The number of patients infected with simian malaria is gradually increasing in many countries of Southeast Asia and South America. The most important risk factor for a zoonotic spillover event of malarial infection is mostly influenced by the interaction between humans, monkeys, and vectors. In this study, we determine the protein expression profile of a wild stump-tailed macaque (Macaca arctoides) from a total of 32 blood samples collected from Prachuap Kiri Khan Province, Thailand. The malarial parasite was analyzed using nested polymerase chain reaction (PCR) assays by dividing the samples into three groups: non-infected, mono-infected, and multiple-infected. The identification and differential proteomic expression profiles were determined using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and bioinformatics tools. A total of 9,532 proteins (total proteins) were identified with the filter-based selection methods analysis, and a subset of 440 proteins were found to be different between each group. Within these proteins, the GhostKOALA functional enrichment analysis indicated that 142 important proteins were associated with either of the organismal system (28.87%), genetic information processing (23.24%), environmental information processing (16.20%), metabolism (13.38%), cellular processes (11.97%), or causing human disease (6.34%). Additionally, using interaction network analysis, nine potential reporter proteins were identified. Here, we report the first study on the protein profiles differentially expressed in the serum of wild stump-tailed macaques between non, mono, and multiple malarial infected living in a natural transmission environment. Our findings demonstrate that differentially expressed proteins implicated in host defense through lipid metabolism, involved with TGF pathway were suppressed, while those with the apoptosis pathway, such as cytokines and proinflammation signals were increased. Including the parasite’s response via induced hemolysis and disruption of myeloid cells. A greater understanding of the fundamental processes involved in a malarial infection and host response can be crucial for developing diagnostic tools, medication development, and therapies to improve the health of those affected by the disease.

Zika virus (ZIKV) is a member of the Flaviviridae virus family and poses a significant global health concern. ZIKV is transmitted by Aedes mosquitoes, and it has been implicated in various neurological conditions associated with fetal brain development. ZIKV has two transmission cycles: a sylvatic cycle in which nonhuman primates are infected via arboreal mosquito bites, and an interhuman (urban) cycle in which the virus is transmitted among primates by Aedes mosquitoes. ZIKV was first discovered in wild macaques, and the danger posed by the virus is increased due to the close proximity between humans and wild animals in modern society. However, data regarding the extent and role of infection in nonhuman primates are limited. Thus, there is an urgent need for improved surveillance, diagnostic methods, and public health interventions to effectively combat ZIKV transmission and its associated health impacts in Southeast Asia. In this study, we used a proteomics and bioinformatics approach to profile serum proteins in wild stump-tailed macaques seropositive for neutralizing antibodies against ZIKV. A total of 9,532 total proteins were identified, and 338 differentially expressed proteins were identified between naïve and seropositive animals. A total of 52 important proteins were used to construct a serum proteomic profile. These 52 important proteins were associated with immune and inflammatory responses (36.54%), neurological damage (23.08%), viral activities (21.15%), the apoptosis signaling pathway (9.61%), and other pathways (9.61%). Our proteomic profile identified proteins that inhibit the apoptosis pathway, intracellular resource competition with the virus, and neurological damage due to ZIKV and the host immune and defense responses.

Bioactive fillers in dental adhesives are designed to release beneficial ions, such as calcium and phosphate, to help in remineralization process, and preventing secondary caries. Their incorporation also aims to improve the longevity of dental restorations. Herein, CaP nanoparticles were synthesized through a water-in-oil emulsion method using Tween80 as a surfactant and methacrylic acid (MAA). MAA officiated as a co-surfactant and reduced the size of CaP nanoparticle, including improved compatibility with the components of dental adhesive. The incorporation of 2% T80/CaP/MAA nanofillers into dental adhesive demonstrated increased efficiency, as evidenced by a higher degree of conversion (%DC) and greater micro-tensile bond strength (μTBS). More importantly, T80/CaP/MAA nanofillers were able to induce remineralization process in the dental structure, which was confirmed by EDX mapping and IR imaging. Finally, a cytotoxicity assessment of the dental adhesive containing T80/CaP/MAA showed no significant adverse effects on human fibroblast cells.

This double-blind, randomized, placebo-controlled trial investigated the effects of oral combined menopausal hormone therapy (MHT) on vaginal microbiota and clinical outcomes in postmenopausal women with genitourinary syndrome of menopause (GSM). Thirty-four healthy postmenopausal women aged 40-60 years with moderate to severe GSM were randomized to receive either oral estradiol 1 mg/dydrogesterone 5 mg (Femoston conti®) or placebo daily for 12 weeks. Vaginal microbiota was assessed at baseline and week 12 using 16S rDNA gene sequencing. Clinical outcomes, including the most bothersome symptom (MBS), vaginal atrophy score (VAS), vaginal pH, vaginal health index (VHI) and vaginal maturation value (VMV), were evaluated at baseline, week 6, and week 12. The MHT group demonstrated significant more improvement in all clinical measures compared with placebo: MBS (p = 0.008), VAS (p < 0.001), vaginal pH (p = 0.001), VHI (p < 0.001) and VMV (p < 0.001). However, no significant differences were observed in microbial composition or diversity between groups after treatment. These findings suggest that while systemic hormone therapy provides meaningful symptom relief and improves vaginal tissue health, these clinical benefits can occur without concurrent, statistically significant changes in the vaginal microbiota. This supports the clinical value of early systemic MHT in managing GSM, even in the absence of microbiome restoration.

Dengue infection is a leading global public health problem, and the virus (DENV) is transmitted by Aedes mosquitoes and causes a wide spectrum of clinical manifestations in humans. Previous studies have shown that wild macaques in Thailand have been exposed to the dengue virus. Accordingly, this study aims to characterize the serum proteomic profiles of wild macaques with (seropositive) and without (naive) dengue virus-neutralizing antibodies, to improve our understanding of differential protein expression and identify candidate biomarkers. We analyzed thirty-two serum samples obtained from wild macaques in Thailand. Out of the 32 samples, 16 macaques (50%) were seropositive for DENV. A total of 9,532 proteins were identified, out of which 582 were differentially expressed (DEPs) and used to generate a proteomic profile. Among the nine identified proteins of interest, three were associated with the nervous system, while the remaining six have been reported to play roles in anti-dengue antiviral mechanisms. These include the induction of interferon responses triggered by cellular stress, degradation of viral RNA, and activation of dendritic cells and Th1-mediated immune responses via IFN-γ and TNF-α. Although these proteins have been implicated in the nervous system and are involved in anti-DENV in several reports, in this study, it was shown that these proteins remain up-regulated even after infection, which requires further study into the dynamics after long-term infection in the future. These findings highlight the need for further longitudinal studies to validate the functional relevance of these proteins.

Background: Therapeutic resistance is a major cause of treatment failure in glioblastoma (GBM), highlighting the need for physiologically relevant models to identify actionable resistance mechanisms. While two‐dimensional (2D) cultures are widely used for target discovery, they poorly represent the tumor microenvironment. In contrast, three‐dimensional (3D) spheroid cultures better recapitulate spatial heterogeneity, hypoxic gradients, and stress‐adaptive signaling observed in tumors. Methods: We applied an integrated 2D–3D quantitative proteomic approach to identify microenvironment‐dependent regulators of chemoresistance in GBM. Proteomic profiling was performed in U87MG and U251MG cells grown as 2D monolayers or 3D spheroids. Differentially expressed proteins were validated by quantitative RT‐PCR, and functional studies were conducted using genetic depletion followed by assessment of temozolomide (TMZ) sensitivity. Results: Comparative analysis identified 13 proteins consistently differentially expressed between 2D and 3D cultures: NDUFB5, RNGTT, MLK4, SYN1, DDX5, EIF2AK2, ITGA1, ZNF33B, ZNF343, WDR19, JPH3, CCT8L2, and FNDC3A. Among these, Mixed Lineage Kinase 4 (MLK4) showed strong and reproducible upregulation in 3D spheroids in both GBM cell lines. Genetic depletion of MLK4 significantly increased TMZ sensitivity without affecting basal cell viability, suggesting a specific role in therapy response. Notably, MLK4 expression was induced only under 3D conditions. Conclusion: MLK4 functions as a microenvironment‐responsive regulator of chemoresistance in GBM. These findings demonstrate that 3D culture systems reveal clinically relevant resistance pathways not detectable in conventional 2D models and highlight 3D proteomic profiling as a powerful strategy for identifying therapeutically actionable targets.