Previous studies have reported that different phenothiazine derivatives have shown a broad spectrum of antibacterial, anticancer and antiplasmid activities. In the present study, we investigated the interactions of phenothiazine (phtz) and its N-methyl-substituted derivative, N-methylphenothiazine (N-Mephtz) with bovine serum albumin (BSA) and calf thymus DNA (ct-DNA) by fluorescence emission spectroscopy to examine their binding affinity towards these biomolecules. Considering that serum albumin is divided into three domains (I–III), with each domain containing two subdomains (A and B), we have also performed fluorescence competition experiments with site markers for BSA to locate the binding site of the investigated compounds to this biomolecule. Eosin Y was used as a marker for site I (subdomain IIA), while ibuprofen was a marker for site II (subdomain IIIA). The obtained results and the values of binding constants (K_A) have indicated that both phtz and N-Mephtz can interact with BSA and ct-DNA, whereby N-Mephtz has higher binding affinity towards these biomolecules. On the other hand, K_A values of both investigated compounds are lower in the presence of eosin Y, while only a slight change was observed in the presence of ibuprofen. These results indicated that the binding of the investigated compounds should be mainly located within site I of BSA, and that the tested compounds had to compete with eosin Y to bind to this protein.

Objective: To investigate the environmental-friendly extracellular biosynthetic technique for the production of the silver nanoparticles (AgNPs) by using leaf extract of Eucalyptus camaldulensis (E. camaldulensis).

Methods: The NP were characterized by color changes and the UV-visible spectroscopy. The cytotoxic effects of prepared AgNPs was detected against four types of pathogenic bacteria, including two Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and two Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) by using agar well diffusion method

Results: A peak absorption value between 400-450 nm for the extract and the color change to dark brown were corresponding to the plasmon absorbance of AgNPs. On the other hand, aqueous extract of E. camaldulensis leaves could be effective against tested microorganisms which showed inhibition zones of 9.0-14.0 mm. Furthermore, biologically synthesized AgNPs had higher ability to suppress the growth of the tested microorganisms (12.0-19.0 mm).

Conclusions: Our findings indicated that extracellular synthesis of AgNPs mediated by E. camaldulensis leaf extract had an efficient bactericidal activity against the bacterial species tested. The exact mechanism of the extracellular biosynthesis of metal NP was not well understood. Further studies are needed to highlight the biosynthesis process of AgNPs and also to characterize the toxicity effect of these particles.

Fetuin-A is a multifactorial glycoprotein predominantly produced by liver but also found in adipose tissue, and tightly regulated by the FBXW7 E3 ubiquitin ligase. Recently, the hepatokine has been implicated in the pathogenesis of insulin resistance and associated metabolic failures in humans through its potent and selective inhibitory effect on tyrosine kinase activity of insulin receptor, however, no reports related to its implication in equine metabolic syndrome onset have been published yet. In this investigation, the effect of FBXW7 E3 ligase on the Fetuin-A/INSR axis has been evaluated. EMS affected liver tissue exhibited significant elevated Fetuin-A levels at protein and mRNA level over lean samples. Moreover, increased Fetuin-A was accompanied by augmentation of circulating levels of IL-1β and TNF-α pro-inflammatory cytokines. Interestingly, liver FBXW7 levels inversely correlated with high Fetuin-A concentrations, and was sensibly downregulated under EMS condition. Treatment of liver explants with exogenous FBXW7 protein resulted in a marked depletion in total Fetuin-A protein expression, which subsequently restored insulin signal transduction via increased INSR phosphorylation. Conclusion: On the whole, EMS affected horses display abnormal high Fetuin-A levels and suppressed FBXW7 expression, which could serve as a new potential therapeutic target for insulin sensitivity restoration in EMS

Equine metabolic syndrome (EMS) is a prevalent endocrine disorder affecting horses, ponies and donkeys, characterized by dysregulated mitochondrial dynamics, chronic low-grade inflammation, and an increased susceptibility to laminitis. EMS poses significant health risks and is associated with obesity, insulin resistance, and diabetes. Recent studies have shown that low serum levels of sex hormone-binding globulin (SHBG) are linked to these metabolic dysregulations. To investigate whether exogenous SHBG could ameliorate mitochondrial dysfunction and inflammation in adipose-derived stromal stem cells from EMS-affected horses (EqASCEMS), we conducted a comprehensive study. EqASCEMS cultures were treated with 50 nM of SHBG for 24 hours. We evaluated various parameters including mitochondrial dynamics and biogenesis as well as the expression of pro-inflammatory and anti-inflammatory markers. Our findings demonstrated that exogenous SHBG treatment significantly enhanced mitochondrial biogenesis. This enhancement was evidenced by increased expression of key genes and proteins associated with mitochondrial dynamics, such as MFN, PARKIN, PINK, Cytochrome C, as well as genes related to mitoribosomes and the mitochondrial oxidative phosphorylation system (NDUFA9, COX4L1, COX4L2 and MTERF4). Furthermore, SHBG exhibited anti-inflammatory properties by reducing the expression of pro-inflammatory cytokine genes (IL-1β, IL-6, TNF-α) and promoting the expression of anti-inflammatory markers (IL-4, IL-10, IL-13) in EqASCEMS. Our study highlights the potential therapeutic value of SHBG in modulating ASC metabolic activities and suggests its importance as a target for the development of effective treatment protocols for EMS. These findings provide valuable insights into mitigating the metabolic and inflammatory consequences of EMS in horses, potentially improving their overall health and well-being.

Gold nanoparticles have been widely used in several areas of human activities, including the biomedical field, due to their small size and unique properties. However, its entrance into the environment may be expected with their increased use. There is thus the need to understand how these nanoparticles used in biomedical applications will affect environmental health, and therefore human health. One of the requirements for using gold nanoparticles is their non-toxic and biocompatible nature to both in vivo and in vitro systems. Some concerns on their possible impact in the environment have been raised with few studies addressing the effects of the particles in marine organisms. This work will present data on the biochemical effects of gold nanoparticles in marine organisms (bivalves and fish), alone and combined with other environmental contaminants. Overall, data show that the particle stability in high ionic strength media and the observed effects are highly dependent on the surface coating. Nonetheless, the studied nanoparticles proved able to induce peroxidative damage both in bivalves and fish, and to promote alterations in neurotransmission. Although the benefits of these nanomaterials are extensively shown, their unintentional release or disposal deserve more care and precaution.

Exogenous exposure to estrogenic compounds is implicated in various health conditions, including reproductive disorders, hormone-related cancers, and metabolic disturbances. In healthy males, activation of the estrogen receptors (ERs) can induce feminization and toxicity. However, screening for estrogenic compounds is limited to uterotrophic assays without an established male analog. Studies have reported that exposure to estrogen significantly enlarged the seminal vesicles (SV). However, screening strictly by the wet weight of the SV could obscure the identification of weakly estrogenic or ER-selective compounds. Therefore, protein-based markers are beneficial to complement the weight-based assessments. In this study, we report a discovery-driven proteomic analysis of 17β-estradiol's (E2) effects on the SV for the first time to identify potential biomarkers for screening estrogenic compounds in males. In our study, orchiectomized mice were subcutaneously injected with E2 or the vehicle daily for five days. Proteins from the SV were extracted and analyzed by shotgun proteomics relying on liquid chromatography-tandem mass spectrometry. Acquired tandem mass spectra were searched against the UniProt Mouse protein database using Proteome Discoverer (Thermo Fisher Scientific) and MaxQuant (Max Planck Institute). Verification of identified proteins and relative quantifications were performed using Scaffold (Proteome Software). Post hoc t-tests were performed to identify differently expressed proteins (DEPs) between groups. DEPs were mapped to protein interaction networks and pathways through Ingenuity Pathway Analysis® (IPA®, Qiagen). Our discovery-driven proteomics identified over 7000 proteins with high confidence from the SV of E2-treated and control mice. In addition, bioinformatics-based analysis using IPA® of the E2-regulated proteins showed enrichment in molecular and cellular functions associated with carbohydrate metabolism, DNA replication, repair, and free radical scavenging. Furthermore, corroborating our treatment, IPA®’s molecule activity predictor tool indicated enhanced upstream activation of the ER-β (ESR2). We identified several preclinical biomarkers participating in androgen receptor activity, ER-mediated transcription, and reproductive system development and function. Targeted proteomics-based validation of these biomarkers is ongoing. Our study aims to provide an in-depth account of the alterations occurring at the protein level in response to E2 exposure for screening and validation of preclinical biomarkers with diagnostic potential.

Over the past few decades, many commonly used antibiotics have become less effective against certain illnesses because many of them produce toxic reactions and drug-resistant. It is essential to investigate new drugs with less resistance and less side effects. Drugs derived from natural sources are cheaper, safe and play a significant role in the prevention and treatment of many human diseases. Thus the present study focus on comparative evaluation of antibacterial activity of volatile oil and crude alkaloids extracted from leaves of Murraya koenigii (Family: Rutaceae) by agar cup method. Volatile oil and crude alkaloids of M. koenigii, both showed dose dependent antibacterial activity against gram positive bacterial strains Staphylococcus aureus, Bacillus subtilis, and gram negative bacterial strains Escherichia coli, Proteus vulgaris. Volatile oil (1%v/v) showed comparable antibacterial activity to the standard drug streptomycin (10mcg/ml) against Staphylococcus aureus and Escherichia coli. Volatile oil (10%v/v) showed significant antibacterial activity to the standard drug streptomycin (10 mcg/ml) against Proteus vulgaris. Crude alkaloids (50 mcg/ml) showed comparable antibacterial activity to the standard drug streptomycin (10 mcg/ml) against Bacillus subtilis. This study reveals that volatile oil have more antibacterial potential compare to alkaloids of M. koenigii leaves. Thus the present study will help to find out active phytoconstituents as well as helps to develop a potent antibacterial herbal formulation using the M. koenigii.

Triple-negative breast cancer(TNBC) with immunohistochemical results negative for estrogen receptor, progesterone receptor and the proto-oncogene Her-2 is the prominent malignant subtype of breast cancer, and targeted therapeutic agents acting on it still need to be investigated for addition. It is well known that natural products with a variety of pharmacological activities can be developed as potential antitumor agents. Among them, pentacyclic triterpenoid 18β-glycyrrhetinic acid(18β-GA) is widely recognized to have a wide range of pharmacological effects such as anti-inflammatory, antibacterial, antitumor and antivirus. In this paper, we showed the potential targets and mechanisms of 18β-GA against TNBC through network pharmacology and molecular docking. Initially, network analysis revealed 85 common targets of 18β-GA in TNBC as shown in the “compound-target-disease” network employing Cytoscape 3.9.1 Further analysis identified TNF, IL-6, MMP9, ALB, PPARG, IGF-1, ESR1, STAT1, CCNA2 and PLK1 as the most crucial hub targets of 18β-GA against TNBC. Moreover, molecular docking simulations and functional enrichment analysis indicated the involvement of multiple signaling pathways in suppressing TNBC.

Protein nanospheres have garnered significant attention in the field of drug delivery and therapeutics due to their remarkable potential as carriers for active substances. These nanoscale structures, primarily composed of biocompatible proteins, offer a versatile platform for encapsulating and delivering a wide range of therapeutic agents, including drugs, peptides, and nucleic acids. This abstract explores the multifaceted role of protein nanospheres as drug delivery carriers, encompassing their fabrication methods, properties, and diverse applications.

One of the key advantages of protein nanospheres lies in their ability to protect and stabilize encapsulated substances, enhancing their bioavailability and pharmacokinetic profiles. Moreover, their tunable surface properties enable specific targeting to disease sites, minimizing off-target effects and reducing systemic toxicity. Various methods of engineering protein nanospheres, such as self-assembly and chemical modification, allow for precise control over their size, shape, and drug release kinetics.

This abstract also addresses the challenges associated with protein nanosphere-based drug delivery, including stability concerns and scale-up issues. Nonetheless, their immense potential in advancing personalized medicine and improving therapeutic outcomes makes protein nanospheres a compelling area of research in the realm of drug delivery.

This research was carried out within the SMART-MAT Functional Materials Science Club of the Faculty of Materials Engineering and Physics of Cracow University of Technology as part of the 3rd edition of the program "Student research clubs create innovation" through the project titled „Transdermal systems in targeted therapy of skin cancer” financed by the Ministry of Science and Higher Education (grant no: SKN 157/568410/2023)

Steroid derivatives are polycyclic compounds with a wide spectrum of biological activities, such as anticancer, antibacterial, antioxidant, antiviral, antiparasitic, and anti-inflammatory. Several studies suggest that steroid derivatives can be effective inhibitors of aldo-keto reductases (AKRs): NADPH-dependent oxidoreductases involved in the biosynthesis and metabolism of steroid hormones, as well as in the detoxification of xenobiotics. Some AKR isoforms are overexpressed in human tumors, such as breast, prostate and colon cancer, glioma, neuroblastoma and acute myeloid leukemia. Substrates for AKRs may include carbonyl-containing cytostatic agents, leading to the inactivation of these drugs and chemoresistance. Specific inhibition of these metabolic enzymes has been established as an attractive strategy for anti-cancer drug development. In this study, the inhibition potential of D-ring modified steroids against the AKR1C4 isoform was evaluated by monitoring the decrease in NADPH fluorescence associated with AKR1C4 activity. His-tagged human recombinant AKR1C4 was expressed in Escherichia coli BL21 cells, purified using immobilized metal affinity chromatography, followed by size-exclusion chromatography. Kinetic characterization of AKR1C4 was performed by a fluorimetric assay using 9,10-phenantrenequinone (PQ) as a substrate. In the presence of several steroid derivatives, reduction of PQ was inhibited, suggesting that these steroid derivatives are promising AKR1C4 inhibitors. These preliminary results could serve as a starting point for the design of novel anticancer drug candidates targeting AKRs with high specificity. It was also shown that hepatic AKR1C4 participates in the reduction of naltrexone, an opioid receptor antagonist used in the treatment of addiction, which further indicates the importance of identifying new inhibitors of this enzyme.