl Methylcellulose Temperature-Sensitive Hydrogel-Containing Inorganic Salts for Forest Fire Suppression

by Yanni Gao, Yuzhou Zhao and Ting Wang

Gels 2024, 10(6), 390; https://doi.org/10.3390/gels10060390 (registering DOI) - 8 Jun 2024

Abstract

Effective forest fire suppression remains a critical challenge, necessitating innovative solutions. Temperature-sensitive hydrogels represent a promising avenue in this endeavor. Traditional firefighting methods often struggle to address forest fires efficiently while mitigating ecological harm and optimizing resource utilization. In this study, a novel [...] Read more.

Effective forest fire suppression remains a critical challenge, necessitating innovative solutions. Temperature-sensitive hydrogels represent a promising avenue in this endeavor. Traditional firefighting methods often struggle to address forest fires efficiently while mitigating ecological harm and optimizing resource utilization. In this study, a novel intelligent temperature-sensitive hydrogel was prepared specially for forest fire extinguishment. Utilizing a one-pot synthesis approach, this material demonstrates exceptional fluidity at ambient temperatures, facilitating convenient application and transport. Upon exposure to elevated temperatures, it undergoes a phase transition to form a solid, barrier-like structure essential for containing forest fires. The incorporation of environmentally friendly phosphorus salts into the chitosan/hydroxypropyl methylcellulose gel system enhances the formation of temperature-sensitive hydrogels, thereby enhancing their structural integrity and firefighting efficacy. Morphological and thermal stability analyses elucidate the outstanding performance, with the hydrogel forming a dense carbonized layer that acts as a robust barrier against the spread of forest fires. Additionally, comprehensive evaluations employing rheological tests, cone calorimeter tests, a swelling test, and infrared thermography reveal the multifaceted roles of temperature-sensitive hydrogels in forest fire prevention and suppression strategies. Full article

(This article belongs to the Section Gel Analysis and Characterization)

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18 pages, 2075 KiB

Open AccessFeature PaperArticle

Impact of Nutritional Strategies to Prevent Post-Weaning Diarrhoea on Performance, Behaviour, and Microbiota in Piglets from Organic Farming

by Marc Bagaria, Yuliaxis Ramayo-Caldas, Olga González-Rodríguez, Lluís Vila, Pino Delàs and Emma Fàbrega

Animals 2024, 14(12), 1730; https://doi.org/10.3390/ani14121730 (registering DOI) - 8 Jun 2024

Abstract

Organic livestock farming is committed to high environmental and animal welfare standards, although pathologies such as post-weaning diarrhoea (PWD) may appear. The main objective of this study was to assess nutritional strategies to prevent PWD in organic piglets. A total of 134 weaned [...] Read more.

Organic livestock farming is committed to high environmental and animal welfare standards, although pathologies such as post-weaning diarrhoea (PWD) may appear. The main objective of this study was to assess nutritional strategies to prevent PWD in organic piglets. A total of 134 weaned piglets were fed one of three diets: high crude protein (17.8%, HCP), low crude protein (16.8%, LCP), and low crude protein supplemented with liquid whey (LCP+W). Piglets were assessed weekly for four weeks on the following parameters: diarrhoea incidence, additional health parameters, average daily gain, and behaviour. Faecal samples were taken to analyse the intestinal microbiota composition. Data were analysed using LMM and GLMM models and Shannon and Whittaker indexes. No significant effect of diet on diarrhoea incidence was found, but the LCP+W diet increased average daily gain. Pigs fed the LCP+W diet presented a lower percentage of drinking and negative social behaviour compared with the HCP diet, and LCP pigs presented higher exploration compared with HCP. In addition, LCP+W piglets showed a higher abundance of the beneficial genus Frisingicoccus. Although liquid whey did not reduce diarrhoea incidence, the benefits found in growth, microbiota composition, and reduced negative social behaviour indicate that it could be an optimal supplement to organic diets. Full article

(This article belongs to the Special Issue Editorial Board Members’ Collection Series: Behavior, Applied Ethology and Welfare of Farmed Animals)

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17 pages, 1814 KiB

Open AccessReview

G9a in Cancer: Mechanisms, Therapeutic Advancements, and Clinical Implications

by Yuchao Ni, Mingchen Shi, Liangliang Liu, Dong Lin, Hao Zeng, Christopher Ong and Yuzhuo Wang

Cancers 2024, 16(12), 2175; https://doi.org/10.3390/cancers16122175 (registering DOI) - 8 Jun 2024

Abstract

G9a, also named EHMT2, is a histone 3 lysine 9 (H3K9) methyltransferase responsible for catalyzing H3K9 mono- and dimethylation (H3K9me1 and H3K9me2). G9a contributes to various aspects of embryonic development and tissue differentiation through epigenetic regulation. Furthermore, the aberrant expression of G9a is [...] Read more.

G9a, also named EHMT2, is a histone 3 lysine 9 (H3K9) methyltransferase responsible for catalyzing H3K9 mono- and dimethylation (H3K9me1 and H3K9me2). G9a contributes to various aspects of embryonic development and tissue differentiation through epigenetic regulation. Furthermore, the aberrant expression of G9a is frequently observed in various tumors, particularly in prostate cancer, where it contributes to cancer pathogenesis and progression. This review highlights the critical role of G9a in multiple cancer-related processes, such as epigenetic dysregulation, tumor suppressor gene silencing, cancer lineage plasticity, hypoxia adaption, and cancer progression. Despite the increased research on G9a in prostate cancer, there are still significant gaps, particularly in understanding its interactions within the tumor microenvironment and its broader epigenetic effects. Furthermore, this review discusses the recent advancements in G9a inhibitors, including the development of dual-target inhibitors that target G9a along with other epigenetic factors such as EZH2 and HDAC. It aims to bring together the existing knowledge, identify gaps in the current research, and suggest future directions for research and treatment strategies. Full article

(This article belongs to the Section Molecular Cancer Biology)

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