Cell and Tissue Journal

Abstract Conclusion: It seems that various factors such as genotype, polyploidy or hybrid formation, slow plant growth (which is common in coniferal), or secondary compounds are the reasons for the low rate of regeneration, especially rooting. Also, the duration of cultivation can be effective in rooting.

Abstract Aim: Lignocellulosic biomass such as agricultural wastes (corn stover, sugar beet pulp and citrus peel) is a widely abundant and attractive source for the production of biofuels and chemicals.
biofuels are sources of clean and renewable energy that are considered as a potential substitute for non-renewable oil fuels. various methods and processes have been tested by scientists and researchers in this field and the most favorable conditions for producing biofuels from biomass. however, this biomass has not been fully exploited in many parts of the world for biofuel production, especially in developing countries, and there is little relation with crop residues and forest and waste in this area. so much work is still needed to replace fossil fuels with biofuels from biomass. lignocellulosic waste biomass such as cassava peels, sugar beet pulp, and Ulva lactuca are suitable materials for bioethanol synthesis.
D-xylose, is the second most abundant sugar in lignocellulosic hydrolysates. Nowadays, considerable efforts have been made to expand microbial cell factories to use D-xylose for the production of value-added chemicals. D-1,2,4-butanetriol (BT) is an extremely important intermediate chemical, is widely used in many fields, such as pharmaceuticals, paper, polymer materials, and military applications.A molecule 1,2,4-butanteriol (BT) is a polyol with unique chemical properties, which has a stereocenter and can be divided into D-BT (the S-enantiomer) and L-BT(the R-enantiomer). BT is widely used in the military industry, medicine, tobacco, polymer.  A synthetic pathway involving four enzymes—D-xylose dehydrogenase (XDH), D-xylonate dehydratase (XD), 2-keto acid decarboxylase (KDC), and aldehyde reductase (ALR)—has been proposed and implemented to produce BT from D-xylose, highlighting its significant role in bioproduction. And because in most studies, the xylonate dehydratase was used in the case of Caulobacter.crescentus, and given the very high genetic similarity between Caulobacter.crescentus and Caulobacter.vibrioides, the aim of this study is to clone and express xylonate dehydratase gene of C.vibrioides in the E.coli. For this purpose, the research was carried out with the aforementioned methods.
Material and methods:  The xylonate dehydratase gene was retrieved from the NCBI database and amplified using PCR with specific primers after extracting the C. vibrioides genome. The piece of the gene was cloned in the pET28 expression vector and then transferred to the E.coli prepared cells using chemical methods. After the induction of the cells, recombinant protein expression was examined using SDS-PAGE. Results: By using restriction enzymes, Colony PCR and sequencing, the cloning process and the entry of the gene into the pET28 expression vector was confirmed. The presence of the recombinant protein was tested by SDS-PAGE gel with a molecular weight of approximately 68 KDa and the expression rate of the recombinant protein, estimated by Image J software, was 54 percent.
Conclusion: The bioproduction of butanetriol requires the construction of a metabolic pathway consisting of several enzymes. The presence of the bacterium E.coli as the target strain and the use of cheap substrate such as xylose-containing biomass and the existence of the enzyme xylonate dehydratase are essential for the production of high-speed and high-volume D-1,2,4 butanetriol.   

Abstract Aim: This study was investigated, the effect of foliar application of different levels of putrescine on physiological, biochemical traits, and essential oil content of Mexican marigold (Tagetes minuta L.) under drought stress.
Material and Methods: The experiment was conducted as a factorial experiment in a completely randomized design with three replications in greenhouse conditions. The first factor was drought stress at three levels (75, 50, and 25% of field capacity) and the second factor was putrescine foliar application at four levels (0, 0.5, 1, and 2 mM). Four seedlings were planted in each pot. Stress was applied when the plants reached a height of about 25 cm. Putrescine was sprayed one week before flowering and every 7 days until the 80% flowering stage.
Results: The results showed that 2 mM foliar application of putrescine was more effective than other concentrations and was able to greatly reduce the harmful effects of drought stress on this plant. With the increasing drought, antioxidant activity, total phenol, proline, and electrolyte leakage also increased, and in contrast, the relative content of leaf water, photosynthetic pigments, and soluble protein decreased. Drought also increased the activity of the antioxidant enzymes guaiacol peroxidase (GPX) and polyphenol oxidase (PPO) and the amount of malondialdehyde in this plant. With increasing drought stress up to 50% of field capacity and foliar application of 1 mM putrescine, the essential oil content of Mexican marigold increased, while under severe stress (25% of field capacity) is decreased. Putrescine decreased by inducing drought resistance. The activity of antioxidant enzymes and the amount of malondialdehyde.
Conclusion: The use of putrescine induced resistance to drought stress in Mexican marigold, and the use of a concentration of 2 mM can be effective and recommended.
 
 
 

Abstract

Abstract Aim: In this study, the alteration of the Leucine-rich repeat kinase 2 (LRRK2) gene, as the most important gene involved in Parkinson's disease, was investigated using CRISPR-Cas editing technology.
Material and methods: Cloning the guide molecules of interest was performed using of gRNA CRISPR cloning kit (Catalog No. C8324K, Zaver Zist Azema). Two sets of forward and reversed gRNA oligonucleotides for exon 41 of the LRRK2 gene were designed using the CHOPCHOP CRISPR guide gRNA designing web software. Double-stranded DNA molecules were made according to standard instructions in the laboratory. 20 base-pair DNA segments were used to generate RNA-Cas-9 enzyme complex and then ligated into the Px459 CRISPR eukaryotic expression vector using Fermentas DNA ligase enzyme. Recombinant plasmids were transformed into E. coli DH5a host competent cells using of heat shock method.
Results: findings by multiple PCR experiments and also DNA sequencing blast analysis showed successful cloning the segments of interest into CRISPR plasmids. Clear PCR bands were in agreement with expected values and DNA sequencing results showed 100% similarity. 
Conclusion: According to the results, development of CRISPR vector system may be used as a wide and powerful tool for editing and alteration of many encountered genes in different diseases such as Parkinson's disease.

Abstract Aim: Organotypic cultures of spinal cord slices from mammalian neonatal and fetal animals are powerful tools for studies of spinal cord injury, neuronal degeneration, and cell death but also motor neuron regeneration. Models in which adult slices are used would be very useful. However, adult spinal cord slices are notoriously difficult to maintain in culture and rapidly deteriorate in vitro. Degeneration of motor neurons in the spinal cord is a critical phenomenon in spinal cord injuries and certain neurodegenerative diseases such as amyotrophic lateral sclerosis, a neurodegenerative disorder in which motor neurons in the spinal cord and motor cortex are lost. A variety of mechanisms have been proposed as having a role in neuronal apoptosis during spinal cord injury. Oxidative stress has been reported as one of the mechanisms involved in the apoptosis of motor neurons in spinal cord injuries and neurodegenerative diseases. This study was conducted to determine whether quercetin, as a potent antioxidant, can delay apoptosis in motor neurons of cultured spinal cords by reducing oxidative stress.
 Material and methods: The thoracic regions of the spinal cord from adult NMRI mice were sliced using a tissue chopper and divided into three groups: 1) 0-hour, 2) control group, and 3) group treated with quercetin (100 µM). Spinal cord slices in the 2 and 3 groups were incubated for 6 hours at 37°C in a Co₂ incubator. The viability of the spinal cord slices was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological features of apoptosis and the number of motor neurons were examined using Hoechst and propidium iodide staining. The malondialdehyde was measured to determine lipid peroxidation while the FRAP (ferric reducing antioxidant power) was assessed to evaluate total antioxidant capacity in fresh and cultured spinal cord slices. Results were expressed as mean±SD. One-way analysis of variance (ANOVA) followed by Tucky’s test was used to assess the statistical significances of the data. In all cases, a statistical probability of p<0.05 was considered significant.
Results: After 6 hours (control group) in culture, the viability of the spinal cord slices, the neuron diameter, and the number of healthy motor neurons significantly decreased compared to the 0-hour group. Also, motor neurons showed the morphological features of apoptosis including cell shrinkage, nuclear and chromatin condensation in the control group. A significant increase in the amount of malondialdehyde and a significant decrease in the total antioxidant capacity was also observed compared with the 0-hour group. After 6 hours, quercetin not only increased the viability and the number of healthy motor neurons in the cultured slices but also reduced the morphological features of apoptosis in the motor neurons compared with the control group. In addition, quercetin significantly reduced the amount of malondialdehyde and increased the total antioxidant power in slices cultured for 6 hours.
Conclusion: Oxidative stress might be considered as one of the mechanisms involved in the apoptosis of motor neurons in cultured spinal cord slices and quercetin, as a potent antioxidant, was able to increase the viability of the cultured spinal cord slices and delay the morphological features of apoptosis in the motor neurons through reducing lipid peroxidation and increasing the total antioxidant capacity.

Abstract Aim: Nowadays, one of the great challenges of mankind is to find a suitable way for the treatment of cancer. Recent developments show that nanotechnology has a significant impact on the prevention, diagnosis and treatment of cancer. In nanotechnology, there are different physical and chemical methods to produce nanoparticles but mostly chemical methods are used. The nanoparticles produced using these methods are harmful to health due to the absorption of chemical species on their surface. In order to eliminate this big problem, biological methods such as the use of fungi, bacteria and plants have replaced other methods for the synthesis of nanoparticles which is called green synthesis. It is cheaper, simpler and non-toxic than other methods. The use of plants for the synthesis of nanoparticles has become more important than other biological methods due to the elimination of the difficult step of cell culture maintenance.
Material and Methods: In this study, gold nanoparticles were formed after treating the HAuCl₄ solution with the aqueous extract of Anarijeh plant with the scientific name Pimpinella affinis. In this research, different amounts of aqueous extracts of leaves and stems of Pimpinella affinis were used for the synthesis of gold nanoparticles. UV-Visible spectroscopy and DLS methods were used to evaluate nanoparticles and determine their average size and stability. Also, SEM method was used to investigate their morphology. FTIR method was used to determine the functional groups of the aqueous extract and synthesized gold nanoparticles; in addition GC/MS device was used to determine the composition of the extract. Finally, MTT analysis was used to investigate the cytotoxic effects of nanoparticles on MCF-7 cell line along with treatments of 24, 48 and 72 hours.
Results: The results of the analysis were showed that the optimal concentration of the Pimpinella affinis extract for the synthesis of gold nanoparticles is 1 mg/ml. The synthesized gold nanoparticles using the mentioned concentration of the extract have an average size of 20 nanometers with a zeta potential of -61 mV which has the best stability. They had a spherical shape with a polydispersity equal to 0.7. The results of FTIR analysis show the presence of hydroxyl, simple carbonyl groups and hydrocarbon functional groups in the compounds of the extract. Experiments showed that phenolic and carboxylic compounds play the most important role in the reduction of gold salt. Also, proteins and lipids have made gold nanoparticles stable. The essential compounds identified in the extract by GC/MS method include Phenolic and flavonoid compounds and terpenes which are confirmed by the FTIR spectrum. The treatment time of 48 hours for MCF-7 cell line is the best time for the toxicity of nanoparticles with IC50 equal to 384.2 µg/ml, which was determined by MTT analysis.
Conclusion: According to the above findings, it can be concluded that Pimpinella affinis is one of the suitable options for the synthesis of gold nanoparticles, and it can be done with more studies on nanoparticles synthesized using this plant, considered it as one of the efficient methods to achieve the goal of proper treatment for breast cancer.

Abstract

Abstract Aim: This study aimed to access the rapid and improved propagation protocol production of Zamifolia plant using tissue culture.
Material and Methods: For callus formation and regeneration, two independent factorial experiments were carried out in a completely randomized design with 3 replications in the tissue culture laboratory of the National Research Institute of Flowers and Ornamental Plants in 2018. In the first experiment, to induce callogenesis, 2,4-D (0, 1, 2, and 4 mg/l) and BA (0, 1 and 2 mg/l) were used. In the second experiment, or regeneration of explants, NAA (0 and 0.5 mg/l) and BA (0, 0.5, and 1 mg/l) were applied.
Results: The results of the callogenesis experiment showed that a modified basal MS medium with concentration of 2 mg/l BA and 1 mg /l 2,4-D produced the best callus with 85% regeneration. At the regeneration stage of the callus, 0.5 mg/l BA and 0.5 mg NAA produced the highest number of seedlings and regenerated tubers (90%). Subsequently, single tuber seedlings were successfully adapted to 80%-90% in the cocopeat and perlite substrates.
Conclusion: The results were successful and could be possible to achieve the callogenesis and regeneration of Zamifolia plant with the formula presented and recommend as practical a protocol for commercial micropropagation of this valuable ornamental plant.
 

Abstract Aim: Thepurpose of this study is the investigation of the effect of aqueous and ehanolic plant extracts including neem, clove, thyme and lavender on the increase storage life via decrease of reactive oxygen and the activity of antioxidant enzymes.
Material and methods: In this study, it was extracted of the neem, clove, thyme and lavender with aqueous and ehanolic solution, then the orange fruits was treated with 2×1000, 4×1000 and 6×1000 concentration of plant extract and chitosan and vax. The treated fruits were stored in the storage with 7C and 80-90% humidity. The enzyme activity and the related genes expression was evaluated per twenty days to 100 days in the orange fruits. In the other section of study, it was done the panel test.
Results: The results showed the activity of catalase was affected with plant extracts. The treated orange fruits with 6×1000 concentration of ethanolic lavender extract showed the least catalase activity, with 2.13; the least peroxidase activity was observed in the fruits treated with aqueous lavender extract with 6×1000 concentration. The ethanolic and aqueous of lavender extract affected on the catalase and peroxidase gene expression with 5.28 and 6.66 respectively.
Conclusion: the results of this study showed the extracts of neem, clove, thyme and lavender have highly effect on the plant physiology and they decreased the enzyme activity and the genes expression.

Abstract Introduction: Titanium dioxide nanoparticles (TiO₂ NPs) are widely used in industry, medicine, food, and cosmetics. TiO₂ NPs are harmful to the environment and human health. Changes in the environment may especially affect the growing neurological system. Therefore, it is impossible to overlook their impact on the development of the embryo and reproductive success. A sensitive and widely used model for evaluating the teratogenic potential and developmental toxicity of different nanoparticles is the chicken embryo. At the outset of embryogenesis, the application of TiO₂ NPs enables the penetration into various tissues, such as brain precursor cells and structures. Furthermore, the organs are unable to remove the nanoparticles from the egg due to their isolation and enclosure from the mother and the environment. Consequently, the embryos are perpetually exposed to TiO₂ NPs during the 20-day embryogenesis period. In recent years, a growing number of studies have been conducted to examine the possible harmful effects of TiO₂ NPs due to worries about inadvertent exposure of NPs on humans.
Aim: In this study, the embryonic toxicity of various dosages of TiO₂ NPs was investigated in the chicken embryo's brain cortex.
Materials and Methods: In this study, 90 fertilized eggs were divided into five groups: control group (untreated group), four treatment groups that received 0 (sham), 12.5, 25, and 50 μg/mL of TiO₂ nanoparticles. The embryos' morphology, weight, and Crown-rump length (CRL) were assessed after 7, 9, and 13 days.   Tissue samples were collected from the cerebrum and cerebellum of the chick embryos. The specimens were immediately fixed in a 10% neutral buffered formalin solution for 48 hours. Subsequently, they were dehydrated through a series of ascending ethanol concentrations (70%, 80%, 90%, and 100%), with each step lasting two hours. The samples were then cleared in xylene for one hour, embedded in paraffin wax, and sectioned at a thickness of 5μm using a microtome. The sections were then mounted on glass slides.  Finally, all tissue sections were stained with hematoxylin and eosin, and the effects of TiO₂ NPs were examined on the histology, pathology, and histomorphometry of the chick embryo cerebrum and cerebellum tissues.
Results: The findings showed that TiO₂ NPs cause embryo death in all days at 25 and 50 μg/mL. In morphological studies, the weight and length of 13-day-old embryos treated with 50 μg/mL of TiO₂ NPs decreased. Counting of cells (neurons and glial cells) in the cerebral cortex of a 13-day-old chick embryo displayed a significant decrease in the experimental group of 50 μg/ml compared to the control and sham groups. The evaluations showed a decrease in the number of Purkinje cells of the cerebellum cortex in 13-day-old embryos treated with 50 μg/ml of TiO₂ NPs.   The group of 13-day-old embryos treated with 50 µg/ml had a considerable capillary hyperemia in the cerebellar cortex.
Conclusion: It was concluded that the in-ovo-administered TiO₂ NPs given immediately before incubation have adverse effects on the developing cerebellum and cerebrum. So that the increase of damage happened in older embryos, and the highest damage occurred on day 13 of incubation.

Abstract Aim: Zinc nanoparticles have gained significant attention due to their wide-ranging applications in various fields, including industries, medicine, and nutrition. These nanoparticles are characterized by their small size, which allows for easy absorption into biological systems, enabling them to interact with different tissues effectively. One of the critical areas of concern regarding the use of zinc nanoparticles is their potential impact on kidney health. The kidneys are vital organs responsible for filtering blood, excreting waste, and regulating essential bodily functions. Due to their high blood flow and the capacity to excrete various compounds, kidneys may be particularly vulnerable to the effects of toxins, including nanoparticles. Therefore, this study aims to investigate the effects of nano zinc oxide on antioxidant activity and histological changes in the kidneys of rats. Understanding these effects is crucial for assessing the safety and potential therapeutic applications of zinc nanoparticles in medical and nutritional contexts.
Material and Methods: In this research, male Wistar rats were utilized as the experimental model. The rats were randomly divided into six groups, with seven rats in each group to ensure statistical validity. Group 1 served as the normal control group, while the remaining groups received varying doses of nano zinc oxide. Specifically, group 2 received 5 mg/kg of nano zinc, group 3 received 10 mg/kg, group 4 received 25 mg/kg, group 5 received 50 mg/kg, and group 6 received 100 mg/kg of nano zinc. The administration of these doses continued for one month to observe both short-term and potential cumulative effects. After this period, the rats were sacrificed, and their kidneys were isolated for further analysis. Various biochemical assays were conducted to assess total antioxidant capacity (TAC), total oxidant capacity (TOS), glutathione levels, and malondialdehyde (MDA) concentration. Additionally, liver enzymes were measured, and kidney histopathology was examined to evaluate any structural changes resulting from nano zinc exposure.
Results: The findings revealed significant alterations in antioxidant activity among the groups that received nano zinc. Specifically, total antioxidant capacity (TAC) and glutathione levels decreased markedly in the rats that received higher doses of nano zinc, particularly at 10, 25, 50, and 100 mg/kg. In contrast, there was a sharp increase in malondialdehyde (MDA) concentrations and total oxidant capacity (TOS) in these groups, indicating heightened oxidative stress. Interestingly, the group that received the lowest dose of 5 mg/kg of nano zinc exhibited no harmful effects on tissue changes or antioxidant capacity, suggesting a threshold below which nano zinc may not pose significant risks. Conversely, in the groups receiving higher doses, observable tissue damage was noted, and these changes appeared to be dose-dependent, emphasizing the importance of dosage in determining the safety and efficacy of nano zinc.
Conclusion:The results of this study underscore the potential harmful effects of nano zinc, particularly at elevated doses, on kidney function and histopathology. The observed increase in oxidative stress markers and accompanying tissue changes highlight the need for caution in the use of zinc nanoparticles, especially in medical and nutritional applications. While nano zinc in low concentrations may offer beneficial effects, such as potential therapeutic properties, the risks associated with higher doses cannot be overlooked. Further research is warranted to explore the mechanisms underlying these effects and to establish safe dosage guidelines for the use of zinc nanoparticles in various applications. Overall, this study contributes valuable insights into the safety profile of nano zinc, particularly concerning kidney health, and emphasizes the need for careful consideration of dosage in therapeutic contexts.

Abstract Aim: The purpose of the present study was to investigate the effect of adding different levels of garlic extract as an antioxidant to diluent on the semen characteristics of Arabi rams under liquid conditions at 5°C.
Material and Methods: Semen samples were collected from 12 Arabi rams weekly for 8 weeks and their semen was immediately mixed, diluted, and divided into the number of experimental treatments and received different levels of garlic extract. Treatments included garlic extract levels (zero, 50, 100, 150 and 200 μl/ml). At different storage times of diluted semen containing experimental treatments (zero, 24, 48, and 72 hours), semen quality parameters were evaluated.
Results: At time zero (immediately after sperm collection and addition of garlic extract), sperm quality parameters in experimental treatments were not statistically significant compared to the control. At 24 hours, 50 μl/ml of garlic extract improved the spermatozoa progressive motility of total motility rates, but 200 μl/ml of extract reduced the total motility of sperm (p < 0.05). At 48 hours, the highest sperm viability was related to the level of 50 μl/ml garlic extracts (p < 0.05). 72 hours after semen collection, the highest progressive motility of spermatozoa was observed at the level of 50 μl/ml garlic extracts (p < 0.05). Malondialdehyde concentration of seminal plasma as an indicator of peroxidation was not affected by experimental treatments.
Conclusion: In general, sperm quality parameters were improved by adding 50 μg/ml garlic extract to the semen diluent of Arabian rams and storage the diluted semen in liquid condition at 5°C.
 

Abstract

Abstract Aim: A multitude of devastating human diseases arise from genetic mutations that lead to cellular dysfunction, as well as from infectious diseases and cell transformation. These diseases have significantly impacted individuals and communities worldwide and led to challenges in medical and biological sciences to address their causes and improve treatment approaches. Since identifying genes as the main heredity unit, generating targeted alterations in specific gene loci for treatment and perception of disease function has been a crucial concern. The discovery of nuclease enzymes revolutionized this field and transformed the concept of genome editing from a dream to a tangible reality.
Currently, targeted gene editing and modification through techniques such as zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and clustered regulatory interspaced short palindromic repeat Cas-associated nuclease (CRISPR/Cas) offers robust methods to assess gene function, also precisely and efficiently manipulate cells behavior.
ZFN and the TALEN were the earliest gene editing approaches that developed, which rely on the FokI enzyme and engineered protein that binds to a specific sequence in the genome and induces double-strand breakage, stimulating the cell's repair mechanisms. Although these techniques led to many significant discoveries and developments in gene editing, they had limitations such as complexity and high prices.
The emergence of CRISPR/Cas technology propelled gene editing to a new stage of efficiency and accessibility. This technique utilizes RNA to guide the Cas enzyme to the target gene sequence and, like the ZFN and TALEN, creates a double-strand breakage, which triggers DNA repair mechanisms. CRISPR/Cas is a versatile, simple, and cost-effective revolutionary gene-manipulating tool that enabled researchers to explore the potential of gene editing in a wide application range.
Base and Prime editors are the latest systems in the genome manipulation area, which, based on the CRISPR/Cas technique, represent the impressive development and improvement in gene editing capabilities. Despite the others, these two technologies induce the single-strand breakage and modify at the nucleotide level in target sequences.
The ability to edit genes indicates new possibilities for treating genetic diseases and even preventing them before they appear. Nowadays, a growing number of clinical trials utilizing genome editing for therapeutic purposes are underway, thanks to the substantial advancements in these tools. These remarkable improvements hold promise in treating various diseases and improving patient’s outcomes.
However, despite the outstanding progress made in gene editing technology, there are still several challenges and obstacles, such as ethical considerations, off-target effects, and safety concerns, which still need more investigation and studies. Consequently, ongoing researches are focused on developing the accuracy and efficacy of these editing tools.
Conclusion: The advancement of gene editing technology has opened up a new way in medical and biological sciences to modify and manipulate the genome, also exploring the various diseases caused.
 

Abstract Aim: The aim of this study was to investigate the apoptotic effect of the alcoholic extract of black seed on HL-60 cancer cells.
Material and Methods: In this study, concentrations of 350, 650, 950 and 1250 µgml
^-1
of Nigella sativa alcoholic extract were added to HL-60 cancer cells for 24 h. MTT assay was applied to find out the portion of living cells. Likewise, PI staining for flow cytometry was performed to determine whether black seed extract inhibits the proliferation of HL-60 cells by modulating cell cycle progression and apoptosis, then AO/EB staining of HL-60 cells was performed to detect apoptosis and necrosis patterns. 
Results: MTT assay showed that in comparison with the control group, alcoholic extract of black seed killed 50% of HL-60 cell in the concentration of 650 µgml
^-1
and based on the concentration, more cell death was caused at the higher concentration. Also, a substantial increase in G0/G1 phase cells was observed from 15. 2% in the control group, up to 51. 65% in the group was treated with 650 µgml
^-1
. It was also found that the concentration of 650 µgml
^-1
induced apoptosis in HL-60 cells, while at higher concentrations of 950 and 1250 µgml
^-1
caused HL-60 cells necrosis compared to the control group.
 
 

Abstract Aim: The aim of this study was the investigation of the effect of chitosan on phenolic compounds, rosmarinic acid content, and  key genes expression involved in the biosynthesis of these compounds in Melissa officinalis L. cell suspension culture.
Material and Methods: A combination of NAA (1mg L^-1), 2, 4-D (1mg L^-1), and kinetin (0.5 mg L^-1) and stem explants were used for callus induction. After initiation of cell suspension culture, chitosan treatment (0, 50, and100 mg L^-1) was performed for one, three, and five days in a factorial experiment based on a completely randomized design with three replications. Phenylalanine ammonia-lyase (PAL) activity, soluble protein, phenolic compound, and rosmarinic acid content were measured. Also, the expressionof phenylalanine ammonia-lyase and rosmarinic acid synthase (RAS) genes was evaluated by real-time PCR.
Results: PAL activity at 50 mg L^-1 chitosan showed the highest increase after three days of treatment, which was accompanied by an increase in phenolic compounds. The relative expression of the PAL gene at 50 mg L^-1 chitosan increased 2.5 fold at three days after treatment compared to control. This increase in the expression of the PAL gene resulted in an increase in enzyme activity. However, the highest increase in expression of the RAS gene was observed at 100 mg L^-1 chitosan in five days after treatment, which led to an increase in rosmarinic acid content.
Conclusion: According to the results, chitosan treatment is recommended to increase of rosmarinic acid content in lemon balm cell culture for five days.
 

Abstract Introduction: Peppermint (Mentha piperita L.) is an economically and medicinally significant plant valued for its essential oil. Silver nanoparticles (AgNPs) act as potent elicitors, triggering defense responses that include the production of reactive oxygen species (ROS) and the accumulation of secondary metabolites. However, the impact of AgNPs on peppermint’s physico-biochemical traits and essential oil performance remains underexplored.
Aims:  This study aimed to evaluate the effects of AgNPs on various physico-biochemical characteristics and essential oil performance in peppermint. The analyzed parameters, including H₂O₂, soluble proteins, phenols, flavonoids, antioxidant enzymes (SOD, CAT, APX), and photosynthetic pigments (chlorophyll and carotenoids), were examined to determine the optimal concentrations for enhanced metabolic activity.
Materials and methods: Rhizomes were planted in pots and grown under controlled greenhouse conditions, including a daytime temperature of 25°C, a photoperiod of 16 h of light and 8 h of darkness, and a relative humidity of 60. At the ten-leaf stage, plants underwent foliar spraying with 0 (control), 1, and 2 mM AgNPs. The AgNPs solution was prepared, characterized via UV-Vis spectroscopy, and SEM (20 nm quasi-spherical particles). A completely randomized design with three replicates was used to assess parameters including H₂O₂ content, soluble proteins, phenols, flavonoids, antioxidant enzymes (SOD, CAT, APX), photosynthetic pigments (chlorophyll, carotenoids), and essential oil performance. Statistical analysis was performed using ANOVA and LSD tests (p<0.05).
Results: AgNPs significantly elevated all measured parameters compared to the control in a generally concentration-dependent manner: Treatment with 1 and 2 mM AgNPs increased H₂O₂ content by 2- and 3-fold, respectively. Treatment with 2 mM AgNPs resulted in a 2.4-fold increase in protein content. 1 and 2 mM AgNPs resulted in a 2.8- and 2.6-fold increase in leaf phenolic content, respectively. Concentrations of 1 and 2 mM AgNPs resulted in a 1.8- and 2.6-fold increase in leaf flavonoid content, respectively. Treatment with 2 mM AgNPs resulted in a 3.3-fold increase in SOD activity. Treatment with 2 mM AgNPs had a significant positive effect (about a 2.2-fold) on CAT enzyme activity. However, treatment with 1 mM AgNPs failed to affect CAT enzyme activity. AgNPs 1 and 2 mM increased APX enzyme activity by 1.7 and 2.5 times, respectively. Treatment with 1 and 2 mM AgNPs increased total chlorophyll concentration by 2.4- and 3-fold, respectively. 1 and 2 mM AgNPs increased carotenoid concentration by 1.8- and 3.5-fold, respectively. Treatment with 1 and 2 mM increased essential oil yield by 2.2- and 2.3-fold, respectively.
Discussion: AgNPs significantly increased the level of H₂O₂, soluble proteins, phenols, flavonoids, antioxidant enzymes (SOD, CAT, APX), chlorophyll, carotenoids, and essential oil yield. Although the mechanisms of plant response to AgNP elicitors are not well understood, we proposed that the improvement in peppermint’s physico-biochemical traits in our study was due to the following series of mechanisms: AgNPs induce ROS (e.g., H₂O₂) generation through NADPH oxidase activation and electron leakage from organelles. This oxidative stress activates MAPK cascades and calcium signaling, which upregulate transcription factors (MYB, bHLH). These transcription factors subsequently enhance the expression of phenylpropanoid pathway enzymes (e.g., PAL) and terpenoid biosynthesis genes, leading to increased essential oil biosynthesis and accumulation. AgNP elicitors also improve antioxidant enzyme activity (SOD, CAT, APX) to mitigate ROS damage. In addition to the enzymatic antioxidant system, AgNPs enhance components of the non-enzymatic antioxidant system, including phenols and phenoloids. Furthermore, these nanoparticles upregulate key chlorophyll biosynthesis genes (HEMA1, CHLH), while suppress degradation-related genes (NYC1, PAO), resulting in elevated photosynthetic pigment concentrations.
Conclusion: Foliar application of 2 mM AgNPs significantly enhances peppermint’s antioxidant capacity, photosynthetic efficiency, and essential oil biosynthesis by modulating ROS-mediated signaling. Therefore, this concentration is recommended for maximizing peppermint’s medicinal and economic potential. Future studies should explore broader concentration ranges and early gene expression responses (6–12 h post-treatment).

Abstract Aim: Infertility is a complex issue that affects many couples worldwide. In vitro fertilization (IVF) has been a groundbreaking technique in addressing this challenge, but it comes with its own set of problems in the development of embryos during the pre-implantation stages. Understanding the processes involved in embryo development is crucial in overcoming these hurdles. During the pre-implantation stage, one of the challenges of treating infertility through in vitro fertilization is the developmental arrest of the embryo. In embryonic arrest, cell division stops for at least 24 hours, which, if it occurs in infertility treatments, leads to failure in ART cycles.
Cell signaling pathways play a vital role in the development and progression of embryos. The phosphatidylinositol 3-kinase (PI3K) pathway, known for its efficiency in regulating the cell cycle and various cellular processes, was the aim of this study. By targeting this pathway, we aimed to explore the effects of specific factors - ITS (Insulin, Transferrin, Selenium), Essential E8 medium (E8), and the CHIR99021 small molecule - on the resumption of development in type I arrested human embryos under in vitro conditions.
Material and Methods: In this study, firstly, after receiving ethics approval and patient consent, day 3 embryos (2-3 cell stage) from the Embryology Department of Royan Research Institute were utilized. The experimental groups consisted of control, CHIR99021, ITS, and E8. The optimum concentrations were chosen 1 for CHIR99021, 0.5% for ITS, and 0.1% for E8 + 10% serum. The culture medium for these groups was prepared and covered with liquid paraffin before being incubated at 37°C and 5% CO2 for 4 hours. Subsequently, the embryos were transferred randomly to either experimental or control groups and cultured in an incubator for 48 to 72 hours. Morphological evaluations of the embryos were conducted using an inverted microscope. Data analysis was performed using SPSS software and chi-square test, with a significance threshold set at P<0.05.
Results: The findings of the study revealed that the rate of arrest in the CHIR99021 and ITS groups showed a significant reduction compared to the control group. Moreover, all three experimental groups (ITS, CHIR99021, and E8) exhibited a notable increase in development rate up to the pre-morula stage when compared to the control group. Interestingly, while none of the embryos in the control group progressed to the blastocyst stage, two embryos in each of the CHIR99021 and ITS groups reached this advanced developmental stage.
Conclusion: In conclusion, the study's outcomes indicate the notable effect of ITS and CHIR99021 in modulating the phosphatidylinositol 3-kinase pathway to stimulate cell cycle progression in type I arrested embryos. ITS factor is probably able to regulate this pathway by activating insulin receptors and small molecule CHIR99021 by inhibiting Glycogen synthase kinase 3 (GSK-3). These findings hold promise for further research and potential applications in improving the success rates of IVF treatments and addressing infertility challenges. Understanding the mechanisms of embryo development is crucial for advancing reproductive medicine. It is important to note that the statistical population used in this study was limited and it is believed that more research is needed. 

Abstract  
Aim: Optimizing cell culture conditions to improve oocyte growth and maturation in vitro culture conditions has been the focus of many researchers today, but the quality improvement mechanism is not fully understood. Apoptosis or programmed cell death is a process to remove old and damaged cells from tissues and plays a major role in the life of follicles and immature oocytes. Most of the defective reproductive cells as well as extra cells are removed from the ovaries through apoptosis. One of the ways to reduce oxidative stress in the laboratory culture of oocyte maturation is the use of antioxidants. Statins are drugs for the treatment of high cholesterol for which antioxidant and anti-apoptotic properties have been reported. Atorvastatin in high doses has side effects for the heart and kidneys, but in low concentrations, it has antioxidant and anti-inflammatory effects for cells, and no side effects have been reported for it.In this study, we investigated the effect of low-dose atorvastatin on the rate of apoptosis in mouse oocytes.
 
Materials and Method: 24 h after PMSG (Pregnant Mare Serum Gonadotropin) inhection, 200 oocytes were obtained from adult female Wistar rats at the age of 4-5 weeks, were from were divided into 2 groups of 100 including the control group (MEM:Minimum Essential Medium culture+ Growth factor) and the atorvastatin group (MEM culture medium + 2 mg/kg atorvastatin+Growth factor) and cultured in the incubator(35 ^oC,CO2 %). After 24 hours of oocyte culture, Matured oocytes that met the standards of a healthy mature oocyte(oocytes that were in the first meiosis and had the first and mature polar body) were isolated the amount of apoptosis in each group was checked using tunnel staining and fluorescent microscope, the data were analyzed by variance analysis.
Results: After 24 hours of oocyte culture, the amount of apoptosis in the group receiving atorvastatin in the culture medium and the control group was investigated. The rate of apoptosis in the atorvastatin group and the control group was 24% and 22%, respectively, In the atorvastatin group, apoptosis increased by 2% compared to the control group butt the difference between the two groups was not statistically significant (p =0.11).
Conclusion: According to the findings of this study, atorvastatin in low doses can have a pro-apoptotic effect and cause partial induction of apoptosis in mouse oocytes in a laboratory environment. Although this study was not conducted on other doses of atorvastatin, it is suggested that the effect of other doses of this drug on the rate of apoptosis induction should be investigated in order to make an evidence-based decision regarding its administration and use.
 

Abstract Aim: This study was performed to evaluate the cytotoxic effects of hydroalcoholic extract of Artemisia sieberi and its effect on the cell cycle in the SKBr3 breast cancer cell line.
Material and Methods: In this study, hydroalcoholic extract of Artemisia‌ was prepared. SKBr3 cells were exposed to different concentrations (1, 10, 100, 1000 µgml^-1) of extract at two different time intervals of 24 and 48 h. We employed MTT assay and flow cytometry analysis to evaluate effects of the prepared extract on the cell cycle characteristics and its cytotoxic properties.
Results: Based on the data obtained from the MTT test, the highest toxicity of the extract observed at the concentration of 1000 µgml^-1 within 48 h after the extract exposure. The IC50 of hydroalcoholic extract was 150 and 50 µgml^-1 at 24 and 48 h, respectively. According to the data obtained from flow cytometry analysis, the extract arrests cell cycle in 24 and 48 h treatment groups.
Conclusion: The hydroalcoholic extract of Artemisia at certain concentrations inhibits the growth of SKBr3 breast cancer cells by ceasing cell cycle step. 
 

Abstract Aim: Long non-coding RNAs (lncRNAs) are a group of RNAs known as key regulators in cancer. The role of these molecules in differenet cellular procecess including cellular growth and differentiation has been documented. Several non-coding RNAs have been identified in the cell. Among the LncRNAs, LnvRNA MIAT is a non-coding RNA whose oncogenic expression is known in different types of cancer. However, uts role in brain tumors in not fully understood. Due to its oncogenic role in other cancers, it is estimated that this molecule to play an important role in brain tumors as well. The aim of this study was to investigate the role of lncRNA MIAT in glioma and identify its regulatory mechanism.
Material and Methods: In this study, the expression of lncRNA MIAT in glioma cancer cells, U87-MG and A172, was evaluated by Quantitative Real-Time PCR (qRT-PCR) method. Then, its expression was inhibited by the RNA interference method, and the expression of CD44 and miRNA-302 target genes was investigated.
Results: The results showed that lncRNA MIAT was highly expressed in glioma cancer cells. Also, after inhibition of the expression of lncRNA MIAT, a decrease in the expression of CD44 and miRNA-302 was observed.
Conclusion: The findings suggested the oncogenic role of lncRNA MIAT in glioma cancer cells through the regulation of CD44/miRNA-302 expression. As a result, lncRNA MIAT could be evaluated as a new biomarker in the prognosis and diagnosis of glioma.

Abstract Aim: Addiction is an important social and health problem in many Middle Eastern countries. Studies show that opium is the most commonly used substance after tobacco in many countries, especially in Iran. Opium is obtained from the seeds of the Papaver somniferum plant and contains more than 40 different alkaloids, the most important of which are morphine, codeine, papaverine, noscapine, and thebaine. Various studies show that morphine, which is one of the most important substances in opium, increases the production of free radicals in the body and reduces the antioxidant capacity. Opium consumption has various adverse health effects on the body. Therefore, long-term use of this combination can be related to some pathological consequences, including neurological disorders, liver toxicity, kidney dysfunction, oxidative stress, and apoptosis. Most drugs are metabolized by liver hepatocytes and excreted by kidney cells. Therefore, opium consumption can directly cause damage to liver cells. Free radicals play an important role in liver diseases. The increase of free radicals and on the other hand the decrease of antioxidants in the body caused liver toxicity. Oxidative stress is caused by an imbalance between the production of free radicals and their neutralization inside the body by antioxidant defense mechanisms. Oxidative stress is produced in various diseases, consumption of toxic substances, and old age, since it is not possible to investigate the effect of opium on human liver cells, and no study has been conducted in this field. The liver is the most important organ that is directly responsible for the metabolism and excretion of opium. Therefore, in this study, the effects of opium on oxidative stress markers in HepG2 cell line were determined.
Material and methods: Human liver cancer cell lines (HepG2) were used in this experimental study. At first, the cell line was placed in a 25 ml flask in the culture medium. DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS (Fetal bovine serum), and 1% penicillin and streptomycin antibiotics were incubated in a 37°C incubator with 5% CO_{2 .} The cells were treated with different concentrations of opium (0-100ug/ml) for 24 hours. Then IC₅₀ was determined and then a lower concentration, IC₅₀, and a higher concentration (three concentrations in total) were used for further studies.
The lipid peroxidation was measured by thiobarbituric acid method. The total oxidant was measured using xylenol orange. The amount of total antioxidants was determined by the FRAP (Ferric Reducing / Antioxidant Power) method. The catalase, glutathione peroxidase, and superoxide dismutase enzyme activities were measured by colorimetry methods. the data was entered into SPSS software (Version 20)  and analyzed using ANOVA and Tukey statistical tests. P less than 0.05 was considered a significant level. Results: Statistical analysis results indicated a significant difference in the amount of oxidative stress factors compared with control (p<0.001). In the group receiving opium with a dose of 60 and 70 mg/ml compared to the control group, the amount of TOS and MDA increased significantly (p<0.001), while the amount of TAC decreased (p<0.001). Also, the activity of catalase, glutathione peroxidase, and superoxide dismutase enzymes decreased in opium-treated groups (p<0.001). Conclusion: The results of this research showed that opium had a lethal effect on HepG2 cells. Also, opium caused an increase in oxidative stress in this cell line, which indicates the vulnerability of the liver against this compound. The results of this study show that although opium has analgesic effects, it can seriously damage the liver tissue. 

Abstract

Abstract Aim: Therefore, this study aimed to investigate the amount of lung tissue damage in trained rats due to biological silver nanoparticles.
Material and Methods: 30 male rats were divided into 6 groups of healthy control, nano-silver, aerobic training, anaerobic training, nano-silver + aerobic training, nano-silver + anaerobic training. First, the training groups trained with the aerobic and anaerobic protocol for 10 weeks. Then, intraperitoneal injection of silver nanoparticles was injected 5 times per 10% of the  bodyweight of each rat, and 48 hours after the last injection, the rats were anesthetized and sampled. The samples were photographed and studied by hematoxylin-eosin staining with a light microscope.
Results: The results showed that anaerobic exercise was significantly effective in weight loss in rats (p = 0.045). Also, oxygen consumption increased in all groups except the group receiving nano-silver compared to the control group (p = 0.000). Injection of biological nano-silver also caused inflammation and hyperemia in the lung tissue of untrained rats. However, in the anaerobic training group, inflammation and hyperemia were less than in the aerobic group.
Conclusion: It seems that injection of biologically produced silver nanoparticles causes structural complications in the lung tissue of male Wistar rats. Also, the results of the present study showed that anaerobic pre-conditioning is more beneficial than aerobic in reducing these effects.