Avicenna journal of medical biotechnology最新文献

Background: Biotechnology is a rapidly developing field, and Iran aims to enhance its position in biosimilars through improved educational frameworks. Successful nations, such as the U.S., UK, Switzerland, China, and India, exemplify the positive impact of targeted educational programs in producing skilled graduates who advance the biotech sector. Research indicates a significant connection between curriculum relevance and graduate employability, suggesting that comprehensive curriculum assessment and reform can enhance student success and faculty involvement. This study investigated the necessity of revising the Master's curriculum in Medical Biotechnology in Iran to align with industry demands and global advancements.

Methods: The current curriculum's effectiveness using the Delphi method was assessed to survey Master's students, professors, and industry stakeholders.

Results: Findings revealed that while the current curriculum moderately aligns with educational objectives, significant gaps exist in practical training and resource availability. Many students reported feeling inadequately prepared for employment, particularly in essential skills such as animal cell culture, vaccine and monoclonal antibody design and production, and human skills like effective communication and teamwork. To address these deficiencies, some new courses focusing on practical experience and interdisciplinary approaches were recommended. Recommendations included enhancing laboratory facilities, integrating internships, and adopting team-based learning methods to improve student engagement and skill acquisition.

Conclusion: Continuous investment in biotechnology education is crucial for maintaining competitive advantages in a globalized market. Overall, this research highlighted the importance of adapting educational programs to meet the dynamic needs of the biotechnology industry, ensuring graduates possess the necessary skills for successful careers in this field.

Background: Acute Respiratory Infections (ARIs) are a leading cause of childhood mortality worldwide, especially in African and Southeast Asian countries. Point of Care Test (POCT) techniques provide faster diagnoses compared to conventional or real-time PCR methods. Recombinase Polymerase Amplification (RPA) offers rapid on-site detection of these infections. Coupling RPA with Enzyme-Linked Immunosorbent Assay (ELISA) (RPA-ELISA) creates a cost-effective alternative, ideal for clinical applications. This study evaluates RPA-ELISA as a rapid diagnostic tool for bacterial respiratory infections.

Methods: From 11 August 2022 to 9 February 2023, respiratory samples were collected and processed using culture methods, biochemical tests, real-time PCR, and RPA assays. The RPA reactions were conducted at 39°C for 30 min, and ELISA was used for detection. Statistical analyses focused on sensitivity, specificity, Positive Predictive Values (PPV), and Negative Predictive Values (NPV).

Results: Forty-two respiratory samples, were collected in this period of which 10 samples showed no growth, and 32 tested positive. Among these positive samples, 15 isolates (35.7%) were identified as Klebsiella pneumoniae (K. pneumoniae), 14 isolates (33.3%) as Streptococcus pneumoniae (S. pneumoniae), and 3 isolates (7.1%) as Moraxella catarrhalis (M. catarrhalis). RPA-ELISA demonstrated 100% sensitivity for all pathogens, comparable to or better than RT-PCR, but had slightly lower specificity and PPV. RT-PCR achieved 100% specificity and PPV for all pathogens, indicating higher accuracy; yet, RPA-ELISA's sensitivity points to its effectiveness as a rapid screening tool.

Conclusion: RPA-ELISA is significantly faster than real-time PCR and culture methods. Its ease of use makes it suitable for on-site diagnoses in resource-limited environments. Limitations include a small sample size for certain bacteria and the necessity for further validation in varied clinical contexts.

Background: RfxCas13d, a key member of the Cas13 family, plays a vital role in CRISPR-based diagnostics for RNA sequence detection and gene silencing. This study aimed to enhance RfxCas13d expression by optimizing key parameters using Response Surface Methodology (RSM).

Methods: The plasmid pET28b-RfxCas13d-His (Addgene 141322) was introduced into BL21 (DE3) and Rosetta™ (DE3) strains. Initial expression tests were conducted, followed by RSM-guided optimization of factors such as isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration, temperature, cell density at induction, and induction time in BL21 (DE3). Protein expression levels were quantified using ImageJ and AlphaEaseFC software to analyze band intensities.

Results: BL21 (DE3) was selected for further optimization based on preliminary results. Analysis of 26 RSM-designed experiments revealed that temperature, induction time, IPTG concentration, and their interactions significantly influenced RfxCas13d expression. Optimal conditions were identified as 0.25 mM IPTG, an OD600 nm of 0.8 at induction, 37°C, and Overnight (ON) of induction. The regression model exhibited high accuracy, with a correlation coefficient of 0.97 and a p-value less than 0.05, confirming a strong linear relationship between predicted and observed values.

Conclusion: This study highlights the significant impact of the four optimized factors on RfxCas13d expression. Under optimized conditions, a soluble protein concentration of 3.6 mg/100 ml cell culture was achieved after purification. It represents the first application of RSM for optimizing RfxCas13d expression, providing a foundation for further refinement of expression conditions. Continued use of RSM in future research will enhance the efficiency of RfxCas13d production for diagnostic and therapeutic applications.

Background: Thyroid cancer is the most common type of cancer affecting the endocrine system. The main treatment approaches consist of surgical procedures and radioiodine therapy. Recently, there has been a heightened interest in investigating alternative treatment options, including probiotics, which could potentially minimize toxicity. Consequently, there is a growing imperative for research aimed at investigating the potential role of probiotics in the management of cancer.

Methods: The B-CPAP cell line was maintained in culture and tested with different dilutions of two bacterial strains. Toxicity evaluations were performed using the MTT assay to identify appropriate concentrations. mRNA was extracted and analyzed via real-time PCR to measure the expression levels of the Bcl-2, Bax, and P53 genes. Furthermore, changes in the cell cycle and the induction of apoptosis were examined using flow cytometry.

Results: The supernatant derived from Lacticaseibacillus casei (L. casei) - ATCC-393 and Lacticaseibacillus rhamnosus (L. rhamnosus)-GG demonstrated a significant inhibitory effect on the growth of B-CPAP cancer cells. The findings indicated that the combination produced a more pronounced anti-cancer effect by enhancing the expression of pro-apoptotic genes while reducing Bcl-2 gene expression in B-CPAP cells.

Conclusion: The findings indicated a notable change in the expression of genes associated with apoptosis and modifications in the cell cycle. This implies that probiotics may enhance the efficacy of chemotherapy in treating thyroid cancer. In particular, L. rhamnosus and L. casei may play a beneficial role in the therapeutic process. Further research is required to investigate the direct impact of probiotics on thyroid function.

Background: Circular RNAs (circRNAs) are important in tumorigenesis and cancer progression, highlighting their potential as biomarkers for diagnosis, prognosis, and treatment monitoring.

Methods: This study consists of experimental and in silico phases. In the experimental phase, the expression of hsa_circ_0052112 in tumor and blood samples from 40 breast cancer women was analyzed, compared to the control group using Sybr Green real-time RT-PCR followed by total RNA extraction and cDNA synthesis. Statistical analysis was performed using the beta-actin gene as a normalizer, compared to the normal control group as a fold change. In the in silico phase, interactions among circRNA, RNA-Binding Proteins (RBPs), and microRNAs (miRNAs) were investigated using Interactome Database. The miRcancer database was utilized to assess breast cancer-related miRNAs linked to hsa_circ_0052112. Target miRNAs were identified with TargetScan and filtered for relevance through DisGeNET. K-means clustering grouped genes by expression patterns, visualized in Cytoscape to illustrate circRNA-miRNA-mRNA relationships. Hub genes underwent pathway enrichment analysis using Reactome database to determine their functional significance.

Results: Data revealed a significant increase in hsa_circ_0052112 expression in both blood and tumour of breast cancer patients. This increase was especially pronounced in patients with estrogen, progesterone, and HER2 receptor positivity, as well as in advanced disease stages with lymph node involvement. Enrichment analysis of hub genes indicates their role in the PI3K/AKT signaling pathway.

Conclusion: hsa_circ_0052112 shows promise as a multifaceted biomarker for breast cancer, enhancing diagnosis and prognosis; while supporting personalized treatment strategies. Further clinical validation is necessary to confirm its utility.

Regenerative medicine is a field of pharmacy and medicine that focuses on stem cells and other methods such as nanoscience and biotechnology to stimulate the body's natural regenerative processes and repairing damaged tissues and organs to improve function and reduce pain. In this review article, focus is on Dental Stem Cells (DSC) and other cells regeneration in human body. The appropriateness of tissue-engineered therapies relying on the multipotent regenerative abilities of DSC is accompanied by significant challenges, as growth factors and epigenetic components are crucial for preserving their multipotency while being susceptible to a range of natural and environmental factors. Current evidence highlights the positive outcomes associated with select regenerative therapies; nevertheless, to provide further support, additional data must be gathered through standardized therapies and further studies. Organoids (3D cell culture) and nano scaffolds are also being explored as potential tools for regenerative therapies. Understanding the mechanisms that determine the behavior of these cells and how they interact will enable future generation therapies. Demonstrating promise, cell therapy is an alternative approach within regenerative medicine. Developmental factors like extracellular vesicle production are thought to mediate the regenerative response through paracrine effects in cell therapy, which is widely recognized.

Non-Alcoholic Fatty Liver Disease (NAFLD) is a spectrum of liver diseases from simple steatosis to the most severe form of hepatocellular carcinoma. Liver injuries resulting from various factors, including viral infections, alcohol consumption, and metabolic disorders, trigger the activation of resident immune cells and the recruitment of circulating immune cells to the liver. This chronic inflammatory environment leads to tissue damage and the progression of liver fibrosis. Macrophages are highly versatile immune cells that play a dual role in fibrosis: they contribute to the progression of fibrosis (M1 and Ly6c^high macrophages) and its resolution (M2 and Ly6c^low macrophages). M1 macrophages and those with high surface expression of Ly6C exhibit pro-inflammatory characteristics, while M2 macrophages and myeloid cells with low expression of Ly6C mitigate inflammation and inhibit fibrosis progression. Environmental stimuli influence the complex mechanisms hepatic macrophages regulate the fibrosis they encounter. Kupffer cells initiate the inflammatory cascade and recruit monocyte-derived macrophages, which modulate the propagation of fibrosis and promote fibrinolysis. Additionally, hepatic macrophages interact with other cell types through exosomes, facilitating the transfer of cellular components that influence the outcome of liver fibrosis. In this review, the critical role of macrophages in inflammation-induced fibrosis and tissue restoration is discussed.

Background: Teucrium persicum (T. persicum) is a well-known Iranian endemic plant that grows in the southern regions of Iran. It is used as a tea to treat abdominal pains, hyperlipidemia, and diabetes in traditional Iranian medicine. It has been previously found that the methanolic extract of T. persicum exerts significant cytotoxicity and inhibitory effects on different cancer cells. This study aimed to investigate the effects of ethyl acetate extract of T. persicum on MCF-7 cells.

Methods: The experiments included MTT, DAPI staining, and investigating the expression of BAX and BCL2 genes. The extract had a significant cytotoxic effect on MCF-7 cells, with an IC₅₀ value of 50 μg/ml for 48 hr.

Results: DAPI staining assays showed that the extract induced morphological changes, chromatin condensation, and nuclear fragmentation. Additionally, the ethyl acetate extract induced the expression of BAX and down-regulated BCL2 genes.

Conclusion: These findings suggest that T. persicum has strong cytotoxic properties and warrants further investigation.