International Journal of Molecular Sciences

13 pages, 1682 KiB

Open AccessArticle

Dihydrocapsaicin Enhances Tumor Necrosis Factor-α-Induced Apoptosis and G1 Cell Cycle Arrest in Human Cervical Cancer Cells Through TAK1-Mediated NF-κB and EGFR Pathways

by Chantana Boonyarat, Hiroaki Sakurai, Yoshihiro Hayakawa, Suchada Chaiwiwatrakul, Rawiwun Kaewamatawong, Teeraporn Sadira Supapaan, Sureewan Duangjit, Benjabhorn Sethabouppha and Pornthip Waiwut

Int. J. Mol. Sci. 2025, 26(11), 5011; https://doi.org/10.3390/ijms26115011 - 22 May 2025

Abstract

Dihydrocapsaicin (DHC), a prominent capsaicinoid derived from red chili peppers, has shown cytotoxic effects against various cancer cell types. However, its role in modulating cytokine-induced survival and apoptotic signaling in cancer cells remains unclear. In this study, we investigated the effects of DHC [...] Read more.

Dihydrocapsaicin (DHC), a prominent capsaicinoid derived from red chili peppers, has shown cytotoxic effects against various cancer cell types. However, its role in modulating cytokine-induced survival and apoptotic signaling in cancer cells remains unclear. In this study, we investigated the effects of DHC on tumor necrosis factor-α (TNF-α)-induced cell cycle arrest and apoptosis in HeLa human cervical cancer cells. Our results demonstrate that DHC significantly enhances TNF-α-induced G1 phase cell cycle arrest and apoptosis by targeting the transforming growth factor-β-activated kinase 1 (TAK1)-mediated prosurvival pathways. DHC inhibited the phosphorylation of TAK1 and downstream effectors including IKKα, NF-κB p65, MAPKs (p38, JNK, ERK), Akt, and EGFR, thereby disrupting key signaling networks that typically confer resistance to TNF-α-induced cytotoxicity. Additionally, DHC suppressed the TNF-α-induced phosphorylation of EGFR at Ser-1046/1047 and Thr-669, sites critical for survival signaling. Co-treatment with DHC and TNF-α led to enhanced apoptotic features, including increased PARP-1 cleavage. These findings suggest that DHC sensitizes cervical cancer cells to cytokine-induced cell death by interfering with TAK1/NF-κB and EGFR signaling axes. Our study positions DHC as a promising candidate for combination therapies aimed at overcoming resistance in cancers with aberrant inflammatory and survival signaling. Full article

(This article belongs to the Special Issue The Role of Natural Compounds in Cancer and Inflammation)

► Show Figures

Figure 1

16 pages, 1085 KiB

Open AccessArticle

Opioid-Induced Regulation of Cortical Circular-Grin2b_011731 Is Associated with Regulation of circGrin2b Sponge Target miR-26b-3p

by Aria Gillespie and Stephanie E. Daws

Int. J. Mol. Sci. 2025, 26(11), 5010; https://doi.org/10.3390/ijms26115010 - 22 May 2025

Abstract

Opioid use induces neurobiological adaptations throughout mesolimbic brain regions, such as the orbitofrontal cortex (OFC), which mediates decision-making and emotional–cognitive regulation. Previously, we showed that a circular RNA (circRNA) species, rno_circGrin2b_011731 (circGrin2b), is upregulated in the OFC of rats [...] Read more.

Opioid use induces neurobiological adaptations throughout mesolimbic brain regions, such as the orbitofrontal cortex (OFC), which mediates decision-making and emotional–cognitive regulation. Previously, we showed that a circular RNA (circRNA) species, rno_circGrin2b_011731 (circGrin2b), is upregulated in the OFC of rats following chronic self-administration (SA) of the opioid heroin. circGrin2b is derived from Grin2b, which encodes the regulatory subunit of the glutamate ionotropic NMDA receptor, GluN2B. However, the upstream regulatory mechanisms of circGrin2b biogenesis and the downstream consequences of circGrin2b dysregulation remain unknown. We hypothesized that opioid-induced elevation of circGrin2b is accompanied by regulation of circRNA biogenesis enzymes, and that circGrin2b may sponge microRNAs (miRNAs), as miRNA sponging is a well-described characteristic of circRNAs. To test these hypotheses, we established an in vitro primary cortical cell culture model to examine alterations in circGrin2b expression following exposure to the opioid morphine. We measured mRNA expression of known circRNA splicing factors and observed significant downregulation of Fused in Sarcoma (Fus), a negative regulator of circRNA biogenesis, following 90 min or 24 h of morphine exposure. Downregulation of Fus at 24 h post-morphine was accompanied by upregulation of circGrin2b and downregulation of miR-26b-3p, a predicted miRNA target of circGrin2b. Luciferase reporter assays confirmed interaction of miR-26b-3p with circGrin2b. Finally, we report a significant negative relationship between circGrin2b and miR-26b-3p expression in the OFC of rats following heroin SA. We conclude that regulation of circGrin2b is an opioid-induced neuroadaptation that may impact downstream signaling of miRNA pathways in the frontal cortex. Full article

(This article belongs to the Special Issue New Advances in Opioid Research)

17 pages, 6118 KiB

Open AccessArticle

Birch Sap Preserves Memory Function in Rats by Enhancing Cerebral Blood Flow and Modulating the Presynaptic Glutamatergic System in the Hippocampus

by Chien-Fen Huang, Tzu-Kang Lin, Chia-Chuan Chang, Ming-Yi Lee, Ching-Yi Lu, Chi-Feng Hung and Su-Jane Wang

Int. J. Mol. Sci. 2025, 26(11), 5009; https://doi.org/10.3390/ijms26115009 - 22 May 2025

Abstract

As the average age of the population increases, memory impairment has become an increasingly prevalent issue. This study investigates the effects of 14 days of oral birch sap administration on memory functions in healthy rats using the Morris water maze (MWM) test and [...] Read more.

As the average age of the population increases, memory impairment has become an increasingly prevalent issue. This study investigates the effects of 14 days of oral birch sap administration on memory functions in healthy rats using the Morris water maze (MWM) test and explores the underlying mechanisms. A compositional analysis revealed that birch soap is rich in polysaccharides, specifically a low-molecular weight polysaccharide (MW 1.29 kDa), and exhibits no hepatotoxicity or renal toxicity at the tested dose. The results from the MWM test demonstrated that the time and distance required to reach the platform were significantly shorter in the birch sap-treated group compared to the control group, suggesting that birch sap supports memory preservation. Moreover, rats treated with birch sap showed improved cerebral blood flow compared to the control rats. Additionally, in hippocampal nerve terminals (synaptosomes), rats treated with birch sap exhibited a significant increase in evoked glutamate release, as well as elevated levels of presynaptic proteins, including vesicular glutamate transporter 1 (VGluT1), synaptophysin, synaptobrevin, synaptotagmin, syntaxin, synapsin I, and the 25 kDa synaptosome-associated protein (SNAP-25). Transmission electron microscopy also revealed a notable increase in the number of synaptic vesicles in hippocampal synaptosomes of the birch-sap-treated rats. These findings suggest that birch sap enhances hippocampal presynaptic glutamatergic functions and cerebral blood flow, contributing to its memory-preserving effects in rats. Full article

(This article belongs to the Special Issue Nutraceuticals for the Maintenance of Brain Health)

► Show Figures

Figure 1

13 pages, 525 KiB

Open AccessBrief Report

Retrospective Observational Study of CSF-Derived HIV-1 Tat and Vpr Amino Acid Sequences in a South African Pediatric Cohort with HIV Subtype C

by Anicia Thirion, Shayne Mason, Du Toit Loots, Regan Solomons and Monray Edward Williams

Int. J. Mol. Sci. 2025, 26(11), 5008; https://doi.org/10.3390/ijms26115008 - 22 May 2025

Abstract

The human immunodeficiency virus (HIV-1) infiltrates the central nervous system (CNS) early in infection, leading to HIV-associated neurocognitive impairments, particularly pronounced in children who exhibit neurodevelopmental delay. Viral proteins, including the transactivator of transcription protein (Tat) and viral protein R (Vpr) are pivotal [...] Read more.

The human immunodeficiency virus (HIV-1) infiltrates the central nervous system (CNS) early in infection, leading to HIV-associated neurocognitive impairments, particularly pronounced in children who exhibit neurodevelopmental delay. Viral proteins, including the transactivator of transcription protein (Tat) and viral protein R (Vpr) are pivotal in HIV-1 neuropathogenesis, with their amino acid sequence variation influencing disease progression. Due to the difficulty of collecting cerebrospinal fluid from children, few studies have examined whether key Tat and Vpr neuropathogenic signatures found in blood are also present in the cerebrospinal fluid (CSF) of children with HIV. We employed Sanger sequencing for Tat and Vpr sequence analysis using retrospectively collected CSF samples from a South African pediatric HIV-1 subtype C cohort (n = 4). We compared our CSF-derived sequences with pediatric blood-derived sequences (n = 43) from various geographical regions, sourced from the Los Alamos database. Neuropathogenic amino acid variants were identified in Tat and Vpr sequences derived from CSF samples of South African pediatric participants No significant differences were found between subtype C sequences from CSF and blood. Regional analysis highlighted unique amino acid signatures. Obtaining pediatric CSF for HIV-1 sequencing is highly challenging. Despite a small sample size, this study offers rare insights into Tat and Vpr sequences in children, improving understanding of the potential HIV-1 brain pathogenesis in pediatric populations. Full article

(This article belongs to the Special Issue Emerging Viral Epidemics)

40 pages, 1569 KiB

Open AccessReview

Cell Type-Specific Expression of Purinergic P2X Receptors in the Hypothalamus

by Jana Cihakova, Milorad Ivetic and Hana Zemkova

Int. J. Mol. Sci. 2025, 26(11), 5007; https://doi.org/10.3390/ijms26115007 - 22 May 2025

Abstract

Purinergic P2X receptors (P2X) are ATP-gated ion channels that are broadly expressed in the brain, particularly in the hypothalamus. As ionic channels with high permeability to calcium, P2X play an important and active role in neural functions. The hypothalamus contains a number of [...] Read more.

Purinergic P2X receptors (P2X) are ATP-gated ion channels that are broadly expressed in the brain, particularly in the hypothalamus. As ionic channels with high permeability to calcium, P2X play an important and active role in neural functions. The hypothalamus contains a number of small nuclei with many molecularly defined types of peptidergic neurons that affect a wide range of physiological functions, including water balance, blood pressure, metabolism, food intake, circadian rhythm, childbirth and breastfeeding, growth, stress, body temperature, and multiple behaviors. P2X are expressed in hypothalamic neurons, astrocytes, tanycytes, and microvessels. This review focuses on cell-type specific expression of P2X in the most important hypothalamic nuclei, such as the supraoptic nucleus (SON), paraventricular nucleus (PVN), suprachiasmatic nucleus (SCN), anteroventral periventricular nucleus (AVPV), anterior hypothalamic nucleus (AHN), arcuate nucleus (ARC), ventromedial hypothalamic nucleus (VMH), dorsomedial hypothalamic nucleus (DMH), tuberomammillary nucleus (TMN), and lateral hypothalamic area (LHA).> The review also notes the possible role of P2X and extracellular ATP in specific hypothalamic functions. The literature summarized here shows that purinergic signaling is involved in the control of the hypothalamic-pituitary endocrine system, the hypothalamic–neurohypophysial system, the circadian systems and nonendocrine hypothalamic functions. Full article

(This article belongs to the Special Issue Ion Channels in the Nervous System)

► Show Figures

Figure 1

21 pages, 2566 KiB

Open AccessArticle

Immune Responses of Mango Callus Infected by Agrobacterium tumefaciens Inhibited Transformation

by Haiyan Shu, Zilhas Ahmed Jewel, Omor Faruk, Luqiong He, Qing Wei, Rulin Zhan and Shenghe Chang

Int. J. Mol. Sci. 2025, 26(11), 5006; https://doi.org/10.3390/ijms26115006 - 22 May 2025

Abstract

Mango is a vital fruit crop in tropical and subtropical regions, yet pests and diseases cause 30–70% production losses. Developing disease-resistant cultivars through transgenic methods could mitigate these issues. Agrobacterium-mediated callus transformation is a common genetic engineering approach, but successful transgenic mango [...] Read more.

Mango is a vital fruit crop in tropical and subtropical regions, yet pests and diseases cause 30–70% production losses. Developing disease-resistant cultivars through transgenic methods could mitigate these issues. Agrobacterium-mediated callus transformation is a common genetic engineering approach, but successful transgenic mango plants from callus remain unreported due to severe browning and necrosis post-infection. We hypothesized that Agrobacterium-induced immune responses trigger callus death, hindering transformation. To improve efficiency, we engineered an Agrobacterium strain carrying the type III secretion system (T3SS) and effector gene AvrPto. Compared to controls, infected calluses exhibited elevated reactive oxygen species (ROS), along with up-regulated ROS-related, gallic acid biosynthesis, and defense genes. Calluses infected with T3SS-AvrPto-harboring Agrobacterium showed delayed browning and necrosis versus those infected with the empty vector (NV). The transformation rate with Agrobacterium (T3SS-AvrPto-EGFP) reached 1.6%, while Agrobacterium (NV-EGFP) failed entirely. These findings demonstrate that T3SS and AvrPto enhance mango transformation efficiency, offering a promising strategy for breeding multi-resistant varieties. Full article

(This article belongs to the Section Molecular Plant Sciences)

30 pages, 2436 KiB

Open AccessReview

Vitamin D in the Prevention and Treatment of Inflammatory Skin Diseases

by Zrinka Bukvić Mokos, Lucija Tomić Krsnik, Kristijan Harak, Danijela Marojević Tomić, Deša Tešanović Perković and Marija Vukojević

Int. J. Mol. Sci. 2025, 26(11), 5005; https://doi.org/10.3390/ijms26115005 - 22 May 2025

Abstract

Vitamin D, a hormone synthesized in the skin through ultraviolet B radiation (UVB), plays a crucial role not only in calcium and phosphate homeostasis but also in regulating skin homeostasis and modulating immune responses. In keratinocytes, vitamin D is converted to its active [...] Read more.

Vitamin D, a hormone synthesized in the skin through ultraviolet B radiation (UVB), plays a crucial role not only in calcium and phosphate homeostasis but also in regulating skin homeostasis and modulating immune responses. In keratinocytes, vitamin D is converted to its active form, 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D), which interacts with the vitamin D receptor (VDR) to regulate gene expression involved in proliferation, differentiation, and antimicrobial defense. Dysregulation of this pathway has been implicated in inflammatory skin diseases such as psoriasis, atopic dermatitis, acne vulgaris, and hidradenitis suppurativa. These conditions are associated with altered epidermal differentiation, immune imbalance, and microbial interactions, where vitamin D plays a modulatory role by suppressing proinflammatory cytokines, enhancing antimicrobial peptide synthesis, and restoring skin barrier integrity. Topical vitamin D analogues have shown therapeutic benefits in psoriasis, while emerging evidence supports the adjunctive role of vitamin D supplementation in acne, hidradenitis suppurativa, and atopic dermatitis. Despite promising associations between low serum vitamin D levels and disease severity, a causal relationship remains uncertain. This review integrates molecular mechanisms with clinical findings, emphasizing the role of vitamin D in cutaneous physiology and pathology, and highlights the need for further research into targeted supplementation strategies in dermatological disorders. Full article

(This article belongs to the Special Issue The Role of Vitamin D in Human Health and Diseases 4.0)

► Show Figures

Figure 1

16 pages, 904 KiB

Open AccessReview

Biological Actions of Bile Acids via Cell Surface Receptors

by Yoshimitsu Kiriyama, Hiroshi Tokumaru, Hisayo Sadamoto and Hiromi Nochi

Int. J. Mol. Sci. 2025, 26(11), 5004; https://doi.org/10.3390/ijms26115004 - 22 May 2025

Abstract

Bile acids (BAs) are synthesized in the liver from cholesterol and are subsequently conjugated with glycine and taurine. In the intestine, bile acids undergo various modifications, such as deconjugation, dehydrogenation, oxidation, and epimerization by the gut microbiota. These bile acids are absorbed in [...] Read more.

Bile acids (BAs) are synthesized in the liver from cholesterol and are subsequently conjugated with glycine and taurine. In the intestine, bile acids undergo various modifications, such as deconjugation, dehydrogenation, oxidation, and epimerization by the gut microbiota. These bile acids are absorbed in the intestine and transported to the liver as well as the systemic circulation. BAs can activate many types of receptors, including nuclear receptors and cell surface receptors. By activating these receptors, BAs can exert various effects on the metabolic, immune, and nervous systems. Recently, the detailed structure of TGR5, the major plasma membrane receptor for BAs, was elucidated, revealing a putative second BA binding site along with the orthosteric binding site. Furthermore, BAs act as ligands for bitter taste receptors and the Leukemia inhibitory factor receptor. In addition, the Mas-related, G-protein-coupled receptor X4 interacts with receptor activity-modifying proteins. Thus, a variety of cell surface receptors are associated with BAs, and BAs are thought to have very complex activities. This review focuses on recent advances regarding cell surface receptors for bile acids and the biological actions they mediate. Full article

(This article belongs to the Section Molecular Biology)

17 pages, 2048 KiB

Open AccessArticle

The MDA-MB-231 Breast Cancer Cell Secretomes Modify Metabolomes of Pseudomonas aeruginosa Breast Microbiome

by Majdoleen AlDawsari, Mysoon M. Al-Ansari, Reem H. AlMalki, Anas M. Abdel Rahman and Monther Al-Alwan

Int. J. Mol. Sci. 2025, 26(11), 5003; https://doi.org/10.3390/ijms26115003 - 22 May 2025

Abstract

Breast cancer (BC) is globally becoming a great challenge, being both the most diagnosed cancer and the leading cause of death in women. In addition to cancer cells, many bacteria co-inhabit BC, which differ in type and number from the resident microbiota found [...] Read more.

Breast cancer (BC) is globally becoming a great challenge, being both the most diagnosed cancer and the leading cause of death in women. In addition to cancer cells, many bacteria co-inhabit BC, which differ in type and number from the resident microbiota found in healthy breast tissue. While many reports have demonstrated the ability of different bacteria to dysregulate BC’s metabolites, the reciprocal effect of these metabolites on the bacterial microbiota has not yet been investigated. Herein, we assess the effect of conditioned media (CM) from a triple-negative BC cell line (MDA-MB-231) on the metabolic profile of Pseudomonas aeruginosa (P. aeruginosa), an important breast resident Gram-negative bacteria that influence oncogenesis. Optical density and scanning electron microscopes were used to assess the impact of MDA-MB-231-CM (BC-CM) on P. aeruginosa growth and morphological changes, respectively. In addition, liquid chromatography–high-resolution mass spectrometry was used to identify metabolic changes in P. aeruginosa and their secretomes in response to the BC-CM. The BC-CM significantly suppressed the growth of P. aeruginosa in the log phase and induced concentration-dependent cytopathological changes in their cell walls. The metabolites of P. aeruginosa were dysregulated considerably depending on the time of exposure to the BC-CM. When treated with the BC-CM, P. aeruginosa induced the purine alkaloid spliceostatin (FR901464), a prominent antitumor metabolite. The BC-CM also promoted other P. aeruginosa metabolites such as amino acids, phosphoribosyl-AMP, 2-aminoacetophenone, pyochelin I, guanosine monophosphate, riboflavin, and terpenoids, which are capable of interfering with oncogenesis. Nine of the significantly identified metabolites from the 0–3 h comparison and four of those identified from the 0–6 h comparison have potential roles in influencing cancer cell behavior. Our findings demonstrate the ability of triple-negative BC-CM not only to alter the growth and morphology of P. aeruginosa but also to modulate their metabolic profile. A better understanding of the influence of BC on certain resident breast microbiomes, such as P. aeruginosa, may open a new therapeutic intervention opportunity for the treatment of cancer. Full article

(This article belongs to the Topic Microbes and Their Products for Sustainable Human Life)

19 pages, 998 KiB

Open AccessReview

Current Knowledge of the Impact of Vitamin D in Coronary Artery Disease

by Freja Esager Jespersen, Daniela Grimm, Marcus Krüger and Markus Wehland

Int. J. Mol. Sci. 2025, 26(11), 5002; https://doi.org/10.3390/ijms26115002 - 22 May 2025

Abstract

Coronary artery disease and vitamin D deficiency are both widespread conditions with a high incidence worldwide. Coronary artery disease is a complex illness with variable manifestation and pathogenesis. It often involves the development of atherosclerosis, and it frequently has serious or even fatal [...] Read more.

Coronary artery disease and vitamin D deficiency are both widespread conditions with a high incidence worldwide. Coronary artery disease is a complex illness with variable manifestation and pathogenesis. It often involves the development of atherosclerosis, and it frequently has serious or even fatal consequences for the patient. Vitamin D receptor expression is found in many tissues throughout the body, which results in a broad effect of the vitamin. Studies have found correlations between vitamin D deficiency and the development of coronary artery disease as well as other cardiovascular diseases, such as hypertension. This review will discuss randomized controlled trials conducted from 2020 forward, aiming to elucidate whether vitamin D supplements have the potential to be used as an add-on treatment for coronary artery disease. The randomized controlled trials all used vitamin D as intervention and tested a population suffering from coronary artery disease or the risk of developing it. Even though animal studies found evidence that vitamin D can regulate inflammation, lipid profile, foam cell formation, vessel reactivity, and blood pressure, which are all mediators in the development of atherosclerosis, the results from the randomized controlled trials were ambiguous. The general older population did not seem to benefit from the treatment, but different subgroups such as patients with type 2 diabetes and patients with more developed coronary artery disease exhibited some positive effects from the treatment. Furthermore, vitamin D showed cardioprotective effects following coronary artery bypass surgery, which make it a possible add-on treatment before invasive coronary intervention. The question in focus still needs further research and a more focused approach on subgroups that may benefit from treatment. Full article

(This article belongs to the Special Issue The Role of Vitamin D in Human Health and Diseases 4.0)

32 pages, 2380 KiB

Open AccessReview

Nanosensors and Microsensors for Body Fluid Monitoring: Various Analyte Detection and Construction Solutions

by Nikola Lenar and Beata Paczosa-Bator

Int. J. Mol. Sci. 2025, 26(11), 5001; https://doi.org/10.3390/ijms26115001 - 22 May 2025

Abstract

This review provides a comprehensive overview of the recent advancements in nanosensors and microsensors for body fluid monitoring. The principles behind sensor technologies, their applications in healthcare, and the types of body fluids that they analyze are described in the scope of this [...] Read more.

This review provides a comprehensive overview of the recent advancements in nanosensors and microsensors for body fluid monitoring. The principles behind sensor technologies, their applications in healthcare, and the types of body fluids that they analyze are described in the scope of this paper. Additionally, this review discusses emerging trends, challenges, and future perspectives in this field. The first two sections explore various body fluids and their diagnostic significance and discuss the fundamentals and classification of nanosensors and microsensors. The main aim of this paper is to highlight recent advancements in nanosensors for body fluid monitoring and to examine the role of microsensors in healthcare diagnostics. Innovative solutions such as microfluidic-based sensors, lab-on-a-chip systems, MEMS-based sensors, and wearable and implantable sensors are discussed in this section. Various construction solutions for microsensors and nanosensors have also been compiled and compared based on their target analytes, which are widely present in body fluids. The following sections review technologies and trends, including AI integration and flexible sensors, and discuss challenges and future perspectives in the development and application of sensors. The conclusion includes a summary of key findings and the future outlook for nanosensors and microsensors in personalized medicine. Full article

(This article belongs to the Special Issue Cutting-Edge Research on Nanosensors and Microsensors)

► Show Figures

Figure 1

18 pages, 2796 KiB

Open AccessArticle

SkinDuo^TM as a Targeted Probiotic Therapy: Shifts in Skin Microbiota and Clinical Outcomes in Acne Patients

by Manuele Biazzo, David Pinzauti and Christine Podrini

Int. J. Mol. Sci. 2025, 26(11), 5000; https://doi.org/10.3390/ijms26115000 - 22 May 2025

Abstract

Acne vulgaris is a common dermatological condition strongly associated with disruptions in the skin microbiota, specifically involving key species such as Cutibacterium acnes and Staphylococcus epidermidis. This study investigates the efficacy of SkinDuo^TM, a topical probiotic containing Lactiplantibacillus plantarum, [...] Read more.

Acne vulgaris is a common dermatological condition strongly associated with disruptions in the skin microbiota, specifically involving key species such as Cutibacterium acnes and Staphylococcus epidermidis. This study investigates the efficacy of SkinDuo^TM, a topical probiotic containing Lactiplantibacillus plantarum, in modulating the skin microbiota and improving clinical outcomes in patients with acne vulgaris. Over a 4-week to 8-week observational study period, microbial composition and diversity shifts were analyzed using full-length 16S rRNA sequencing. Patient responses were categorized into “good” responders (showing significant clinical improvement) and “no_change” responders (with minimal or no improvement). SkinDuo^TM treatment resulted in lower post-treatment Cutibacterium acnes abundance in the “good” group compared to the “no_change” group. The “good” group maintained a stable level of alpha diversity following treatment. In contrast, the “no_change” group exhibited a marked reduction in microbial diversity. Beta diversity analysis revealed distinct clustering patterns associated with improved clinical outcomes. These findings suggest that the preservation of microbial richness and evenness may serve as a potential biomarker for positive response to probiotic therapy. This study highlights the potential of SkinDuo^TM to restore microbial balance and alleviate acne symptoms, contributing to the growing body of evidence supporting microbiome-based therapeutic strategies in dermatology. Full article

(This article belongs to the Special Issue Molecular Advances in Skin Diseases: 3rd Edition)

► Show Figures

Figure 1

21 pages, 1963 KiB

Open AccessArticle

Effects of Resveratrol Derivatives on Melanogenesis and Antioxidant Activity in B16F10 Cells

by Soyeon Kim, Changho Jhin, Sullim Lee, Ho Sik Rho and Chan Yoon Park

Int. J. Mol. Sci. 2025, 26(11), 4999; https://doi.org/10.3390/ijms26114999 - 22 May 2025

Abstract

Excessive melanogenesis causes abnormal pigmentation and a higher risk of skin disorders (e.g., melanoma). Resveratrol (RSV), a natural polyphenol, exerts antioxidant and anti-aging effects. However, the effects of RSV and its derivatives on melanogenesis remain unclear. This study investigated their effects on melanogenesis [...] Read more.

Excessive melanogenesis causes abnormal pigmentation and a higher risk of skin disorders (e.g., melanoma). Resveratrol (RSV), a natural polyphenol, exerts antioxidant and anti-aging effects. However, the effects of RSV and its derivatives on melanogenesis remain unclear. This study investigated their effects on melanogenesis and antioxidant activity in B16F10 cells. After measuring cell viability, B16F10 cells were incubated with 50 µM of RSV, dihydroresveratrol (DIRSV), and other RSV derivatives for 24 h. The relative melanin content and tyrosinase activity were quantified. The protein and mRNA levels of melanogenesis-related genes (MITF, CREB, TYR, and TRP) and the binding affinity of RSV derivatives to their target proteins were measured. The antioxidant activity was evaluated using ABTS and DPPH assays. RSV and DIRSV (50 µM) significantly reduced melanin content and tyrosinase activity, respectively. However, other derivatives had no significant effects. RSV, DIRSV, and other derivatives significantly suppressed MITF and CREB levels. Additionally, DIRSV significantly reduced p-CREB and TYR protein levels and showed a higher affinity for CREB than RSV, despite no significant changes in MITF, TYR, or TRP mRNA levels. In the antioxidant assays, RSV and DIRSV exhibited significant ABTS and DPPH radical scavenging activities. DIRSV, like RSV, inhibits melanogenesis and exhibits antioxidant effects in B16F10 cells. However, RSV derivatives demonstrate partial antioxidant activity and inhibit melanogenesis-related proteins but do not significantly affect melanogenesis. DIRSV’s practical applications as a skin-protective and -whitening agent warrant further exploration. Full article

(This article belongs to the Special Issue Potential Health Benefits of Resveratrol: Laboratory-Confirmed Mechanisms of Action)

21 pages, 4078 KiB

Open AccessArticle

The Effects and Mechanisms of Continuous 7-Day Hypobaric Hypoxia Exposure on Sleep Architecture in Rats

by Fang Li, Xianxie Zhang, Anping Ye, Ling Qi, Tianke Huang, Xitai Chen, Maoxing Li, Chengrong Xiao, Yuguang Wang, Yue Gao and Zengchun Ma

Int. J. Mol. Sci. 2025, 26(11), 4998; https://doi.org/10.3390/ijms26114998 - 22 May 2025

Abstract

High-altitude environments pose significant risks for insomnia development, which severely compromises both physiological health and occupational performance. To elucidate the mechanisms underlying altitude-induced sleep disruption and establish a validated animal model for therapeutic intervention development, we exposed Sprague-Dawley rats to hypobaric hypoxia (5500 [...] Read more.

High-altitude environments pose significant risks for insomnia development, which severely compromises both physiological health and occupational performance. To elucidate the mechanisms underlying altitude-induced sleep disruption and establish a validated animal model for therapeutic intervention development, we exposed Sprague-Dawley rats to hypobaric hypoxia (5500 m altitude equivalent: 308 mmHg, 20.37% O₂, PiO₂ 8.0 kPa) for 7 days. We employed continuous wireless telemetry to monitor EEG/EMG signals, with concurrent analysis of physiological parameters, blood biochemistry, histopathology, transcriptomics, and protein expression. Quantitative analyses demonstrated decreased caloric intake, transient body mass reduction, and immune-metabolic disturbances. While total sleep duration showed no significant variation, sleep architecture displayed elevated wakefulness periods, reduced active wakefulness, a decreasing trend of slow-wave sleep (SWS), and increased paradoxical sleep (PS) accompanied by attenuated circadian oscillations. The duration of SWS episodes was significantly shortened, indicating a sleep homeostasis imbalance that peaked on day 3. Biochemical profiling revealed reduced levels of antioxidant enzymes, elevated pro-inflammatory cytokines, and hypothalamic–pituitary–adrenal axis activation. Transcriptomic analyses identified the critical involvement of serotonergic/glutamatergic synaptic regulation, lipid metabolism, IL-17 signaling, and cortisol synthesis pathways. Western blot analyses confirmed OX2R upregulation, 5-HT1AR downregulation, and circadian gene dysregulation. Our findings demonstrate that hypobaric hypoxia induces sleep disruption via coordinated mechanisms involving oxidative stress, inflammatory activation, HPA axis hyperactivity, neurotransmitter imbalance, and circadian clock dysfunction, providing a robust preclinical model for mechanistic exploration and therapeutic target identification. Full article

(This article belongs to the Section Biochemistry)

► Show Figures

Figure 1

20 pages, 10181 KiB

Open AccessArticle

Encapsulation of Transforming Growth Factor-β3 in Poly(hydroxybutyrate-co-hydroxyvalerate) Nanoparticles for Enhanced Cartilage Tissue Engineering

by Ana Isabel Rodríguez-Cendal, José Señarís-Rodríguez, María Piñeiro-Ramil, Loreto Cabarcos-Mouzo, María del Carmen Veiga-Barbazán, Rosa María Mejide-Faílde, Francisco Javier de Toro-Santos, Isaac Manuel Fuentes-Boquete and Silvia María Díaz-Prado

Int. J. Mol. Sci. 2025, 26(11), 4997; https://doi.org/10.3390/ijms26114997 - 22 May 2025

Abstract

Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) is a naturally occurring biopolymer belonging to the polyhydroxyalkanoate (PHA) family. Due to its excellent properties (biocompatible, biodegradable, and non-toxic), this biopolymer is presented as a very suitable option for use in regenerative therapy as a drug delivery system (DDS). The [...] Read more.

Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) is a naturally occurring biopolymer belonging to the polyhydroxyalkanoate (PHA) family. Due to its excellent properties (biocompatible, biodegradable, and non-toxic), this biopolymer is presented as a very suitable option for use in regenerative therapy as a drug delivery system (DDS). The protein encapsulated in this study is transforming growth factor β3 (TGF-β3), which plays a key role in the chondrogenic differentiation of mesenchymal stem cells (MSCs). The main objective of this work is to evaluate the efficacy of PHBV nanoparticles (NPs) produced from a dairy by-product (whey) as a DDS of TGF-β3 for cartilage regeneration and extracellular matrix (ECM) synthesis and to reduce the complications associated with multiple high doses of TGF-β3 in its free form. For this purpose, biopolymer cytotoxicity, factor release, cell viability, cell proliferation, and differentiation were analyzed. The results showed that the biomaterial purified with chloroform and ethanol, either by single or double precipitation, was not toxic to cells. A sustained release profile was observed, reaching its maximum around day 4. The TGF-β3 NPs promoted the differentiation of MSCs into chondrocytes and the formation of ECM. In conclusion, PHBV demonstrated its potential as an optimal material for DDSs in cartilage regenerative therapy, effectively addressing the key challenge of the need for a single delivery method to reduce complications associated with multiple high doses of TGF-β3. Full article

(This article belongs to the Special Issue Bone and Cartilage Injury and Repair: Molecular Aspects)

► Show Figures

Figure 1

16 pages, 1686 KiB

Open AccessArticle

Mechanistic Insights into Anti-Nectin4-VcMMAE-Induced Ocular Toxicity: From Cellular Uptake Pathways to Molecular Modification

by Jialing Zhang, Meng Li, Weiyu Li, Yuxuan Yang, Gang Wu, Peng Guo, Chuanfei Yu and Lan Wang

Int. J. Mol. Sci. 2025, 26(11), 4996; https://doi.org/10.3390/ijms26114996 - 22 May 2025

Abstract

Antibody–drug conjugates (ADCs) represent a novel approach to cancer treatment. Enfortumab vedotin (PADCEV), as a prominent example, has demonstrated remarkable clinical efficacy. However, its ocular toxicity has raised concerns. This study aimed to explore the molecular mechanisms underlying PADCEV—induced ocular toxicity. SD rats, [...] Read more.

Antibody–drug conjugates (ADCs) represent a novel approach to cancer treatment. Enfortumab vedotin (PADCEV), as a prominent example, has demonstrated remarkable clinical efficacy. However, its ocular toxicity has raised concerns. This study aimed to explore the molecular mechanisms underlying PADCEV—induced ocular toxicity. SD rats, whose ocular structures are similar to those of humans, were selected to establish an ocular toxicity model to mimic the human response. In vitro experiments were conducted using human primary corneal epithelial cells, HCE-T. The results confirmed that nectin-4 plays a crucial role in the cellular uptake of PADCEV, and non-specific pinocytosis is also involved. Additionally, a variant was obtained by introducing point mutations in the Fc region of PADCEV, which was found to reduce corneal epithelial toxicity. The findings of this study not only deepen our understanding of ADC-induced ocular toxicity but also provide new insights into optimizing ADC design and enhancing treatment safety. Full article

(This article belongs to the Section Molecular Toxicology)

22 pages, 996 KiB

Open AccessArticle

DNA Methylation Changes Reflect Aluminum Stress in Triticale and Epigenetic Control of the Trait

by Agnieszka Niedziela, Renata Orłowska and Piotr Tomasz Bednarek

Int. J. Mol. Sci. 2025, 26(11), 4995; https://doi.org/10.3390/ijms26114995 - 22 May 2025

Abstract

Aluminum (Al) stress is typical for acidic soils and may affect cereals’ yield. Al tolerance in triticale is mostly affected by the aluminum-activated malate transporter (ALMT) gene (7R) and some other QTLs (3R, 5R, and 6R). The trait is heritable in about 36% [...] Read more.

Aluminum (Al) stress is typical for acidic soils and may affect cereals’ yield. Al tolerance in triticale is mostly affected by the aluminum-activated malate transporter (ALMT) gene (7R) and some other QTLs (3R, 5R, and 6R). The trait is heritable in about 36% of cases, indicating that epigenetic factors may impact the phenomenon. This study demonstrates that utilizing different methods to quantify DNA methylation changes induced by Al stress results in detail differences, and the results evaluated should be compared critically. The Common and the basic General approaches are sufficient if general information is needed. The General (extended variant) approach may deliver data on methylation changes affecting symmetric sequence contexts. The markers assigned to DN-CG, DM-CG, and DN-CHG were suggested as the most important in explaining Al tolerance in triticale. Analysis of the maps constructed based on root tips and leaf tissues showed different densities of the epigenetic markers but reflected the comparable patterns of their distribution, supporting the hypothesis that Al stress could be transmitted to other plant tissues due to somatic memory. Methylation changes occur throughout the genome and are not associated with specific genes related to aluminum stress. Full article

(This article belongs to the Special Issue Plant Responses to Abiotic and Biotic Stresses)

20 pages, 1415 KiB

Open AccessArticle

Evidence for a Putative Regulatory System Consisting of an ECF σ^E-Type Factor, LIC_12757, and a FecR-like σ Factor Regulator, LIC_12756, in the Pathogenic Spirochaetes Leptospira interrogans

by Sabina Kędzierska-Mieszkowska, Barbara Kędzierska, Laura Pardyak and Zbigniew Arent

Int. J. Mol. Sci. 2025, 26(11), 4994; https://doi.org/10.3390/ijms26114994 - 22 May 2025

Abstract

ECF σ factors, which constitute the most abundant and diverse group of the σ⁷⁰-family, are important signal response regulatory proteins in bacterial adaptative responses to harsh environmental changes and for bacterial survival. Their activity is commonly controlled by specific and reversible [...] Read more.

ECF σ factors, which constitute the most abundant and diverse group of the σ⁷⁰-family, are important signal response regulatory proteins in bacterial adaptative responses to harsh environmental changes and for bacterial survival. Their activity is commonly controlled by specific and reversible interactions with their cognate anti-σ factors (soluble or transmembrane proteins), which directly or indirectly sense the environmental signals and transmit them to their partner σ factor. The genome of pathogenic L. interrogans is predicted to encode 11 ECF σ^E-type factors and more than 30 regulators predicted as anti-σ factors, anti-anti-σ factors, and regulators of anti-anti-σ factors. We have recently demonstrated that one of the L. interrogans ECF σ factors, i.e., LIC_12757, indeed functions as a transcriptional factor and is autoregulated at the transcriptional level. This study is a next step towards determining key aspects of LIC_12757 functioning in Leptospira. By using genetic and proteomic approaches, we provide strong evidence that the LIC_12757 activity is controlled via interactions with its putative FecR-like regulator, LIC_12756. We also demonstrate that LIC_12756 exhibits not only an anti-σ activity but also acts as a positive regulator of LIC_12757 in the presence of specific environmental cues. Interestingly, we found that the nutrient-limiting conditions, including iron deficiency, may act as specific signals for the LIC_12757 activation. In conclusion, we identified the L. interrogans regulatory system consisting of an ECF σ factor, LIC_12757, and a FecR-like regulator, LIC_12756, which is most likely involved in the response of pathogenic Leptospira to iron and nutrient limitation, and thus also likely involved in their response to host-induced stress. Full article

(This article belongs to the Section Molecular Biology)

22 pages, 965 KiB

Open AccessReview

Integrating Proteomics into Personalized Medicine for Inflammatory Bowel Disease—Reality or Challenge?

by Horia Minea, Ana-Maria Singeap, Manuela Minea, Simona Juncu, Stefan Andrei Chiriac, Catalin Victor Sfarti, Carol Stanciu and Anca Trifan

Int. J. Mol. Sci. 2025, 26(11), 4993; https://doi.org/10.3390/ijms26114993 - 22 May 2025

Abstract

Inflammatory bowel diseases (IBD) represent chronic conditions with etiopathogenic mechanisms incompletely elucidated despite extensive research efforts. Therefore, it is essential for clinical monitoring of the implementation of personalized medicine, enabling risk stratification and the selection of therapies with the highest likelihood of a [...] Read more.

Inflammatory bowel diseases (IBD) represent chronic conditions with etiopathogenic mechanisms incompletely elucidated despite extensive research efforts. Therefore, it is essential for clinical monitoring of the implementation of personalized medicine, enabling risk stratification and the selection of therapies with the highest likelihood of a favorable response. Multi-omics approaches have emerged as an excellent opportunity for the prevention, clinical phenotype differentiation, and prediction of IBD development. Proteomics has gained significant enthusiasm in medical practice, primarily due to its focus on studying the composition and dynamic expression of various cellular and tissue structures. This approach provides critical insights into their impact on signaling pathways, post-translational modifications, and the development of sequence variations. Hence, it could provide the foundation for developing biomarkers with the potential to assess mucosal healing and predict prognostic variability among patients, facilitating the implementation of a personalized therapeutic approach. This review focuses on the recent research regarding the possibility of implementing proteomics technologies into clinical practice, given the challenges and limitations, and the advantages of increasing the quality of life in patients with IBD. Full article

(This article belongs to the Special Issue New Research on Anti-Inflammatory Molecules in Diseases)

► Show Figures

Figure 1

14 pages, 802 KiB

Open AccessReview

VEGF in Diabetic Retinopathy and Age-Related Macular Degeneration

by Andrew Callan, Justin Heckman, Giani Tah, Samantha Lopez, Laura Valdez and Andrew Tsin

Int. J. Mol. Sci. 2025, 26(11), 4992; https://doi.org/10.3390/ijms26114992 - 22 May 2025

Abstract

Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis throughout the human body, influencing countless physiological and pathological processes, including tumor growth, preeclampsia, and retinal diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). In DR, VEGF promotes retinal [...] Read more.

Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis throughout the human body, influencing countless physiological and pathological processes, including tumor growth, preeclampsia, and retinal diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). In DR, VEGF promotes retinal neovascularization and intraretinal fluid accumulation, leading to complications like diabetic macular edema (DME) and proliferative diabetic retinopathy (PDR). Regular intravitreal anti-VEGF injections are commonly used to manage PDR and DME, though repeated treatments are often required, and efficacy can be limited. AMD, a major cause of vision loss in older adults, is characterized by either dry or wet forms. While the dry form has not been shown to be influenced by VEGF, the choroidal neovascularization of wet AMD has strong associations with VEGF. Current treatment for wet AMD consists primarily of anti-VEGF injections, the gold standard of care, but is limited by varying patient responses, as treatments are often repeated every 4-8 weeks indefinitely. This review explores the pathogenic role of VEGF in both DR and AMD, discussing the molecular mechanisms underlying these diseases and the therapeutic approaches targeting VEGF. Despite advancements, the variability in treatment responses highlights the need for continued research to develop more effective therapies to prevent vision loss and blindness associated with these retinal diseases. Full article

(This article belongs to the Special Issue Advances in the Pathophysiology and Treatment of Diabetic Retinopathy)

► Show Figures

Figure 1

27 pages, 1593 KiB

Open AccessArticle

TPP-Based Nanovesicles Kill MDR Neuroblastoma Cells and Induce Moderate ROS Increase, While Exerting Low Toxicity Towards Primary Cell Cultures: An In Vitro Study

by Silvana Alfei, Carola Torazza, Francesca Bacchetti, Marco Milanese, Mario Passalacqua, Elaheh Khaledizadeh, Stefania Vernazza, Cinzia Domenicotti and Barbara Marengo

Int. J. Mol. Sci. 2025, 26(11), 4991; https://doi.org/10.3390/ijms26114991 - 22 May 2025

Abstract

Neuroblastoma (NB) is a malignant childhood tumour, which originates from neuroblasts with an incidence of approximately 15,000 new cases per year worldwide. Therapy-induced secondary tumorigenesis and the emergency of drug resistance in its high-risk (HR-NB) forms drive to a survival rate of <50%, [...] Read more.

Neuroblastoma (NB) is a malignant childhood tumour, which originates from neuroblasts with an incidence of approximately 15,000 new cases per year worldwide. Therapy-induced secondary tumorigenesis and the emergency of drug resistance in its high-risk (HR-NB) forms drive to a survival rate of <50%, despite aggressive treatments. Our recent research is focused on testing in vitro the effects of synthetized triphenyl phosphonium (TPP)-based bola amphiphilic nanovesicles (BPPBs) against both drug-sensitive and multi-drug-resistant (MDR) cancer cell lines. In the present study, BPPB demonstrated sub-micromolar IC₅₀ values (0.4–0.9 µM) towards drug-sensitive HTLA 230, while 1.20–1.35 µM IC₅₀ were determined on MDR HTLA ER. Noteworthily, we have demonstrated that BPPB triggers apoptosis of both NB cell populations. Additionally, since MDR NB cells (HTLA ER) are equipped with higher levels of antioxidants than sensitive ones (HTLA 230), the potential involvement of reactive oxygen species (ROS) in the cytotoxic action of BPPB was also investigated. Then, a novel analytical approach was applied to the results of cell viability and ROS monitoring for their better interpretation. Proper dispersion graphs and their best fitting nonlinear regression models were used to verify if the cytotoxic effects of BPPB could depend on BPPB concentrations, exposure times, and/or ROS generation, and if ROS increase could depend on BPPB concentrations and/or exposure times. A ROS-dependent mechanism was found in 24 h and 24/48 h treatments of HTLA ER and HTLA 230, respectively. Furthermore, the potential clinical development of BPPB as a new curative option for children affected by HR-NB was assessed by testing BPPB on astrocyte and neuron primary cell cultures, and analytical correlation studies were used to interpret the results. Notably, BPPB administration was sufficiently and well tolerated by neurons and astrocytes, respectively, allowing selectivity index values of up to 23.7. These in vitro results, associated with the low haemolytic activity of BPPB, pave the way for future in vivo investigations and, upon confirmation, for the possible development of BPPB as a novel therapeutic strategy to treat MDR HR-NB. Full article

(This article belongs to the Special Issue New Molecular Mechanisms and Advanced Therapies for Solid Tumors)

► Show Figures

Graphical abstract

17 pages, 5866 KiB

Open AccessArticle

RANKL Drives Bone Metastasis in Mammary Cancer: Protective Effects of Anti-Resorptive Treatments

by Evi Gkikopoulou, Christos-Chrysovalantis Syrigos, Ioanna Mantogiannakou, Chrysa-Eleni Petraki, Melina Stathopoulou, Melina Dragolia, Vagelis Rinotas, Vasileios Ntafis, Martina Rauner and Eleni Douni

Int. J. Mol. Sci. 2025, 26(11), 4990; https://doi.org/10.3390/ijms26114990 - 22 May 2025

Abstract

Receptor activator of nuclear factor-κB ligand (RANKL) is essential for osteoclast formation and bone resorption, in osteolytic conditions such as osteoporosis and bone metastases. However, its role in metastasis progression remains incompletely understood. Herein, we examined whether the overexpression of human RANKL in [...] Read more.

Receptor activator of nuclear factor-κB ligand (RANKL) is essential for osteoclast formation and bone resorption, in osteolytic conditions such as osteoporosis and bone metastases. However, its role in metastasis progression remains incompletely understood. Herein, we examined whether the overexpression of human RANKL in transgenic mice (TgRANKL) affects their susceptibility to breast cancer bone metastasis compared to their wild-type (WT) littermates. Bone metastasis was induced by injecting EO771 mouse mammary adenocarcinoma cells into the caudal artery of syngeneic WT and TgRANKL mice. RANKL overexpression led to an earlier onset and increased burden of bone metastasis in EO771-bearing TgRANKL mice compared to WT mice. It also exacerbated the bone destruction caused by metastasis-associated osteolysis. The prophylactic inhibition of RANKL activity with denosumab, a monoclonal antibody targeting human RANKL, prevented osteolysis and significantly reduced the incidence and progression of bone metastases in TgRANKL mice. However, the therapeutic denosumab treatment had no effect on metastasis incidence or tumor burden, although it alleviated osteolysis. The treatment with zoledronic acid, an anti-resorptive agent inhibiting osteoclast activity, yielded results similar to those of denosumab. These findings emphasize the significance of initiating early treatment with anti-resorptive agents such as denosumab or zoledronic acid to reduce the risk of bone metastasis in patients at high risk. Full article

(This article belongs to the Section Molecular Oncology)

► Show Figures

Figure 1

17 pages, 3199 KiB

Open AccessArticle

The Splice Variant of the NCOR2 Gene BQ323636.1 Modulates ACSL4 Expression to Enhance Fatty Acid Metabolism and Support of Tumor Growth in Breast Cancer

by Ho Tsoi, Chan-Ping You, Koei Ho-Lam Cheung, Yin-Suen Tse and Ui-Soon Khoo

Int. J. Mol. Sci. 2025, 26(11), 4989; https://doi.org/10.3390/ijms26114989 - 22 May 2025

Abstract

BQ323636.1 (BQ), a splice variant of NCOR2, is associated with endocrine therapy resistance and poorer prognosis in ER-positive breast cancer. This study investigates the role of BQ in modulating lipid metabolism to support tumor growth. RNA sequencing of BQ-overexpressing breast cancer cells revealed [...] Read more.

BQ323636.1 (BQ), a splice variant of NCOR2, is associated with endocrine therapy resistance and poorer prognosis in ER-positive breast cancer. This study investigates the role of BQ in modulating lipid metabolism to support tumor growth. RNA sequencing of BQ-overexpressing breast cancer cells revealed significant enrichment of fatty acid metabolism pathways (hsa01212 and hsa00061; p < 0.05), with ACSL4 identified as a key target. We show that BQ disrupts the NCOR2-PPARγ interaction, leading to ACSL4 upregulation, which enhances fatty acid oxidation (FAO), acetyl-CoA by 1.8-fold, and ATP production by 2.5-fold to fuel tumor proliferation. BQ also upregulates FASN and SCD, increasing lipids. A metabolites study with mass spectrometry indicated that BQ overexpression increases the fatty acid amount from 47.97 nmol/10⁶ cells to 75.18 nmol/10⁶ cells in MCF7 and from 56.19 nmol/10⁶ cells to 95.37 nmol/10⁶ cells in ZR-75. BQ activates NRF2, which mitigates ROS-induced stress, promoting cell survival. Targeting ACSL4 with the inhibitor PRGL493 reduced ATP production and suppressed tumor growth in vitro and in vivo, without inducing apoptosis, suggesting a cytostatic effect. PRGL493 treatment can reduce BQ overexpressing tumors by 40% in the xenograft model. These results highlight BQ can serve as a transcriptional hub driving lipid metabolism via ACSL4 in breast cancer. Our findings suggest that ACSL4 inhibition could be a novel therapeutic strategy to overcome treatment resistance in high-BQ expressing ER-positive breast cancer. Full article

(This article belongs to the Special Issue Hormone Signaling in Cancers and Cancer-Promoting Pathologies)

► Show Figures

Figure 1

18 pages, 2496 KiB

Open AccessArticle

IgA Antibodies to Bovine Serum Albumin in Adult Patients with Celiac Disease

by Elena Savvateeva, Marina Yukina, Nurana Nuralieva, Svetlana Bykova, Ivan Abramov, Vera Polyakova, Natalia Bodunova, Maxim Donnikov, Lyudmila Kovalenko, Elena Mazurenko, Elizaveta Pavlova, Elena Kulagina, Ekaterina Troshina and Dmitry Gryadunov

Int. J. Mol. Sci. 2025, 26(11), 4988; https://doi.org/10.3390/ijms26114988 - 22 May 2025

Abstract

This study investigated the IgA antibodies targeting bovine serum albumin (BSA) in 27 adult celiac disease (CD) patients adhering to a gluten-free diet (GFD), compared to 123 controls (including individuals with autoimmune disorders, those with gastrointestinal cancers, and healthy donors). Serum samples were [...] Read more.

This study investigated the IgA antibodies targeting bovine serum albumin (BSA) in 27 adult celiac disease (CD) patients adhering to a gluten-free diet (GFD), compared to 123 controls (including individuals with autoimmune disorders, those with gastrointestinal cancers, and healthy donors). Serum samples were evaluated using a multiplex assay based on a microarray comprising 66 immobilized antigens, including autoantigens associated with autoimmune diseases, different albumins, cytokines, and inflammatory markers. Elevated IgA-BSA levels were detected in 22% of CD patients versus 3.25% of controls. IgA-BSA did not cross-react with milk proteins like casein, β-lactoglobulin, and γ-globulin, nor with autoantigens and human albumin, ruling out autoimmunity against self-proteins. The observed cross-reactivity with porcine albumin suggests that antibodies target epitopes shared by bovine and porcine albumin. Increased IgA-BSA levels may interfere with immunoassays performed using BSA as a stabilizer, necessitating protein-free buffers to avoid false results when testing CD patients. Elevated IgA-BSA levels may reflect ongoing gut barrier dysfunction in CD patients on a GFD, allowing dietary proteins like BSA to trigger immune responses. This study identifies a novel immune response in CD patients on a GFD, emphasizing the need for tailored diagnostic approaches (BSA-free assays) and further research into the clinical and dietary implications of IgA-BSA elevation. Full article

(This article belongs to the Special Issue Autoimmune Diseases: From Pathogenesis to Treatment and Prognostic Markers)

► Show Figures

Figure 1

18 pages, 6620 KiB

Open AccessReview

Extracellular-Matrix-Mimetic Hydrogels by Using Nanomaterials

by Do Gyun Kim and Gi Doo Cha

Int. J. Mol. Sci. 2025, 26(11), 4987; https://doi.org/10.3390/ijms26114987 - 22 May 2025

Abstract

Matrigel, a tumor-derived basement membrane extract, has been commercially used in the field of cell culture and tissue engineering due to its extracellular-matrix-mimetic (ECM-mimetic) properties. However, its batch-to-batch variability and limited mechanical tunability hinder reproducibility and clinical translation. To overcome these issues, synthetic [...] Read more.

Matrigel, a tumor-derived basement membrane extract, has been commercially used in the field of cell culture and tissue engineering due to its extracellular-matrix-mimetic (ECM-mimetic) properties. However, its batch-to-batch variability and limited mechanical tunability hinder reproducibility and clinical translation. To overcome these issues, synthetic ECM-mimetic hydrogels have been developed to improve reproducibility and biocompatibility. While they are effective in mimicking ECMs, these materials must go beyond passive replication by implementing the complex functionalities of the ECM. The integration of nanomaterials with hydrogel could address this need by reinforcing mechanical properties, enabling various functionalities, and featuring dynamic responsiveness. In this review, we present the evolution from Matrigel to ECM-mimetic hydrogels and ECM-mimetic hydrogel nanocomposites, exploring their key advancements and challenges. We will discuss the advantages and disadvantages of the transition from Matrigel to ECM-mimetic hydrogels and ECM-mimetic hydrogel nanocomposites, along with cases that have addressed Matrigel’s limitations and added new functionalities. Furthermore, we discuss future directions for the design of the ECM-mimetic hydrogels, emphasizing how nanotechnology strategies can drive innovation in tissue engineering and regenerative medicine. Full article

(This article belongs to the Special Issue Functional Nanomaterials and Nanocomposites for Biomedical Application)

► Show Figures

Figure 1

17 pages, 712 KiB

Open AccessArticle

Association of Functional Gene Variants in DYSF–ZNF638, MTSS1 and Ferroptosis-Related Genes with Multiple Sclerosis Severity and Target Gene Expression

by Tamara Djuric, Ana Djordjevic, Jovana Kuveljic, Milan Stefanovic, Evica Dincic, Ana Kolakovic and Maja Zivkovic

Int. J. Mol. Sci. 2025, 26(11), 4986; https://doi.org/10.3390/ijms26114986 - 22 May 2025

Abstract

Multiple sclerosis (MS) is a chronic inflammatory, neurodegenerative disease with yet-unresolved mechanisms of progression. To address MS severity and neurological deficits, we analyzed seven potentially functional genetic variants and their haplotypes in 845 MS patients. Based on our previous results of targeted RNAseq [...] Read more.

Multiple sclerosis (MS) is a chronic inflammatory, neurodegenerative disease with yet-unresolved mechanisms of progression. To address MS severity and neurological deficits, we analyzed seven potentially functional genetic variants and their haplotypes in 845 MS patients. Based on our previous results of targeted RNAseq on ferroptosis-related genes in distinctive MS phenotypes, we selected putative regulatory variants in the top three DEGs (CDKN1A, MAP1B and EGLN2) and investigated their association with gene expression, plasma/serum parameters and disease severity (EDSS, MSSS, gARMSS). The study included 604 patients with relapsing–remitting (RR) and 241 with progressive (P) MS. The variants CDKN1A rs3176326 and rs3176336, EGLN2 rs111833532, MAP1B rs62363242 and rs1217817 with the previously reported DYSF-ZNF638 locus rs10191329, and MTSS1 rs9643199 were genotyped using TaqMan^®, and the HLA-DRB1*15:01 status was also determined. Significant association of the rare MAP1B rs62363242 allele with PMS in females, independent of HLA-DRB1*1501, was found. The A allele-containing genotypes were associated with molecular components of iron metabolism. CDKN1A haplotypes were significantly associated with CDKN1A mRNA levels in RRMS and SPMS patients. RAB4B-EGLN2 locus rs111833532 and DYSF-ZNF638 locus rs10191329 showed significant associations with EDSS, MSSS and gARMSS. We detected haplotypes associated with the expression of CDKN1A, a part of the p53-p21 axis known to affect T cell activation/proliferation. RAB4B-EGLN2, an oxygen sensor and critical regulator of the response to hypoxia, variant rs111833532, along with DYSF-ZNF638 locus rs10191329, was associated with clinical severity. The indicated, novel, sex-specific association of MAP1B rs62363242 with the course of MS remains to be validated in larger studies. Full article

(This article belongs to the Special Issue Molecular Research and Treatment in Multiple Sclerosis)

► Show Figures

Figure 1

13 pages, 2311 KiB

Open AccessArticle

Immunotherapy Platform That Conjugates Antigen to Complement C3-Targeted Liposomes Induces a Robust Adaptive Immune Response

by R. G. Barber, Steven Cherry, Sydney Stephens, Kristine Mann, Holly A. Martinson and Max Kullberg

Int. J. Mol. Sci. 2025, 26(11), 4985; https://doi.org/10.3390/ijms26114985 - 22 May 2025

Abstract

The activation of immunosuppressed antigen-presenting cells (APCs) in the tumor microenvironment is a key goal in modern cancer immunotherapy. Our laboratory utilizes a liposome-based immunotherapy platform that binds endogenous complement to deliver antigen, adjuvant, and therapeutic agents to APCs in vivo. The liposomes [...] Read more.

The activation of immunosuppressed antigen-presenting cells (APCs) in the tumor microenvironment is a key goal in modern cancer immunotherapy. Our laboratory utilizes a liposome-based immunotherapy platform that binds endogenous complement to deliver antigen, adjuvant, and therapeutic agents to APCs in vivo. The liposomes contain external linker groups, which readily bind complement protein C3, and mediate liposomal uptake via complement receptor 3 into APCs. To test the ability of a model antigen to bind to these external linker groups on C3-liposomes and elicit a robust adaptive immune response, we conjugated a modified ovalbumin peptide (OVA-C) to the liposomes and incorporated a toll-like receptor (TLR) 4 agonist, monophosphoryl lipid A (MPLA), in the liposomal membrane. Adaptive immune responses from C57BL/6 mice were analyzed by ELISA and ELISpot. Mice vaccinated with OVA-C liposomes elicited significantly greater humoral and cellular adaptive responses relative to controls. Furthermore, female mice vaccinated with MPLA + OVA-C liposomes produced significantly more IgG antibodies than males vaccinated with the same liposomes. In conclusion, antigen binding on the exterior of C3-liposomes markedly improves antigen loading efficiency and still allows for complement C3-targeted delivery to APCs. These data demonstrate the initiation of a robust cellular and humoral immune response using a new liposomal delivery platform. Full article

(This article belongs to the Special Issue Nanomedicine in Gene Therapy and Immunotherapy)

► Show Figures

Figure 1

25 pages, 1118 KiB

Open AccessReview

Induced Pluripotent Stem Cells in Cardiomyopathy: Advancing Disease Modeling, Therapeutic Development, and Regenerative Therapy

by Quan Duy Vo, Kazufumi Nakamura, Yukihiro Saito, Satoshi Akagi, Toru Miyoshi and Shinsuke Yuasa

Int. J. Mol. Sci. 2025, 26(11), 4984; https://doi.org/10.3390/ijms26114984 - 22 May 2025

Abstract

Cardiomyopathies are a heterogeneous group of heart muscle diseases that can lead to heart failure, arrhythmias, and sudden cardiac death. Traditional animal models and in vitro systems have limitations in replicating the complex pathology of human cardiomyopathies. Induced pluripotent stem cells (iPSCs) offer [...] Read more.

Cardiomyopathies are a heterogeneous group of heart muscle diseases that can lead to heart failure, arrhythmias, and sudden cardiac death. Traditional animal models and in vitro systems have limitations in replicating the complex pathology of human cardiomyopathies. Induced pluripotent stem cells (iPSCs) offer a transformative platform by enabling the generation of patient-specific cardiomyocytes, thus opening new avenues for disease modeling, drug discovery, and regenerative therapy. This process involves reprogramming somatic cells into iPSCs and subsequently differentiating them into functional cardiomyocytes, which can be characterized using techniques such as electrophysiology, contractility assays, and gene expression profiling. iPSC-derived cardiomyocyte (iPSC-CM) platforms are also being explored for drug screening and personalized medicine, including high-throughput testing for cardiotoxicity and the identification of patient-tailored therapies. While iPSC-CMs already serve as valuable models for understanding disease mechanisms and screening drugs, ongoing advances in maturation and bioengineering are bringing iPSC-based therapies closer to clinical application. Furthermore, the integration of multi-omics approaches and artificial intelligence (AI) is enhancing the predictive power of iPSC models. iPSC-based technologies are paving the way for a new era of personalized cardiology, with the potential to revolutionize the management of cardiomyopathies through patient-specific insights and regenerative strategies. Full article

(This article belongs to the Special Issue Myocardial Disease: Molecular Pathology and Treatments)

► Show Figures

Figure 1

12 pages, 3789 KiB

Open AccessArticle

Phenylalanine Ammonia-Lyase GhPAL9 Confers Resistance to Verticillium Wilt in Cotton

by Chuanzong Li, Guoshuai Zhang, Guanfu Cheng and Qi Wang

Int. J. Mol. Sci. 2025, 26(11), 4983; https://doi.org/10.3390/ijms26114983 - 22 May 2025

Abstract

Verticillium wilt (VW), induced by the soil-borne fungus Verticillium dahliae, represents a significant threat to global cotton production. Phenylalanine ammonia-lyase (PAL) is an essential enzyme in lignin metabolism that helps plants defend themselves against pathogenic fungal. Nonetheless, its role in imparting resistance [...] Read more.

Verticillium wilt (VW), induced by the soil-borne fungus Verticillium dahliae, represents a significant threat to global cotton production. Phenylalanine ammonia-lyase (PAL) is an essential enzyme in lignin metabolism that helps plants defend themselves against pathogenic fungal. Nonetheless, its role in imparting resistance to V. dahliae in cotton required further investigation. This study identified the GhPAL9 (GH_D04G1247) as a crucial gene in cotton resistance to V. dahliae via RNA-seq analysis, demonstrating significant upregulation in the resistant variety Xinluzao84. Bioinformatics analysis revealed the conserved evolutionary relationship of GhPAL9 with PAL homologs across various species and highlighted stress-responsive cis-elements in its promoter region. The expression of GhPAL9 was rapidly activated in roots, stems, and leaves following infection with V. dahliae, peaking between 2 and 8 h post inoculation (hpi). Silencing GhPAL9 through virus-induced gene silencing (VIGS) technology intensified disease symptoms, elevated relative fungal biomass, and diminished lignin accumulation, thereby affirming its function in cotton resistance to V. dahliae. The overexpression of GhPAL9 in Arabidopsis improved resistance to V. dahliae, and the OE-GhPAL9 transgenic lines demonstrated reduced disease severity and diminished relative fungal biomass. The results gave us new information about how VW resistance at the molecular level, which established that GhPAL9 acted as a positive regulator to increase resistance to VW via lignin accumulation. Full article

(This article belongs to the Section Molecular Plant Sciences)

► Show Figures

Figure 1

30 pages, 1033 KiB

Open AccessReview

Targeting Aging Hallmarks with Monoclonal Antibodies: A New Era in Cancer Immunotherapy and Geriatric Medicine

by Michele Dal Bo, Marta Gambirasi, Idris Vruzhaj, Erika Cecchin, Abbas Pishdadian, Giuseppe Toffoli and Amin Safa

Int. J. Mol. Sci. 2025, 26(11), 4982; https://doi.org/10.3390/ijms26114982 - 22 May 2025

Abstract

Aging is characterized by a progressive deterioration in physiological function and an increased susceptibility to age-related diseases, such as cancer. Monoclonal antibodies (mAbs) constitute a novel therapeutic approach aimed at addressing aging mechanisms such as cellular senescence, inflammaging, and immunosenescence. This text presents [...] Read more.

Aging is characterized by a progressive deterioration in physiological function and an increased susceptibility to age-related diseases, such as cancer. Monoclonal antibodies (mAbs) constitute a novel therapeutic approach aimed at addressing aging mechanisms such as cellular senescence, inflammaging, and immunosenescence. This text presents an overview of mAb methods aimed at the markers of aging and their potential application in cancer treatment. The mAbs can be categorized into senolytics, senescence-associated secretory phenotype (SASP) neutralizers, and immune checkpoint inhibitors, each targeting fewer aging-related pathways relevant to cancer therapeutic enhancement than the last. Translating promising preclinical evidence into enhanced efficacy and safety in cancer therapy presents challenges, particularly in older populations. This study examines the therapeutic efficacy of mAbs in the treatment of cancer and age-related disorders, focusing on their current and future roles in oncology practice. Full article

(This article belongs to the Special Issue New Insights in Antibody Therapy)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Int. J. Mol. Sci., Volume 26, Issue 11 (June-1 2025) – 50 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI