Biology

16 pages, 2972 KiB

Open AccessArticle

Protective Effects of N-Acetylcysteine in Alleviating Cocaine-Mediated Microglial Activation and Neuroinflammation

by Uma Maheswari Deshetty, Abiola Oladapo, Yazhini Mohankumar, Elias Horanieh, Shilpa Buch and Palsamy Periyasamy

Biology 2025, 14(7), 893; https://doi.org/10.3390/biology14070893 (registering DOI) - 20 Jul 2025

Abstract

Cocaine misuse induces microglial activation and neuroinflammation, contributing to neurodegeneration and behavioral impairments. Prior studies have shown that cocaine induces mitochondrial dysfunction, dysregulated mitophagy, and lysosomal impairment in microglia. Here, we investigated the therapeutic potential of N-acetylcysteine (NAC) in mitigating cocaine-induced microglial activation [...] Read more.

Cocaine misuse induces microglial activation and neuroinflammation, contributing to neurodegeneration and behavioral impairments. Prior studies have shown that cocaine induces mitochondrial dysfunction, dysregulated mitophagy, and lysosomal impairment in microglia. Here, we investigated the therapeutic potential of N-acetylcysteine (NAC) in mitigating cocaine-induced microglial activation and neuroinflammation. Mouse primary microglial cells (MPMs) were pretreated with NAC (5 mM) for 1 h prior to cocaine exposure (10 µM, 24 h) and analyzed for markers of microglial activation, mitophagy, and lysosomal integrity using Western blot, Seahorse assays, lysosomal pH, and membrane potential measurements. In vivo, C57BL/6N mice received NAC (200 mg/kg, i.p.) 1 h before daily cocaine injections (20 mg/kg, i.p.) for 7 days. Behavioral assays (open field, novel object recognition) and brain biomarker analyses (frontal cortex, hippocampus) were performed. Cocaine exposure elevated CD11b, mitophagy markers (PINK1, PARK, and DLP1), and autophagy proteins (Beclin1, and p62), while impairing mitochondrial and lysosomal functions. NAC pretreatment restored mitochondrial and lysosomal function, reduced reactive oxygen species, and normalized protein expression. In vivo, NAC also alleviated cocaine-induced microglial activation and behavioral deficits. These findings highlight NAC as a promising therapeutic agent to counteract cocaine-mediated neuroinflammation and neurotoxicity. Full article

(This article belongs to the Section Biochemistry and Molecular Biology)

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20 pages, 2031 KiB

Open AccessReview

Anti-Inflammatory Pathways Mediating tDCS’s Effects on Neuropathic Pain

by Haipeng Zhang, Xinyan Zheng and Binn Zhang

Biology 2025, 14(7), 892; https://doi.org/10.3390/biology14070892 (registering DOI) - 20 Jul 2025

Abstract

Neuropathic pain (NP) is a prevalent clinical condition resulting from diseases or injuries affecting the somatosensory system. Conventional analgesics often exhibit limited efficacy, leading to suboptimal therapeutic outcomes. The pathogenesis of NP is complex and involves multiple mechanisms. The existing evidence suggests that [...] Read more.

Neuropathic pain (NP) is a prevalent clinical condition resulting from diseases or injuries affecting the somatosensory system. Conventional analgesics often exhibit limited efficacy, leading to suboptimal therapeutic outcomes. The pathogenesis of NP is complex and involves multiple mechanisms. The existing evidence suggests that maladaptive neuronal plasticity plays a central role in NP development. Additionally, emerging research highlights the contribution of neuroinflammatory responses mediated by glial cells in the onset of NP and associated sensory hypersensitivity. Among non-invasive neuromodulation techniques, transcranial direct current stimulation (tDCS) has gained prominence as a potential treatment for NP. Numerous studies have demonstrated its analgesic effects; however, the precise regulatory mechanisms remain unclear. The current evidence indicates that tDCS may alleviate NP by enhancing glial–neuronal interactions, which suppress nociceptive signaling pathways and reduce pain sensitivity. The reciprocal modulation between tDCS-mediated anti-inflammatory actions, as evidenced by decreased levels of pro-inflammatory cytokines and increased levels of anti-inflammatory mediators, and its facilitation of adaptive neural plasticity represents a particularly compelling therapeutic axis. This review elucidates inflammatory regulation by tDCS as a fundamental mechanism for NP alleviation, while delineating important unresolved questions regarding these complex interactions. Full article

(This article belongs to the Section Immunology)

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20 pages, 4686 KiB

Open AccessArticle

Genome-Wide Identification of the SRS Gene Family in Cucurbitaceae: Clade Identification and Expression Analysis of CmSRS Genes Under Drought and Salt Stress

by Haozhe Min, Kexiang Wang, Yao Guo, Junyan Yang, Xuhui Wang, Miao He, Tao Lin, Jiancai Mao and Zhengying Xuan

Biology 2025, 14(7), 891; https://doi.org/10.3390/biology14070891 (registering DOI) - 20 Jul 2025

Abstract

Background: The short strand-related sequence (SRS) gene family is a class of plant-specific transcription factors related to a group of genes known as the short internode (SHI) or SRS/STY gene family, which plays important roles in regulating plant growth and development and [...] Read more.

Background: The short strand-related sequence (SRS) gene family is a class of plant-specific transcription factors related to a group of genes known as the short internode (SHI) or SRS/STY gene family, which plays important roles in regulating plant growth and development and stress responses. Although the SRS genes have been studied in many plants, in cucurbit crops, they have thus far only been identified in cucumber. Methods: In the Cucurbitaceae database from melon (Cucumis melo), cucumber (Cucumis sativus), watermelon (Citrullus lanatus), bottle gourd (Lagenaria siceraria), wax gourd (Benincasa hispida), moschata pumpkin (Cucurbita moschata), and pumpkin (Cucurbita maxima), a total of 60 SRS genes were identified in seven Cucurbitaceae crops, which were classified into three subfamilies. Results: The same subfamily showed conserved motifs and gene structures. The differences in the number of SRS genes in different Cucurbitaceae crops implied likely gene loss or duplication events during evolution. Analysis of promoter cis-regulatory elements indicated that these SRS genes may be involved in hormone response, growth and development, and biotic and abiotic stress responses in plants. Most of the CmSRS genes in melons were expressed in the roots, with a few expressed in the leaves and ovaries. In addition, CmSRS expression was induced by biotic (wilt and powdery mildew) and abiotic (drought and salt) stresses. Subcellular localization of CmSRS proteins showed predominant expression in the nucleus. Conclusions: A total of 60 Cucurbitaceae SRS genes are present in the genomes of seven Cucurbitaceae crops. These cucurbit SRS genes seem to have maintained similar characteristics and functions during the evolutionary process. These results lay the foundation for the study of biological functions of SRS genes in Cucurbitaceae crops. Full article

22 pages, 5920 KiB

Open AccessArticle

The V5-Epitope Tag for Cell Engineering and Its Use in Immunohistochemistry and Quantitative Flow Cytometry

by Katja Fritschle, Marion Mielke, Olga J. Seelbach, Ulrike Mühlthaler, Milica Živanić, Tarik Bozoglu, Sarah Dötsch, Linda Warmuth, Dirk H. Busch, Arne Skerra, Christian Kupatt, Wolfgang A. Weber, Richard E. Randall, Katja Steiger and Volker Morath

Biology 2025, 14(7), 890; https://doi.org/10.3390/biology14070890 (registering DOI) - 20 Jul 2025

Abstract

Synthetic biology has fundamentally advanced cell engineering and helped to develop effective therapeutics such as chimeric antigen receptor (CAR)-T cells. For these applications, the detection, localization, and quantification of heterologous fusion proteins assembled from interchangeable building blocks is of high importance. The V5 [...] Read more.

Synthetic biology has fundamentally advanced cell engineering and helped to develop effective therapeutics such as chimeric antigen receptor (CAR)-T cells. For these applications, the detection, localization, and quantification of heterologous fusion proteins assembled from interchangeable building blocks is of high importance. The V5 tag, a 14-residue epitope tag, offers promising characteristics for these applications but has only rarely been used in this context. Thus, we have systematically evaluated the murine anti-V5 tag antibody mu_SV5-Pk1 as well as its humanized version, hu_SV5-Pk1, to analyze cells expressing V5-tagged receptors in samples from various in vitro and in vivo experiments. We found that the V5 tag signal on cells is affected by certain fixation and detachment reagents. Immunohistochemistry (IHC) on formalin-fixed paraffin-embedded (FFPE) mouse tissue samples was performed to sensitively detect cells in tissue. We improved IHC by applying the hu_SV5-Pk1 monoclonal antibody (mAb) to avoid cross-reactivity within and unspecific background signals arising on fixed mouse tissue. Conversely, the absence of unspecific binding by the mu_SV5-Pk1 mAb was evaluated on 46 human normal or cancer tissues. Our findings present a robust toolbox for utilizing the V5 tag and cognate antibodies in synthetic biology applications. Full article

(This article belongs to the Section Biochemistry and Molecular Biology)

25 pages, 2029 KiB

Open AccessArticle

Germination Enhances Phytochemical Profiles of Perilla Seeds and Promotes Hair Growth via 5α-Reductase Inhibition and Growth Factor Pathways

by Anurak Muangsanguan, Warintorn Ruksiriwanich, Pichchapa Linsaenkart, Pipat Tangjaidee, Korawan Sringarm, Chaiwat Arjin, Pornchai Rachtanapun, Sarana Rose Sommano, Korawit Chaisu, Apinya Satsook and Juan Manuel Castagnini

Biology 2025, 14(7), 889; https://doi.org/10.3390/biology14070889 (registering DOI) - 20 Jul 2025

Abstract

Seed germination is recognized for enhancing the accumulation of bioactive compounds. Perilla frutescens (L.) Britt., commonly known as perilla seed, is rich in fatty acids that may be beneficial for anti-hair loss. This study investigated the hair regeneration potential of perilla seed extracts—non-germinated [...] Read more.

Seed germination is recognized for enhancing the accumulation of bioactive compounds. Perilla frutescens (L.) Britt., commonly known as perilla seed, is rich in fatty acids that may be beneficial for anti-hair loss. This study investigated the hair regeneration potential of perilla seed extracts—non-germinated (NG-PS) and germinated in distilled water (0 ppm selenium; G0-PS), and germinated with 80 ppm selenium (G80-PS)—obtained from supercritical fluid extraction (SFE) and screw compression (SC). SFE extracts exhibited significantly higher levels of polyphenols, tocopherols, and fatty acids compared to SC extracts. Among the germinated groups, G0-PS showed the highest bioactive compound content and antioxidant capacity. Remarkably, treatment with SFE-G0-PS led to a significant increase in the proliferation and migration of hair follicle cells, reaching 147.21 ± 2.11% (p < 0.05), and resulted in complete wound closure. In addition, its antioxidant and anti-inflammatory properties were reflected by a marked scavenging effect on TBARS (59.62 ± 0.66% of control) and suppressed nitrite amounts (0.44 ± 0.01 µM). Moreover, SFE-G0-PS markedly suppressed SRD5A1-3 gene expression—key regulators in androgenetic alopecia—in both DU-145 and HFDPCs, with approximately 2-fold and 1.5-fold greater inhibition compared to finasteride and minoxidil, respectively. Simultaneously, it upregulated the expression of hair growth-related genes, including CTNNB1, SHH, SMO, GLI1, and VEGF, by approximately 1.5-fold, demonstrating stronger activation than minoxidil. These findings suggest the potential of SFE-G0-PS as a natural therapeutic agent for promoting hair growth and preventing hair loss. Full article

(This article belongs to the Special Issue Nature's Pharmacy: Ethnobotanical Approaches to Medicinal Plant Research for the Discovery and Development of Potential Phytochemicals)

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31 pages, 2257 KiB

Open AccessArticle

Contrasted Ethnobotanical and Literature Knowledge of Anti-Mosquito Plants from Guadeloupe

by Yolène Duchaudé, Laura Brelle, Muriel Sylvestre, Anubis Vega-Rúa and Gerardo Cebrián-Torrejón

Biology 2025, 14(7), 888; https://doi.org/10.3390/biology14070888 (registering DOI) - 19 Jul 2025

Abstract

The Aedes aegypti mosquito, vector of dengue, is a major public health threat in the Caribbean. In Guadeloupe, where dengue outbreaks occur frequently, traditional plant-based remedies are part of the local heritage but remain poorly documented. This study aimed to evaluate the anti-mosquito [...] Read more.

The Aedes aegypti mosquito, vector of dengue, is a major public health threat in the Caribbean. In Guadeloupe, where dengue outbreaks occur frequently, traditional plant-based remedies are part of the local heritage but remain poorly documented. This study aimed to evaluate the anti-mosquito potential of 38 Guadeloupean plants through an ethnobotanical survey. A semi-structured online questionnaire was conducted over five months, targeting the plant knowledge of residents. Inclusion/exclusion criteria were applied to identify and validate relevant species. Ethnobotanical indices such as Frequency of Citation (FC), Fidelity Level (FL), and Relative Frequency of Citation (RFC) were calculated. Out of the 38 surveyed plants, 22 were confirmed for their traditional anti-mosquito uses. The most cited species included Cymbopogon citratus (93.3%), Artocarpus altilis (25%), and Pimenta racemosa (18.3%). Comparative analysis with existing literature showed that 12 of these plants had not been previously reported for vector control. This highlights the value of ethnobotanical approaches for discovering alternative, eco-friendly vector control options and the importance of preserving traditional knowledge. The study reveals both the high potential of Guadeloupean flora and the risk of cultural erosion, supporting further research into the bioactive compounds of the most cited species. Full article

(This article belongs to the Special Issue Young Researchers in Plant Sciences)

16 pages, 2100 KiB

Open AccessReview

Romanian Dendrocoelidae Hallez, 1892 (Platyhelminthes, Tricladida, Dendrocoelidae) Revisited: A Tribute to Radu Codreanu and Doina Balcesco

by Anda Felicia Babalean

Biology 2025, 14(7), 887; https://doi.org/10.3390/biology14070887 (registering DOI) - 19 Jul 2025

Abstract

This paper presents the current state of knowledge on the Romanian Dendrocoelidae as part of the European/Palearctic Dendrocoelidae, emphasizing the contributions of the Romanian zoologists Radu Codreanu and Doina Balcesco. The main objective of this work was to identify the knowledge gaps for [...] Read more.

This paper presents the current state of knowledge on the Romanian Dendrocoelidae as part of the European/Palearctic Dendrocoelidae, emphasizing the contributions of the Romanian zoologists Radu Codreanu and Doina Balcesco. The main objective of this work was to identify the knowledge gaps for future alignment with current standards. This article presents the species inventory and a short historical overview of the classical phylogenetic system and discusses some morphological characters used in the systematics of the group. This study also analyzes the arguments (and hypotheses) put forward by Codreanu, Balcesco, and other authors regarding the phylogenetic value of various factors, including (a) the position of the oviducts between the male atrium and the bursal canal (typical for Paradendrocoelum); (b) the eyes and the penial flagellum in relation to the palaeogeographical context governed by the Quaternary Glaciation; and (c) the point of view of Codreanu and Balcesco on the origin and composition of the actual Romanian Dendrocoelidae fauna. The major key finding is that the Dendrocoelidae species in Romania should be reinvestigated in an integrative way, and specific research needs and future directions are suggested. Full article

(This article belongs to the Section Zoology)

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16 pages, 3609 KiB

Open AccessArticle

Will Wind Turbines Affect the Distribution of Alashan Ground Squirrel? Insights from Large-Scale Wind Farms in China

by Yuan Wang, Wenbin Yang, Qin Li, Min Zhao, Ying Yang, Xiangfeng Shi, Dazhi Zhang and Guijun Yang

Biology 2025, 14(7), 886; https://doi.org/10.3390/biology14070886 (registering DOI) - 19 Jul 2025

Abstract

The wind energy resources in the northwestern desert and semi-desert grassland regions of China are abundant. However, the ramifications of large-scale centralized wind farm operations on terrestrial rodents remain incompletely understood. In May and September 2024, we employed a grid sampling method combined [...] Read more.

The wind energy resources in the northwestern desert and semi-desert grassland regions of China are abundant. However, the ramifications of large-scale centralized wind farm operations on terrestrial rodents remain incompletely understood. In May and September 2024, we employed a grid sampling method combined with burrow counting and kernel density analysis to investigate the spatial distribution of Alashan ground squirrel (Spermophilus alashanicus) burrows in different wind turbine power zones (control, 750 kW, 1500 kW, 2000 kW, and 2500 kW) at the Taiyangshan wind farm in China. Using generalized additive models and structural equation models, we analysed the relationship between burrow spatial distribution and environmental factors. The results revealed no significant linear correlation between burrow density and turbine layout density, but was significantly positively correlated with turbine power (p < 0.05). The highest burrow density was observed in the 2500 kW zone, with values of 24.43 ± 7.18 burrows/hm² in May and 21.29 ± 3.38 burrows/hm² in September (p < 0.05). The squirrels exhibited a tendency to avoid constructing burrows within the rotor sweeping areas of the turbines. The burrow density distribution exhibited a multinuclear clustering pattern in both May and September, with a northwest–southeast spatial orientation. Turbine power, aspect, and plan convexity had significant positive effects on burrow density, whereas vegetation height had a significant negative effect. Moreover, vegetation height indirectly influenced burrow density through its interactions with turbine power and relief degree. Under the combined influence of turbine power, topography, and vegetation, Alashan ground squirrels preferred habitats in low-density, high-power turbine zones with shorter vegetation, sunny slopes, convex landforms, and minimal disturbance. Full article

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31 pages, 2323 KiB

Open AccessArticle

Alpha-Lipoic Acid and Metformin Combination Therapy Synergistically Activate Nrf2-AMPK Signaling Pathways to Ameliorate Cognitive Dysfunction in Type 2 Diabetic Encephalopathy: A Preclinical Study

by Abdulmajeed F. Alrefaei and Mohamed E. Elbeeh

Biology 2025, 14(7), 885; https://doi.org/10.3390/biology14070885 - 18 Jul 2025

Abstract

Diabetic encephalopathy affects over 40% of diabetic patients globally, yet effective treatments remain critically limited. This study investigated the synergistic neuroprotective potential of alpha-lipoic acid (ALA) and metformin through the coordinated activation of Nrf2 and AMPK signaling pathways in type 2 diabetes mellitus [...] Read more.

Diabetic encephalopathy affects over 40% of diabetic patients globally, yet effective treatments remain critically limited. This study investigated the synergistic neuroprotective potential of alpha-lipoic acid (ALA) and metformin through the coordinated activation of Nrf2 and AMPK signaling pathways in type 2 diabetes mellitus (T2DM)-induced encephalopathy. Using a clinically relevant streptozotocin-nicotinamide-induced T2DM rat model, sixty male Sprague–Dawley rats were randomly assigned to five groups: control, diabetic, ALA-treated (300 mg/kg), metformin-treated (50 mg/kg), and combination-treated groups over eight weeks. Combination therapy produced statistically validated synergistic effects with significant interaction terms (p < 0.01) across all evaluated parameters. Nuclear Nrf2 translocation increased 3.9-fold and AMPK phosphorylation rose 3.2-fold compared to monotherapies, surpassing mathematical additivity. Mitochondrial function was remarkably restored, with ATP production increasing to 92% of control levels. Cognitive performance was normalized, with spatial memory approaching control values. Combination index analysis (CI < 1.0) confirmed true synergistic interactions across molecular, cellular, and behavioral endpoints. These findings establish a novel convergent mechanism providing compelling evidence for combination ALA–metformin therapy as an innovative treatment strategy for diabetes-associated neurodegeneration. Full article

(This article belongs to the Special Issue Animal Models of Neurodegenerative Diseases)

22 pages, 1821 KiB

Open AccessArticle

Effects of Temperature Increase on Microbiome of Carnivorous Plant Utricularia vulgaris L. in Peat Bog Ecosystems

by Aleksandra Bartkowska-Bekasiewicz and Tomasz Mieczan

Biology 2025, 14(7), 884; https://doi.org/10.3390/biology14070884 - 18 Jul 2025

Abstract

Climate change can have a direct impact on the decomposition of organic matter, as well as indirect effects on peatland vegetation (including carnivorous plants) and the microbial communities associated with this environment. The activity of microbes varies depending on the type of peatland [...] Read more.

Climate change can have a direct impact on the decomposition of organic matter, as well as indirect effects on peatland vegetation (including carnivorous plants) and the microbial communities associated with this environment. The activity of microbes varies depending on the type of peatland they inhabit. Because some microorganisms are highly sensitive, they can be used as indicators of climate change. However, there is still little knowledge of how changes in the temperature of the environment can affect the microbiome of carnivorous plants. The study was conducted to test the following hypotheses: (1) The effect of rising water temperature on the qualitative and quantitative structure of the microbiome of carnivorous peatland plants depends on the type of peatland; (2) habitats with a higher trophic status stimulate the development of microbial communities in the water, but are an unfavourable habitat for the development of the microbiome of plant traps. Irrespective of the type of peatland, the species richness of microorganisms was much higher in the water than in the traps. As the temperature increased, there was an increase in the abundance of bacteria, heterotrophic flagellates, and testate amoebae, which was much more pronounced in the peat bog than in the carbonate fen. Full article

(This article belongs to the Section Microbiology)

13 pages, 1279 KiB

Open AccessArticle

Transcriptome Sequencing-Based Analysis of Premature Fruiting in Amomum villosum Lour.

by Yating Zhu, Shuang Li, Hongyou Zhao, Qianxia Li, Yanfang Wang, Chunyong Yang, Ge Li, Yanqian Wang and Lixia Zhang

Biology 2025, 14(7), 883; https://doi.org/10.3390/biology14070883 - 18 Jul 2025

Abstract

Amomum villosum Lour., a perennial medicinal plant in the Zingiber genus, usually requires approximately 3–4 years of vegetative growth from seed germination to first fruiting, resulting in high initial investment costs and a prolonged revenue cycle, which pose significant challenges to the industry’s [...] Read more.

Amomum villosum Lour., a perennial medicinal plant in the Zingiber genus, usually requires approximately 3–4 years of vegetative growth from seed germination to first fruiting, resulting in high initial investment costs and a prolonged revenue cycle, which pose significant challenges to the industry’s sustainable development. Our research team observed a distinct premature fruiting phenomenon in A. villosum. We investigated the regulatory mechanisms underlying premature fruiting in A. villosum by identifying the key differentially expressed genes (DEGs) and metabolic pathways governing the premature fruiting (Precocious) and typical plants (CK) of the ‘Yunsha No.8’ cultivar. Transcriptomic sequencing (RNA-seq) and bioinformatic analyses were performed using the DNBSEQ^TM platform. The sequencing generated 29.0 gigabases (Gb) of clean data, and 115,965 unigenes were identified, with an average length of 1368 bp. Based on the sequencing results, 1545 DEGs were identified. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were annotated for these DEGs. This study identifies phytohormone signaling, carbohydrate and lipid metabolism, and polysaccharide degradation as critical pathways controlling premature fruiting in A. villosum. Six randomly selected DEGs were validated using real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the results corroborated the transcriptome data, confirming their reliability. This study lays the foundation for the elucidation of the molecular mechanisms and metabolic pathways driving premature fruiting in A. villosum. Full article

(This article belongs to the Special Issue Young Investigators in Biochemistry and Molecular Biology)

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23 pages, 39249 KiB

Open AccessArticle

Single-Cell Atlas of Spleen Remodeling Reveals Macrophage Subset-Driven ASFV Pathogenesis

by Liyuan Wang, Shouzhang Sun, Lei Liu, Yun Chen, Haixue Zheng and Zhonglin Tang

Biology 2025, 14(7), 882; https://doi.org/10.3390/biology14070882 - 18 Jul 2025

Abstract

African swine fever virus (ASFV) causes global swine outbreaks, but its cellular pathogenesis is poorly understood. Using single-cell RNA data from ASFV-infected pig spleens across four timepoints, we identified macrophages as the primary viral reservoir, with infection driving lymphoid depletion and myeloid expansion. [...] Read more.

African swine fever virus (ASFV) causes global swine outbreaks, but its cellular pathogenesis is poorly understood. Using single-cell RNA data from ASFV-infected pig spleens across four timepoints, we identified macrophages as the primary viral reservoir, with infection driving lymphoid depletion and myeloid expansion. We characterized four functionally distinct macrophage subsets, including a metabolically reprogrammed SusceptibleMac population serving as the major viral niche and an AntiviralMac subset rapidly depleted during infection. Viral gene expression analysis revealed E165R as a central hub in viral replication networks, while host transcriptomics uncovered disruption of Netrin signaling pathways that may facilitate immune evasion. Pseudotime analysis revealed dynamic macrophage state transitions during infection. These findings provide a high-resolution cellular atlas of ASFV pathogenesis, revealing macrophage subset-specific responses that shape disease outcomes and identifying potential targets for therapeutic intervention. Full article

(This article belongs to the Special Issue Viral Infections in Animals: Pathogenesis and Immunity)

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14 pages, 2193 KiB

Open AccessArticle

Neighboring Patch Density or Patch Size? Which Determines the Importance of Forest Patches in Maintaining Overall Landscape Connectivity in Kanas, Xinjiang, China

by Zhi Wang, Lei Han, Luyao Wang, Hui Shi and Yan Luo

Biology 2025, 14(7), 881; https://doi.org/10.3390/biology14070881 - 18 Jul 2025

Abstract

The precise identification of priority areas for conservation based on connectivity can significantly enhance protection efficacy and mitigate biodiversity loss in fragmented landscapes. Priority area selection efforts are typically conducted in landscapes with a limited number of patches or simplified to focus on [...] Read more.

The precise identification of priority areas for conservation based on connectivity can significantly enhance protection efficacy and mitigate biodiversity loss in fragmented landscapes. Priority area selection efforts are typically conducted in landscapes with a limited number of patches or simplified to focus on large patches, while landscapes with numerous patches are rarely explored. In this paper, we used a forest in Kanas, Xinjiang, China, as a case study to explore priority patches for conservation according to their contribution to maintaining overall landscape connectivity, as well as to assess how structural factors influence patch importance in connectivity, based on graph theory. We found that the rank of patches varied with patch importance indices (which can be used to calculate the contribution of individual patches to maintaining overall landscape). Dispersal distances were selected, as they placed different emphasis on the size and topological location of patches, and different types of links (binary or probabilistic connection) were used. One critical and seven important connected patches were identified as priority patches for conservation after taking multiple connectivity indices and dispersal distances into comprehensive consideration. In addition, neighboring patch density was the dominant factor that influenced patch importance for species with 50 and 100 m dispersal distances, while patch size contributed most for species with 200 m and longer dispersal distances; therefore, we suggested that neighboring patch density and patch size could be used to support efforts to identify priority patches. Overall, our results provide a unique perspective and a more simplified process for the selection of priority protected sites in patch-rich landscapes, allowing us to highlight which action is suitable for optimizing landscape connectivity and biodiversity conservation. Full article

(This article belongs to the Section Conservation Biology and Biodiversity)

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13 pages, 1649 KiB

Open AccessArticle

Assessing the Population Demographic History of the Tsushima Leopard Cat and Its Genetic Divergence Time from Continental Populations

by Hideyuki Ito, Nobuyoshi Nakajima, Manabu Onuma, Takushi Kishida and Miho Inoue-Murayama

Biology 2025, 14(7), 880; https://doi.org/10.3390/biology14070880 - 18 Jul 2025

Abstract

The Tsushima leopard cat (Prionailurus bengalensis euptilurus), an endangered feline endemic to Tsushima Island, Japan, faces critical threats due to its small and isolated population. Understanding its demographic history and genetic differentiation from continental populations is essential for conservation planning. In [...] Read more.

The Tsushima leopard cat (Prionailurus bengalensis euptilurus), an endangered feline endemic to Tsushima Island, Japan, faces critical threats due to its small and isolated population. Understanding its demographic history and genetic differentiation from continental populations is essential for conservation planning. In this study, we performed whole-genome sequencing of four Tsushima individuals and applied demographic inference methods, including pairwise sequentially Markovian coalescent (PSMC) and Sequentially Markovian Coalescent (SMC++), to reconstruct the historical effective population size (Ne) and estimate divergence times. PSMC revealed a population expansion between 200,000 and 100,000 years ago, followed by a long-term decline. SMC++ inferred a continuous decline and estimated that the divergence from the Korean leopard cat population occurred approximately 30,000–20,000 years ago. Genetic diversity analysis showed that the Tsushima population has significantly lower heterozygosity and higher inbreeding levels than continental populations. Genetic clustering based on genome-wide SNPs indicated that the Tsushima population is genetically closest to the Korean population, forming a northern cluster distinct from southern populations, such as Borneo and the Malay Peninsula. These findings provide valuable insights into the evolutionary history and genetic status of the Tsushima leopard cat and contribute critical data for the design of future conservation strategies targeting this unique insular lineage. Full article

(This article belongs to the Special Issue Genetic Variability within and between Populations)

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16 pages, 2268 KiB

Open AccessArticle

Epichloë Endophyte Alters Bacterial Nitrogen-Cycling Gene Abundance in the Rhizosphere Soil of Perennial Ryegrass

by Munire Maimaitiyiming, Yanxiang Huang, Letian Jia, Mofan Wu and Zhenjiang Chen

Biology 2025, 14(7), 879; https://doi.org/10.3390/biology14070879 - 18 Jul 2025

Abstract

Perennial ryegrass (Lolium perenne), an important forage and turfgrass species, can establish a mutualistic symbiosis with the fungal endophyte Epichloë festucae var. lolii. Although the physiological and ecological impacts of endophyte infection on ryegrass have been extensively investigated, the response of [...] Read more.

Perennial ryegrass (Lolium perenne), an important forage and turfgrass species, can establish a mutualistic symbiosis with the fungal endophyte Epichloë festucae var. lolii. Although the physiological and ecological impacts of endophyte infection on ryegrass have been extensively investigated, the response of the soil microbial community and nitrogen-cycling gene to this relationship has received much less attention. The present study emphasized abundance and diversity variation in the AOB-amoA, nirK and nosZ functional genes in the rhizosphere soil of the endophyte–ryegrass symbiosis following litter addition. We sampled four times: at T₀ (prior to first litter addition), T₁ (post 120 d of 1st litter addition), T₂ (post 120 d of 2nd litter addition) and T₃ (post 120 d of 3rd litter addition) times. Real-time fluorescence quantitative PCR (qPCR) and PCR amplification and sequencing were used to characterize the abundance and diversity of the AOB-amoA, nirK and nosZ genes in rhizosphere soils of endophyte-infected (E+) plants and endophyte-free (E−) plants. A significant enhancement of total Phosphorus (P), Soil Organic Carbon (SOC), Ammonium ion (NH₄⁺) and Nitrate ion (NO₃⁻) contents in the rhizosphere soil was recorded in endophyte-infected plants at different sampling times compared to endophyte-free plants (p ≤ 0.05). The absolute abundance of the AOB-amoA gene at T₀ and T₁ times was higher, as was the absolute abundance of the nosZ gene at T₀, T₁ and T₃ times in the E+ plant rhizophere soils relative to E− plant rhizosphere soils. A significant change in relative abundance of the AOB-amoA and nosZ genes in the host rhizophere soils of endophyte-infected plants at T₁ and T₃ times was observed. The experiment failed to show any significant alteration in abundance and diversity of the nirK gene, and diversity of the AOB-amoA and nosZ genes. Analysis of the abundance and diversity of the nirK gene indicated that changes in soil properties accounted for approximately 70.38% of the variation along the first axis and 16.69% along the second axis, and soil NH₄⁺ (p = 0.002, 50.4%) and soil C/P ratio (p = 0.012, 15.8%) had a strong effect. The changes in community abundance and diversity of the AOB-amoA and nosZ genes were mainly related to soil pH, N/P ratio and NH₄⁺ content. The results demonstrate that the existence of tripartite interactions among the foliar endophyte E. festucae var. Lolii, L. perenne and soil nitrogen-cycling gene has important implications for reducing soil losses on N. Full article

(This article belongs to the Collection Plant Growth-Promoting Bacteria: Mechanisms and Applications)

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18 pages, 751 KiB

Open AccessArticle

Effects of Salinity, Temperature, and Diet on the Biological Characteristics of Brachionus plicatilis Müller, 1786

by Quynh-Anh Tran-Nguyen, Truong Nhat Phan, Quang-Anh Tran, Hong Thi Mai, Thao Linh Phan Thi, Dang Doan Phan and Mau Trinh-Dang

Biology 2025, 14(7), 878; https://doi.org/10.3390/biology14070878 - 18 Jul 2025

Abstract

This study investigates the effects of salinity, temperature, and diet on the biological characteristics of the rotifer Brachionus plicatilis, an essential live feed in aquaculture. The results indicate that environmental factors have a significant influence on reproductive traits and survival. At a [...] Read more.

This study investigates the effects of salinity, temperature, and diet on the biological characteristics of the rotifer Brachionus plicatilis, an essential live feed in aquaculture. The results indicate that environmental factors have a significant influence on reproductive traits and survival. At a salinity of 5 ppt, B. plicatilis achieved the highest fecundity (25.50 ± 0.58 inds.), while the longest lifespan (273.00 ± 72.52 h) was observed at 35 ppt. The temperature had a strong influence on developmental rates, with the shortest juvenile period recorded at 35 °C (8.00 ± 0.00 h) and the longest lifespan at 20 °C (270.62 ± 30.38 h). The diet also played a critical role, with Chlorella vulgaris supporting maximum fecundity, whereas mixed diets prolonged lifespan to 290.50 ± 62.83 (h). These findings provide valuable insights into optimizing rotifer culture systems to improve aquaculture productivity and sustainability. Full article

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14 pages, 2425 KiB

Open AccessReview

Immunological Factors in Recurrent Pregnancy Loss: Mechanisms, Controversies, and Emerging Therapies

by Efthalia Moustakli, Anastasios Potiris, Athanasios Zikopoulos, Eirini Drakaki, Ioannis Arkoulis, Charikleia Skentou, Ioannis Tsakiridis, Themistoklis Dagklis, Peter Drakakis and Sofoklis Stavros

Biology 2025, 14(7), 877; https://doi.org/10.3390/biology14070877 - 17 Jul 2025

Abstract

Immunological factors have gained growing recognition as key contributors to recurrent pregnancy loss (RPL) after in vitro fertilization (IVF), representing a major challenge in reproductive medicine. RPL affects approximately 1–2% of women trying to conceive naturally and up to 10–15% of those undergoing [...] Read more.

Immunological factors have gained growing recognition as key contributors to recurrent pregnancy loss (RPL) after in vitro fertilization (IVF), representing a major challenge in reproductive medicine. RPL affects approximately 1–2% of women trying to conceive naturally and up to 10–15% of those undergoing IVF, where overall success rates remain around 30–40% per cycle. An imbalance in maternal immunological tolerance toward the semi-allogeneic fetus during pregnancy may lead to miscarriage and implantation failure. IVF-related ovarian stimulation and embryo modification offer additional immunological complications that can exacerbate existing immune dysregulation. Recent advances in reproductive immunology have significantly deepened our understanding of the immune mechanisms underlying RPL following IVF, particularly highlighting the roles of regulatory T cells (T regs), natural killer cells, cytokine dysregulation, and disruptions in maternal–fetal immune tolerance. In order to better customize therapies, this evaluation incorporates recently discovered immunological biomarkers and groups patients according to unique immune profiles. Beyond conventional treatments like intralipid therapy and intravenous immunoglobulin, it also examines new immunomodulatory medications that target certain immune pathways, such as precision immunotherapies and novel cytokine modulators. We also discuss the debates over immunological diagnostics and therapies, such as intralipid therapy, intravenous immunoglobulin, corticosteroids, and anticoagulants. The heterogeneity of patient immune profiles combined with a lack of strong evidence highlights the imperative for precision medicine to improve therapeutic consistency. Novel indicators for tailored immunotherapy and emerging treatments that target particular immune pathways have encouraging opportunities to increase pregnancy success rates. Improving management approaches requires that future research prioritize large-scale clinical trials and the development of standardized immunological assessments. This review addresses the immunological factors in RPL during IVF, emphasizing underlying mechanisms, ongoing controversies, and novel therapeutic approaches to inform researchers and clinicians. Full article

(This article belongs to the Section Immunology)

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16 pages, 2412 KiB

Open AccessArticle

Dynamic Network Driver Analysis Identifies Master Factors Associated with Progression of Solar Lentigines

by Deyu Cai, Hong Zhang, Chengming Zhang, Xue Xiao, Xiao Cui, Xuelan Gu and Luonan Chen

Biology 2025, 14(7), 876; https://doi.org/10.3390/biology14070876 - 17 Jul 2025

Abstract

Solar lentigines, commonly caused by prolonged ultraviolet exposure, raise the risk of skin disorders and remain challenging to manage due to their complex mechanisms. Understanding the molecular mechanisms driving the progression of solar lentigines is crucial for developing effective protective strategies. In this [...] Read more.

Solar lentigines, commonly caused by prolonged ultraviolet exposure, raise the risk of skin disorders and remain challenging to manage due to their complex mechanisms. Understanding the molecular mechanisms driving the progression of solar lentigines is crucial for developing effective protective strategies. In this study, we introduced a novel method, Dynamic Network Driver (DND), which identifies upstream regulators that drive disease progression by integrating the Dynamic Network Biomarker (DNB) approach with network control theory. By applying DND to multi-omics data from solar lentigines subjects, we (1) identified the key drivers associated with solar lentigo progression, with their functions involved in differentiation and dermal–epidermal junction; and (2) highlighted ARNT2 and TBX2 as significant master factors supported by in vitro validation in melanocytes and pigmented 3D living skin equivalent models. These results demonstrate the potency of DND for uncovering the molecular mechanisms behind solar lentigines and informing therapeutic strategies. In summary, the DND approach identified novel drivers of solar lentigo progression, acting as new markers for spot mitigation in 3D spot mimic models. Full article

(This article belongs to the Special Issue Physiology and Pathophysiology of Skin)

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18 pages, 1449 KiB

Open AccessReview

Three-Dimensional Culture System: A New Frontier in Cancer Research, Drug Discovery, and Stem Cell-Based Therapy

by Guya Diletta Marconi, Antonella Mazzone, Ylenia Della Rocca, Oriana Trubiani, Jacopo Pizzicannella and Francesca Diomede

Biology 2025, 14(7), 875; https://doi.org/10.3390/biology14070875 - 17 Jul 2025

Abstract

Two-dimensional culture systems have been used for a long time in the research field but their disadvantages make it difficult to reproduce the in vivo environment. Three-dimensional culture systems overcome these limitations, simulating the physiological context of an organism, from the molecular level [...] Read more.

Two-dimensional culture systems have been used for a long time in the research field but their disadvantages make it difficult to reproduce the in vivo environment. Three-dimensional culture systems overcome these limitations, simulating the physiological context of an organism, from the molecular level to the cellular, tissue, and organ complexity levels. This review focuses on 3D cellular models, such as spheroids and tumoroids, which reproduce tumor heterogeneity and microenvironments. It also includes 3D cultures of mesenchymal stem cells (MSCs), particularly those derived from teeth. In conclusion, 3D models are profoundly impacting the biomedical field by offering more accurate in vitro platforms for drug development and disease modeling, thereby significantly reducing the reliance on animal testing and leading to the advancement of personalized and regenerative medicine. Full article

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