Rahil Mashhadi

Background: Silver nanoparticles (AgNPs) trigger apoptosis in cancer cells, while albumin nanoparticles enable effective drug delivery. This study compares the antitumor and cytotoxic effects of albumin-coated AgNPs (AgNPs-Alb) versus AgNPs on human prostate cancer cell lines. Method: AgNPs-Alb were synthesized and tested against PC3 and LNCaP prostate cancer cell lines. Characterization via Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), and Ultraviolet-Visible (UV-Vis) spectroscopy confirmed their properties. IC50 values were determined using MTT assay, with apoptosis assessed by Annexin-V/PI staining. DNA cell cycle was analyzed by PI staining. Migration, proliferation, and nuclear morphology were evaluated through scratch-wound, colony-forming, and Hoechst staining assays. Gene expression of Snail, E-cadherin, VEGF-C, VEGF-A, Bcl2, Bax, and P53 was analyzed using real-time PCR. Results: The IC50 values for AgNPs and AgNPs-Alb were 48 μM and 32 μM in PC3 cells, and 110 μM and 95 μM in LNCaP cells, respectively. AgNPs-Alb significantly inhibited PC3 cell migration compared to AgNPs (p < 0.001) and Bicalutamide (p < 0.0001). In both cell lines, AgNPs-Alb significantly reduced colony formation compared to AgNPs and Bicalutamide (p < 0.05). Flow cytometry revealed a higher percentage of apoptotic cells in PC3 with AgNPs-Alb treatment compared to AgNPs and Bicalutamide. In LNCaP cells, AgNPs-Alb induced a significantly higher percentage of Sub-G1 cells. AgNPs-Alb treatment caused greater mRNA suppression of VEGF-A and a higher Bax/Bcl2 ratio in PC3 and LNCaP cells (p < 0.05). Additionally, a significant increase in P53 and E-cadherin, alongside a decrease in VEGF-C expression in LnCAP cells, was observed (p < 0.05). Conclusions: This study suggests that AgNPs-Alb have stronger anticancer and cytotoxic effects compared to AgNPs alone against PCa cell lines and higher effects were observed on PC3 cells compared to LnCAP cells.

Background: Considering the hepatotoxicity and other side effects associated with flutamide as a first‐line treatment for prostate cancer (PCa), this study aimed to evaluate metformin as a potential anticancer agent to reduce the required dose of flutamide and thereby minimize its adverse effects. Method: We assessed the influence of metformin, flutamide, and metformin–flutamide combination therapy on LNCaP, DU145, and PC3 cell lines, which represent human PCa. The tests include wound‐healing assay, colony formation assay (CFA), analysis of apoptosis (programmed cell death) and cell cycle by flow cytometry, gene expression at the mRNA level by real‐time PCR ( BAX/BCL2 , E-cadherin , Snail , HIF1α , VEGFC , and KLK3 genes), and assessment of the treatments’ hepatotoxicity potential via measuring AST and ALT values. Result: To determine the IC50 (half‐maximal inhibitory concentration) values, cell lines were treated with different concentrations of the drugs. The IC50 values for metformin (800 μM) in the three cell lines and for flutamide (12 μM for PC3 and 10 μM for LNCaP/DU145), as determined by MTT assay, were confirmed by flow cytometry, indicating significant cell cycle arrest at the G0/G1 phase. The combination of metformin and flutamide significantly increased the BAX/BCL2 mRNA ratio in all three cell lines ( p < 0.0001) and downregulated the expression of KLK3 ( p < 0.01), HIF1α ( p < 0.01), VEGFC ( p < 0.001), and EMT pathway genes in PC3 and LNCaP ( p < 0.01). Liver injury assessment reported a reduction in flutamide’s hepatotoxicity in combination with metformin. Conclusion: Metformin in combination with flutamide reduced its dose and increased the sensitivity of PCa cells to treatment. Additionally, it mitigated the hepatotoxic effects of flutamide. Therefore, this combination may represent a new treatment strategy for PCa.

Background: Prostate cancer is the second most commonly diagnosed cancer worldwide. Although androgen deprivation therapy initially demonstrates clinical benefit, disease relapse with more aggressive phenotypes frequently occurs. The acidic tumor microenvironment in solid tumors may alter drug responsiveness. This study investigates how extracellular pH influences the cytotoxic effects of bicalutamide in human prostate cancer cell lines. Methods: PC3 and LNCaP cells were exposed to bicalutamide at varying concentrations at pH 7.4 and pH 6.8. IC50 values were determined using the MTT assay. Cell migration, apoptosis, and cell cycle distribution were evaluated by wound‐healing assay, annexin V/PI staining, and DNA content analysis, respectively. The expression of BAX , BCL2 , E-cadherin , N-cadherin , SNAI1 , AR , and VEGF-C was quantified by qPCR. Results: Bicalutamide (140 μg/mL) reduced PC3 cell viability to 39.62% at pH 7.4 compared with 51.36% at pH 6.8. In LNCaP cells, viability declined to 33.64% at pH 7.4% and 56.09% at pH 6.8. Treated PC3 cells exhibited significantly greater migration at pH 6.8 ( p < 0.01). Early apoptosis in treated LNCaP cells was significantly reduced at pH 6.8 ( p < 0.001). Both cell lines demonstrated enhanced S phase accumulation and reduced G1‐phase distribution at pH 6.8. The BAX / BCL2 ratio was significantly decreased at pH 6.8, indicating the suppression of proapoptotic signaling. Additionally, genes associated with epithelial–mesenchymal transition (EMT) were upregulated, and VEGF-C and AR expression increased at pH 6.8 ( p < 0.05). Conclusion: The efficacy of bicalutamide in prostate cancer cells is significantly influenced by extracellular pH. The drug exerts stronger cytotoxic, antimigratory, and proapoptotic effects at physiological pH (7.4) compared with acidic conditions (6.8).

Previous studies showed that the bladder extracellular matrix (B‐ECM) could increase the differentiation efficiency of mesenchymal cells into smooth muscle cells (SMC). This study investigates the potential of human amniotic membrane‐derived hydrogel (HAM‐hydrogel) as an alternative to xenogeneic B‐ECM for the myogenic differentiation of the rabbit adipose tissue‐derived MSC (AD‐MSC). Decellularized human amniotic membrane (HAM) and sheep urinary bladder (SUB) were utilized to create pre‐gel solutions for hydrogel formation. Rabbit AD‐MSCs were cultured on SUB‐hydrogel or HAM‐hydrogel‐coated plates supplemented with differentiation media containing myogenic growth factors (PDGF‐BB and TGF‐β1). An uncoated plate served as the control. After 2 weeks, real‐time qPCR, immunocytochemistry, flow cytometry, and western blot were employed to assess the expression of SMC‐specific markers (MHC and α‐SMA) at both protein and mRNA levels. Our decellularization protocol efficiently removed cell nuclei from the bladder and amniotic tissues, preserving key ECM components (collagen, mucopolysaccharides, and elastin) within the hydrogels. Compared to the control, the hydrogel‐coated groups exhibited significantly upregulated expression of SMC markers ( p ≤ .05). These findings suggest HAM‐hydrogel as a promising xenogeneic‐free alternative for bladder tissue engineering, potentially overcoming limitations associated with ethical concerns and contamination risks of xenogeneic materials.

Introduction: Kidney transplantation is the optimal treatment strategy for some end-stage renal disease (ESRD); however, graft survival and the success of the transplantation depend on several elements, including the genetics of recipients. In this study, we evaluated exon loci variants based on a high-resolution Next Generation Sequencing (NGS) method. Methods: We evaluated whole-exome sequencing (WES) of transplanted kidney recipients in a prospective study. The study involved a total of 10 patients (5 without a history of rejection and 5 with). About five milliliters of blood were collected for DNA extraction, followed by whole-exome sequencing based on molecular inversion probes (MIPs). Results: Sequencing and variant filtering identified nine pathogenic variants in rejecting patients (low survival). Interestingly, in five patients with successful kidney transplantation, we found 86 SNPs in 63 genes 61 were variants of uncertain significance (VUS), 5 were likely pathogenic, and five were likely benign/benign. The only overlap between rejecting and non-rejecting patients was SNPs rs529922492 in rejecting and rs773542127 in non-rejecting patients’ MUC4 gene. Conclusions: Nine variants of rs779232502, rs3831942, rs564955632, rs529922492, rs762675930, rs569593251, rs192347509, rs548514380, and rs72648913 have roles in short graft survival.

Aim. Accurate diagnosis of prostate cancer (PCa) has a fundamental role in clinical and patient care. Recent advances in diagnostic testing and marker lead to standardized interpretation and increased prescription by clinicians to improve the detection of clinically significant PCa and select patients who strictly require targeted biopsies. Methods. In this study, we present a systematic review of the overall diagnostic accuracy of each testing panel regarding the panel details. In this meta-analysis, using a structured search, Web of Science and PubMed databases were searched up to 23 September 2019 with no restrictions and filters. The studys outcome was the AUC and 95 confidence interval of prediction models. This index was reported as an overall and based on the WHO region and models with/without MRI. Results. The thirteen final articles included 25,691 people. The overall AUC and 95 CI in thirteen studies were 0.78 and 95 CI: 0.730.82. The weighted average AUC in the countries of the Americas region was 0.73 (95 CI: 0.700.75), and in European countries, it was 0.80 (95 CI: 0.720.88). In four studies with MRI, the average weighted AUC was 0.88 (95 CI: 0.860.90), while in other articles where MRI was not a parameter in the diagnostic model, the mean AUC was 0.73 (95 CI: 0.700.76). Conclusions. The present studys findings showed that MRI significantly improved the detection accuracy of prostate cancer and had the highest discrimination to distinguish candidates for biopsy.

The incidence of urolithiasis has increased in both the developed and the developing countries during the past decades. Economically, the increase of urolithiasis contributes to the rise of the healthcare burden everywhere. Moreover, this increase has been associated with a change in the epidemiology of urolithiasis in terms of age and sex distribution, and also the location and type of calculi. We searched the MEDLINE for relevant literature dating back to 1980. This review compared the trends in epidemiological factors affecting urolithiasis in the developed and the developing countries during the past decades. People in the developing countries are more likely to contract kidney calculi at a younger age than in the developed countries. Although calculus disease is still more prevalent in men than in women, the latter are increasingly affected in both worlds. Uric acid calculi are more prevalent in the developing than in industrialized countries. There is a progressive increase in the frequency of calcium oxalate and calcium phosphate calculi in the developing countries where these used to be less frequent. The incidence and prevalence of urinary calculi is increasing globally. Many factors including aging of the population, changes in diet, global warming, and employment of more accurate diagnostic tools seem to be involved in this increase. An increasing affluence and adaptation of Western diet habits in many developing countries seem likely to contribute to the changes.

Pneumococcal autolysins are enzymes involved in cell wall turnover and cellular division physiologically. They have been found to be involved in the pneumococcus pathogenesis. The aim of this study was to identify the autolytic activity of Spr1754 as a novel protein of Streptococcus pneumoniae. Moreover, the binding of the recombinant protein to plasma proteins was also determined. The spr1754 gene was amplified by PCR and cloned into the pET21a(+) prokaryotic expression vector. The constructed pET21a(+)/spr1754 recombinant plasmid was transformed into E. coli Origami (DE3) and induced using IPTG. The recombinant protein of Spr1754 was purified by Ni-NTA affinity chromatography and confirmed by SDS-PAGE and Western blot analysis using anti-His tag monoclonal antibody. Autolytic activity and the ability of the recombinant protein in binding to plasma proteins were performed using zymogram analysis and western blot, respectively. The spr1754 with expected size was cloned and overexpressed in Escherichia coli Origami (DE3), successfully. After purification of the Spr1754 recombinant protein, the autolytic activity was observed by zymography. Of the four plasma proteins used in this study, binding of lactoferrin to Spr1754 recombinant protein was shown. The Spr1754 recombinant protein has a bifunctional activity, i.e., as being autolysin and lactoferrin binding and designated as Aap (autolytic/ adhesion/ pneumococcus). Nevertheless, characterization of the Aap needs to be followed using gene inactivation and cell wall localization.

Purpose: To determine serum fetuin-A pattern after renal transplantation and its association with graft outcome. Materials and Methods: In 41 renal transplant recipients, serum pretransplant fetuin-A levels and serum fetuin-A concentrations on days 7 and 30 after transplantation were measured using the enzyme-linked immunosorbent assay (ELISA) method. Also, the association between serum fetuin-A levels-with clinical and laboratory parameters was evaluated. Results: A significant decrease in serum fetuin-A levels was noted in the first week after transplantation (P < .001). Subsequently, it started to increase and surpass pretransplant values during the first month (P < .001). Pretransplant fetuin-A levels did not differ among patients with different diethylenetriamine pentaacetic acid (DTPA) results. In addition, serum fetuin-A levels did not significantly correlate with metabolic parameters. Conclusion: In this prospective study there was no increase in serum fetuin-A levels during the first month and pretransplant fetuin-A levels are not predictive for allograft outcome in renal transplant recipients.

Purpose: To compare the expression rate of sex steroid hormone receptors of estrogen (ER), progesterone (PR) and androgen (AR) in normal urothelium and urothelial bladder cancer (UBC) and to evaluate the possible associations of these receptors expression with cancer progression and patient's survival. Materials and Methods: We evaluated the clinical data and tumor specimens of 120 patients with pathologically confirmed primary UBC with 132 normal healthy controls. Both patients and controls selected from list of subjects who have been referred to Sina Urology clinic, and had a minimum of one year follow-up duration. Data collected from medical cords. For evaluation of expression, immunohistochemistry was performed on paraffm-embedded tissue sections using a monoclonal antibody for androgen, estrogen and progesterone receptors. Presence of at least 10% positive cells defined as positive expression. Results: None of the control subjects showed AR expression, while 22% of the patients were AR-positive. ER/PR expressions were observed in 4.2%/ and 2.5% of the cases and in 2.3% and 1.5% of the controls, respectively. A statistically significant correlation was found between AR expression and tumor stage and grade (P<.001). AR-positive patients showed a significantly poorer prognosis than AR-negative cases (log-rank test, P=.02, hazard ratio = 2.12; 95% confidence interval: 1.36-4.65). Conclusion: AR expression was significantly associated with higher grade and poorly differentiated tumors with unfavorable outcome. AR expression test might be useful as a diagnostic tool for determining the malignancy and outcome of UBC patients.