Development Of A Plant Regeneration System And Analysis Of 101 Heat Shock Protein In Strawberry Cv. Camarosa Following Gene Bombardment

The aims of this study were to develop in vitro regeneration system and to confirm the transient expression of HSP101 gene via protein analysis in strawberry cv. Camarosa. In the in vitro study, two types of explants which were shoot tips derived from runner tips and leaves were used. Different types of cytokinins such as BAP, TDZ and zeatin at different concentrations were assessed for shoot induction, while the auxins IBA and NAA also at different concentration were used in the root induction experiment. The experiments were conducted in a Randomized Complete Block Design (RCBD). In the shoot induction experiment using shoot tips cultured on different concentrations and combinations of TDZ and BAP, MS medium supplemented with 4 μM BAP in combination with 2 μM TDZ was optimum for strawberry shoot proliferation. In the shoot induction experiment from shoot tips using zeatin, the highest percentage of explant producing shoots and number of shoots formed per explant were obtained on MS medium containing 4 μM zeatin. High frequency of shoot regeneration from strawberry leaves using different concentrations and combinations of BAP and TDZ was achieved on MS medium containing 4 μM TDZ, without BAP. In the rooting study, MS medium containing 1 μM NAA, MS medium containing 1 μM IBA and MS medium without auxins, were most suitable in inducing the highest number of roots per explant, highest percentage of root formation and the longest root, respectively. Biolistic method of gene transfer has the advantage of allowing a fast and rapid analysis, and was therefore selected for transient expression of HSP101 gene in strawberry via protein analysis. In this study, in vitro leaf explants of strawberry were used. Transient gene expression assays of the AtHSP101 gene showed that this gene can be transiently expressed in strawberry plants. An additional faint protein band of approximately 100 kD was observed on SDS polyacrylamide gel electrophoresis after bombardment of the leaf explant with plasmid, which most probably corresponded to the HSP101 encoded product. In the study on total protein assay using Bradford method, the amount of total protein after bombardment of leaf explant with plasmid containing HSP101 gene increased in comparison with bombardment without plasmid and with non bombarded explants. This result also confirmed that this gene can be transiently expressed in strawberry plants. Therefore by using the regeneration protocol obtained in this study and HSP101 gene which can be transiently expressed, genetic engineering of strawberry cv. Camarosa for heat tolerance can be achieved

An efficient Agrobacterium-mediated transformation of strawberry cv. camarosa by dual plasmid system

An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet

Examining Family Cohesion during the Virtual Education of Students in the Course of the COVID-19 Pandemic

Introduction: The COVID-19 pandemic is linked to the development of virtual education for students at home and a shift in parental role. The family's cohesion and structure can be affected by a change in parental roles. This study was designed and implemented with the aim of investigating the effects of virtual education system designed for students on family cohesion during the COVID-19 pandemic. Materials and Methods:A cross-sectional descriptive-analytical study was conducted with 374 parents of public school students in Khaf city in 2021. Samani's family cohesion questionnaire was utilized to collect the required data. The data were then analyzed through SPSS version 26, more specifically by using Tukey's one-way analysis of variance and post hoc statistical tests. The level of significance was set at p<0.05. Results: Mothers played the biggest role in the virtual education process of their children (69%). The majority of the participants (47.9%) were parents of elementary school students. The mean family cohesion score among the parents was 81.69±23.66, indicating a moderate level. There was a significant difference (p<0.05) in the mean score of family cohesion among the studied parents based on the students’ education stage, the parents’ education levels, occupations, and monthly income. Conclusion: The results showed that the virtual education of students in the corona pandemic has affected the cohesion of the family. The average level of family cohesion during virtual education is a source of danger for the foundation of the family. More attention should be paid to the issue of the family. The excellence and cohesion of the family requires accurate and practical programs in order to recognize the problems faced by the family during the virtual education of students

Evolution of the Magnetized, Neutrino-Cooled Accretion Disk in the Aftermath of a Black Hole Neutron Star Binary Merger

Black hole-torus systems from compact binary mergers are possible engines for gamma-ray bursts (GRBs). During the early evolution of the post-merger remnant, the state of the torus is determined by a combination of neutrino cooling and magnetically-driven heating processes, so realistic models must include both effects. In this paper, we study the post-merger evolution of a magnetized black hole-neutron star binary system using the Spectral Einstein Code (SpEC) from an initial post-merger state provided by previous numerical relativity simulations. We use a finite-temperature nuclear equation of state and incorporate neutrino effects in a leakage approximation. To achieve the needed accuracy, we introduce improvements to SpEC's implementation of general-relativistic magnetohydrodynamics (MHD), including the use of cubed-sphere multipatch grids and an improved method for dealing with supersonic accretion flows where primitive variable recovery is difficult. We find that a seed magnetic field triggers a sustained source of heating, but its thermal effects are largely cancelled by the accretion and spreading of the torus from MHD-related angular momentum transport. The neutrino luminosity peaks at the start of the simulation, and then drops significantly over the first 20\,ms but in roughly the same way for magnetized and nonmagnetized disks. The heating rate and disk's luminosity decrease much more slowly thereafter. These features of the evolution are insensitive to grid structure and resolution, formulation of the MHD equations, and seed field strength, although turbulent effects are not fully convergedComment: 17 pages, 18 figure

Antifungal effect of sesame medicinal herb on Candida Species: original study and mini-review

The aim of this study was to evaluate the antifungal susceptibility patterns of three antifungals, methanolic extracts and N-hexane oil of sesame seeds on C. albicans and C. glabrata, isolated from oral cavity of liver transplant recipients. The results were compared with other reports to develop a mini review as well. Candida species were isolated from liver transplant recipients. To evaluate the antifungal activity of sesame seed oil and methanolic extract, fluconazole, caspofungin and nystatin, the corresponding minimum inhibitory concentrations were determined by CLSI M27-A3 standard method. Minimum fungicidal concentration was also evaluated. The most prevalent species was C. albicans, followed by C. glabrata. Findings indicated sensitivity to antifungal agents and resistance to methanolic extract and N-hexane oil for all C. albicans and C. glabrata isolates. The rate of Candida colonization in the oral cavity of liver transplant recipients was high. Our results revealed that the methanolic and N-hexan extracts of sesame seeds are not effective on C. albicans and C. glabrata species, isolated from the patients. The sesame seed oil pulling and mouthwash cannot effectively cleanse and remove the Candida species in the mouth. Investigation of other medicinal plants or other parts of sesame like leaves and roots are suggested

Micropropagation of strawberry cv. camarosa: prolific shoot regeneration from in vitro shoot tips using thidiazuron with N6-benzylamino-purine

An efficient micropropagation system for strawberry cv. Camarosa was developed. Sterilized runner tips were cultured on hormone-free Murashige and Skoog (MS) medium with 3% sucrose, 1 mL L-1 Plant Preservative Mixture, and solidified using 0.25% phytagel to produce in vitro stock plants. Shoot tips derived from the in vitro stock plants were cultured on MS media containing 0, 2, 4, and 8 mM thidiazuron (TDZ) and 0, 4,9,18, and 27 mM N6-benzylamino-purine (BAP) for shoot induction. Shoots produced on the best shoot induction medium were rooted on MS media containing 1, 2, 3, and 5 mM of either indole-3-butyric acid (IBA) or naphthaleneacetic acid (NAA). Results showed that MS medium with 2 mM TDZ and 4 mM BAP was optimum for shoot multiplication from the shoot tips. The most suitable medium for inducing the highest number of roots per explant, the highest percentage of explant with roots, and the highest mean root length were 1 mM NAA, 1 mM IBA, and hormone-free MS medium, re-spectively. Plantlets were transplanted into substrate consisting of perlite + vermiculite + cocopeat (2:1:2 v/v/v) resulting in 90% survival. After 1 month, plants were irrigated using Hoagland's solution and runners were produced after 3 months

Breaking-off tissue specific activity of the oil palm metallothionein-like gene promoter in T1 seedlings of tomato exposed to metal ions

Metallothioneins (MTs) are cysteine-rich metal-binding proteins that are involved in cell growth regulation, transportation of metal ions and detoxification of heavy metals. A mesocarp-specific metallothionein-like gene (MT3-A) promoter was isolated from the oil palm (Elaeis guineensis Jacq). A vector construct containing the MT3-A promoter fused to the β-glucuronidase (GUS) gene in the pCAMBIA 1304 vector was produced and used in Agrobacterium-mediated transformation of tomato. Histochemical GUS assay of different tissues of transgenic tomato showed that the MT3-A promoter only drove GUS expression in the reproductive tissues and organs, including the anther, fruit and seed coat. Competitive RT-PCR and GUS fluorometric assay showed changes in the level of GUS mRNA and enzyme activity in the transgenic tomato (T0). No GUS mRNA was found in roots and leaves of transgenic tomato. In contrast, the leaves of transgenic tomato seedlings (T1) produced the highest GUS activity when treated with 150 μM Cu2+ compared to the control (without Cu2+). However, Zn2+ and Fe2+ treatments did not show GUS expression in the leaves of the transgenic tomato seedlings. Interestingly, the results showed a breaking-off tissue-specific activity of the oil palm MT3-A promoter in T1 seedlings of tomato when subjected to Cu2+ ions

Vitis Elegan as a Promising Antidiabetic Herb: Phytochemical and Pharmacological Assessment

In this research, we investigated the antidiabetic activity of Vitis elegans rhizome, which is used as traditional treatment for diabetes mellitus. The methanol, chloroform, petroleum ether, and hexane extracts of the plant root were obtained through serial exhaustive extraction and were analyzed by Thin Layer Chromatography (TLC).  Glycogen phosphorylase (GP) assay was done to determine the inhibitory effects of respective extracts on GP enzyme. Total phenol content was measured using the Folin-Ciocalteu method and brine shrimp test was done to evaluate the toxicity of the extracts. Evaluation of TLC plates alone and after spraying with different reagents indicated that terpenoid was the major component of the sample followed by alkaloid and phenol. Chloroform extract applied more inhibitory effects on GP enzyme activity with percentages of 39.65 in concentration of 2.5 mg/ml. This suppression effect was higher than glucose, as a standard inhibitory agent in the body. The highest amount of phenol was found in the methanol extract, equal to 49 mg GAE g-1. Petroleum ether, chloroform and methanol extracts were considered as non-toxic solvents with LC50 values of 8.9, 3.5 and 3.7 mg/ml respectively. While hexane with 0.089 mg/ml LC50 value was classified as toxic extract. Based on the results of this study, we concluded that Vitis elegans rhizome, has the potential to be further studied for anti-hyperglycemic properties

AGS cell line xenograft tumor as a suitable gastric adenocarcinoma model: growth kinetic characterization and immunohistochemistry analysis

Objective(s): Gastric cancer is the third leading cause of cancer-related death worldwide. The overall survival rate of patients is poor because gastric cancers are usually diagnosed at the late stages. Therefore, further research is needed and appropriate research tools are required to develop novel therapeutic approaches.Materials and Methods: Eight female athymic nude mice with a C57BL/6 background were used in this study. AGS cells were inoculated into the flank. The tumor volumes were calculated and growth curves were drawn. When the volume of the tumors reached 1000 mm3, the animals were humanely euthanized with CO2 gas. After harvesting, tumors were analyzed with Hematoxylin and Eosin (H&E) and immunohistochemistry (IHC). A pathologist confirmed tumor entity through H&E staining. Tumors were evaluated for expression of HER-2, P53, Ki-67, CD34, cytokeratin 8 (CK8), vimentin, estrogen receptor (ER), and progesterone receptor (PR) utilizing immunohistochemistry.Results: The tumor take rate was 62.5%, mean doubling time was 40.984 d, and the latency period was 30.62 days. The H&E staining results showed highly malignant hyperchromatin epithelial cells. IHC assessment showed the mutation status of P53 gene. The expression score of the CK8 protein in the tumor cells was +3. Vimentin protein was not expressed and changes in mesenchymal phenotype were not observed. Ki-67 IHC indicated that the proliferation rate was >43% and angiogenesis was defined as high MVD.Conclusion: The respective AGS xenograft model provides an opportunity to understand the pattern of tumor growth as well as to evaluate new gastric cancer therapies in in vivo studies

Detection of tetracycline resistance genes, aminoglycoside modifying enzymes, and coagulase gene typing of clinical isolates of Staphylococcus aureus in the Southwest of Iran

Objective(s): The aim of the present study was to determine the aminoglycoside modifying enzymes (AMEs) encoded genes, tetracycline resistance genes, and the coa based typing of Staphylococcus aureus isolates in the Southwest of Iran. Materials and Methods: Antimicrobial susceptibility of isolates was carried out by agar disk diffusion methods. Two sets of multiplex PCR mixture were used for detection of AME genes and tet genes.  All of the isolates were typed with the coagulase gene typing method. Of the 121 isolates, 29.75% and 47.93% were resistant to at least one aminoglycosides and tetracyclines, respectively. Results: The aac(6')-Ie-aph(2'') was the most frequent gene (97.22%), and aph (3')-IIIa and ant (4')-Ia genes were detected in 61.11% and 11.11% of aminoglycoside resistant isolates, respectively. The tetK and tetM genes were detected in 82.75% and 56.9% of tetracycline resistant isolates, respectively. Overall 31.4% of isolates were MRSA. Totally 17 distinct coa gene RFLP patterns, numbered C1 to C17, were observed.  The C5 was the most frequent coa type with 31 isolates. Conclusion: The aac(6')-Ie-aph(2'') and aph (3')-IIIa genes were the most important genes contributing to aminoglycosides resistance, while resistance to tetracyclines was mediated by tetK and tetM genes. Interestingly all S. aureus with C5 as the most prevalent coa-type were resistant to at least one of the aminoglycoside antibiotics and tetracycline simultaneously. Moreover, 30 out of 31 isolates with this coa type were MRSA, indicating the importance of the C5 coa-type in MRSA strains and also in isolates that were resistant to aminoglycosides and tetracycline