Spermatogonial stem cells (SSCs) play a crucial role in preserving male fertility, and they are increasingly recognised as essential tools in conservation biology. However, the molecular identity of SSCs in domestic cats (Felis catus) remains insufficiently characterised. This study aimed to investigate the immunohistochemical expression patterns of three candidate markers Fibroblast Growth Factor Receptor 3 (FGFR3), Integrin β1 (ITGB1), and Chemokine Receptor Type 4 (CXCR4) in prepubertal and pubertal cat testes. Testicular tissues from five prepubertal and five pubertal cats were collected and classified based on histological evaluation of spermatogenic cells. Cryosections were obtained and analysed using immunofluorescence staining protocols combined with α-SMA and DAPI counterstaining. The expression patterns were evaluated on a fluorescence microscope, and quantitative fluorescence intensities were measured using ImageJ. Our results reveal that ITGB1 expression is robust and uniformly distributed throughout seminiferous tubules in both developmental stages, with no significant variation between groups (p > 0.05). In contrast, CXCR4 expression is significantly elevated in prepubertal testes compared to pubertal samples (p < 0.05), showing restricted localisation to basal compartments post-puberty. FGFR3 expression was primarily observed in gonocytes during the prepubertal period and localised to basal membranes and sperm tail regions during puberty, with no significant difference between pubertal and prepubertal groups (p > 0.05). These findings provide novel insights into the molecular characteristics of feline SSCs and offer a valuable foundation for the development of advanced reproductive technologies to preserve genetic diversity and support the conservation of endangered felid species.

The dog ovary undergoes dynamic and metabolic remodeling during prepubertal development and throughout the estrous cycle. Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear transcription factors that regulate lipid metabolism and cellular differentiation; however, their stage-specific spatial regulation in the dog ovary remains poorly defined. This study characterized the compartment-specific localization and semi-quantitative abundance patterns of PPARα, PPARδ, and PPARγ in prepubertal and cyclic sexually mature dogs. Ovarian tissues were obtained from 10 prepubertal bitches and 40 adult bitches representing proestrus, estrus, diestrus, and anestrus. Immunofluorescence staining was performed, and expression in granulosa and luteal cells was evaluated using a semi-quantitative scoring approach. All three PPARs were present based on cell-type and stage-dependent nuclear immunoreactivity within granulosa and luteal tissues. Granulosa PPARγ exhibited greater abundance in estrus (p ≤ 0.05) and diestrus (p ≤ 0.001) than in anestrus, and luteal PPARγ nuclear immunoreactivity was increased in diestrus (p ≤ 0.01). Granulosa PPARα levels were lower in prepubertal ovaries than in estrus and diestrus (p ≤ 0.05), whereas luteal PPARα levels were higher during diestrus compared with anestrus (p ≤ 0.001). Granulosa cell PPARδ reached its highest levels during estrus relative to prepubertal and anestrus stages (p ≤ 0.05-0.01), and remained elevated during diestrus compared with anestrus (p ≤ 0.05). Collectively, these findings demonstrate PPAR-specific, stage-dependent, and tissue-specific regulation of PPARs in the canine ovary. The pronounced diestrus-associated nuclear enrichment of PPARγ, the metabolically coherent modulation of PPARα, and the sustained distribution of PPARδ suggest coordinated metabolic adaptation during ovarian follicular maturation and prolonged luteal function in the dog, while the detection of all PPARs in prepubertal ovaries indicates that PPAR-mediated regulatory networks are established prior to endocrine cyclicity during the dog reproductive cycle.

Nosema infection negatively affects colony health, productivity, and survival. The most commonly used drug for treating Nosema infection is fumagillin, but due to its side effects and being banned in some regions, alternative approaches need to be explored. This study aimed to examine the effects of an infusion prepared from the aerial parts of wormwood and the buds of European red pine on bee health and Nosema infection. The effects of the infusion on bee health were determined by a chronic toxicity test (10-day toxicity), locomotor activity, and histopathological examination of the midgut. Also, the anti-Nosema activity of the infusion was determined by comparison with fumagillin under laboratory conditions. The 10-day toxicity test showed that the infusion was well-tolerated, maintaining a high survival rate of 99.2

Myocardial infarction remains a leading cause of heart failure owing to the limited regenerative capacity of adult cardiac tissue, underscoring the need for biomimetic therapeutic platforms that combine structural support with biological functionality. Accordingly, this study aimed to develop a multifunctional electrospun cardiac patch by integrating decellularized neonatal porcine myocardial extracellular matrix (dECM), gelatin, and microbiota-derived postbiotics for cardiac tissue engineering. The fabricated patches were comprehensively characterized in terms of their morphology, mechanical properties, biodegradation behavior, antibacterial activity, antioxidant capacity, and in vitro biocompatibility. Postbiotics derived from Lactiplantibacillus plantarum EIR/IF-1 exhibited potent antimicrobial activity against methicillin-resistant Staphylococcus aureus, strong antioxidant capacity, and significant anti-inflammatory activity through the suppression of pro-inflammatory mediators and upregulation of IL-10 expression. Moreover, they protected H9c2 cardiomyoblasts from oxidative stress, promoted COL1A1 expression, and supported ECM remodeling. The fabricated electrospun cardiac patches exhibited a homogeneous nanofibrous architecture, mechanically suitable properties (Young’s modulus ~4 MPa), controlled biodegradation over 7 days, favorable cell viability, and maintained the biological functionality of the incorporated postbiotics. Overall, the synergistic integration of tissue-specific dECM and microbiota-derived postbiotics yielded a multifunctional biohybrid cardiac patch with favorable structural and biological properties, supporting its potential as a promising platform for myocardial regeneration and next-generation cardiac tissue engineering.

This study investigated the expression of three ovarian remodeling-associated markers, fibroblast growth factor receptor 3 (FGFR3), C-X-C chemokine receptor type 4 (CXCR4), and integrin beta-1 (ITGB1) in the canine ovary across both the prepubertal stage and the distinct phases of the estrous cycle. Ovarian tissues were obtained from 50 dogs (n = 10 per group: prepubertal, proestrus, estrus, diestrus, and anestrus) and analyzed using hematoxylin and eosin staining to confirm stage-specific ovarian morphology, alongside immunofluorescence to evaluate marker localization. FGFR3 immunoreactivity in granulosa cells progressively increased from prepuberty to estrus (p < 0.05) but declined sharply in anestrus (p < 0.0001). CXCR4 showed stage-dependent variation, with interstitial cell expression peaking in proestrus (p < 0.05) and luteal positivity restricted to diestrus (p < 0.01 vs. anestrus). ITGB1 was already detectable in prepubertal ovaries and broadly expressed across compartments, including oocytes in diestrus, but decreased significantly in anestrus compared with proestrus (p < 0.05) and estrus (p < 0.01). These results demonstrate that FGFR3, CXCR4, and ITGB1 display stage and cell-type-specific expression in the canine ovary, with baseline expression already evident in prepubertal tissue. Collectively, the findings underscore the involvement of growth factor, chemokine, and integrin pathways in follicular growth, luteal function, and ovarian quiescence, providing novel insights into canine reproductive biology and offering comparative perspectives for ovarian physiology in other species.

The biodistribution profile is a critical determinant of the success of nanomaterial applications, governing both therapeutic efficacy and systemic safety in emerging photothermal therapy (PTT) agents. Despite the promise of PTT, the clinical translation of many nanomaterial-based photothermal agents is impeded by intrinsic toxicity and insufficient tumor specificity. Moreover, off-target biodistribution of photosensitizers after systemic administration can increase systemic exposure, reduce tumor delivery, and ultimately compromise therapeutic efficacy, underscoring the need for advanced material delivery concepts that enhance selective tumor accumulation and therapeutic outcome. Here, we present a targeted MXene-based PTT platform designed to address these challenges. Among the tested MXenes (Ti3C2, Nb2C, and V2C), Ti3C2 was identified as the most biocompatible and safest candidate, demonstrating minimal cytotoxicity, negligible impact on cell-cycle progression, and no significant impairment of long-term proliferative capacity. Ti3C2 MXene was subsequently coated with polydopamine (PDA) and conjugated to an anti-human-CEACAM1 (hCC1) antibody (Ab) to enable antigen-dependent tumor targeting. In a murine melanoma model bearing human CC1-positive (hCC1+) and CEACAM1-negative (hCC1-) tumors, we evaluated the outcomes of both intravenous (IV) and intratumoral (IT) administration of the Ti3C2-PDA-anti-hCC1 nanoconjugate. Quantification of titanium by inductively coupled plasma atomic emission spectrometry (ICP-AES), in combination with histological analysis, revealed preferential accumulation of the targeted nanoconjugate in hCC1+ tumors compared with hCC1- controls. Notably, after IV delivery, Ti3C2-PDA-anti-hCC1 conjugate demonstrated enhanced deposition in the tumor and lower non-specific uptake in clearance organs, including the liver, lungs, and spleen, compared with non-antibody-functionalized MXene. Collectively, these results support the Ti3C2-PDA-anti-hCC1 conjugates as a tumor-selective biodistribution platform with reduced clearance-organ burden, supporting further therapeutic validation of MXene-based targeted PTT.

A 22-week trial was conducted to assess the effects of replacing inorganic phosphorus (P) with two levels of a hybrid bacterial 6-phytase in low-energy diets for laying hens, from 23 to 44 weeks of age. The study focused on hen performance, egg quality and bone health of laying hens. For this purpose, Lohmann Brown Classic hens (n = 432) were randomly allocated to four dietary groups, each comprising nine replicates of 12 birds. The groups included: (1) positive control (PC), a standard diet containing 3.7% calcium, 0.38% non-phytate phosphorus (nPP) and 2730 kcal/kg metabolizable energy (ME), (2) negative control (NC), a diet similar in nutritional specifications to the PC but with reduced nPP (0.12%) and ME (2630 kcal/kg), (3) NC300 and (4) NC600, where NC diets were supplemented with 300 and 600 phytase unit (FTU) per kg feed, respectively. All diets were provided as mash and formulated using corn, soybean meal and sunflower meal as the main ingredients. The NC diet significantly impaired hen performance compared to the PC diet (p < 0.05). Specifically, the NC diet led to deterioration in egg production (p < 0.001), egg weight (p = 0.001), egg mass (p < 0.001), feed intake (p < 0.001), feed conversion ratio (p = 0.002), body weight (p < 0.001), and livability (p = 0.036). Additionally, the NC diet increased the incidence of cracked (p < 0.001) and shell-less eggs (p < 0.001) and lowered eggshell breaking strength (p = 0.005). Bone health was also adversely affected by the NC diet, as indicated by reduced tibia ash content (p < 0.001), stiffness (p = 0.005), and maximum load-bearing capacity (p = 0.040). Moreover, with NC diet, there was a decrease in osteoprotegerin (OPG) expression (p < 0.001) and an increase in receptor activator of nuclear factor kappa-B ligand (RANKL) expression (p < 0.001) in tibia, resulting in a greater RANKL/OPG ratio (p < 0.001). Supplementing the NC diet with bacterial 6-phytase at both levels (300 and 600 FTU/kg) effectively mitigated all adverse effects of P and ME deficiency on the aforementioned parameters, bringing them to levels comparable to those of the PC. Notably, the 600 FTU/kg supplementation provided slightly better results in terms of egg weight and eggshell breaking strength than the 300 FTU/kg level. Overall, this study suggests that supplementing the hybrid bacterial 6-phytase (300-600 FTU/kg) to P-deficient (0.12% nPP) and low energy (-100 kcal/kg) diets can fully replace inorganic P without compromising laying performance, egg quality, or bone health. Further research is recommended to determine the optimal levels of hybrid bacterial 6-phytase in P-deficient diets for laying hens throughout laying cycle of the birds with other nutrient matrices (energy, amino acids, calcium) to optimize layer feed formulations.

Inflammatory bowel disease (IBD) is a chronic, relapsing disease that poses significant challenges in treatment. This study aimed to develop silk fibroin-based mesalazine and chitosan:TNF-α siRNA polyplex-loaded, 3D bioprinted hydrogels for the oral treatment of IBD. For this purpose, bioink formulations composed of silk fibroin, hyaluronic acid, and sodium alginate were optimized. A chitosan:TNF-α siRNA polyplex was also formulated at a 40:1 chitosan-to-siRNA ratio. Hydrogel formulations were fabricated using 3D bioprinting and characterized in terms of compatibility, thermal stability, swelling behavior, degradation, mechanical properties, and mucoadhesion to both healthy and IBD-induced colon tissues. The optimized oral hydrogel (H-M-P12) demonstrated a swelling index of 366±67 % and underwent 31.2 % degradation after 24 h in vitro. Mesalazine and TNF-α siRNA exhibited a sustained release profile from the hydrogels. Cytotoxicity studies confirmed the biocompatibility of the hydrogels and a TNF-α gene silencing efficiency of 46.53 % was obtained. In vivo studies in a Balb/c mouse model of IBD revealed significant improvements in physiological parameters, macroscopic and microscopic morphology, and biochemical markers following treatment with the developed hydrogels. These findings suggest that silk fibroin-based hydrogels incorporating mesalazine and chitosan/TNF-α siRNA polyplex, produced via 3D bioprinting, hold promise as an effective therapeutic approach for IBD.

This study presents the development of an advanced three-dimensional (3D) bioprinted skin scaffold integrating sodium alginate (SA), gelatin (Gel), human skin-derived decellularized extracellular matrix (dECM), and microbiota-derived postbiotics. To ensure a biocompatible and functional ECM source, human skin samples collected during elective aesthetic surgical procedures were utilized. Following enzymatic treatment, the dermal layer was carefully separated from the epidermis and subjected to four different decellularization protocols. Among them, Protocol IV emerged as the most suitable, achieving significant DNA removal while maintaining the structural and biochemical integrity of the ECM, as confirmed by Fourier-transform infrared spectroscopy. Building on this optimized dECM-4, microbiota-derived postbiotics from Limosilactobacillus reuteri EIR/Spx-2 were incorporated to further enhance the scaffold’s bioactivity. Hybrid scaffolds were then fabricated using 7% Gel, 2% SA, 1% dECM-4, and 40 mg/mL postbiotics in five-layered grid structures via 3D bioprinting technology. Although this composition resulted in reduced mechanical strength, it exhibited improved hydrophilicity and biodegradability. Moreover, antimicrobial assays demonstrated inhibition zones of 16 mm and 13 mm against methicillin-resistant Staphylococcus aureus (MRSA, ATCC 43300) and Pseudomonas aeruginosa (ATCC 27853), respectively. Importantly, biocompatibility was confirmed through in vitro studies using human keratinocyte (HaCaT) cells, which adhered, proliferated, and maintained normal morphology over a 7-day culture period. Taken together, these findings suggest that the engineered hybrid scaffold provides both regenerative support and antimicrobial protection, making it a strong candidate for clinical applications, particularly in the management of chronic wounds.

AbstractHost metabolic fitness is a critical determinant of infectious disease outcomes. Obesity, aging, and other related metabolic disorders are recognized as high-risk disease modifiers for respiratory infections, including coronavirus infections, though the underlying mechanisms remain unknown. Our study highlights fatty acid-binding protein 4 (FABP4), a key regulator of metabolic dysfunction and inflammation, as a modulator of SARS-CoV-2 pathogenesis, correlating strongly with disease severity in COVID-19 patients. We demonstrate that loss of FABP4 function, by genetic or pharmacological means, reduces SARS-CoV2 replication and disrupts the formation of viral replication organelles in adipocytes and airway epithelial cells. Importantly, FABP4 inhibitor treatment of infected hamsters diminished lung viral titers, alleviated lung damage and reduced collagen deposition. These findings highlight the therapeutic potential of targeting host metabolism in limiting coronavirus replication and mitigating the pathogenesis of infection.

The increasing global demand for animal protein necessitates sustainable poultry production strategies prioritizing poultry flocks' health and welfare. This study investigates the effects of in-ovo administration of probiotic consortia and their corresponding postbiotics on the intestinal health of broiler chickens. Probiotic strains, first isolated from the cecal contents of the healthy, untreated broiler chickens and survived under gastrointestinal tract conditions, were amplified by the 16S rDNA gene sequencing method and using the database of NCBI GenBank (National Center for Biotechnology Information), multiple alignments were done with some selected and related sequences using MEGA 12 software. Selected strains for animal experiments were combined to form a consortium and their corresponding postbiotics. A total of 480 fertilized Ross 308 broiler eggs (n = 120 per group) were divided into four groups: a negative control, a positive control injected with phosphate-buffered saline, a postbiotic group injected with 0.5 mL of cell-free supernatant from a probiotic consortium, and a probiotic group injected with 0.5 mL of the probiotic consortium itself (108 CFU/mL). All injections were administered in ovo on day 17 of incubation, and samples were collected on day 21 of incubation and day 1 post-hatch for analysis. Histological and immunohistochemical analyses were performed on intestinal tissues to assess goblet cell activity, cellular proliferation, and the expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90). Results indicated that both probiotics and postbiotics significantly enhanced goblet cell numbers (p ≤ 0.05) and PCNA expression (p ≤ 0.01), particularly in the duodenum. The postbiotic group demonstrated increased expression of heat shock proteins (p ≤ 0.05) and higher PCNA levels (p ≤ 0.01) compared to the probiotic group in the duodenum, indicating a stronger cellular stress response and proliferative activity. These findings highlight the potential of postbiotics as functional biological agents to enhance early gut development and stress adaptation, offering promising applications in poultry production systems aimed at improving intestinal health and reducing reliance on conventional inputs.

Breast cancer remains among the most prevalent malignancies affecting women globally. Current treatment approaches, including mastectomy, chemotherapy, and radiotherapy, often fail to prevent cancer recurrence and can result in substantial tissue damage, esthetic concerns, and diminished quality of life. Three‐dimensional (3D) bioprinting, stem cell‐based technologies, and MXene nanomaterials show promise in tissue repair and cancer treatment. However, there is a lack of strategies that can offer multiple effects, preventing both breast tissue regeneration and tumor recurrence. In this study, we developed 3D hydrogel scaffolds incorporating stem cells and MXene quantum dots (MQDs) for in vivo application in a mouse model of breast cancer. We compared cellular, acellular, cellular MQD, and acellular MQD scaffolds transplanted into mouse after tumor resection and mastectomy. Notably, the acellular MQD group showed no tumor recurrence by day 14. It demonstrated superior tissue regeneration, confirmed by histological and immunostaining analyses. As a result, we offer a nanotechnological 3D scaffold based on hydrogel with dual functionality in preventing tumor recurrence and facilitating tissue regeneration. This innovative approach has the potential to revolutionize breast cancer treatment by reducing dependence on chemotherapy and radiotherapy. Thus, it offers a promising alternative for improving patient treatment outcomes.

Exosomes, nanoscale extracellular vesicles, have emerged as promising carriers in drug delivery due to their ability to bypass biological barriers, low toxicity, high stability, and intrinsic targeting capabilities. Mesenchymal stem-cell-derived exosomes (EXOMSC), with their natural tropism toward the tumor microenvironment, offer an ideal platform for enhancing therapeutic cargo delivery. In this study, we demonstrate an approach where red-emission chlorophyll-based carbon dots (Chl-CDs) were encapsulated within EXOMSC through a cell-driven uptake mechanism, creating CD@EXOMSC. These exosomes achieved superior photodynamic therapy (PDT) efficacy, requiring 40 times less nanomaterial compared to freestanding Chl-CDs. Mechanistic insights from glioblastoma cell miRNA profiling revealed that the enhanced efficacy was mediated by the regulation of efflux transporter genes, oxidative stress responses, and endocytosis pathways. This work highlights the synergistic potential of combining photosensitizers and miRNA-rich exosomes to achieve targeted and sustained therapeutic delivery, paving the way for a multifaceted approach in cancer therapy.

Age determination is an important part of forensic investigations and is used for assessment of population dynamics for animals and humans, understanding environmental conditions and so on. Age determinations using cementum annuli have been utilized for wild animals but have never been used for domestic dogs. The aim of this study was to test the accuracy of the method for domestic dogs and demonstrate the presence of cementum annuli. Ten domestic dogs were used for this experiment. The teeth were extracted from skulls via boiling and both mandibular and maxillary teeth were utilized. All teeth were decalcified using 10 % formic acid and 10 % formaldehyde solution. The decalcified teeth were embedded in paraffin and cut in 15 µm thickness. After staining with hematoxylin, the annuli were counted manually. The results obtained from this study suggest that domestic dogs indeed have cementum annuli and the annuli are countable, the number of annuli in teeth is compatible with the ages of animals, and canines of the same dogs show the same results meaning all canine teeth can be used for the age determination.

This study aimed to develop and evaluate a CpG oligodeoxynucleotide (CpG ODN)-adjuvanted trivalent inactivated Salmonella vaccine including S. enterica subsp. Enterica serovar Typhimurium, Salmonella enterica subsp. Enterica serovar Enteritidis, and Salmonella enterica serotype Infantis, for its immunogenic efficacy in chickens. The immunomodulatory effects of various CpG ODNs were assessed based on proinflammatory cytokine secretion and the expression levels of CD80, CD86, and MHC-II in the chicken cell lines HD11 and DT40. According to the results, CpG ODNs D35 3CG PO, D35 3CG MB, 1466 Acore PO, 1466 Acore MB, and K3 which exhibited non-cytotoxicity in both HD11 and DT40 cell lines, were selected for vaccine formulation. To evaluate their effects under in vivo conditions, chicks (n = 25) were randomly assigned to fourteen groups (G1: only sterile pyrogen-free saline solution, G2: only inactivated vaccine, G3: inactivated vaccine with 150 mg/dose of ALUM, G4: commercial Salenvac T vaccine, G5-G14: various experimental vaccine formulations which included different CpG ODNs combined with inactivated bacterial strains, with or without ALUM). Immune responses were analyzed through serological assays for antigen-specific antibody titers and ex vivo splenocyte cultures for cytokine secretion. Flow cytometry was performed to assess T-cell activation and IFN-γ production. The results demonstrated that the CpG ODNs-adjuvanted vaccine formulations significantly enhanced both humoral and cellular immunity compared to the commercial vaccine. Specifically, the Vac#5+ ALUM formulation, which included the K3 CpG ODN, induced robust antibody responses against Salmonella antigens and significantly increased IFN-γ secretion, nearly two-fold higher than the commercial vaccine. This effect was primarily mediated by CD4+ helper and CD8+ cytotoxic T cells. These findings highlight the potential of CpG ODNs as effective vaccine adjuvants in poultry. To the best of our knowledge, this is the first study to investigate the use of CpG ODNs as adjuvants in inactivated Salmonella vaccine formulations. Future studies should focus on evaluating the long-term protective efficacy of this vaccine formulation and its ability to provide cross-protection against a broader spectrum of Salmonella serovars.

This study evaluated the effects of two phytogenic blends on broiler performance, intestinal histomorphology, CD4+ (cluster of differentiation) and CD8+ T-cell numbers, and mRNA abundances of several cytokines in broilers. For this purpose, a total of 300 Ross 308 male broiler chicks that were 1 d old were randomly allocated to five experimental groups. The control group was fed a basal diet without any additives, and there were two phytogenic supplement groups (blend A, mainly comprising extracts of Thymus vulgaris and Filipendula ulmaria, and blend B, consisting of Ginkgo biloba and Silybum marianum) with two dosage regimens each (100 and 200 mg kg−1 (denoted A100 and A200) and 100 and 300 mg kg−1 (B100 and B300) of the diet, respectively). Over the total growing period, body weight gain and feed intake were unchanged among the groups, although phytogenic blend B showed a dose-dependent improvement in feed conversion ratio. Both phytogenic blends did not affect carcass characteristics. Jejunal morphology (villus height, crypt depth, and their ratio) was modified depending on both the composition and the dosage levels of the selected phytogenics. Also, both phytogenic blends linearly increased the CD4+ and CD8+ T-cell numbers in the jejunum. Moreover, no major treatment effects were observed on mRNA abundances of cytokines (IL-1β, IL-6, and TNFα). However, across the two phytogenic additives employed, a positive linear dose response in IL-1β abundance was noted on day 21 in broilers fed phytogenic blend B. Overall, dietary phytogenic blend B improved the intestinal health and growth performance of chickens compared to blend A. Further studies are suggested to elucidate the effects of the tested phytogenic blends on gut microbiome and on oxidative stress in broiler chickens.

The appeal of carbon dots (CDs) has grown recently, due to their established biocompatibility, adjustable photoluminescence properties, and excellent water solubility. For the first time in the literature, copper chlorophyllin-based carbon dots (Chl-D CDs) are successfully synthesized. Chl-D CDs exhibit unique spectroscopic traits and are found to induce a Fenton-like reaction, augmenting photodynamic therapy (PDT) efficacies via ferroptotic and apoptotic pathways. To bolster the therapeutic impact of Chl-D CDs, a widely used cancer drug, temozolomide, is linked to their surface, yielding a synergistic effect with PDT and chemotherapy. Chl-D CDs' biocompatibility in immune cells and in vivo models showed great clinical potential.Proteomic analysis was conducted to understand Chl-D CDs' underlying cancer treatment mechanism. The study underscores the role of reactive oxygen species formation and pointed toward various oxidative stress modulators like aldolase A (ALDOA), aldolase C (ALDOC), aldehyde dehydrogenase 1B1 (ALDH1B1), transaldolase 1 (TALDO1), and transketolase (TKT), offering a deeper understanding of the Chl-D CDs' anticancer activity. Notably, the Chl-D CDs' capacity to trigger a Fenton-like reaction leads to enhanced PDT efficiencies through ferroptotic and apoptotic pathways. Hence, it is firmly believed that the inherent attributes of Chl-CDs can lead to a secure and efficient combined cancer therapy. In a groundbreaking development, copper chlorophyllin-based carbon dots (Chl-D CDs) are synthesized, displaying unique red emissive traits, and enhancing photodynamic therapy (PDT) by harnessing Fenton-like reaction. Chl-D CDs' ability to enhance PDT through ferroptotic and apoptotic pathways offers a secure and efficient approach to combined cancer therapy, making them an intriguing topic for exploration.image

The envelope (env) protein of SARS-CoV-2, a pivotal component of the viral architecture, plays a multifaceted role in viral assembly, replication, pathogenesis, and ion channel activity. These features make it a significant target for understanding virus–host interactions and developing vaccines to combat COVID-19. Recent structural studies provide valuable insights into the conformational dynamics and membrane topology of the SARS-CoV-2 env protein, shedding light on its functional mechanisms. The strong homology and highly conserved structure of the SARS-CoV-2 env protein shape its immunogenicity and functional characteristics. This study examines the ability of the recombinant SARS-CoV-2 env protein to stimulate an immune response. In this study, recombinant envelope proteins were produced using the baculovirus expression system, and their potential efficacy was evaluated in both in vivo and in vitro models. Our results reveal that the env protein of SARS-CoV-2 stimulates humoral and cellular responses and highlight its potential as a promising vaccine candidate for combating the ongoing pandemic.

Nanomedicine plays an essential role in the development of tumor treatment modalities. However, tumors have a complex structure that includes a variety of immune cell types, which, along with tumor cells, comprise the heterogeneous tumor microenvironment (TME). Although nanoparticles that overcome the limitations of other classical therapeutics and navigate heterogeneous biological barriers have been developed, many unknown players within the TME still exist. The role of immune cell populations and signaling pathways in the TME, which can affect the nanoparticle distribution in the tumor or the overall outcome of nanotherapeutic treatments, are poorly described. In this study, we used spatial transcriptomics to determine in situ gene expression and identified immune cells, other than tumor-associated macrophages, that play vital roles following nanoparticle exposure. In this proof-of-concept study, a recently explored type of nanoparticle, Ti3C2Tx MXene quantum dots (MQDs), with a single particle diameter of <= 10 nm, were administered to orthotopic breast cancer model mice. Thanks to their red-emitting fluorescence properties, we could track the distribution of MQDs in the tumor. Whole-transcriptome analysis and immunofluorescence staining suggested that the heterogeneous distribution of MQDs results in different tumor and immune cell responses in situ. We observed a more tumor-suppressive phenotype at tumor regions with high MQD accumulation compared to regions with decreased MQD accumulation or na & iuml;ve tumors. Based on pathway analysis and cell deconvolution, we also identified other immune cell types altered by MQDs, including B cells and neutrophils. Specifically, the application of MQDs recruited and activated B cells and resulted in neutrophil degranulation and NETosis in vivo. Using spatial transcriptomics technology, we defined the fundamental molecular and cellular changes that occur in situ in the TME following MQD administration. Future spatial omics studies involving different nanoparticles with other material characteristics will help design more effective nanotherapeutics.

The placental cholinergic system; known as an important factor in intracellular metabolic activities, regulation of placental vascular tone, placental development, and neurotransmission; can be affected by persistent organic pesticides, particularly organochlorine pesticides(OCPs), which can influence various epigenetic regulations and molecular pathways. Although OCPs are legally prohibited, trace amounts of the persistent dichlorodiphenyltrichloroethane(DDT) are still found in the environment, making prenatal exposure inevitable. In this study, the effects of 2,4’-DDT and 4,4’-DDT; and its breakdown product 4,4’-DDE in the environment on placental cholinergic system were evaluated with regards to cholinergic genes. 40 human placentas were screened, where 42,50% (17 samples) were found to be positive for the tested compounds. Average concentrations were 10.44μg/kg; 15.07μg/kg and 189,42μg/kg for 4,4’-DDE; 2,4’-DDT and 4,4’-DDT respectively. RNA-Seq results revealed 2396 differentially expressed genes in positive samples; while an increase in CHRM1,CHRNA1,CHRNG and CHRNA2 genes at 1.28, 1.49, 1.59 and 0.4 fold change were found(p<0,028). The increase for CHRM1 was also confirmed in tissue samples with immunohistochemistry. In vitro assays using HTR8/SVneo cells; revealed an increase in mRNA expression of CHRM1, CHRM3 and CHRN1 in DDT and DDE treated groups; which was also confirmed through western blot assays. An increase in the expression of CHRM1,CHRNA1, CHRNG(p<0,001) and CHRNA2(p<0,05) were found from the OCPs exposed and non exposed groups.The present study reveals that intrauterine exposure to DDT affects the placental cholinergic system mainly through increased expression of muscarinic receptors. This increase in receptor expression is expected to enhance the sensitivity of the placental cholinergic system to acetylcholine.