Multivariate analysis of the phytochemical composition and antioxidant properties in twenty-five accessions across three Achillea species.

This study explored the chemical composition, antioxidant activity, and total phenol content of aerial parts from 25 accessions of three Achillea species (Achillea wilhelmsii C. Koch, Achillea vermicularis Trin., and Achillea tenuifolia Lam.). The plants were collected from various natural habitats across Iran, encompassing regions such as Central, Western, Southern, Northern, Western, and Northwestern parts of the country. Subsequently, they were grown together under field conditions. The study revealed significant variation in essential oil yields among accessions of A. wilhelmsii, ranging from 0.01 to 0.107%, A. vermicularis with a range of 0.075 to 1.5%, and A. tenuifolia showing a variation of 0.1 to 2%. The study utilized Gas Chromatography-Mass Spectrometry (GC-MS) analysis, revealing 75, 49, and 75 compounds in the essential oils of A. wilhelmsii, A. tenuifolia, and A. vermicularis, respectively. Major components included camphor, 1,8-cineole, anethole, α-pinene, and phytol in A. wilhelmsii, 1,8-cineole, camphor, levo-carvone, and δ-terpinene in A. vermicularis, and ß-cubebene, elixene, ß-sesquiphellandrene, 1,8-cineole, camphor, and δ-terpinene in A. tenuifolia. The essential oil compositions of A. wilhelmsii and A. vermicularis were predominantly characterized by oxygenated monoterpenes, whereas that of A. tenuifolia was characterized by sesquiterpenes. Cluster analysis grouped accessions into three clusters, with A. tenuifolia forming a distinct group. Principal Component Analysis (PCA) triplot (62.21% of total variance) confirmed these results and provided insights into compound contributions. Furthermore, total phenolic content and antioxidant activity of the accessions of three species were assessed over 2 years. A. tenuifolia exhibited the highest levels in both categories, with statistically significant linear regression between antioxidant activity and total phenol content for A. tenuifolia and A. wilhelmsii. These findings emphasize significant phytochemical diversity within Achillea species, positioning them as promising natural sources of antioxidants. Further exploration and selection of specific accessions within each species are crucial for unlocking their medicinal potential and supporting cultivation and conservation efforts.

Successful use of Nicotiana tabacum hairy roots for the recombinant production of Cecropin A peptide.

Cecropin A, as an antimicrobial peptide (AMP), is possible to use in medical and agricultural fields as a new and safe biocontrol agent. Therefore, it is highly necessary to find a cost-effective and scalable approach to generate a large scale of it. In this research, the Agrobacterium rhizogenes strain ATCC 15834 was used to transfer the Cecropin A gene to the Nicotiana tabacum. After confirmation of transgenic hairy roots, the antibacterial activity of purified Cecropin A peptide was measured using the agar gel diffusion method. Successful transforming of Cecropin A was confirmed at the RNA and protein levels in hairy root cells using RT-PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The highest Cecropin A amount was detected in line 4 of the transgenic lines using ELISA in comparison with the nontransgenic line. Subsequently, the antimicrobial activity of Cecropin A extracted from line 4 showed the highest inhibition activity against Aspergillus niger. Besides, this activity was stable against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans pathogens after 7 days. The recombinant production of Cecropin A AMP had a yield of 63.81 µg/g of fresh weight. According to a significant yield, this system can be used to produce the Cecropin A peptide for pharmacological and food science applications.

Genotypic diversity of 17 cacti species and application to biosynthesis of gold nanoparticles.

The genotypic diversity of 17 cacti species were examined and grouped in four clusters using seven inter simple sequence repeat (ISSR) markers. Group representatives (five species) were chosen for AuNPs synthesis in the cacti syrups. To synthesize the Gold nanoparticles (AuNPs), reducing and capping potential of five species of cacti rich in the polyphenolics were explored. Based on the synthesized AuNPs traits (concentration, pH, temperature, and synthesis time), Opuntia pycnacantha with the highest absorption peak at 540 nm was chosen for further characterizations. Varieties of diffraction peaks confirmed the successful synthesis of AuNPs. AuNPs functionalization with the phenolic compounds was confirmed by Fourier transform infrared (FTIR) spectroscopy. At the optimum conditions (pH = 5.0 and T = 60 °C), both dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed more than 87% of AuNPs to be 2.5 nm in size with Zeta potential to be equal to -19.9 mV.

VARIATION SIMULATION OF RADON CONCENTRATION IN THE TISSUES AND ORGANS OF THE BODY BY AN ELECTRICAL CIRCUIT.

In this study, a new model based on electric circuit theory has been introduced to simulate the dynamics of radioactive chemically inert gases in the human body. For this manner, it is assumed that inert gas is transported through the body to various organs via the blood stream. In this simulation, a voltage source is equivalent to gas generation in the atmosphere, the conductivity is equivalent to the cardiac output of the organ, the capacitor capacitance is equivalent to the volume of blood or tissue and voltage across a capacitor is equivalent to the gas concentration in air or blood or a tissue. This simulation can be used to study the dynamics of any inert gas whose partition coefficients are known. We use this simulation to study the dynamics of radon in human body. The physiologically based pharmacokinetic (PBPK) model that describes the fate of radon in systemic tissue has been used for this simulation. Using this simulation, the effective dose equivalent resulting from inhalation of radon has been estimated. The calculated values agree with the previously reported value. Also, using the model, it has been shown that after inhalation of radon gas, absorbed dose has been decreased in different tissues by increasing the inhalation rate without radon. So that, by doubling the inhalation rate and the rate of cardiac output, the value of the absorbed dose has been decreased 11.88% in the adipose tissue, 25.49% in the red marrow tissue and 20.3% in the liver organ.

Phaseolus vulgaris genome possesses CAMTA genes, and phavuCAMTA1 contributes to the drought tolerance.

Calmodulin-binding transcription activators (CAMTAs) are a family of transcription factors that play an important role in plants' response to the various biotic and abiotic stresses. The common bean (Phaseolus vulgaris L.) is one of the most important crops in the world and plays a pivotal role in sustainable agriculture. To date, the composition of CAMTA genes in genomes of Phaseolus species and their role in resistance to drought stress are not known. In this study, five PhavuCAMTA genes were characterized in common bean genome through bioinformatics analysis, the morphological and biochemical response of 23 Ph.vulgaris genotypes to different levels of drought stress were evaluated and the expression patterns of PhCAMTA1 in the leaf tissues of sensitive and tolerant genotypes were analysed. Gene structure, protein domain organization and phylogenetic analyses showed that the CAMTAs of Phaseolus were structurally similar and clustered into three groups as other plant CAMTAs. Genotypes showeda differential response to drought stress. Thus, the plant height, number of nodes and flower, total chlorophyll and total protein content, and activity of antioxidant enzymes (ascorbate peroxidase and catalase) in plants significantly influenced by genotype and drought stress interaction. Moreover, the resistant and susceptible genotypes were identified according to three-dimensional plots and the expression patterns of PhavuCAMTA1 gene were studied using real-time quantitative polymerase chain reaction. The results of the present study serve as the basis for future functional studies on the Phaseolus CAMTA family.

Practical analysis of aflatoxin M1 reduction in pasteurized Milk using low dose gamma irradiation.

Milk and dairy products can be exposed to potential dangers such as aflatoxin M1 (AFM1). Various factors affect the formation of aflatoxin, which can be due to environmental changes and the lack of suitable substrate for healthy livestock feeding. The goal of this study is to reduce the toxin in pasteurized milk to a level below the European Codex Alimentarius Commission standard. For this purpose, the proper structure of the radioactive granite stone was first designed as a low level gamma irradiation (LLGI) without contact with pasteurized milk, and and the pasteurized milk containing AFM1 that placed in this structure is measured and compared with the control sample values using Association of Official Analytical Chemists (AOAC) method. Then, the reduction of the resulting aflatoxin in the milk and the LLGI dose rate are obtained. The LLGI dose rate is calculated using the Monte Carlo N-Particle Transport Code (MCNP). For simulation, in addition to the spectrum of gamma radiation emitted by radioactive granites, weight percent of each composition of the pasteurized milk and its component elements are also calculated. The results showed a 51.5% reduction of aflatoxin in pasteurized milk after 4 days and 99% reduction after 8 days compared to the control sample. The LLGI dose rate in milk is 0.39 mGy per day. According to the international atomic energy agency (IAEA) report and pervious results, this dose rate level does not significantly affect chemical and sensory quality of milk, but can extend the shelf-life and provide a safer milk. Therefore, the structure constructed using radioactive granite in this study can be considered as one of the suitable methods for reducing aflatoxin.

An investigation of the HMGR gene and IPI gene expression in black caraway (Bunium persicum).

Black caraway is of great importance for its terpene compounds. Many genes are involved in the biosynthesis of secondary metabolites in medicinal plants. For this study, black caraway seeds were collected from five different regions, i.e. [Isfahan; Kerman (Khabr); Semnan; Kerman (Sirch); and Hormozgan]. The black caraway seed oil was extracted and analyzed by means of the gas chromatography method. There was a negatively significant correlation (p ≤ 0.05) observed between cuminaldehyde and gammaterpinene compounds. 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and isopentenyl pyrophosphate isomerase (IPI) play an important role in the biosynthesis of secondary metabolites. Appropriate primers were designed for these genes based on the conserved regions in other plants. Amplified fragments were then sequenced. Blastn results indicated the similarity of the high RNA sequences between new sequences and other HMGR and IPI gene sequences in GenBank, and it also identified the HMGR and IPI gene sequences of B. persicum. A fragment of the HMGR gene with KJ143741 number was recorded in the gene bank. Quantitative PCR showed that the relative expression of two genes in different growth stages of B. persicum was significantly different between the germination stage and the multi-leaf stage, and also between the germination stage and the flowering stage (p < 0.05); however, there was no significant difference observed between the flowering stage and the multi-leaf stage. The results indicated that the expression of HMGR increased from the germination stage to the adult plant, and then it got stable until the flowering stage; in the same vein, the expression of IPI increased continuously from the germination stage to the flowering stage. The expression of HMGR and IPI genes occurred differently at the germination stage of five ecotypes. The Hormozgan ecotype showed the least expression rate.

Anti-carcinogenic and anti-angiogenic properties of the extracts of Acorus calamus on gastric cancer cells.

OBJECTIVE: Acorus calamus (A. calamus) has been used as a medicinal plant in Asia for its effects on digestive system for the last 2000 years. To investigate the anti-cancer activity of rhizome of A. calamus, the ethanolic and methanolic extracts and essential oil of the rhizome were prepared and their effects were assessed on human gastric cancer cell line (AGS). MATERIALS AND METHODS: The viability of cells which were treated with the extracts and the essential oil was assessed by MTT assay. To evaluate the anti-angiogenic property of the extracts, in vitro tube formation assay was done. Cell cycle distribution and the expression of Oct4 and Nucleostemin, after treatments, were checked by flowcytometry and quantitative RT-PCR, respectively. Furthermore, analysis of essential oil from A.calamus was done by GC-MS. RESULTS: Our results showed that the growth of AGS cells was inhibited by the extracts and essential oil and the extracts inhibited the angiogenesis in HUVEC cells. Our data revealed that the extracts and essential oil of A. calamus caused G1 arrest in AGS cells and downregulation of Oct4 and NS after treatment. By GC-MS analysis, we found new compounds such as epiprezizaene, valencene and isocyclocitral in essential oil of A. CONCLUSION: All together, our results showed that the extracts of A. calamus have anti-proliferative and anti-angiogenic effects on cancer cells.

Optimum location of external markers using feature selection algorithms for real-time tumor tracking in external-beam radiotherapy: a virtual phantom study.

In external-beam radiotherapy, using external markers is one of the most reliable tools to predict tumor position, in clinical applications. The main challenge in this approach is tumor motion tracking with highest accuracy that depends heavily on external markers location, and this issue is the objective of this study. Four commercially available feature selection algorithms entitled 1) Correlation-based Feature Selection, 2) Classifier, 3) Principal Components, and 4) Relief were proposed to find optimum location of external markers in combination with two "Genetic" and "Ranker" searching procedures. The performance of these algorithms has been evaluated using four-dimensional extended cardiac-torso anthropomorphic phantom. Six tumors in lung, three tumors in liver, and 49 points on the thorax surface were taken into account to simulate internal and external motions, respectively. The root mean square error of an adaptive neuro-fuzzy inference system (ANFIS) as prediction model was considered as metric for quantitatively evaluating the performance of proposed feature selection algorithms. To do this, the thorax surface region was divided into nine smaller segments and predefined tumors motion was predicted by ANFIS using external motion data of given markers at each small segment, separately. Our comparative results showed that all feature selection algorithms can reasonably select specific external markers from those segments where the root mean square error of the ANFIS model is minimum. Moreover, the performance accuracy of proposed feature selection algorithms was compared, separately. For this, each tumor motion was predicted using motion data of those external markers selected by each feature selection algorithm. Duncan statistical test, followed by F-test, on final results reflected that all proposed feature selection algorithms have the same performance accuracy for lung tumors. But for liver tumors, a correlation-based feature selection algorithm, in combination with a genetic search algorithm, proved to yield best performance accuracy for selecting optimum markers.

High-level extracellular secretion of organophosphorous hydrolase of Flavobacterium sp. in Escherichia coli BL21(DE3)pLysS.

Organophosphorus (OPs) compounds are widely used in many pesticides, insecticides, and chemical nerve agents. These compounds are hazardous for humans and the environment. There are many reports on detoxification of these compounds, among them enzymatic cleavage of these compounds with organophosphorus hydrolase (OPH) has been taken into more consideration. Several studies have been performed to improve OPH secretion in Escherichia coli by different signal peptides, but have not been successful. In this study, to achieve the extracellular secretion of OPH in E. coli, the complete opd gene along with its native signal peptide was codon optimized and expressed in E. coli BL21(DE3)pLysS. The culture medium showed OPH activity after 2, 4, and 6 H of induction time. The extracellular secretion of OPH was also confirmed by SDS-PAGE and Western blot analysis. The effects of different factors in growth medium were also investigated regarding expression and extracellular secretion of OPH. It appears that the secretion of OPH into the extracellular medium is highly affected by culture conditions. Therefore, our results revealed that the recombinant OPH was successfully secreted into the extracellular medium. This secretion system can be considered as a high efficiency biocatalyst for detoxification of OPs compounds.

Genetic analysis of an F2 intercross between two strains of Japanese quail provided evidence for quantitative trait loci affecting carcass composition and internal organs.

The purpose of this study was to identify genomic regions, quantitative trait loci (QTL), affecting carcass traits on chromosome 1 in an F2 population of Japanese quail. For this purpose, two white and wild strains of Japanese quail (16 birds) were crossed reciprocally and F1 generation (34 birds) was created. The F2 generation was produced by intercrossing of the F1 birds. Phenotypic data including carcass weight, internal organs and carcass parts were collected on F2 animals (422 birds). The total mapping population (472 birds) was genotyped for 8 microsatellite markers on chromosome 1. QTL analysis was performed with interval mapping method applying the line-cross model. Significant QTL were identified for breast weight at 0 (P < 0.01), 172 (P < 0.05) and 206 (P < 0.01), carcass weight at 91 (P < 0.05), carcass fatness at 0 (P < 0.01), pre-stomach weight at 206 (P < 0.01) and uropygial weight gland at 197 (P < 0.01) cM on chromosome 1. There was also evidence for imprinted QTL affecting breast weight (P < 0.01) on chromosome 1. The proportion of the F2 phenotypic variation explained by the significant additive, dominance and imprinted QTL effects ranged from 1.0 to 7.3%, 1.2 to 3.3% and 1.4 to 2.2%, respectively.

Expression of the truncated tissue plasminogen activator (K2S) gene in tobacco chloroplast.

As because the plant plastid genome is highly polyploid, the transformation of chloroplasts permits the introduction of thousands of copies of foreign genes per plant cell and generates extraordinarily high levels of recombinant protein. Human tissue-type plasminogen activator is one of the most important pharmaceutical proteins involved in the breakdown of blood clots in brain and heart blood vessels. We report the introduction and expression of the truncated human tissue plasminogen activator (K2S) gene in tobacco chloroplasts. The K2S-containing vector pKCZK2S was successfully transferred to tobacco plastomes using the biolistic delivery procedure. Transplastomic plants were selected on RMOP medium containing spectinomycin (500 mg/l). In order to achieve homoplasmy, several rounds of selection and regeneration were performed. The presence, site-specific integration, homoplasmy, expression and activity assay of the transgene were confirmed in the transplastomic plants by PCR, Southern-blot, RT-PCR, SDS-PAGE, ELISA, Dot-blot, Western-blot and zymography analysis. Our results show that the tissue plasminogen activator (K2S form) protein to be expressed in tobacco chloroplasts in active form.

Enzyme activity and biochemical changes of three date palm cultivars with different softening pattern during ripening.

The physicochemical composition changes during palm ripening were studied. The activities of invertase and cell wall-modifying enzymes, namely pectin methylesterase (PME), ß-galactosidase (ß-Gal), endo-1,4-ß-D-glucanase (EGase), and cellulase were monitored during growth and ripening of Shahani, Piarom and Deiry cultivars with different textures. Also, we estimated the concentrations of six organic acids by HPLC. Reducing sugars, most of organic acid, pH and TSS increased up to the full-ripe stage of all date types. On the contrary, moisture, macro and microelements and proteins decreased during the same period. Despite a considerable rise in invertase activity (200 units) during ripening of Shahani, no significant trend could be discerned in Deiry cultivar (8 units) at different stages. Our results also showed that cell wall enzymes were increased in activity during ripening and these increases were coincident with fruit softening. Furthermore, our results showed that the composition and the variation of the chemical compositions mainly depended on the cultivar and maturity stage.

Assessment of bile salt effects on s-layer production, slp gene expression and some physicochemical properties of Lactobacillus acidophilus ATCC 4356.

In many conditions, bacterial surface properties are changed as a result of variation in growth medium and conditions. This study examined the influence of bile salt concentrations (0-0.1%) on colony morphotype, hydrophobicity, H2O2 concentration, S-layer protein production and slpA gene expression in Lactobacillus acidophilus ATCC 4356. It was observed that two types of colonies (R and S) were in the control group and the stress condition. When the bile level increased in the medium, the amount of S type was more than the R. A stepwise increment in the bile concentration resulted in a stepwise decline in the maximum growth rate. The results showed that hydrophobicity was increased in 0.01%-0.02% bile but it was decreased in 0.1% bile. Treatment by bile (0.01%- 0.1%) profoundly decreased H2O2 formation. S-layer protein and slpA gene expression was also altered by stress condition. S-protein expression was increased in stress condition. slpA gene expression increased in 0.01%-0.05% bile and it decreased in 0.1% bile. However, we found that different of bile salt concentrations influence on morphology and some surface properties of L. acidophilus ATCC 4356. These changes were very different in the 0.1% bile. It appears that the bacteria respond abruptly to 0.1% bile.