Recent advancements in breast cancer research suggest that potassium channel blocker peptides from scorpion venom could serve as innovative therapies. Ion channels, particularly Voltage-Gated Potassium Channels (VGKCs), are therapeutic targets. They are regularly overexpressed in breast cancer tissues and play critical roles in proliferation, tumor progression, angiogenesis, and apoptosis. VGKCs also affect voltage-sensitive Ca2+ channels by regulating membrane potential and Ca2+ influx, which is vital for processes associated with cancer cell behavior. Scorpion venoms, rich in peptide toxins, specifically target these potassium channels, with the α-KTx family being notable for their high binding affinity and significant influence on therapeutic outcomes. Understanding the mechanisms of these toxins is crucial, primarily because potassium channel dysfunction is associated with various conditions, including cancer. Traditional treatments often prove ineffective for TNBC due to the deficiency of specific receptor expression, highlighting the need for alternative approaches like scorpion venom peptides. The distinctive properties of ion channels, especially the modifiable Kv10.1 and Kv1.3 channels, make them therapeutic targets, as their blockers exhibit anti-proliferative effects. VGKCs play a significant role in cell proliferation and apoptosis, and their modulation by scorpion venom peptides can impact the survival and growth of cancer cells. Additionally, the peptide's interaction with ion channels can alter membrane potential and calcium influx, thereby altering cancer cell behavior. This research lays the groundwork for future studies examining the mechanisms and specificity of scorpion venom-derived peptides, paving the way for translating these findings into clinical advancements in breast cancer therapies.

Neuropeptide Y (NPY) is a highly conserved 36–amino acid neuropeptide broadly distributed throughout the central and peripheral nervous systems, where it classically regulates appetite, stress responses, and circadian rhythms. Increasing evidence now positions NPY as a critical mediator at the interface of neural and immune signaling within the tumor microenvironment (TME). In cancer, NPY is released not only from tumor-innervating sympathetic fibers but also, in some contexts, directly from tumor cells, thereby establishing autocrine and paracrine signaling circuits that support tumor progression. Acting through its G protein–coupled receptors (Y1, Y2, Y4, Y5, and Y6), NPY exerts pleiotropic effects on both malignant and immune cell populations. Activation of Y1R and Y2R has been associated with enhanced tumor cell proliferation, angiogenesis, and vascular remodeling, whereas Y5R links stress-associated neuroendocrine signaling to accelerated tumor growth. Importantly, within the immune compartment, NPY promotes macrophage polarization toward an M2-like immunosuppressive phenotype, suppresses natural killer cell cytotoxicity, and dampens T cell activation, collectively fostering a tolerogenic and immune-evasive TME. These convergent neural and immunological effects highlight NPY as a dual-function neuromodulator and immunoregulator in cancer. In this review, we propose that NPY signaling represents a previously underappreciated neuro-immune checkpoint that integrates stress signals with tumor immune suppression. Targeting the NPY–receptor axis may therefore offer novel opportunities to reprogram the neuro-immune landscape of tumors and enhance the efficacy of cancer immunotherapy, particularly in stress-responsive malignancies.

Lung cancer remains the most commonly diagnosed malignancy worldwide and is a leading cause of cancer-related mortality. Conventional therapies, including chemotherapy and radiotherapy, are often limited by treatment resistance and disease recurrence, highlighting the need for alternative approaches such as immunotherapy. In this study, we investigated the pro-apoptotic effects of activated T cells on A549 lung cancer cells, with a particular focus on TRPV4-mediated calcium signaling. A549 cells were co-cultured with Jurkat T cells in the presence of calcium to evaluate T cell-mediated cytotoxicity. Cell viability was assessed using the MTT assay at 24 and 72 h, while apoptosis and necrosis were quantified by Annexin V staining and flow cytometry. T cell activation was confirmed through immunophenotyping for CD3 expression. We further evaluated oxidative stress markers, including total antioxidant capacity (TAC) and total oxidant status (TOS), alongside TRPV4 protein expression. Activated T cells significantly decreased A549 cell viability in a dose-dependent manner. Notably, calcium alone enhanced cell viability, whereas its combination with activated T cells markedly increased cytotoxicity and necrosis. TRPV4 expression was upregulated in cells treated with both calcium and activated T cells, suggesting a key role in mediating calcium influx and subsequent apoptosis. Additionally, combinatorial treatment led to decreased TAC and elevated TOS levels, indicating enhanced oxidative stress. These findings reveal a novel immunotherapeutic mechanism whereby activated T cells induce apoptosis through TRPV4-dependent calcium signaling, offering a potential strategy to improve lung cancer treatment efficacy. Further studies are warranted to elucidate the modulation of calcium pathways in T cell-driven anti-tumor responses.

Epithelial cell adhesion molecule (EpCAM, CD326) is a transmembrane glycoprotein that plays important roles in both normal epithelial tissues and epithelial malignancies. Its frequent overexpression in several cancers, including pancreatic, colorectal, and prostate carcinomas, has made it an attractive therapeutic target. In this study, we designed and evaluated a novel EpCAM-binding diphtheria toxin fusion construct, DT-SNFYMPL, in which the EpCAM-binding peptide SNFYMPL was fused to a truncated diphtheria toxin. The recombinant construct was cloned, expressed in Escherichia coli BL21 cells, purified, refolded, and characterized in vitro. Structural modeling and validation predicted a stable three-dimensional structure for the designed protein. Binding analysis demonstrated interaction of DT-SNFYMPL with recombinant EpCAM in an ELISA-based assay. Cytotoxicity studies revealed dose- and time-dependent inhibition of MCF-7 and TC-1 cell proliferation, whereas no significant effect was observed in HUVEC cells under the conditions tested. DT-SNFYMPL also reduced the migration of cancer cells in Transwell assays; however, this effect is likely attributable, at least in part, to toxin-mediated cytotoxicity rather than direct inhibition of migratory pathways. Although the findings demonstrate biological activity of the DT-SNFYMPL construct and support its potential as an EpCAM-binding therapeutic candidate, the present study does not establish EpCAM-dependent internalization, receptor-mediated cytotoxicity, or therapeutic selectivity. Further mechanistic and preclinical studies are required to evaluate its mode of action, safety, and therapeutic potential in EpCAM-expressing malignancies.

Breast cancer, particularly invasive and epithelial–mesenchymal transition (EMT)–associated phenotypes, remains a major therapeutic challenge. MeuKTx, a Kv1.3-targeting peptide derived from Mesobuthus eupeus venom, was nanoformulated using chitosan to improve peptide stability and cellular delivery. This study presents the first nano-encapsulation of MeuKTx (NP-MeuKTx) for anticancer evaluation. The resulting nanoparticles exhibited uniform morphology, nanoscale size, and high encapsulation efficiency. NP-MeuKTx exhibited significantly enhanced cytotoxicity compared with free MeuKTx. The IC₅₀ value of free MeuKTx in MCF-7 cells was greater than 400 pM, whereas NP-MeuKTx exhibited a markedly lower IC₅₀ of 25.3 pM. In MCF-7-EMT cells, the IC₅₀ of free MeuKTx was 85.32 pM, which was substantially reduced to 23.27 pM following NP-MeuKTx treatment. Migration assays demonstrated that NP-MeuKTx markedly suppressed cell migration in both MCF-7 and MCF-7-EMT cells, with significantly greater inhibition than free MeuKTx (p ≤ 0.0022). Flow cytometric analysis revealed that NP-MeuKTx induced the highest levels of apoptosis in both cell models, significantly exceeding the effects of free peptide (p = 0.0001). Furthermore, NP-MeuKTx strongly inhibited angiogenesis in HUVEC tube formation assays, significantly reducing branch points, total tube length, and mesh area compared with controls and free MeuKTx (p < 0.0001). Overall, these findings indicate that chitosan-based nanoformulation improves the functional anticancer activity of MeuKTx in breast cancer models.

BACKGROUND:Breast cancer has become the most commonly diagnosed cancer worldwide and represents a major burden to public health. Advances in understanding ferroptosis pathways and identifying new therapeutic targets raise hope for using ferroptosis modulators to treat untreatable diseases. METHODS:In this study, BALB/c mice were divided into several groups: model, Doxorubicintreated, FINO2-treated, Pirfenidone-treated, and a combined Pirfenidone + FINO2 group. After treatment, we assessed iron content in cancer cells, fibrosis area, CD34 expression, and mRNA levels of solute carrier family 7 member 11(SLC7A11) and heme oxygenase 1 (HMOX1). RESULTS:Results showed that the average tumor size in the Pirfenidone + FINO2 group was significantly smaller than in the doxorubicin group. Treatments with FINO2, Pirfenidone, or their combination significantly increased iron content in cancer cells and reduced the fibrosis area. Cotreatment with FINO2 and Pirfenidone also led to notable decreases in CD34 expression and mRNA levels of SLC7A11 and HMOX1. CONCLUSION:These findings suggest that FINO2 ferroptosis agonists, when combined with other anticancer agents like Pirfenidone, can enhance ferroptosis and reduce tumor fibrosis. Additionally, the overexpression of SLC7A11 and HMOX1 in breast cancer model mice is associated with increased tumor growth and reduced metastasis, indicating that targeting these proteins with specific inhibitors may be a promising strategy for breast cancer treatment.

Testicular ischemia–reperfusion (TIR) injury is a major cause of male infertility, driven by excessive oxidative stress and activation of apoptotic pathways. This study investigated the protective effects of a niosomal formulation containing hesperidin-rich Citrus sinensis peel extract against testicular ischemia–reperfusion (I/R) injury in rats. Adult male Wistar rats underwent 720° torsion for 2 h, followed by detorsion. Animals received either free hesperidin or niosomal hesperidin (100 mg/kg/day, orally) for 30 days. Oxidative stress (MDA), sperm parameters, histopathology, and p53 and Caspase-3 expression were assessed using biochemical assays, light microscopy, and qPCR. Niosomes were characterized by DLS and FESEM. TIR markedly increased MDA levels, disrupted seminiferous tubule architecture, and elevated p53 and Caspase-3 expression. Free hesperidin attenuated these effects, whereas niosomal hesperidin produced significantly greater improvements, including reduced lipid peroxidation, higher sperm motility and viability, increased germ cell counts, and more substantial downregulation of p53 and Caspase-3. Niosomal hesperidin-rich Citrus peel extract provides superior protection against TIR-induced oxidative and apoptotic injury compared with free hesperidin, likely due to enhanced stability. These findings support the use of nanocarrier-based delivery systems to improve the therapeutic potential of natural antioxidants in reproductive medicine.

Septicemia is a significant threat to newborn calves, often due to inadequate colostrum intake in the first day of life. The study aimed to assess the effects of a newly developed herbal formulation on septicemia induced by Escherichia coli strain O111:H8. Ten Holstein-Friesian calves aged 8-10 days were divided into two groups. Experimental septicemia was induced for all calves (n=10). The treatment group (n=5) received a herbal formulation containing extracts from Rosa canina, Urtica dioica, Tanacetum vulgare, selenium, flavonoids, and carotenes, in addition to antibiotics. The control group (n=5) received a placebo (5% dextrose) along with antibiotics for five days. The animals were monitored for 14 days. Blood samples were analyzed for cytokines, cardiac enzymes, renal function, and total antioxidant capacity before and after treatment. The treatment group had non-significantly higher CD4+ counts compared to the control. The serum level of IL-6 increased after treatment, with a considerable difference between the groups at 72 h (p=0.0014). The herbal formulation positively impacted renal and cardiac function evidenced by decreased cardiac troponin I levels and increased total antioxidant capacity (TAC). Lactate dehydrogenase (LDH) levels changed significantly over time (p<0.05), with a positive correlation between ECG changes and peak LDH levels (p<0.05). The increased cytokines beside ameliorative effects on heart and kidney functions suggest that the herbal drug may possess immunomodulatory and anti-inflammatory properties that aid in managing the inflammatory response during sepsis. These findings support the use of this herbal-based drug as an adjunctive treatment in veterinary practices for managing septicemia in calves.

Neuropeptide Y (NPY) and the voltage-gated potassium channel Kv1.3 are closely associated with breast cancer progression and apoptosis regulation, respectively. NPY receptors (NPYRs), which are overexpressed in breast tumors, contribute to tumor growth, migration, and angiogenesis. In parallel, Kv1.3 plays a pivotal role in mitochondrial-mediated apoptosis, and its inhibition can induce cancer cell death. To exploit these mechanisms, we developed and characterized a novel niosomal drug delivery system encapsulating margatoxin (MgTx), a potent Kv1.3 inhibitor, and functionalized with NPY for targeted breast cancer therapy. Niosomes were synthesized via a modified thin-film hydration method and decorated with NPY peptides to enable selective binding to NPYR-overexpressing cancer cells. Physicochemical analyses using dynamic light scattering (DLS), atomic force microscopy (AFM), and field emission scanning electron microscopy (FESEM) confirmed a nanoscale size range (134–161 nm), spherical morphology, and successful surface modification. The system demonstrated high encapsulation efficiency, prolonged stability at 4°C, and sustained MgTx release over 72 h. In vitro cytotoxicity studies revealed that NPY-decorated MgTx-loaded niosomes significantly reduced the viability of MCF-7 and MDA-MB-231 breast cancer cells while exerting minimal toxicity on non-tumorigenic MCF-10A cells. qRT-PCR analysis indicated upregulation of pro-apoptotic genes (Bax, Caspase-3) and downregulation of anti-apoptotic Bcl2, confirming induction of apoptosis in treated cancer cells. These findings highlight the potential of NPY-functionalized niosomes as an effective and selective nanoplatform for targeted breast cancer therapy.

Following the worldwide spread of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there is a vital requirement for safe and effective vaccines against Coronavirus disease 2019 (COVID-19). Therefore, several vaccine-candidate platforms have been designed, tested, and developed. Based on guidelines, preclinical studies are recommended to assess the safety and potency of COVID-19 vaccines in appropriate in vitro and in vivo settings. These studies provide essential information to describe the potential toxic properties of a vaccine and the formulation of vaccine agents during the preclinical trial phase. In toxicology studies, several factors must be considered, such as the appropriate animal species and strains, dosing timetable, mode of administration, time of sampling for biochemistry and antibody evaluation, and necropsy. Pharmacokinetic/ biodistribution studies are not usually required for infectious disease prophylaxis vaccines unless the vaccine contains a novel substance. Evaluating their biodistribution is crucial for newly developed vaccines, such as lipid nanoparticles -messenger RNA (LNP-mRNA), DNA, and Viral vectors in non-replicated (VVnr), or recombinant virus vaccines. The review highlights the importance of preclinical studies in assessing the safety and efficacy of vaccine candidates. This guidance is essential for researchers and manufacturers to design effective vaccines that can progress to clinical trials safely.

Rheumatoid arthritis (RA) is a prevalent autoimmune disorder affecting millions of individuals worldwide, leading to chronic joint inflammation, progressive tissue damage, and substantial impairment of quality of life. In this study, we explored the therapeutic potential of chitosan nanoparticles encapsulating MeuKTx, a potassium channel inhibitory peptide targeting Kv1.3, in modulating articular tissue alterations in a neonatal rat model of RA. The peptide, alpha-KTx 3.13, was synthesized using solid-phase peptide synthesis, followed by purification and characterization through high-performance liquid chromatography (HPLC) and mass spectrometry to ensure its quality and bioactivity. Neonatal Wistar rats were assigned to various experimental groups, including a healthy control, an untreated RA model, and groups treated with methotrexate or peptide-based formulations delivered via chitosan nanoparticles. The treatment groups demonstrated significant reductions in serum malondialdehyde and rheumatoid factor (RF) levels, indicating a decrease in oxidative stress and systemic autoimmune activity. At the molecular level, expression of pro-inflammatory mediators, including transforming growth factor-beta (TGF-β) and monocyte chemoattractant protein-1 (MCP-1/CCL2), as well as Caspase-8 protein levels, were markedly decreased in peptide-treated groups, correlating with improved histopathological outcomes in joint tissues. Notably, the chitosan nanoparticle formulation enhanced the bioavailability and therapeutic efficacy of MeuKTx, suggesting that targeted delivery to Kv1.3 channels can modulate immune and inflammatory responses more effectively than free peptide administration. Collectively, these findings highlight the potential of MeuKTx-loaded chitosan nanoparticles as a novel therapeutic strategy for RA, offering a promising approach to reduce joint inflammation, oxidative stress, and immune dysregulation while potentially minimizing reliance on conventional chemical drugs. This study provides a foundation for further exploration of peptide-based nanotherapies in autoimmune and inflammatory diseases.

Four significant influenza outbreaks have occurred over the past 100 years, and the 1918 influenza pandemic is the most severe. Since influenza viruses undergo antigenic evolution, they are the pathogens most likely to trigger a new pandemic shortly. Intranasal vaccination offers a promising strategy for preventing diseases triggered by respiratory viruses by eliciting an immunoglobulin A (IgA) response, limiting virus replication and transmission from the respiratory tract more efficiently than intramuscular vaccines. Combining intranasal administration and mRNA-lipid nanoparticles can be an ideal strategy for limiting the extent of the next flu pandemic. This study explored the immunogenicity of intranasally delivered mRNA encapsulated in mannose-histidine-conjugated chitosan lipid nanoparticles (MHCS-LNPs) as a vaccine against influenza A (H1N1) in BALB/c mice. Intranasal administration of mRNA-MHCS-LNPs resulted in the generation of influenza A (H1N1) hemagglutinin-specific neutralizing antibodies in vaccinated animals. The enzyme-linked immunosorbent assay (ELISA) results indicated a notable increase in the quantity of immunoglobulin G (IgG) and IgA antibodies in serum and the bronchoalveolar lavage fluid (BALF), respectively, and exhibited influenza A-specific IFN-γ secretion in vaccinated mice, as well as a noticeable alteration in IL-5 production. Overall, this study demonstrated an effective immunogenic response against respiratory viral infections through intranasal delivery of an mRNA-MHCS-LNP vaccine.

Cyclodextrins (CDs), characterized by their hydrophobic inner surface, form inclusion complexes with various organic and inorganic lipophilic compounds. These unique structures enhance the solubility, stability, and bioavailability of poorly soluble drugs. Chemical modifications of the parent CDs (α-CD, β-CD, and γ-CD) improve their suitability for parenteral administration, enabling a wide range of pharmaceutical applications, including drug delivery, cancer therapy, gene delivery, and biosensing. β-CDs, in particular, are instrumental in controlled-release formulations, protecting labile compounds from degradation, and enhancing the solubility and permeability of active pharmaceutical ingredients. Recent advancements have integrated CDs, especially β-CDs, into nanoparticle-based drug delivery systems (NPs), further broadening their functionality. Β-CD-conjugated NPs (β-CD-NPs) exhibit improved drug solubility, targeted delivery to specific sites, such as cancer cells, and reduced off-target toxicity. CDs often surpass traditional NPs in encapsulation efficiency and drug-loading capabilities. This review critically explores the diverse applications of CDs in NP-based drug delivery, focusing on their roles in inclusion complexes, polymeric NPs, magnetic and metallic NPs, and nanosponges. Additionally, the synergistic impact of β-CDs on modifying NP properties is discussed, emphasizing their potential as versatile and efficient carriers in advanced therapeutic and diagnostic systems.

PURPOSE:This study aimed to assess the effectiveness of ozone therapy in treating Diabetes-related Foot Ulcer (DFU) and its outcomes. METHODS:A systematic search was conducted in PubMed/MEDLINE, Scopus, Web of Science, and ProQuest databases for published studies evaluating the use of ozone as an adjunct treatment for DFU, from inception to December 21, 2022. The primary outcome measure was the change in wound size after the intervention compared to pretreatment. Secondary outcomes included time to complete ulcer healing, number of healed patients, adverse events, amputation rates, and hospital length of stay. Quantitative data synthesis for the meta-analysis was performed using a random-effects model and generic inverse variance method, while overall heterogeneity analysis was conducted using a fixed-effects model. Interstudy heterogeneity was assessed using the I2 index (>50%) and the Cochrane Q statistic test. Sensitivity analysis was performed using the leave-one-out method. RESULTS:The meta-analysis included 11 studies comprising 960 patients with DFU. The results demonstrated a significant positive effect of ozone therapy on reducing foot ulcer size (Standardized Mean Difference (SMD): -25.84, 95% CI: -51.65 to -0.04, p = 0.05), shortening mean healing time (SMD: -38.59, 95% CI: -51.81 to -25.37, p < 0.001), decreasing hospital length of stay (SMD: -8.75, 95% CI: -14.81 to -2.69, p < 0.001), and reducing amputation rates (Relative Risk (RR): 0.46, 95% CI: 0.30-0.71, p < 0.001), compared to standard treatment. CONCLUSION:This meta-analysis indicates that ozone therapy has additional benefits in expediting complete DFU healing, reducing the amputation rates, and decreasing hospital length of stay, though its effects do not differ from standard treatments for complete ulcer resolution. Further research is needed to address the heterogeneity among studies and to better understand the potential beneficial effects of ozone therapy.

Chlorpyrifos(CPF) is a well-known hepatotoxic agent that has side effects on several organs. On the contrary, hepatic macrophages are crucial in maintaining liver tissue integrity. The main objective of this study was to evaluate the effects and possible mechanisms of niosomal hesperidin (Nio + Hesp), a flavanone glycoside found in citrus fruits, on M1-M2 liver macrophage polarization and inflammatory cells in the brain, liver, and ovarian tissues. Forty C57 mice were divided into CPF(3 mg/kg), Sham(Dimethyl sulfoxide 40 mu L/kg), CPF + Hesp(100 mg/kg), and CPF + Nio + Hesp (100 mg/kg) groups. The activity of sera superoxide dismutase (SOD) and malondialdehyde (MDA), brain, liver, and ovary tissues changes, and M1-M2 liver macrophage polarization were evaluated by examining the expression of CD163 and CD68 genes. Hepatic lesions consisting of sporadic foci of coagulation necrosis, inflammatory cell reaction, and regenerative fibrosis were seen following CPF injection, reflected by significant overexpression of CD163 and CD68 genes. In comparison, Nio + Hesp declined the amount of cell apoptosis in the liver and downregulated CD163 and CD68 gene expression. Both Nio + Hesp and Hesp alleviated CPF-induced hepatotoxicity, however, Nio + Hesp was superior to hesperidin in the down-regulation of the CD163 and CD68 gene expression. Even though a significant difference between hesperidin and Nio + Hesp was observed in the number of Graafian follicles, corpus luteum, and peri-antral follicles, no sub-stantial difference was observed in primary follicles. The ameliorative effects of Hesp and Nio + Hesp may be at least in part due to their antioxidant and anti-inflammatory properties. These findings showed that both M1-and M2-macrophages contributed to the development of hepatic lesions induced by CPF and provided information about macrophage activation, indicating the importance of analysis of macrophage phenotypes for hepatotoxicity based on M1/M2-polarization which can be downregulated by niosomal nesperidin.

The immune system relies on a delicate balance between attacking harmful pathogens and preserving the body’s own tissues, a balance maintained by immune checkpoints. These checkpoints play a critical role in preventing autoimmune diseases by restraining excessive immune responses while allowing the immune system to recognize and destroy abnormal cells, such as tumors. In recent years, immune checkpoint inhibitors (ICIs) have become central to cancer therapy, enabling the immune system to target and eliminate cancer cells that evade detection. Traditional antibodies, such as IgGs, have been widely used in immune therapies but are limited by their size and complexity. Nanobodies (Nbs), derived from camelid heavy-chain-only antibodies, offer a promising alternative. These small, stable antibody fragments retain the antigen-binding specificity of traditional antibodies but have enhanced solubility and the ability to target otherwise inaccessible epitopes. This review explores the use of Nbs as ICIs, emphasizing their potential in cancer immunotherapy and other immune-related treatments. Their unique structural properties and small size make Nbs highly effective tools for modulating immune responses, representing a novel approach in the evolving landscape of checkpoint inhibitor therapies.

BACKGROUND:The role of probiotics and micronutrients in improving immune system function and response to vaccination has been proven. Hence, this study aimed to investigate the effects of probiotics enriched with micronutrients on the immunogenicity of PastoCovac® vaccine. METHODS:The probiotic supplement BioBoost® and PastoCovac® vaccine, which contain six expressed Receptor- binding Domains (RBD) and conjugated with tetanus toxin, were administered concurrently. The safety and efficacy were assessed by determining Immunoglobulin G (IgG) antibody titers to RBD and cytokines, mRNA expression of Toll-like Receptors (TLRs) 5, and clinical symptoms. RESULTS:Results revealed that the administration of the probiotics enriched with micronutrients and vitamins for 14 days before the first vaccine dose, followed by continued supplementation for 14 days after the first dose, and in conjunction with the second vaccine dose, yielded the most significant elevation in Interleukin 4 (IL-4), Tumor Necrosis Factor-alpha (TNF alpha), Interferon-gamma (IFN-gamma), and anti-SARS-CoV-2 RBD IgG levels within the supernatant samples collected from spleen cultures with the highest expression of TLR5 genes in intestinal samples, compared to the control group. CONCLUSION:Our results indicated that the inclusion of probiotics enriched with micronutrients and vitamins significantly enhanced the immunogenicity of the PastoCovac® vaccine. Based on the recommendation to administer third and fourth vaccine doses, particularly for vulnerable and elderly individuals, the utilization of supplements containing probiotics is expected to favorably influence immune responses.

The use of mRNA and ribonucleoproteins (RNPs) as therapeutic agents is a promising strategy for treating diseases such as cancer and infectious diseases. This review provides recent advancements and challenges in mRNA- and RNP-based therapies, focusing on delivery systems such as lipid nanoparticles (LNPs), which ensure efficient delivery to target cells. Strategies such as microfluidic devices are employed to prepare LNPs loaded with mRNA and RNPs, demonstrating effective genome editing and protein expression in vitro and in vivo. These applications extend to cancer treatment and infectious disease management, with promising results in genome editing for cancer therapy using LNPs encapsulating Cas9 mRNA and single-guide RNA. In addition, tissue-specific targeting strategies offer potential for improved therapeutic outcomes and reduced off-target effects. Despite progress, challenges such as optimizing delivery efficiency and targeting remain. Future research should enhance delivery efficiency, explore tissue-specific targeting, investigate combination therapies, and advance clinical translation. In conclusion, mRNA- and RNP-based therapies offer a promising avenue for treating various diseases and have the potential to revolutionize medicine, providing new hope for patients worldwide.

Drug resistance refers to the reduction in the effectiveness of a drug in treating a disease or improving the stability of symptoms. It can occur in various types of pathogens, including bacteria, parasites, viruses, fungi, and cancer cells. This experimental study was conducted between 2018 and 2019 in an area with an annual mean rainfall of 130mm. The sowing date was September 10th, and 2-3 seeds were planted per cell. MTT assays (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) were used to determine the percentage of viability in adenocarcinomic human alveolar basal epithelial cells (A549) and Medical Research Council cell strain 5 (MRC5) cell lines incubated with methanolic extract and valerenic acid for 48 hr. The methanol extract was prepared by adding 1000 mg of rhizomes to 100 mL of methanol, followed by sonication for 30 minutes, stirring, and centrifugation at 4000 rpm for 10 minutes. Minimum inhibitory concentration (MIC) and agar gel diffusion were used to assess the antimicrobial activity of the methanol extract of valerian against two important pathogenic microorganisms, Staphylococcus aureus and Candida albicans. However, valerenic acid did not reveal antimicrobial activity at doses of 200, 100, 50, 25, 12.5, and 6.25 µg/mL. The methanolic extract of V. officinalis contains high quantities of sesquiterpenes, specifically valerenic acid, which did not show cytotoxic effects on A549 and MRC5 cell lines as assessed by the MTT assay. In vivo evaluation of the extract in mice and guinea pigs did not reveal any toxic effects based on histopathological and clinical symptom assessments. Our study confirms that Valeriana officinalis has dose-dependent potential to improve existing treatment approaches for Staphylococcus aureus and Candida albicans infections.