farahnaz zare

 This research investigated the development of short hairpin RNA (shRNA) molecules designed to target specific regions of the human respiratory syncytial virus (HRSV) M and F genes. The study aimed to assess the therapeutic potential of these shRNAs and evaluate the effectiveness of Tat peptide-mediated delivery in enhancing their functionality.

 We acquired isolates from pediatric patients experiencing respiratory illness then cultured in HEp-2 cells. We constructed plasmids expressing shRNAs. Tat peptide as a facilitator for shRNA plasmid delivery was used. The cytotoxicity of ribavirin, shRNA constructs, and control agents was assessed using the MTT assay. The transfection efficiency of Tat peptide-mediated shRNA delivery with that of lipofectamine 3000™ were compared. Finally, real-time PCR was employed to quantify HRSV replication in the treated cells.

 Tat peptide-mediated delivery of shRNA plasmids significantly suppressed the expression of the M and F genes of HRSV compared to lipofectamine 3000™. This suppression was evident in both short-term experiments and scenarios involving stable shRNA expression. Furthermore, the combination of ribavirin with shRNA treatment resulted in a substantial reduction in viral load. Notably, the most pronounced antiviral effect was observed when both shRNAs were employed simultaneously.

 Our findings suggest that Tat peptide-mediated delivery of shRNA plasmids holds significant potential for achieving stable suppression of HRSV genes. This approach warrants further investigation as a potential gene therapy strategy for HRSV. By demonstrating promising results in vitro, this study highlights the need for future in vivo studies to comprehensively evaluate the therapeutic potential of this approach in a clinical setting.

Chemotherapy, the primary treatment for acute lymphoblastic leukemia (ALL), often yields inadequate responses. Epigallocatechin gallate (EGCG) has been shown to significantly affect tumor cells through various mechanisms, including cell cycle arrest, apoptosis, and autophagy.

The objective of this study is to explore the impact of EGCG on autophagy, apoptosis, and the interplay between them in NALM-6, a pre-B-ALL cell line.

NALM-6 cells were subjected to various concentrations of EGCG for 24 and 48 hours. Additionally, NH4Cl 10 mM was used as an autophagy inhibitor to examine this mechanism. The EGCG effect on cell viability and apoptosis was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), trypan blue exclusion assay, and flow cytometry. Moreover, western blot analysis and real-time PCR were performed to investigate autophagy.

Our findings demonstrated that EGCG significantly affected cell proliferation and viability. It reduced cell viability by 55.2 ± 8.4% (P < 0.0001) while inducing apoptosis by 51 ± 1.9% (P = 0.006). Furthermore, in the presence of NH4Cl, EGCG led to a 3.9 ± 1.8-fold increase in LC3 protein level (P = 0.001). It also resulted in an approximately 1.34 ± 0.34-fold enhancement in DRAM1 mRNA expression levels (P = 0.013), while reducing of LC3B by 33.3 ± 30.5% (P = 0.008), P62/SQSTM1 by 46.5 ± 28.3% (P < 0.001), and Atg2B by 45.5 ± 16.3% (P < 0.001). However, the inhibition of autophagy did not alter the apoptosis rate in either untreated or EGCG-treated cells.

Overall, our findings suggest that EGCG can trigger apoptosis and autophagy in NALM-6 cell line. However, blockage of autophagy does not appear to impact apoptosis in this cell line.

The behavioral intentions of a Muslim person in different dimensions, including the intention to buy products, are influenced by Islamic values, and Islamic values are also formed based on Islamic beliefs; Considering the growing number of Muslim consumers around the world as the target market of businesses, this research has been conducted with the aim of analyzing the action dimension of Muslim consumer's purchase intention. The paradigm that governs this research is interpretive and exploratory research based on qualitative content analysis through interviews with experts in the field and university to conceptualize data and analyze information with open, central and selective coding to explain the active nature of Muslim consumer's purchase intention. Paid. The analysis of the text of the interviews yielded 60 codes in the form of 12 general categories indicating the action dimension of the Muslim consumer's purchase intention and is a guide for business managers who have chosen Muslim consumers as the target market and the path to achieving their marketing goals. based on strengthening the consumer's purchase intention through advertising and effective communication.

Antigen presentation using bacterial surface display systems, on one hand, has the benefits of bacterial carriers, including low-cost production and ease of manipulation. On the other hand, the bacteria can help in stimulating the immune system as an adjuvant. For example, using bacterial surface display technology, we developed a hepatitis C virus (HCV) multiple antigens displaying bacteria's surface and then turned it into a bacterial ghost.

The HCV core and NS3 proteins' conserved epitopes were cloned into the AIDA gene plasmid as an auto transporter. The recombinant plasmid was then transformed into Escherichia coli (E. coli) Bl21 (DE3). Recombinant bacteria were then turned into a bacterial ghost, an empty cell envelope. Whole-cell ELISA, flow cytometry, and Western blot techniques were used for monitoring the expression of proteins on the surface of bacteria.

A fusion protein of HCV core-NS3-AIDA was successfully expressed on the E. coli Bl21 (DE3) surface and confirmed by western blotting, Enzyme-Linked Immunosorbent Assay (ELISA), and flow cytometry detection techniques.

The presence of HCV antigens on non-pathogen bacteria surfaces holds promise for developing safe and cost-benefit-accessible vaccines with optimal intrinsic adjuvant effects and exposure of heterologous antigens to the immune system.

In various cancers, Ganoderic Acid A (GAA), an active triterpenoid derived from Ganoderma lucidum, has been proved to show potent anti-tumor effects. However, the possible impacts of GAA on the human leukemia cell line (Nalm-6) are not fully elucidated. Therefore, this research aimed to study the antineoplastic effect of GAA on Nalm-6 cells.

In this laboratory trial study, Nalm6 cells were cultured in vitro and treated with different doses of GAA (25, 50, 100, 200, and 400 μg/mL) for 24, 48, and 72 hours. The optimal treatment concentration of GAA was determined by the MTT assay. Flow cytometry was used to determine the death of Nalm-6 cells caused by GAA treatment by utilizing FITC-conjugated propidium iodide (PI) and annexin V staining. After incubation, the expression levels of miR-17-5p and miR-181b were monitored using real-time polymerase chain reaction (PCR).

Based on the half-maximal inhibitory concentration (IC50) measurements of the MTT assay, the optimal treatment concentration of GAA was 140 μg/mL (in a dose and time-dependent manner, p<0.0001). The GAA treatment was selectively toxic to the leukemia Nalm-6 cells and could remarkably induce cell apoptosis (p<0.0001). Besides, GAA downregulated the expression of miR-17-5p and miR-181b in the Nalm-6 cells compared with the untreated cells (P=0.0067 and P=0.0014, respectively).

Based on the present findings, GAA merits further investigation as a promising natural reagent for treating hematologic malignancies.

Short hairpin RNA (shRNA) has proven to be a powerful tool to study genes’ function through RNA interference mechanism. Three different methods have been used in previous studies to produce shRNA expression vectors including oligonucleotide-based cloning, polymerase chain reaction (PCR)-based cloning, and primer extension PCR approaches. The aim of this study was designing a reliable and simple method according to the primer extension strategy for constructing four shRNA vectors in order to target different regions of Metadherin (MTDH) mRNA in human leukemic cell line Jurkat.

Oligonucleotides for construction of four shRNA vectors were designed, synthesized and fused to U6 promoter. Each U6-shRNA cassette was cloned into a pGFP-V-RS vector. MTDH shRNAs were transfected into the Jurkat cell line by using the electroporation method. The ability of shRNAs to knock down MTDH mRNA was analyzed through qRT-PCR. Apoptosis assay was used to evaluate the effect of down regulation of MTDH expression on cell integrity.

A significant reduction (about 80%) in the expression levels of MTDH mRNA and an increase in the percentages of apoptotic cells (about 20%) were observed in the test group in comparison with control.

MTDH shRNA constructs effectively inhibited gene expression. However, simplicity and inexpensiveness of the method were additional advantages for its application.

Ionizing radiation plays a significant role in cancer treatment. Despite recent advances in radiotherapy approaches, the existence of irradiation-resistant cancer cells is still a noteworthy challenge. Therefore, developing novel therapeutic approaches are still warranted in order to increase the sensitivity of tumor cells to radiation. Many types of research rely on the role of mitochondria in radiation protection.

Here, we aimed to target the mitochondria of monocyticleukemia (THP-1) radio-resistant cell line cells by a mitochondrial disrupting peptide, D (KLAKLAK)2, and investigate the synergistic effect of Gamma-irradiation and KLA for tumor cells inhibition in vitro.

In this experimental study, KLA was delivered into THP-1 cells using a Cell-Penetrating Peptide (CPP).The cells were then exposed to gamma-ray radiation both in the presence and absence of KLA conjugated with CPP. The impacts of KLA, ionizing radiation or combination of both were then evaluated on the cell proliferation and apoptosis of THP-1 cells using MTT assay and flow cytometry, respectively.

The MTT assay indicated the anti-proliferative effects of combined D (KLAKLAK)2 peptide with ionizing radiation on THP-1cells. Moreover, synergetic effects of KLA and ionizing radiation reduced cell viability and consequently enhanced cell apoptosis.

Using KLA peptide in combination with ionizing irradiation increases the anticancer effects of radio-resistant THP-1 cells. Therefore, the combinational therapy of (KLAKLAK)2 and radiation is a promising strategy for cancer treatment the in future.

Despite all the efforts for discovery of efficient anti-cancer therapeutics, cancer is stilla major health concern worldwide. p28 is a bacterial small peptide which has been widelyinvestigated due to its preferential cell internalization and anti-cancer activities. Intracellularly,p28 offers its anti-cancer traits by impeding the degradation of tumor-suppressor protein "p53".In this study, we investigated the potency of p28 in inducing apoptosis or decreasing cellviability in p53-null "HeLa" cell line.

The coding sequence for p28 peptide was obtained from Pseudomonas aeruginosaby PCR amplification of the p28 gene. The coding gene was cloned in pET-28a vector andtransformed into E. coli bacterial host. Subsequently, the expressed peptide was purified usingNi-NTA chromatography system and introduced into the target cells. The anti-proliferative andapoptotic activity of p28 on HeLa and HEK-293 cells were investigated using MTT and PEAnnexinV Flowcytometry assays.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Westernblotting confirmed the expression of p28 peptide in the bacterial host. Bradford assay revealeda concentration of 0.05 mg/mL for the purified p28. MTT assay of cells treated with p28 atconcentrations of 0, 0.5, 1, 2 and 2.5 μM indicated 24h viability values of 97%, 89%, 88%,87% and 84% for HeLa cells, respectively. Data obtained from flowcytometry analyses revealed24h apoptosis rate of 7.17%, 8.05%, 8.63% and 8.84% for HeLa cells treated with 0, 0.5, 1,and 2 μM p28, respectively.

MTT and flowcytometry apoptosis assays suggest no statistically significant effectof p28 on the viability and apoptosis status of p53-null HeLa cells when results compared todata obtained from HEK-293 cells (P > 0.05). These results imply that anti-cancer efficacy of p28is directly dependent on the presence of p53, suggesting p28 as an inappropriate therapeuticagent for treatment of cancers with negative p53 status.

The p28 is a small-sized cell-penetrating peptide derived from bacterial protein azurin and can function as a cancer-specific anti-proliferative agent. It can penetrate cancer tissues easily without involving the immune system, and increase the intracellular concentration of p53. In this study, we have expressed and purified recombinant p28, then evaluated its anti-proliferative and pro-apoptotic effects on Raji cancer cell line. The p28 gene was amplified and cloned into pTZ57R cloning vector and was sequenced subsequently. Afterward, it was transformed into E. coli BL21 bacterial host by using pET-28a expression vector. Peptide purification was carried out using Ni-NTA ‎chromatography system. Bradford, SDS-PAGE, and western blotting assays were applied to assess the concentration and expression level of the recombinant peptide. The proficiency ‎of p28 in inhibition of tumor growth and induction of apoptosis in cancerous cells was investigated by evaluating the Raji and HEK-293 cells treated with different concentrations of p28. The overexpression of the p28 peptide in the bacterial host was confirmed by SDS-PAGE and western blotting. Moreover, Bradford assay revealed desirable concentrations of the recombinant p28 before and after dialysis. The MTT and PE-Annexin V apoptosis assays indicated the specific function of p28 in impeding the proliferation of cancerous cells and triggering the apoptosis. The p28 induces apoptosis in cancerous cells but not in normal control cells. ‎In summary, p28 is a non-immunogenic small peptide that can penetrate cancerous cells ‎preferentially, impede the cell proliferation, and induce the apoptosis. Overall, these ‎findings suggest p28 as‎ a promising anticancer drug.

Selective therapy has always been the main challenge in cancer treatments. Recently, it has been shown that Human Gyrovirus-derived protein apoptin (HGV-Apoptin) has selective cytotoxic effects on cancer cells similar to its homologue, Chicken Anemia Virus-derived Apoptin (CAV-Apoptin). However, apoptotic effects of Human Gyrovirus apoptin have been only evaluated on a few cancerous cell lines and need to be further investigated. In this study, we have evaluated the apoptotic effects of HGV-Apoptin and CAV-Apoptin expression on lung cancer (A549) and normal (HEK-293) cell lines, in order to provide more information about the specificity of these proteins on cancerous cells. Target cells were transfected by the calcium-phosphate precipitation method with constructed plasmids expressing HGV-Apoptin and CAV-Apoptin proteins as well as the control plasmid. Transfection efficiency was followed and imaged by fluorescence microscopy. Quantification of apoptosis was performed by flow cytometry. Measurements were compared by paired Student t-test. Cells were successfully transfected with control and constructed plasmids. Flowcytometry analysis showed that A549 cells transfected with HGV-Apoptin and CAV-Apoptin expressing plasmids, undergone the apoptosis compared to A549 cells transfected with control plasmid (P<0.001). None of the plasmids could induce apoptosis in HEK-293 cells. Human Gyrovirus-derived apoptin (HGV-Apoptin) similar to its homologue, chicken anemia virus derived Apoptin (CAV-Apoptin) can induce apoptosis in Non-small-cell lung carcinoma cell line A549, but not in normal human embryonic kidney cell line HEK-293, which can be introduced as a promising novel specific antitumor agent.

There are few studies indicating the post-neonatal HBV vaccination level of anti-HBs antibody in first-year enrolled university students in Iran. In addition, anti- HBc antibody detection, which is a good indicator of virus exposure, has not been reported in vaccines. Hence, this study was conducted to determine the level of anti-HBs and anti-HBc antibodies in the serum sample of medical laboratory students who had received primary infantile HBV vaccination.
This study was conducted on first-year students enrolled in the department of laboratory sciences at Shiraz University of Medical Sciences, Iran. For determining anti-HBs and anti-HBc titers, 5 mL of venous blood was aseptically collected. Anti-HBs and anti-HBc antibody levels were determined by enzyme-linked immunosorbent assay. HBV DNA was also performed on DNA extracted from individuals positive for an anti-HBc antibody test.
Of the 257 vaccinated individuals (188 females and 69 males) who participated in this study, 36.2% showed a non-protective anti-HBs response (anti-HBs Our results indicate that a substantial number of our study population vaccinated against HBV during childhood showed non-protective anti-HBs antibody level. Therefore, a booster dose of vaccine needs to be scheduled for students with anti-HBs level