Development of a bioluminescent homogenous nanobody-based immunoassay for the detection of prostate-specific antigen (PSA).

Prostate cancer is the most prevalent cancer in men. At present, the diagnosis and screening of prostate cancer rely on the essential biomarker known as prostate-specific antigen (PSA). The main purpose of this study was to develop a novel immunoassay for the detection of PSA based on a tri-part split-nanoluciferase system and a nanobody targeting PSA. In our approach, two small components of the split-nanoluciferase, referred to as ß9 and ß10, were individually fused to two anti-PSA nanobodies, N7 and N23. When these proteins bind to PSA and in the presence of the third nanoluciferase component, called Δ11S, the split-nanoluciferase components are brought into close proximity, facilitating the reassembly of the active nanoluciferase and activation of luminescence. These proteins were expressed in a bacterial expression system, purified, and employed for the intended immunoassay. The developed immunoassay demonstrated the capability to sensitively detect PSA within a linear range from 1.0 to 20.0 ng/mL with LOD of 0.4 ng/mL, and the results obtained through this immunoassay agreed with those derived from the ELISA. Our study indicates that the homogeneous immunoassay developed with nanobodies exhibits remarkable specificity for PSA and can serve as a reliable, fast, and user-friendly test for detecting PSA.

Oral Papillary Squamous Cell Carcinoma and Oral Squamous Cell Carcinoma: A Histopathological and Immunohistochemical Comparative Study.

PURPOSE: The aim of the study is to investigate the immunohistochemical expression of both Alpha smooth muscle actin and Transforming Growth Factor beta and compare their expression in oral papillary squamous cell carcinoma with their expression in different histological grades of oral squamous cell carcinoma. A correlation between these immuno-histochemical expressions and histological findings will then be performed. The research question is "Do the percentages of α-SMA and TGF-ß immune-expression in OPSCC differ from that in the conventional OSCC?". METHODS: This will be achieved by collecting archival blocks of oral papillary squamous cell carcinoma and different grades of oral squamous cell carcinoma, staining the specimens with Transforming Growth Factor beta and alpha smooth muscle actin, then measuring the mean staining index of expression in each group and the area percent of both markers. RESULTS: Results revealed that transforming growth factor beta expression in the epithelium was high in all cases of well-differentiated squamous cell carcinoma, most oral papillary squamous cell carcinoma, and poorly differentiated oral squamous cell carcinoma. On the other hand, different grades of oral squamous cell carcinoma showed a high staining index of alpha smooth muscle actin expression in the stroma. While cases of oral papillary squamous cell carcinoma were either moderate or low-staining. CONCLUSIONS: Oral papillary squamous cell carcinoma has a favourable prognosis compared to different histological grades, and the prognosis does not depend only on histological grade but also on other prognostic factors.

Assessment of the Effect of Surface Modification of Metal Oxides on Silver Nanoparticles: Optical Properties and Potential Toxicity.

Silver nanoparticles (AgNPs) have garnered significant interest due to their distinctive properties and potential applications. Traditional fabrication methods for nanoparticles often involve high-energy physical conditions and the use of toxic solvents. Various green synthesis approaches have been developed to circumvent these issues and produce environmentally benign nanoparticles. Our study focuses on the green synthesis of AgNPs using L-ascorbic acid and explores the modification of their properties to enhance antibacterial and anticancer effects. This is achieved by coating the nanoparticles with Zinc oxide (ZnO) and Silica oxide (SiO2), which alters their optical properties in the visible spectrum. The synthesized formulations-AgNPs, zinc oxide-silver nanoparticles (Ag@ZnO), and silica oxide-silver nanoparticles (Ag@SiO2) core/shell nanoparticles-were characterized using a suite of physicochemical techniques, including Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), Zeta potential measurement, UV-Vis spectroscopy, Refractive Index Measurements, and Optical Anisotropy Assessment. TEM imaging revealed particle sizes of 11 nm for AgNPs, 8 nm for Ag@ZnO, and 400 nm for Ag@SiO2. The Zeta potential values for Ag@ZnO and Ag@SiO2 were measured at -17.0 ± 5 mV and -65.0 ± 8 mV, respectively. UV-Vis absorption spectra were recorded for all formulations in the 320 nm to 600 nm wavelength range. The refractive index of AgNPs at 404.7 nm was 1.34572, with slight shifts observed for Ag@ZnO and Ag@SiO2 to 1.34326 and 1.37378, respectively. The cytotoxicity of the nanocomposites against breast cancer cell lines (MCF-7) was assessed using the MTT assay. The results indicated that AgNPs and Ag@ZnO exhibited potent therapeutic effects, with IC50 values of 494.00 µg/mL and 430.00 µg/mL, respectively, compared to 4247.20 µg/mL for Ag@SiO2. Additionally, the antibacterial efficacy of AgNPs was significantly enhanced under visible light irradiation. Ag@ZnO demonstrated substantial antibacterial activity both with and without light exposure, while the Ag@SiO2 nanocomposites significantly reduced the inherent antibacterial activity of silver. Conversely, the Ag@ZnO nanocomposites displayed pronounced antibacterial and anticancer activities. The findings suggest that silver-based nanocomposites, particularly Ag@ZnO, could be practical tools in water treatment and the pharmaceutical industry due to their enhanced therapeutic properties.

Reparative potential of mesenchymal stem cells and platelet-rich plasma on irradiated submandibular glands of male albino rats.

OBJECTIVE: To appraise and compare the reparative role of bone marrow-mesenchymal stem cells (BM-MSCs) and platelet-rich plasma (PRP) against irradiation damage on albino rats' submandibular gland. DESIGN: Seventy four male albino rats were used, one for BM-MSCs harvesting, 10 for PRP preparation, seven as control group (Group 1). The remaining 56 rats were subjected to single dose (6 Gy) gamma irradiation and were divided into equal four groups; (Group 2): received no treatment, (Group 3): each rat was injected with 1 × 105 BM-MSCs, (Group 4): each rat was injected with 0.5 ml/kg PRP, and (Group 5): each rat was injected with 1 × 105 BM-MSCs and 0.5 ml/kg PRP. Each group was further subdivided into two subgroups in which rats sacrificed after one and two weeks from irradiation. Any structural changes were examined histopathologically, immunohistochemically using proliferating cell nuclear antigen (PCNA) and CD31 primary antibodies and histochemically using picrosirius red (PSR) stain, then analyzed statistically. RESULTS: Histopathological examination of Group 2 showed atrophied acini, with nuclear changes and signs of degeneration in duct systems. Treated groups revealed signs of regeneration in form of uniform acini and regenerated duct systems especially in Group 5 and in a time depended manner. Immunohistochemical examination revealed increased immunoexpression of PCNA and CD31, while histochemical examination showed decreased PSR in all treated groups in relation to the irradiated group and this was proved statistically. CONCLUSIONS: BM-MSCs and PRP are effective as treatment for irradiation-induced submandibular gland damage. However, the combined therapy is recommended over each one separately.

Recent progress in biologically active indole hybrids: a mini review.

The indole moiety is one of the most widespread heterocycles found in both natural products and biological systems. Indoles have important biological activities including anticancer, antioxidant, anti-inflammatory, antifungal, anticholinesterase, and antibacterial properties. Scientists are therefore interested in the synthesis of biologically active indole-based hybrids such as indole-coumarin, indole-chalcone, indole-isatin, indole-pyrimidine and so on, with the aim of improving activity, selectivity, and mitigating side effects. This review will discuss the newly synthesized indole-based hybrids along with their biological activity which will be useful in drug discovery and development.

Insights into the Role of Gremlin-1, a Bone Morphogenic Protein Antagonist, in Cancer Initiation and Progression.

The bone morphogenic protein (BMP) antagonist Gremlin-1 is a biologically significant regulator known for its crucial role in tissue differentiation and embryonic development. Nevertheless, it has been reported that Gremlin-1 can exhibit its function through BMP dependent and independent pathways. Gremlin-1 has also been reported to be involved in organ fibrosis, which has been correlated to the development of other diseases, such as renal inflammation and diabetic nephropathy. Based on growing evidence, Gremlin-1 has recently been implicated in the initiation and progression of different types of cancers. Further, it contributes to the stemness state of cancer cells. Herein, we explore the recent findings on the role of Gremlin-1 in various cancer types, including breast, cervical, colorectal, and gastric cancers, as well as glioblastomas. Additionally, we highlighted the impact of Gremlin-1 on cellular processes and signaling pathways involved in carcinogenesis. Therefore, it was suggested that Gremlin-1 might be a promising prognostic biomarker and therapeutic target in cancers.

Multifunctional nanoparticles in stem cell therapy for cellular treating of kidney and liver diseases.

The review gives an overview of the mechanisms of internalization and distribution of nanoparticles in stem cells this is achieved via providing analysis of the methods used in exploring the migration routes of stem cells, and their reciprocity. In addition, exploring microenvironment target in the body, and tracking the fate of exogenously transplanted stem cells by using innovative and non-invasive techniques will also be discussed. Such techniques like magnetic resonance imaging (MRI), multimodality tracking, optical imaging, and nuclear medicine imaging, which were designed to follow up stem cell migration. This review will explain the various distinctive strategies to enhance homing of labeled stem cells with nanoparticles into damaged hepatic and renal tissues, this purpose was obtained by inducing a specific gene into stem cells, various chemokines, and applying an external magnetic field. Also, this work illustrates how to improve nanoparticles uptake by using transfection agents or covalently binding an exogenous protein (i.e., Human immunodeficiency virus-Tat protein) or conjugating a receptor-specific monoclonal antibody or make modifications to iron coat. It contains stem cell labeling methods such as extracellular labeling and internalization approaches. Ultimately, our review indicates trails of researchers in nanoparticles utilization in stem cell therapy in both kidney and liver diseases.

Apigenin sensitizes colon cancer cells to antitumor activity of ABT-263.

Apigenin is an edible plant-derived flavonoid that shows modest antitumor activities in vitro and in vivo. Apigenin treatment resulted in cell growth arrest and apoptosis in various types of tumors by modulating several signaling pathways. In the present study, we evaluated interactions between apigenin and ABT-263 in colon cancer cells. We observed a synergistic effect between apigenin and ABT-263 on apoptosis of colon cancer cells. ABT-263 alone induced limited cell death while upregulating expression of Mcl-1, a potential mechanism for the acquired resistance to ABT-263. The presence of apigenin antagonized ABT-263-induced Mcl-1 upregulation and dramatically enhanced ABT-263-induced cell death. Meanwhile, apigenin suppressed AKT and ERK activation. Inactivation of either AKT or ERK by lentivirus-transduced shRNA or treatment with specific small-molecule inhibitors of these pathways enhanced ABT-263-induced cell death, mirroring the effect of apigenin. Moreover, the combination response was associated with upregulation of Bim and activation of Bax. Downregulation of Bax eliminated the synergistic effect of apigenin and ABT-263 on cell death. Xenograft studies in SCID mice showed that the combined treatment with apigenin and ABT-263 inhibited tumor growth by up to 70% without obvious adverse effects, while either agent only inhibited around 30%. Our results demonstrate a novel strategy to enhance ABT-263-induced antitumor activity in human colon cancer cells by apigenin via inhibition of the Mcl-1, AKT, and ERK prosurvival regulators.