Oncostatic treatment effect of triple negative breast cancer cell line with copper (I)-nicotinate complex.

The treatment of triple-negative breast cancer (TNBC) remains a major challenge. The present study aimed to throw more light on the role of copper (I)-nicotinate complex (CNC) as an antitumor as well as a proapoptotic agent. In this study, the HCC-1806 cell line was used as a model for TNBC. Cell cycle, apoptosis assay, and programmed cell death protein-1 were investigated by flowcytometry. Besides, the comet assay was performed using a fluorescence microscope. The enzyme-linked immunosorbent assay technique was used for the detection of phospho-Chk1 at ser 317 and caspase-3. Moreover, the gene expression of survivin was identified by real-time polymerase chain reaction. Finally, superoxide dismutase (SOD) was calorimetrically assayed. The viability of HCC-1806 cells treated with CNC was decreased in a dose-dependent manner. The tendency for apoptotic machinery was observed through the increase in the sub G0 peak, the percentage of early and late apoptotic phases, and the elevation in caspase-3 levels associated with a downregulation of the survivin gene expression. The antioxidant property of the complex, reflected by elevated SOD activity, may contribute to mediate the cell death pathways. Low concentrations of CNC were found to favor the apoptotis-mediated mechanism. However, one cannot neglect the abundance of cell necrosis-mediated death of cells via CNC, especially at higher concentrations.

Role of mesenchymal stem cells versus angiotensin converting enzyme inhibitor in kidney repair.

AIM: The current study sought to clarify the role of bone marrow derived mesenchymal stem cells (BM-MSCs) and adipose tissue derived mesenchymal stem cells (AD-MSCs) in repressing nephropathy in the experimental model. Moreover, the aim of this work was extended to compare between stem cells role and angiotensin converting enzyme inhibitor in kidney repair. METHODS: Isolation and preparation of MSCs culture, flow cytometry using CD34, CD44 and CD105 cell surface markers, biochemical analyses for determination of serum creatinine, urea, transforming growth factor ß (TGF-ß), cystatin C (CYS-C) and urinary N-Acetyl-ß-D-Glucosaminidase (UNAG), and histopathological investigation of kidney tissue sections were performed. RESULTS: The results of the present study revealed that single intravenous infusion of MSCs either derived from bone marrow or adipose tissue was able to enhance renal reparative processes through significantly decreased serum creatinine, urea, TGF-ß and CYS-C levels as well as UNAG level and significantly increase glomerular filtration rate. Additionally, the histopathological investigations of kidney tissues showed that MSCs have significant regenerative effects as evidenced by the decrease in focal inflammatory cells infiltration, focal interstitial nephritis and congested glomeruli as well as degenerated tubules. CONCLUSION: The current data provided distinct evidence about the favourable impact of AD-MSCs and BM-MSCs in attenuation of cyclosporine-induced nephropathy in rats through their ability to promote functional and structural kidney repair via transdifferentiation.

Apoptosis and cell proliferation: correlation with BCL-2 and P53 oncoprotein expression in human hepatocellular carcinoma.

BACKGROUND/AIM: Occurrence and biological characteristics of tumors are related not only to over-proliferation of carcinoma cells but also to decrease of apoptosis. The present study was suggested to evaluate the correlation between P53 and Bcl-2 oncoprotein expression with apoptosis and cell proliferative activity in HCC patients. METHODOLOGY: P53 and Bcl-2 protein expression were estimated in the sera and in liver tissues of 45 HCC cases using ELISA and immunohistochemistry. Apoptosis was estimated as apoptotic index (AI) and cell proliferative activity was detected using AgNORs. RESULTS: Serum p53 antigen in HCC patients (0.46±0.331ng/ml) showed significant elevation than healthy individuals (0.24±0.11ng/ml, p<0.05). P53 protein was immunostained in 41% of HCC; 37.5% of these positive cases were in diffuse pattern representing the mutant p53. Serum Bcl-2 was elevated in HCC cases (50.28±25.83u/ ml) than healthy individuals (26.65±8.63u/ml, p<0.05). Bcl-2 was immunohistochemically localized in 35.9% of HCC and the positivity was inversely proportional with the histological grade (47.4%, 25%, 25% in grade I,II,III respectively). Bcl-2 showed a positive linear correlation with p53 in the sera of carcinoma patients (p<0.05). CONCLUSION: Bcl-2 may play a role in hepatocarcinogenesis as an inhibitor of apoptosis. However, a positive linear correlation was found between bcl-2 and p53 suggesting that bcl-2/p53 co-expression pattern may be of value in the development of more effective medical therapies in HCC.

Experimental and computational studies of silver(I) dibenzoylmethane-based complexes, interaction with DNA/RNA/BSA biomolecules, and in vitro cytotoxic activity.

Two silver(I) complexes of composition [Ag2(L)2] (1) and [Ag(L)(PPh3)2](2) (HL = dibenzoyl- methane, PPh3 = triphenylphosphine) were synthesized and characterized by elemental analysis, FTIR, NMR, XRPD, and UV-visible spectra. The molecular structures of the studied ligands and Ag(I) complexes have been characterized using Density Function Theory (DFT) calculations. This analysis has enabled us to determine the reactivity and the coordination site(s) for each ligand. Ag(I) ion is found to be coordinated with the ligand's oxygens in almost a linear fashion in complex (1), while in complex (2) it adopts a tetrahedral geometry. The interaction compounds with biomolecules; calf thymus (ct DNA), yeast-tRNA, and bovine serum albumin (BSA) were investigated using both absorption and fluorescence spectroscopy. The in vitro cytotoxic studies of the complexes against normal human lung fibroblast (WI38), cancerous breast (MDA-MB-231), mammary gland breast (MCF7), hepatocellular (HePG2), and prostate (PC3) cell lines indicated that the complexes are highly toxic to the cancer cells but less toxic towards the normal one when compared with the ligand. Flow cytometric results showed that complex (1) induced cell cycle arrest at the G2/M phase, and complex (2) at G2/M and S phases. Moreover, the results of apoptotic genes (caspase3 and p53) and anti-apoptotic (Bcl2) led us to suggest an apoptotic killing mechanism of cells rather than a necrotic one.

Controlled synthesis of in-situ gold nanoparticles onto chitosan functionalized PLGA nanoparticles for oral insulin delivery.

Chitosan-based nanoparticles (chitosan nanoparticles (ChNps), chitosan gold Nps (ChAuNps), and chitosan gold Nps functionalized with poly lactic-co-glycolic acid (PLGA) (ChAuNps/PLGA)) were prepared as nanocarriers for insulin to improve its oral uptake. The emulsion solvent diffusion method was employed to functionalize the Nps with PLGA. TEM, SEM, DLS, and zeta potential were conducted to characterize the Nps. The morphological analysis confirmed the formation of spherical Nps with hydrodynamic particle sizes of 138±23, 16±2.2, and 50±9.3 nm for ChNps, ChAuNps, and ChAuNps/PLGA, respectively. Zeta potential measurements indicated two types of Nps, regardless of insulin entrapment, positively charged, (ChNps (+36 ± 4.2, +31 ± 2.2mv)) and ChAuNps (+37 ± 4.3, +33 ± 2.5mv) and negatively charged (ChAuNps/PLGA (-31 ± 2.7, -26 ± 2.1 mv)). The in vitro studies were assessed by measuring the entrapment efficiencies (EE%) and the release profiles of insulin at different pH values. EE% for ChNps, ChAuNps, and ChAuNps/PLGA were 97 ± 1.5, 98.4 ± 1.9, and 99 ± 1.2%, respectively. At an acidic medium, a significant level of insulin retention was observed (96 ± 0.08%) for ChAuNps/PLGA. While a high amount was released at higher pH values over an extended period of time. In vivo studies, diabetic rats treated with insulin-loaded Nps had reduced blood glucose level (BGL) (38 ± 2.8, 35 ± 6.5, and 27 ± 5.6%) for ChNps ChAuNps and ChAuNps/PLGA, respectively. The pharmacological availability (PA%) and bioavailability (FR%) for insulin-loaded ChAuNps/PLGA were 15.8 ± 0.71% and 7.7 ± 0.93%, respectively. Altogether, emphasize the role of biocompatible Nps and their efficiency in the convenient delivery of insulin, thus lowering the BGL in a safe condition.

Detection of a Helicobacter pylori antigen in cerebrospinal fluid of patients with meningitis.

Meningitis is a common life threatening disease which may be caused by a bacterium, fungus, or virus. Here, the presence of a Helicobacter pylori antigen was investigated in serum and CSF samples from 173 individuals with meningitis. The influence of H. pylori infection on CSF levels of Thl/Th2 cytokines was also evaluated. H. pylori antigen was detected using ELISA and Western blot based on specific anti-H. pylori antibody. Thl/Th2 cytokines (IFN-gamma & IL-10, respectively) were also determined. A target epitope of 58-kDa was detected in selected CSF and serum samples using Western blot. H. pylori antigen was detected in the CSF samples of 75% of meningitis patients showing H. pylori antigen in their sera. A significant correlation (p < 0.001, r = 0.21) was shown between serum and CSF levels of 58-kDa H. pylori antigen. Only the levels of Thl cytokine (IFN-gamma) were significantly higher (p < 0.05) in CSF of meningitis patients positive for H. pylori antigen than negative patients with meningitis. In conclusion, the 58-kDa H. pylori antigen crossed the blood brain barrier and entered the CSF of patients with meningitis. A significant upregulation of Thl response may be associated with H. pylori infection in patients with meningitis.