Cell and Tissue Journal

Abstract Aim: Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. This enzyme catalyses the first step of flavonoid biosynthesis pathway. The purpose of this study was to identify chalcone synthase gene in Calotropis procera.
Material and Methods: The genomic DNA was extracted from fresh leaves and used as template in PCR. Primers were designed according to the conserved regions of this gene in other plants. The PCR product was purified and sequenced. Multiple sequence alignment and sequence homology analyses were carried out with DNAMAN software using CHS sequences retrieved from Genbank. A neighbor-joining phylogenetic tree based on CpCHS and other CHS sequences were constructed using MEGA6 software.
Results: PCR results indicated the existence of this gene and amplification of 570 nucleotides fragment that belong to exon2 of CHS gene. This gene was denominated as CpCHS and deposited at the GenBank accession KM878672. Comparison analysis based on nucleotide sequence alignment revealed that CpCHS shared a high degree of similarity to CHS from Arabidopsis thaliana (59%), Nicotiana tabacum (63%), Olea europaea (63%) and Petunia X hybrida (64%). Maximum similarity was observed between CpCHS and CHS from Catharanthus roseus (67%). Phylogenetic analysis showed that CpCHS was grouped into a branch with Catharanthus roseus.
Conclusion: These results suggest that CpCHS may play a similar role as other CHS in regulation of flavonoids biosynthesis pathway. Identification of this gene is an effective step which may lead to increase in the accumulation of useful medicinal extract in this plant.

Abstract Aim: In this study, we explored the effect of epigenetic modifications on planarian regeneration process by treating with small molecule epigenetic modifiers
Material and Methods: Planaria worms were cut into 3 sections including head, trunk, and tail regions and treated with 6 small molecules with epigenetic modification effects. All samples were evaluated morphologically for 20 days to monitor the effects on regeneration process.
Results: Morphological analysis of samples treated with selected six small molecules with different epigenetic modification functions revealed that treating with two of them including Sodium butyrate and Valporic acid will result in delayed head, trunk, and tail fragments regeneration, and delayed eye formation in trunk and tail fragments, respectively. In addition, samples treated with other four small molecules including BIX01294, RG108, SAHA, and Tranylcypromine with other epigenetic modification functions, regenerated normally.
Conclusion: These results indicating that different epigenetic levels and activity of histone deacetylases play a key role in planarian regeneration and inhibition of histone deacetylases activity will result in abnormal regeneration. However, further work is needed to determine the exact mechanism of histone deacetylases activity effect on planarian regeneration process. Finally, we hope to understand more about the underlying mechanism of planarian and vertebrate’s regeneration inhibition and activation using these findings.

Abstract Aim: The aim of the current study was to isolate microglial cells from neonatal rat brain, activate the cells using lipopolysaccharide (LPS) and examine the effect of the inflammatory factors that they express on dopaminergic (DAergic) neurons.
Materials and Methods: mixed glial cells were isolated from 1-3 day rat neonatal brains and then microglial cells were extracted from them. Upon treatment of the cells with LPS, their conditioned media (CM) were collected. DAergic SH-SY5Y cell line was seeded in 96-well plates and fed with the collected CM. The rate of viability and apoptosis of the neuronal cells was examined using MTT and apoptosis tests and the data were analyzed statistically.
Results: Ten to thirteen days after isolation, mixed glial cells were composed of three types: astroglia, oligodendrocytes and microglia. Microglia were floating while semi-attached to the surface. Twenty four hours post-isolation, these cells were transferred to and cultured in separate dishes where they formed spindled and ramified phenotypes. At this stage, the cells were treated with LPS to be activated. This activation was demonstrated by morphological changes from spindle-like form to amoeboid form, detection of increase in NO expression using Griess test and induction of inflammatory iNOS and TNF-α gene expression. Also teatment of SH-SY5Y cell line with conditioned medium of activated microglia led to both apoptotic and necrotic cell death.
Conclusion: LPS-mediated activation of microglia results in overexpression of neuroinflammatory markers. The overproduction of these markers results in both apoptotic and necrotic cell death amongst DAergic neurons via neuroinflammation.

Abstract Aim: In this study, the effect of salinity stress on quantitative gene expression of sucrose synthase1, relative water content and the uptake ratio of sodium to potassium in the leaves of wheat were evaluated.
Material and Methods: Wheat seedlings treated with 100 and 200 mM sodium chloride and at times zero, 6, 12, 24 and 36 hours after treatment relative water content, the uptake ratio of sodium to potassium in the leaves and quantitative gene expression of Sus1 by qRT-PCR method were studied. 
Results: Results showed a significant decrease in relative water content, increase in the uptake ratio of sodium to potassium by leaves and increase in the relative expression of sucrose synthase1 gene at treated plants compared to control group.
Conclusion: Generally, increase in the expression of sucrose synthase 1 gene could be one of the possible mechanisms in the process of tolerance to salinity stress.

Abstract Aim: The goal of the present research was evaluation of the effects of industrial area air pollution of Arak Aluminum Company on Robinia pseudoacacia (acacia) and tree-of-heaven anatomical changes.
Material and methods: Information from the Environmental Protection Agency of Markazi province, Aluminum park as the infected area and Haftad-Ghole in 35 kilometers of Arak as the area clean were used. Leaf samples of Robinia and Alianthus were harvested in August 2013 simultaneously. Density and length of trichomes, stomatal density and opening, length of the palisade cells and diameter of the spongy cells of leaf were measured. Data analysis were performed by SPSS 16 and Excel software and means comparison by T-Test.
Results: Under air pollution, trichome density in abaxial epidermis and trichome length in both surfaces of epidermis of acacia leaf increased significantly. Also in tree-of-heaven leaf, trichome density decreased and trichome length increased on abaxial epidermis significantly. Stomatal density on abaxial epidermis of acasia and tree-of-heaven leaves exposed to air pollution decreased and increased, respectively. Stomatal opening of Robinia and Ailanthus leaves had no significant effect. Length of the palisade cells of leaf in infected area decreased in acacia and increased in tree-of-heaven as compared with clean area, diameter of the spongy cells had no significant change in acacia leaf but decreased in tree-of-heaven leaf.
Conclusion: Results showed the anatomical changes in both plants for further compromise with regional air pollution.

Abstract Aim: This study aimed to investigate the effect of Bisphenol­ A on the cell viability, morphologic changes and induction of apoptosis in bone marrow mesenchymal stem cells of an adult rat. Material and Methods: The bone marrow mesenchymal stem cells were extracted using flashing-out method. At the end of the third passage, cells were divided into 11 groups of control and experimental. Experimental cells were treated with the different doses of Bisphenol­ A (1, 5, 10, 50, 100, 250, 500, 1000, 2000 and 4000 nM) for four periods of 5, 10, 15 and 21 days in the osteogenic media containing 10% of fetal bovine serum. The cell viability, DNA damage, expression of genes and the morphologic changes of the cells were then investigated during the procedure of osteogenesis. The study data was analyzed using one-way ANOVA and T-Test setting the significant P value at p < strong>Results: Within Bisphenol­ A treated cells, the mean viability, the mean of nuclei diameter and the expression of anti-apoptotic Bcl-2 gene of the mesenchymal stem cells treated with Bisphenol­ A significantly decreased (p < 0.05), compared to the control group. In addition the DNA damage and the expression of apoptotic Bax gene of the cells treated with Bisphenol­ A significantly increased, compared to the control group (p < 0.05). Conclusion: Our data suggest that BPA decreases cell viability and induces apoptosis in mesenchymal stem cells, in a dose dependant manner.

Abstract Aim: The aim of this study is to compare sperm parameters (concentration, motility and morphology), DNA damage, percentage and intensity of ROS (Reactive Oxygen Species) and relationship between these parameters in the fertile and infertile men with varicocele.
Material and Methods: In this study, the semen samples from 83 individuals with varicocele and 32 fertile men were studied. Sperm parameters according WHO (World Health Organization) guidelines, protamine deficiency (chromomycin A3 staining), sperm DNA damage (TUNEL assay), percentage and intensity of ROS (DCFH-DA staining) were assessed in fertile and infertile men with varicocele.
Results: The results of this study showed that sperm concentration, percentage of motility and normal morphology were significantly lower in individuals with varicocele compared to fertile individuals. In addition, percentage of sperm DNA damage, protamine deficiency and ROS positive were significantly higher in infertile men with varicocele.
Conclusion: The increase of stress oxidative and testicular temperature in infertile men with varicocele, lead to decrease the quality of sperm parameters as well as negative effects on sperm chromatin integrity. Therefore the process of spermatogenesis and the rate of fertility were affected in these individuals.

Abstract Aim: The aim of this study was to assess side effects of clinical dose of busulfan on testis and epididymal sperm of adult male mouse. Material and Methods: Male adult NMRI mice (25-35 g) were divided into two groups of sixteen each. Control and busulfan treated group were administered 100 μL DMSO (Dimethyl Sulfoxide) and 3.2 mg/kg busulfan for 4 days, respectively. The animals were sacrificed 35 days after starting treatment. The histological change on germinal epithelium, sperm quality parameters (count and normal morphology), viability and DNA fragmentation on sperm were analyzed by light microscopy, CASA (Computer–aided Sperm Analyzer), MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5 Diphenyltetrazolium Bromide) and TUNEL assays, respectively. Results: Busulfan administration significantly decreased parameters of sperm (count and normal morphology) and increased germinal epithelium destruction. The head, mid piece and tail abnormalities of treated group were increased significantly versus control. The lower levels of MTT in treated group were significant compared with control group. Higher levels of TUNEL positive cells that were found in treated groups demonstrated the increasing of DNA fragmentation in sperms following busulfan treatment. Conclusion: The results showed that clinical dose of busufan induces genotoxic, pre-apoptotic and cytotoxic effects on spermatogenesis. Also, busulfan can increase cytotoxicity on testis tissue of adult male mouse.

Abstract Aim: The aim of the present study was to investigate the effect of lead concentrations on the growth, antocyanin, chlorophyll, carotenoid, total soluble sugars, reducing sugars and free amino acids of harmal (Peganum harmala L.) populations.  
Material and methods: Plants of two different Peganum harmala populations (metallicolous and non metallicolous) were grown in hydroponic condition. Then plants were exposed to 0, 5, 10, 25 and 50 ppm Pb for 14 days. Finally, parameters were measured and calculated by spectrophotometer.
Results: Results showed that the increase of Pb concentrations in the nutrient solution reduced root length and there were significant difference between two populations. Chlorophylls and carotenoid pigments were generally decreased by Pb concentrations, especially under the highest Pb treatment (50 ppm) in both populations. Anthocyanin, total soluble sugars and free amino acids were increased in both populations.
Conclusion: The higher amounts of anthocyanin, total soluble sugars and free amino acids in metallicolous population reflecting the higher degree of tolerance to Pb in this population.

Abstract Aim: The aim of this study was to determine the effect of Carob seedextract as natural antioxidant on quality cryopreserved Farahani rams breeding sperm.
Material and Methods: In this study semen was collected from five mature ram (weight: 60±5 kg) twice a week using an artificial vagina and the ejaculates were pooled in order to eliminate the individual effect of rams. Different levels of carob seed extracts (0, 0/05, 0/1, 0/15 and 0/2 mL) were added to diluent based tris-egg yolk. After cooling, filling and sealing of the samples, they were frozen and with nitrogen vapor and immersed in liquid nitrogen and were stored until evaluation time. Thereafter and after thawing and incubation for5 min, sperm quality parameters includingmotility, progressive motility, viability (Nigrosine–eosin staining), membrane integrity with Hypoosmotic (Host) and morphology abnormality (Hancock test).
Results: Results indicated that the level of 0/05 mL carob seedextract significantly improved some parameters including motility, viability, plasma membrane and morphology integrity compared to control (p < 0.01).
Conclusion: Results indicated that so added 0/05 of carob seed extract peel to Tris based extender was beneficial in storage in sperm Farahani ram breeding after freeze-thawing.
 

Abstract

Abstract Aim: The aim of this study was the effect of topotecan on the activity of nitric oxide synthase in the tumor cell line Hela in comparison with the normal Hek cell line.
Material and Methods: For this purpose, Hela and Hek cells were randomly divided into the control groups and treatment groups exposed to 7.8, 15.6,31.25, 62.5,125, and 250 µg/ml of topotecan for 24,48,72 hr. Then, the data analysis of plate supernatant was evaluated the number of live cells using the MTT method, grease reaction as well as Real-Time PCR method and the data were compared using the one-way ANOVA statistical method between groups.
Results: The viability of Hela cells exposed to 250, 125, and 61.5 µg/ml of topotecan significantly decreased in comparison to the control group in 24, 48, and 72 h (p≤0.05). Also, the level of nitric oxide in Hela cells that were exposed to 250 µg/ml drug increased ½ times in comparison to the control group.
Conclusion: Topotecan has an inhibitory effect on cervical cancer cells. Also, topotecan increased the level of nitric oxide in Hela cells and this amount of nitric oxide kills the cells in cervical cancer cells.

Abstract Aim: Oxidative stress is an imbalance between oxidants and antioxidants at the cellular level which leads to infertility in males. For several decades, reactive oxygen species have been known as destructive and damaging agents to cells and tissues. Cells produce small and controlled amounts of ROS to control their physiological activities. Under normal conditions, ROS produced in semen are continuously deactivated by antioxidants in semen, Unsaturated fatty acid chains in sperm plasma membrane are vulnerable to oxidative stress conditions, and thus spermatozoa depend on extracellular antioxidant systems to overcome oxidative stress conditions. Another reason for creating oxidative stress conditions for spermatozoa, in addition to the low level of antioxidants in semen, is excessive production of ROS by spermatozoa with abnormal morphology. so one of the reasons for the creation of oxidative stress in semen is due to the imbalance between ROS production and its inactivation by antioxidants. Exposure to high concentrations of ROS causes disruption of the mitochondrial membrane, plasma membrane, and also chromosome fragmentation, which reduces sperm motility and viability. Increased formation of ROS is associated with decreased sperm motility. There is a possibility that the increase in ROS production will ultimately cause a decrease in the phosphorylation of axonemic proteins and sperm motility. This condition leads to a decrease in fluidity of the membrane, which in turn is necessary for sperm-oocyte fusion. The use of antioxidants such as silymarin can prevent the effects of oxidative stress.Milk thistle is the most well-known English common name for this species and other names include Holy thistle, Mary thistle, St. Mary’s thistle, Marian thistle, Lady’s thistle, Christ’s crown, Venus thistle, Heal thistle, Variegated thistle, Pig leaves, Royal thistle, Snake milk, Sow thistle, and Wild artichoke. The milk thistle medicinal plant is widely used in the traditional medicine of China and most European countries in the treatment of liver and biliary disorders. The seed extract of this medicinal plant, which is known as silymarin, protects the liver against a variety of poisonings. Silymarin contains a collection of flavonoids and other compounds with antioxidant, anti-inflammatory and cellular glutathione-increasing properties. Among the various flavonoid compounds found in the Silybum marianum, are silybin, silychristian and silydianin, which are collectively called silymarin. In many cases, the antioxidant properties of Silymarin are considered to be responsible for its protective actions. Oxidative stress-induced apoptosis in spermatozoa may lead to male infertility. Environmental pollutants and heavy metals cause harmful effects on the reproductive system and sperm parameters through the induction of oxidative stress. Silymarin, as a potent antioxidant, is able to inhibit oxidative stress. Silymarin with its antioxidant properties can have reduced the effects of compounds such as; aluminum, cadmium, lithium, nickel, benzopyrene, doxycycline, tetracarbon chloride, nicotine, methotrexate, arsenic and lead acetate on the sperm. And improve the damaged sperm parameters resulting from the mentioned compounds. Silymarin is also effective in reducing the harmful effects of varicocele and radiation therapy on sperm parameters.
 
 
 

Abstract Aim: The field of targeted drug delivery is a rapidly advancing area of pharmaceutical research that focuses on optimizing the efficacy and safety of therapeutic agents. The primary goal is to enhance the concentration of a drug at its desired site of action while minimizing its presence in non-target areas. This selective accumulation not only improves the therapeutic outcomes but also significantly reduces the adverse effects often associated with drug therapies. By employing various carriers and mechanisms, targeted delivery systems can exert precise control over the rate and duration of drug release. This is particularly beneficial in chronic treatments where maintaining therapeutic drug levels is crucial. Moreover, the reduction in systemic exposure allows for lower dosages, which inversely correlates with increased bioavailability and drug effectiveness. Consequently, this leads to decreased healthcare costs and less frequent dosing schedules, improving patient compliance and quality of life. One innovative approach in this field is the utilization of nanotopography, which involves the replication of natural surface patterns at the nanometer scale onto drug carrier systems. These nature-inspired designs are not merely aesthetic; they play a functional role in modulating the interactions between the drug delivery vehicle and the biological target. For instance, surfaces mimicking the intricate patterns found in natural environments can influence cellular behavior, enhance adhesion, or even provide antimicrobial properties. Exploring the nuances of nano-engineered surfaces, this scholarly work reviews how nature-inspired topographies impact drug delivery and efficacy in pharmaceutical applications. It discusses the various methods of fabricating nanoscale topographies and their implications for drug delivery systems. The review highlights how these microscopic landscapes can affect biological functions such as cell adhesion, tissue barrier dynamics, and drug absorption rates. Surface modification techniques at the nanoscale are diverse, ranging from simple chemical treatments to sophisticated lithography and microcasting. These methods allow for the creation of specific patterns that can be tailored to enhance the performance of drug carriers. For example, In the context of drug delivery systems, the use of virus-inspired particles has facilitated drug absorption. These bioinspired designs enhance therapeutic outcomes. The role of nanotopography in drug delivery is multifaceted. It can facilitate the formation of bioadhesive interfaces that ensure the drug remains at the site of action for extended periods. This is particularly useful in mucosal drug delivery, where rapid clearance by bodily fluids can be a challenge. Additionally, the rearrangement of cellular barriers can be induced by these topographies, allowing for more efficient drug penetration in areas like the blood-brain barrier. Antimicrobial interfaces created through nanotopography can prevent the colonization of pathogens on drug delivery devices, reducing the risk of infections. This is especially important in implantable devices where biofilm formation can lead to device failure and serious health complications. In a nutshell, the review article provides a comprehensive overview of how nature-inspired nanotopography can revolutionize the field of drug delivery. By mimicking the wisdom of natural designs, pharmaceutical scientists can develop more effective and safer drug delivery systems that promise to improve therapeutic outcomes and patient care.
 

Abstract Aim: Folate, also known as Vitamin B9, is a water-soluble nutrient present in natural food sources. It plays a crucial role in DNA synthesis and repair, as well as in methylation reactions. Folic acid (FA) is the synthetic form of folate found in enriched foods and supplements. The effects of folate deficiency after birth have not been extensively studied. MicroRNAs (miRNAs) are a type of non-coding RNAs that play essential roles in regulating gene expression. In recent years, miRNAs have been associated with various aspects of brain development, including neurogenesis, neuronal migration, axon and dendrite formation, and synaptogenesis. Additionally, altered expression and dysregulation of miRNAs have been linked to neurodevelopmental disorders. Methionine synthase (Mtr) is involved in amino acid metabolism and is responsible for converting the amino acid homocysteine to methionine. Mtr is crucial for brain development and is associated with myelin content. This project aimed to investigate the impact of maternal FA supplementation during pregnancy on the expression of mmu-miR-103-1-5p and Mtr in the cerebral cortex of mouse pups.
Materials and Methods: A total of 45 pregnant mice were divided into three groups, with 15 mice in each group. The first and second groups received FA at doses of 2 and 40 mg/kg of body weight daily by gavage throughout their entire pregnancy, respectively. The third group did not receive FA and served as the control. Subsequently, the cerebral cortex of the offspring was collected immediately after birth for the analysis of mmu-miR-103-1-5p and Mtr expression using real-time PCR. Statistical analysis was conducted using GraphPad Prism software.
Results: In the FA-treated group, the expression of mmu-miR-103-1-5p at a dose of 2 mg/kg body weight was 0.46±0.01, and at a dose of 40 mg/kg body weight, it was 0.27±0.02, whereas in the control group, it was 1.0007±0.03. Statistical analysis revealed a significant decrease in mmu-miR-103-1-5p expression at both the 2 mg/kg and 40 mg/kg doses compared to the controls (p

Abstract Introduction: Concern over fossil energy costs and environmental deterioration, along with energy security, has created a strong motivation for research and development of routes to provide sustainable, renewable fuels. In recent years, the use of biomass to produce highly valued chemicals has attracted widespread attention. lignocellulosic biomass, as a promising renewable resource for biofuel production, has distinct advantages in terms of economic and environmental aspects. The conversion of renewable raw materials to hydrocarbon fuels is an attractive alternative to fossil fuels from an economic and environmental point of view. The production process of lignocellulosic biomass mainly consists of biomass accumulation, biomass decomposition, simple sugars, and conversion of sugars to biofuel. One of the crucial steps in the economic success of lignocellulosic biofuels depends on the inhibition of competitive metabolism in microorganisms to achieve high productivity. To date, a growing focus is on the use of S. cerevisiae and E. coli as cell lines. These two cellular factories have the benefits that are well known.

Aim: The organic compound D-1,2,4-Butanetriol is a valuable chemical with wide-ranging applications in various fields. The chemical synthesis routes for BT have many drawbacks. By genetically modifying microorganisms, the metabolic pathway for producing many substances, including BT, can be engineered. The organic compound D-1,2,4-butanediol (BT) is an important intermediate chemical widely used in fields such as pharmaceuticals, paper, polymer materials, and military applications. When D-xylose sugar is provided to the bacterium, it is first converted to an intermediate compound called xylonolactone. This compound itself slowly converts into xylonate through a non-enzymatic reaction. To produce xylonate, the engineered bacteria have received xylose and initially, by a dehydrogenase reaction by the xylose dehydrogenase enzyme, that converts it into an intermediate substance: xylonolactone. The xylonolactone is converted slowly and in a nonenzymatic reaction to xylonate. Xylonate is a five-carbon organic acid. Over the past few years, xylonate has been increasingly being considered as an important chemical due to its potential as an important chemical component. Xylonate has many applications that can be used in the food, chemical, and pharmaceutical industries. Specifically, xylonate can act as a precursor for the synthesis of D-1,2,4-Butanetriol and a decrease in concrete water. E.coli due to fast growth in cheap culture medium, having two enzymes for the bto synthesis and product production in lower 24 hours of fermentation was chosen as the target strain of genetic engineering and metabolism. This study aimed to clone and express xylose dehydrogenase from Caulobacter vibrioides in E.coli. Materials and Methods: At first, to access the bacterial gene sequence, the genome of the target bacterium was extracted. Then, to create a strain expressing the enzymes xylose dehydrogenase and xylonolactonase, the genes for these proteins were amplified from Caulobacter vibrioides CB1 and transferred to E. coli. For this purpose, the target genes were amplified using specifically designed primers via the Polymerase Chain Reaction (PCR) method and initially cloned into a pTZ57cloning vector and then subcloned into pET 26b expression vector. At the final step, the expression of the enzyme was assessed by SDS-PAGE, and the other confirmation was the reduction of NAD+ to NADH, which was used as an activity indicator of the enzyme, as investigated by a change in NADH absorbance at 340 nm. Results: Confirmatory tests were performed to ensure the presence of the gene in the vectors (using restriction enzymes and colony PCR for gene amplification). The expression and activity of the enzyme were analysed. The recombinant protein's presence was confirmed by SDS-PAGE for the xylose dehydrogenase gene, with a molecular weight of 52.2 kDa. The estimated recombinant protein expression levels were approximately 25%. Conclusion: The objective of this research was solely to establish the metabolic pathway for xylonate production in E. coli by surface expression of enzymes in this pathway (xylose dehydrogenase). The results obtained in this study confirm that half of the pathway is active at the cell surface, but further experiments are required to determine the precise production levels and complete the pathway. This study aimed to create a metabolic pathway for producing xylonate in E.coli.

Abstract Introduction: Concerns about fossil energy costs, environmental deterioration, and energy security has created strong motivation for the research and development of routes to provide sustainable and renewable fuels. In recent years, the use of biomass to produce highly valued chemicals has attracted widespread attention. Lignocellulosic biomass, as a promising renewable resource for biofuel production, has distinct advantages in terms of economic and environmental benefits. The conversion of renewable raw materials to hydrocarbon fuels is an attractive alternative to fossil fuels from economic and environmental perspectives. The production process of lignocellulosic biomass mainly consists of biomass accumulation, biomass decomposition, simple sugars, and conversion of sugars to biofuel. One of the crucial steps for the economic success of lignocellulosic biofuels depends on the inhibition of competitive metabolism in microorganisms to achieve high productivity. To date, there has been a growing focus on the use of S. cerevisiae and E. coli as cell lines. These two cellular factories have well known advantages. They are genetically transmissible and several tools are available for genetic manipulation. In order to produce xylonate, the engineered xylose is first converted by a dehydrogenase into the intermediate xylonolactone, which is then slowly converted to xylonate in a nonenzymatic reaction.
Aim: The organic compound D-1,2,4-Butanetriol (BT) is a valuable chemical with wide-ranging applications in various fields such as pharmaceuticals, paper, polymer materials, and military applications. However, the chemical synthesis routes for BT have many drawbacks. By genetically modifying microorganisms, the metabolic pathway for producing many substances, including BT, can be engineered. When D-xylose is supplied to the bacterium, it is first converted into an intermediate compound called xylonolactone. This compound slowly converts into xylonate through a non-enzymatic reaction. To produce xylonate, the engineered bacteria receive xylose, which is initially converted by a dehydrogenase reaction catalysed by the xylose dehydrogenase enzyme into an intermediate compound, xylonolactone. Xylonolactone is slowly converted to xylonate in a nonenzymatic reaction. Xylonate is a five-carbon organic acid. Over the past few years, xylonate has increasingly been considered as an important chemical due to its potential as an important chemical component. Xylonate has many applications in the food, chemical, and pharmaceutical industries. Specifically, xylonate can act as a precursor for the synthesis of D-1,2,4-Butanetriol and as a concrete water reducing agent. E. coli was chosen as the target strain for genetic and metabolic engineering due to its fast growth in inexpensive culture media, the presence of two enzymes for BT synthesis, and product formation in less than 24 hours of fermentation. This study aimed to clone and express xylose dehydrogenase from Caulobacter vibrioides in E.coli.
Materials and Methods: At first, to access the bacterial gene sequence, the genome of the target bacterium was extracted. Then, to create a strain expressing the enzymes xylose dehydrogenase and xylonolactonase, the genes for these proteins were amplified from Caulobacter vibrioides CB1 and transferred into E. coli. For this purpose, the target genes were amplified using specifically designed primers via the Polymerase Chain Reaction (PCR) method and initially cloned into a pTZ57cloning vector and then subcloned into pET 26b expression vector. At the final step, the expression of the enzyme was assessed by SDS-PAGE, and the other confirmation was the reduction of NAD+ to NADH, which was used as an activity indicator of the enzyme, as investigated by a change in NADH absorbance at 340 nm.
Results: Confirmatory tests were performed to ensure the presence of the gene in the vectors (using restriction enzymes and colony PCR for gene amplification). The expression and activity of the enzyme were analyzed. The recombinant protein's presence was confirmed by SDS-PAGE for the xylose dehydrogenase gene, with a molecular weight of 52.2 kDa. The estimated expression level of the recombinant protein was approximately 25%.
Conclusion: The objective of this research was solely to establish the metabolic pathway for xylonate production in E. coli by surface expression of enzymes in this pathway (xylose dehydrogenase). The results obtained in this study confirm that half of the pathway is active at the cell surface, but further experiments are required to determine the precise production levels and complete the pathway.

Abstract

Abstract Aim: Cancer is one of the leading causes of death worldwide, and its treatment is always associated with side effects such as drug resistance. So, there is a strong tendency for the identification of new herbal anti-cancer compounds.
Material and Methods: In this study, a range of 0.5-12 µg/mL of hydroalcoholic extract of Tamarindus indica kernel and 5-Fluorouracil was applied to prostate cancer (LNCaP), colon cancer (HT-29) and normal fibroblast (HSkMC) cells for 24 hours. The cytotoxic effect of the extracts was measured by the MTT method. The rate of DNA synthesis and incidence of apoptosis was measured by BrdU and TUNEL assays, respectively.
Results: Following treatment with the highest amount of the extract, the viability of prostate, colon, and fibroblast cells was reduced to 4.8, 65.1, and 60.5%, respectively. The IC₅₀ for the three cell lines was 4.60, 17.0 and 13.79 μg/mL respectively. The rate of DNA synthesis also reduced by 32, 37 and 15% for prostate, colon and fibroblast cancer cells, respectively. The rate of apoptosis in LNCaP and HT-29 cells was 31 and 4%, respectively.
Conclusion: Collectively, the toxicity of the extract was higher for LNCaP cells than for the other two cells (p-value ˂0.01). Further studies in vivo and analysis of compounds in tamarind can lead to the identification of anti-cancer compounds from this plant.

Abstract Aim: The purpose of this research was aimed to evaluate the protective effects of crocin, as an antioxidant agent on Mast cells, blood vessels and biochemical changes of ovary and serum in Busulfan-induced oxidative stress in Mice.                                                                                                                                
Material and Methods: Thirty mature 6-8 weeks aged female NMRI mice in the weight of 22-25 g were randomly divided into 6 groups, and treated for 21 days. The control group only received solvent of Busulfan (BSF) (0.1 ml) intraperitoneally, and BSF group received only Busulfan (10 mgkg^-1, IP/single dose). The experimental groups no. 1, 2, 3 received BSF (10 mgkg^-1 /single dose) with crocin (100, 200, 400 mgkg^-1 /day, IP) and positive group only received crocin (400 mgkg^-1, IP/day). At the end of treatment period, animals were euthanized and left ovary were studied for Mast cells, ovary blood vessels, and Serum, and right ovary for biochemical evaluations. Data was subjected to one‒way analysis of variance (ANOVA) and Tukey to determine if significant difference (P≤0.01) existed among the observed results using SPSS.
Results: Busulfan significantly (P≤0.01) increased mast cells and MDA, while decreased ovary blood vessels and SOD rate, significantly (P=0.000) in comparison to control group. However, crocin in all the used doses, especially in the dose of 200 mgkg^-1, significantly decreased the adverse effects of Busulfan.
Conclusion: The results indicated that crocin can protect ovaries against Busulfan induced damages, and it can be considered as a suitable drug for reducing the toxic effects of Busulfan in chemotherapy.
 

Abstract Aim: Salinity stress is one of the most important environmental stress in the world and one of the important factors in reducing growth in many plants, especially in arid regions of the world. Salinity stress and increased sodium ion lead to the induction of oxidative stress and consequent cell death. Melatonin is a multiple function molecule spread in different plant and triggers several physiologic responses to different environmental stress. Exogenous application of melatonin to several plants can improve crop growth and development in response to many abiotic and biotic stresses with adjusting the antioxidant system of plants. Current studies reported that, the exogenous melatonin can increase plants' stress resistance by regulating both the enzymatic and non-enzymatic antioxidant defense process. In this study, the effect of melatonin on alfalfa roots under tissue culture condition was investigated. The aim of the present study was to investigate how melatonin regulated the antioxidant system and non-enzymatic antioxidants such as reduced glutathione and ascorbate under salt stress
Materials and methods: In this study, alfalfa seeds (Medicago sativa) of the Isfahani variety were used and we studied the effect of melatonin and salinity stress on the alfalfa root. In order to sterilize the seeds, they were placed in a 70% ethanol solution for one minute and then in a 20%  sodium hypochlorite solution for 20 minutes. After disinfecting the seeds, seeds were placed in each culture dish containing MS (Murashige and Skoog) culture medium. After germination, alfalfa seeds were transferred to MS culture medium containing concentrations of 0, 0.1, 10, and 15 micromolar melatonin and concentrations of 0, 150 and 200 mM salt. After 10 days of growth, total antioxidant capacity,
the activity level of catalase, ascorbate peroxidase, superoxide dismutase, guaiacol peroxidase, and glutathione reductase, ascorbate and glutathione levels in alfalfa roots were measured.
 
Results: Also, in the salinity stress, melatonin treatment significantly increased the total antioxidant power, while no significant difference was observed between different concentrations of melatonin, 0.1µM melatonin 82 درصد  and 62 درصد  raised antioxidant activity under 150 and 200 mM NaCl, so melatonin can reduce the levels of reactive oxygen species by scavenging of them through antioxidant enzyme or non- antioxidant system. Based on the results Melatonin treatment caused a significant increase in antioxidant power, the activity of CAT, APX, POD, SOD, GR enzymes, and antioxidant compounds in the glutathione-ascorbate cycle including DHA, ASC/DHA, GSH, and GSH/GSSG. along with increasing salinity concentration. On the other hand, salt stress increased oxidized compounds including DHA and GSSG in alfalfa roots. The data were carried out by two-way analysis of variance (ANOVA), followed by Duncan’s multiple range tests.




Conclusion: In addition to its direct role in clearing free radicals, melatonin activates the antioxidant defense system of the root, i.e. both antioxidant enzymes and antioxidant compounds in the ascorbate-glutathione cycle, which increases the resistance to damage oxidative effects caused by salinity stress in alfalfa root. These findings proposed that exogenous melatonin utilization dramatically activated ROS scavenging systems including enzymatic and non-enzymatic antioxidants to keep a relatively low amount of ROS and increased the tolerance of alfalfa root against salinity stress.

Abstract Aim: Dates are the second most important crops in Iran after pistachios, which show different structural and textural adaptations to drought and high temperatures. Acquiring the necessary knowledge about various structural and physiological changes from flowering to fruit ripening plays a significant role in obtaining high-quality and marketable fruit. Date fruit is a berry characterized by exocarp, fleshy mesocarp and membrane endocarp around the seed. This berry is formed from a fertilized carpel present in the female flower, while the other two carpels do not grow and decay. The fruit development period is long and lasts about 7 months. The sweetness and texture of date fruit is closely related to the maturity stage of the fruit. During the growth period of date fruit, several changes in the color and chemical composition of the fruit are observed. Mozafati is one of the most important commercial varieties of dates in Iran especially in Bam city. In the present study, the morphology and histology of the date fruit (Phoenix dactylifera L.) of the Mozafati cultivar were investigated during the developmental stages.
Material and Methods: Samples were collected from a commercial garden located in Bam city. Five adult 17-year-old date palms of Mozafati cultivar derived from offshoots grown in similar environmental conditions were used as female parents. The pollen used in this experiment is the same pollen that is normally used by local gardeners for pollination. Sample collection was done in five different developmental stages including Hababouk, Kimri, Khalal, Sarkhal and Rutab (4, 12, 19, 24 and 25 weeks after pollination, respectively). In this research, the local Iranian names such as Sarkhal and Rutb were used for the last two stages. The collected fruits, after cleaning and sorting, were packed and stored at 4ºC. The hand sections prepared from the fresh samples at each developmental stage were studied with an optical microscope after staining and then photographed.
Results: An anatomical study of fruits in different stages of development revealed several general characteristics in their pericarp structure: 1) Pericarp differentiation starts in the early stages of development (immediately after pollination) and its development continues progressively and regularly. 2) The exocarp contains four differentiated layers in all developmental stages, from outside to inside, includes layers of the epidermis, hypodermis, skin parenchyma, and layer of stone cells. 3) The mesocarp (which is the largest region forming the pericarp) has a large number of layers consisting of parenchymal cells and is divided into two outer and inner regions, which are separated by a layer of parenchymal cells specialized to store tannins (idioblast). Vascular vessels with different sizes are distributed in the outer and inner mesocarp, which are less abundant in the inner parts. 4) The endocarp can be recognized as an unspecialized single-layered epidermis only in the early stages of development and eventually forms the seed coat together with the inner layers of the mesocarp.
Conclusion: Organized protection and defense mechanisms, including the presence of thick cuticle, stone layer, and specialized tannin layer (tannin idioblasts) in all stages of pericarp development, were among the interesting histological features of this cultivar, which showed its adaptive and evolutionary role in specific habitats

Abstract Introduction: Gene therapy involves transferring genetic material into target cells to correct mutations or introduce new biological functions. Among delivery systems, lentiviral vectors are considered efficient and reliable tools due to their ability to integrate stably into the host genome and transduce both dividing and non-dividing cells. This property provides long-term gene expression, which is highly valuable for therapeutic and experimental applications. The PDX1 (Pancreatic and Duodenal Homeobox 1) gene plays a central role in pancreatic organogenesis and the regulation of insulin-producing beta cells. It acts as a transcription factor controlling genes critical for endocrine differentiation and insulin secretion. Chick embryos are a useful experimental model due to their accessibility, rapid development, and the responsiveness of their fibroblast and germ cells to gene transfer systems. These features make them suitable for studying gene delivery efficiency and expression stability.
Aim: The aim of this study was to construct and produce recombinant lentiviral vectors carrying the PDX1 gene in HEK293T-LentiX cells and evaluate their transfer efficiency in chick embryonic fibroblast and germ cells. This work was conducted to assess the potential of lentiviral systems for stable gene delivery in avian cells.
Materials and Methods: HEK293T-LentiX cells were selected as producer cells due to their high transfection efficiency and viral packaging capability. They were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin, and streptomycin under standard incubation conditions (37°C, 5% CO₂). Lentiviral particles were generated using a three-plasmid packaging system, including a transfer vector containing the PDX1 gene and two helper plasmids. Transfection was carried out using the calcium phosphate method. After 48–72 hours, the viral-containing supernatant was collected, filtered, and concentrated. Viral titers were determined by evaluating GFP expression in target cells through fluorescence microscopy and flow cytometry. Chick embryonic fibroblast and primordial germ cells were isolated and infected with various viral concentrations in the presence of 8 µg/ml polybrene to enhance infection. After incubation, cells were examined for GFP signal as evidence of successful gene transfer.
Results: High-titer recombinant lentiviral particles were successfully produced in HEK293T cells. Fluorescence microscopy revealed strong GFP expression, confirming the presence of functional viral particles. Flow cytometry analysis provided quantitative confirmation of high viral titers. Following transduction, chick embryonic fibroblast and germ cells exhibited clear GFP expression, indicating efficient infection and gene transfer. The PDX1 gene was successfully delivered and expressed within target cells. Although transduction efficiency varied slightly between cell types, the overall results demonstrated that the lentiviral system provided stable and effective gene delivery to chick embryo-derived cells.
Discussion: The study confirmed that lentiviral vectors carrying the PDX1 gene could be efficiently produced and used to achieve stable gene transfer in chick embryonic cells. This system’s ability to integrate permanently into the host genome ensures consistent gene expression over time without repeated transfection. For functional genes like PDX1, this stability is crucial for maintaining insulin-related pathways and pancreatic cell differentiation. Chick embryos serve as an advantageous model because their cells are easily accessible, grow rapidly, and respond well to viral vectors. Such characteristics make them ideal for investigating genetic regulation during early development. Evaluation of viral titers using fluorescence microscopy and flow cytometry provided reliable data confirming efficient vector production. The integration of new tools such as CRISPR/Cas9 can further enhance lentiviral design precision, allowing targeted modification of specific genes. Combining these technologies may open promising avenues for studying metabolic disorders and for gene-based therapies.
Conclusion: Recombinant lentiviral vectors carrying the PDX1 gene were successfully generated and used to transduce chick embryonic fibroblast and germ cells. The system exhibited high production efficiency, stable gene expression, and suitability for in vitro studies. These findings demonstrate that lentiviral vectors represent a powerful and versatile platform for gene transfer and experimental modeling in avian systems. Moreover, coupling lentiviral vectors with genome editing technologies could expand future applications in regenerative medicine and genetic engineering.

Abstract Introduction: Breast cancer is the most important and widespread type of cancer in women's population.
Aim:This study was conducted to synthesize zinc oxide nanoparticles conjugated with gingerol (ZnO@CPTMS-Gingerol) and evaluate their anticancer effects on breast cancer cells.
Materials and Methods: To synthesize ZnO@CPTMS-Gingerol nanoparticles, one gram of ZnO@CPTMS nanoparticles was dispersed in 30 ml of dry toluene. One gram of gingerol and 10 ml of triethylamine were added to the reaction mixture and refluxed for 24 hours. The product was washed twice with a mixture of distilled water and ethanol (1:1), and the final product was dried at 100°C for 24 hours. Physicochemical properties of ZnO@CPTMS-Gingerol nanoparticles were studied by FT-IR, XRD, DLS, EDS, zeta potential measurement, and electron microscope imaging. The inhibitory effects of different concentrations of ZnO@CPTMS-Gingerol nanoparticles on MCF-7 breast cancer cells and HEK293, as normal cells, were evaluated by the MTT test. To perform this experiment, cells were prepared in 96-well cell culture plates with a density of 104 cells/well, and then were treated with concentrations of 15.625, 31.25, 62.5, 125, 250, and 500 μg/mL of ZnO@CPTMS-Gingerol. After incubating the cells for 24 hours at 37°C, 0.2 ml of MTT solution was added to each well. The wells without nanoparticles treatment were considered as controls. After incubation for 4 hours, the supernatant was removed, and 100 μl of DMSO solution was added to each well. After pipetting, the optical density was read at 570 nm using an ELISA Reader. To determine the percentage of apoptotic and necrotic cells, 5x10⁵ cells were treated with ZnO@CPTMS-Gingerol nanoparticles for 24 hours with half inhibitory concentration (IC50). Then, the treated and control cells were stained with annexin V and propidium iodide (PI) dyes. Finally, cell analysis was done by a flow cytometer. Data analysis was done using device software and dividing the points recorded in the two-dimensional curve into four regions including Q1 to Q4. The experiments were performed in three replicates, and the results were expressed as mean ± standard deviation. Statistical analysis including t-tests, and one-way ANOVA was performed using SPSS. A p

Abstract