Soumya Ranjan Jena

The anticancer mechanism of DOX/MET co-loaded GA–GT nanohydrogel in MCF-7 cells, showing enhanced intracellular drug delivery, PI3K/AKT/mTOR inhibition, suppression of TNT-mediated intercellular communication, and subsequent induction of apoptosis and inhibition of cell migration.

Idiopathic male infertility (IMI), which accounts for nearly 50% of male infertility cases, remains a major clinical challenge due to the inability of standard semen analysis to reveal underlying molecular defects. Post-translational modifications such as lysine acetylation are increasingly recognized as key regulators of sperm function, affecting antioxidant defense, energy metabolism, and spermatogenesis. In this study, global acetyl-proteomic profiling of spermatozoa from idiopathic infertile patients (IIP) and fertile donors (FD) was performed using immunoprecipitation-based enrichment followed by high-resolution LC–MS/MS. Bioinformatics analyses, including STRING, Cytoscape, and Ingenuity Pathway Analysis (IPA), identified 718 differentially acetylated proteins (DAPs), with significant enrichment in pathways related to redox homeostasis, molecular transport, glycolysis, and mitochondrial metabolism. Hub proteins such as SOD1, PARK7, and PRKACA emerged as key regulators linking oxidative stress with defective motility and impaired sperm physiology. Western blot validation confirmed the downregulation of these hub proteins in IIP, supporting their role in redox imbalance and mitochondrial dysfunction. Our findings highlight dysregulated lysine acetylation as a defining molecular feature of IMI and suggest that targeting acetylation-associated pathways may provide novel diagnostic biomarkers and therapeutic strategies to improve sperm function and male reproductive outcomes.

Cancer remains the leading cause of death worldwide. Despite decades of continuous research, limitations persist in existing therapeutic approaches. Conventional strategies such as surgery, chemotherapy, and radiotherapy, though advanced, face challenges including poor bioavailability, toxic side effects, inadequate targeting of cancer cells, and limited survival benefits. The major issue lies in the inability of improved drug formulations to effectively reach cancer cells. Emerging approaches such as photodynamic therapy (PDT) and immunotherapy have shown greater promise, offering reduced side effects and higher treatment efficiency compared to traditional methods. Various natural and synthetic nanocarriers, including exosomes, liposomes, solid lipid nanoparticles (SLNs) and micelles have been explored as drug delivery vehicles in these therapies. Among them, exosomes, being natural secretory vesicles, have shown unique potential as independent delivery systems. However, challenges and limitations remain in their application for precise cancer targeting. A combinational strategy, integrating exosomes with other lipid-based drug delivery systems (LBDDS), while preserving their intrinsic properties and engineering their surface to carry photosensitizers (PS) or immune modulators, could overcome these barriers. Such well-designed natural cargos may enhance therapeutic efficacy, modulate the tumor microenvironment, and address current shortcomings in cancer therapy. This review highlights the individual applications of PDT and immunotherapy using exosomes and LBDDS, and explores their potential synergistic use for more effective and targeted cancer treatment.

Background: Extracellular vesicles, particularly exosomes, play a crucial role in cell–cell communication and as carriers of biomarkers. However, their use in clinical settings is limited due to a lack of standardized isolation and characterization. Ultracentrifugation (UC) is considered a gold standard for exosome isolation but presents several limitations. Size-exclusion chromatography (SEC) has recently gained attention as a superior method, which offers better yield, purity, and protection of exosome physical properties. This study focused on optimizing the SEC method for isolation of exosomes from seminal plasma and comparing yield, quality, and proteome profiles with those obtained by UC. Methods: In this SEC method, seminal plasma (0.5 mL) was loaded onto a SEC column and collected in 13 fractions of 0.4 mL each. The physical and molecular characterization of exosomes was carried out using a ZetaView analyzer and Western blot, respectively. Further, SEC-isolated exosomes were used for proteomic profiling and functional bioinformatic analysis. Results: The second and third fractions had the highest concentration of exosomes with uniform size and strong expression of exosome markers. Also, comparative proteomic analysis identified 3315 proteins in SEC-isolated exosomes and 931 in UC-isolated exosomes, with 709 proteins in common. SEC-isolated exosomes showed greater overlap with Vesiclepedia’s and ExoCarta’s top 100 lists than UC-isolated exosomes (Vesiclepedia: 91 vs. 77 proteins, ExoCarta: 94 vs. 79). Proteins from SEC- and UC-isolated exosomes showed similar enrichment profiles across all three gene ontology categories. Conclusions: Overall, this optimized SEC protocol is a reliable alternative method to isolate seminal exosomes with high purity, supporting its potential applications in clinical and basic research.

Objectives: Widely used organophosphorus pesticides, methyl parathion (MePa), alter the reproductive functions in various animals and humans by induction of oxidative stress on augmented release of reactive oxygen species (ROS). MePa affects semen quality by inducing DNA damage through spermatogenic stages. Several heat shock proteins (HSPs) are expressed in response to environmental stressors particularly the redox-active ones for regulation of protein turnover. Since oxidative stress and sperm motility are implicated in MePa toxicity, studying the expression of HSP90b will unravel the mechanism behind its noxiousness. Material and Methods: Spermatozoa isolated from healthy donors were subjected to various concentrations of MePa (50, 250, 500, and 750 μM) for studying its effect on sperm motility, ROS generation, sperm chromatin integrity, and expression of stress responsive molecular chaperone HSP90b. Results: In vitro exposure of MePa at concentrations ≥500 μM results in a decline in sperm motility and an increased generation of ROS, DNA damage, and HSP90b expression. Conclusion: ROS-mediated modulation of HSP90b expression may affect the structural integrity of client proteins and oxidative injury to membrane lipid, along with DNA integrity resulting in declined sperm motility in response to MePa.

Respiratoryepithelialadenomatoid hamartoma (REAH) - JDPO- Print ISSN No: - 2581-3714 Online ISSN No:- 2581-3706 Article DOI No:- 10.18231/j.jdpo.2022.037, IP Journal of Diagnostic Pathology and Oncology-IP J Diagn Pathol Oncol

Oxidative stress (OS) is implicated in 80% of idiopathic male infertility (IMI) patients where exposure to redox active environmental toxicants such as polyaromatic hydrocarbons (PAHs) may play. In the present study the seminal exposome of various PAH was analyzed in two separate cohorts including 43 fertile donors and 60 IMI patients by HPLC and receiver operator characteristic curve was applied to find out the cut-off limits. Furthermore, spermatozoa from both the groups were subjected to label free liquid chromatography mass spectroscopy (LC-MS/MS) followed by bioinformatics analysis to elucidate the molecular mechanism(s) involved and the key proteins from the affected pathways were validated by western blot along with oxidative modification of proteins. Of the 16 standard PAH 13 were detected in the semen. Receiver Operator Character (ROC) Curve analysis (AUC _ROC ) revealed the PAHs having most significant effect on fertility are of the following order Anthracene<benzo(a)pyrene<benzo[b]fluoranthene<Fluoranthene<benzo(a)anthracene<indol (123CD)pyrene<pyrene<naphthalene<dibenzo(AH)anthracene<fluorene<2bromonaphthalene <chrysene<benzo(GH1)perylene. Benzo[a] pyrene is invariably present in all infertile patients while naphthalene is present in both fertile and IMI group. Of the total 773 detected proteins (Control: 631 and PAH: 717); 71 were differentially expressed (13 underexpressed, 58 overexpressed) in IMI patients resulting in impaired mitochondrial dysfunction and oxidative phosphorylation, DNA damage, Aryl hydrocarbon receptor (AHR) signalling, xenobiotic metabolism and induction of NRF-2 mediated oxidative stress response (increased in 4-hydroxynonenal and nitrosylated protein adduct formation, and declined antioxidant defence). The increased GSH/GSSG ratio in patients may be an adaptive response to metabolize the xenobiotics via conjugation as evidenced by overexpression of AHR and Heat shock protein 90 beta (HSP90β) in patients. Seminal PAH concentrations, oxidative protein modification along with protein markers (e.g. AHR and HSP90β) may help in better prediction and management of IMI. Contribution of environment borne PAH in semen should not undermined in infertility evaluation.

Recent evidence entail paternal factors as plausible contributors in spontaneous recurrent pregnancy loss (RPL). Seminal extracellular vesicles secreted from cells of male reproductive tract carry regulatory proteins and RNAs. They are proposed to regulate sperm maturation and function while their fusion to endometrial stromal cells helps in decidualization. Nevertheless, the mechanism(s) involved in these processes are poorly understood. This study aims at elucidating the molecular basis of paternal contribution by comparative proteomics (label‐free LC‐MS/MS) of isolated seminal extracellular vesicles from fertile men and partners of patients with RPL (n = 21 per group). Bioinformatics analysis revealed the identified differentially expressed proteins to be involved in DNA replication, recombination and repair, gene expression, cellular assembly and organization, cell death, and survival. Major disease pathways affected were identified as developmental, hereditary, and immunological disorders. Of the three identified hub genes regulating the above disease pathways, two (HNRNPC and HNRNPU) are overexpressed while RUVBL1 is underexpressed along with over expression of HIST1H1C, DDX1, surmising defective chromatin packaging, and histone removal in spermatozoa resulting in improper expression in paternal genes thereby leading to abnormal embryo development. Besides, alteration in GSTP1 expression points oxidative predominance in RPL group. Differential expression of C3, C4a/C4b, CFB, and GDF 15 may be involved in altered maternal immune response to paternal antigens resulting in impaired decidualization.

A series of C₃‐symmetricaltriazine‐cored small dendritic molecules containing three to nine peripheral o‐carborane clusters were synthesized through Cu(I)‐catalyzedazide–alkyne cycloaddition reactions. The newly synthesized molecules containing multiple o‐carborane moieties were characterized using nuclear magnetic resonance and matrix‐assisted laser desorption/ionization‐time of flight mass spectral analysis. The biological evaluation of these three to nine cage dendrimers was performed using breast cancer cells (Michigan Cancer Foundation 7). All these dendritic compounds showed cytotoxicity toward breast cancer cells, and the toxicity increased as the number of peripheral o‐carboranes increased. The 9‐cage molecule showed the highest cytotoxicity, and the half maximal inhibitory concentration (IC₅₀) value was found to be 80.67 ng/ml. Its cytotoxicity was significantly higher than the common chemotherapy agent cisplatin. As expected, the boron‐richo‐carborane‐appended molecules showed high thermal stability. The thermal stability increased as the number of peripheral o‐carborane moieties increased.