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Aquaculture by-products have been of great interest for producing protein hydrolysates with multiple biological activities. The present experiment was carried out to evaluate dietary effects of a low-molecular fraction (10 kDa) from shrimp waste hydrolysate in forms of unprotected and nanocapsulated on growth and immunity of rainbow trout. Therefore, six diets were designed including a control diet (no supplementation), D1 (1 g kg

Eggplant (Solanum melongena L.) is an economically important solanaceous crop in Iran with fruits used for food and traditional medicine. Despite the importance of Alternaria leaf spot and blight disease of solanaceous crops which is commonly seen in the fields, our knowledge about the causal agents on eggplant is limited. In this study, a set of large-spored Alternaria isolates was recovered from eggplant with leaf spot and blight symptoms in Somehsara region, Guilan province, Iran. All recovered isolates shared conspicuous morphological characteristics e.g. production of large, solitary conidia with several transverse disto- and eusepta and long tapering filamentous beak resemble those seen in the members of Alternaria section Porri. Multi-locus phylogenetic analyses based on the internal transcribed spacer region of nrDNA (ITS-rDNA) and parts of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), second largest subunit of RNA Polymerase II (RPB2), translation elongation factor 1-alpha (TEF1-α) and Alternaria major allergen (Alt a 1) gene sequences provided further evidence supporting not only their exact placement in Alternaria sect. Porri, but also in a distinct lineage representing a new species. The new species was named, described and illustrated herein as Alternaria guilanica sp. nov.. The phylogenetic and morphological comaprisions of the new species with other closely related species were also provided. Pathogenicity test conducted for the new strains revealed that they were capable to induce disease symptoms on eggplant leaves under greenhouse conditions, and re-isolation of the inoculated isolates confirmed Koch’s postulates.

Fungal protoplast fusion is an approach to introduce novel characteristics into industrially important strains. Cellulases, essential enzymes with a wide range of biotechnological applications, are produced by many species of the filamentous fungi Trichoderma. In this study, a collection of 60 natural isolates were screened for Avicel and carboxymethyl cellulose degradation, and two cellulase producers of Trichoderma virens and Trichoderma harzianum were used for protoplast fusion. One of the resulting hybrids with improved cellulase activity, C1-3, was fused with the hyperproducer Trichoderma reesei Rut-C30. A new selected hybrid, F7, was increased in cellulase activity 1.8 and 5 times in comparison with Rut-C30 and C1-3, respectively. The increases in enzyme activity correlated with an upregulation of the cellulolytic genes cbh1, cbh2, egl3, and bgl1 in the parents. The amount of mRNA of cbh1 and cbh2 in F7 resembled that of Rut-C30 while the bgl1 mRNA level was similar to that of C1-3. AFLP (amplified fragment length polymorphism) fingerprinting and GC–MS (gas chromatography – mass spectrometry) analysis represented variations in parental strains and fusants. In conclusion, the results demonstrate that a 3-interspecific hybrid strain was isolated, with improved characteristics for cellulase degradation and showing genetic polymorphisms and differences in the volatile profile, suggesting reorganizations at the genetic level.

Sclerotinia sclerotiorum, a destructive fungal pathogen with an extensive host range infecting more than 400 plant species, causes lettuce drop on the leafy green lettuce that potentially have an enormous economic impact on lettuce cultivation worldwide. To gain insights into how lettuce regulates its defense pathways, gene expression profiles of five defense-related genes (LsPRB1, LsSOD, LsERF1, LsLTC1, and LsHPL1) triggered following infection of susceptible Mazandaran line 1 (ML1) and tolerant Jahrom (Jah) lettuce accessions by the S. sclerotiorum were compared by the real-time quantitative RT-PCR (RT-qPCR) approach. In the current study, we observed temporal and quantitative gene expression fluctuations between two examined accessions of L. sativa in response to S. sclerotiorum attack. All genes, except LsHPL1, were up-regulated earlier (24 hours after inoculation) in the Jah accession compared with the susceptible one. This data implies strong defense responses established in the tolerant accession to arrest the fungal growth, but it resulted in restricting lesion development rather than in preventing infection. This research contributes to a better understanding of the kinetics of lettuce reactions induced following S. sclerotiorum infection and may be employed to develop effective strategies to manage lettuce drop.

In a survey of pyricularia-like fungi on different plants, especially on sedges in the north of Iran, we identified two typical isolates characteristics of the genus Pseudopyricularia. Phylogeny analysis based on DNA sequences of internal transcribed spacer regions (ITS-rDNA), the largest subunit of the RNA polymerase II (RPB1), calmodulin (CAL), and large subunit of the ribosomal RNA (LSU) gene regions, and morphological criteria confirmed the new species of Pseudopyricularia. Pseudopyricularia hedjaroudii sp. nov., isolated from the leaves of Cyperus sp., is described and illustrated. Conidia of the Ps. hedjaroudii are obclavate, obpyriform, and 1-septate distinct from other allied species in size and shape.

Split luciferase complementary assay has been used to investigate the effect of WD domain deletion on Apaf-1 oligomerization. Apaf-1 is an adaptor molecule in formation of apoptosome that activates caspase-9, an activation that is a key event in the mitochondrial cell death pathway. Structural studies suggest that normally Apaf-1 is held in an inactive conformation by intramolecular interactions between Apaf-1's nucleotide binding domain and one of its WD40 domains (WD1). In the prevailing model of Apaf-1 activation, cytochrome c binds to sites in WD1 and in Apaf-1's second WD40 domain (WD2), moving WD1 and WD2 closer together and rotating WD1 away from the nucleotide binding domain. This allows Apaf-1 to bind dATP or ATP and to form the apoptosome, which activates caspase-9. This model predicts that cytochrome c binding to both WD domains is necessary for apoptosome formation and that an Apaf-1 with only WD1 will be locked in an inactive conformation that cannot be activated by cytochrome c. Here we investigated the effect of removing one WD domain (Apaf-1 1-921) on Apaf-1 interactions and caspase activation. Apaf-1 1-921 could not activate caspase-9, even in the presence of cytochrome c. These data show that a single WD domain is sufficient to lock Apaf-1 in an inactive state and this state cannot be altered by cytochrome c.

International audience; Blast disease is a notorious fungal disease leading to dramatic yield losses on major food crops such as rice and wheat. The causal agent, Pyricularia oryzae, encompasses different lineages, each having a different host range. Host shifts are suspected to have occurred in this species from Setaria spp. to rice and from Lolium spp. to wheat. The emergence of blast disease on maize in Iran was observed for the first time in the north of the country in 2012. We later identified blast disease in two additional regions of Iran: Gilan in 2013 and Golestan in 2016. Epidemics on the weed barnyard grass (Echinochloa spp.) were also observed in the same maize fields. Here, we showed that P. oryzae is the causal agent of this disease on both hosts. Pathogenicity assays in the greenhouse revealed that strains from maize can infect barnyard grass and conversely. However, genotyping with simple sequence repeat markers and comparative genomics showed that strains causing field epidemics on maize and on barnyard grass are different, although they belong to the same previously undescribed clade of P. oryzae. Phylogenetic analyses including these strains and a maize strain collected in Gabon in 1985 revealed two independent host-range expansion events from barnyard grass to maize. Comparative genomics between maize and barnyard grass strains revealed the presence or absence of five candidate genes associated with host specificity on maize, with the deletion of a small genomic region possibly responsible for adaptation to maize. This recent emergence of P. oryzae on maize provides a case study to understand host range expansion. Epidemics on maize raise concerns about potential yield losses on this crop in Iran and potential geographic expansion of the disease.

Sclerotinia sclerotiorum is one of the most destructive lettuce diseases, restricting lettuce cultivation globally. Here, we measured the activity of three antioxidant enzymes in two contrastingly ...

Bene oil (pistacia atlantica), as a plant source, is rich in phenolic and tocopherol compounds and has significant antioxidant, therapeutic and antimicrobial effects. Encapsulation of hydrophobic compounds in liposome system is an ideal solution for protecting them against destruction during storage. An important advantage of liposomes is the encapsulation of hydrophilic, hydrophobic and amphiphilic compounds by using natural phospholipids, such as lecithin, with beneficial effects. The aim of this study is to encapsulate the bene kernel oil in the form of microliposomes. For this purpose, the effect of composition of liposomes based on lecithin and cholesterol was studied using the Mozafari method. Liposomes are prepared using lecithin and cholesterol in the ratios of 60:0, 50:10, 40:20 and 30:30. Particle size, size distribution, zeta potential and encapsulation efficiency were characterised. According to the result, the size of liposomes was dependent on their composition, but the wasaffected significantly affected by adding cholesterol (P < 0.05). Average diameter of the particles was between 4 and 9 µm. Liposome with a ratio of 40:20 had the smallest size. By applying cholesterol, zeta potential increased from 16.4 mV to 32.39 mV, which indicates electrostatic stability of liposomes. In general, with encapsulation efficiency of 84.33%, the ratio of 40:20 is considered as an ideal concentration in the formulation of microliposomes. Based on the results, bene oil-loaded liposomes with a lecithin: cholesterol formulation ratio of :as 40:20 was chosen as an optimal formulation because of its smaller particle size, higher zeta potential and suitable stability, which can be used in trapping, delivering and releasing hydrophilic, adipose-friendly and amphiphilic compounds (dual-friendly).

Innovative thin-walled structures, bio-inspired by the microstructure of Morpho wings, were proposed as energy absorbing devices in this study. A finite element model, experimentally validated, was...

Gut microbiota composition is unique in every individual, it impacts on organ functions that produce hormones. Gut microbiota composition balance is directly related to our general health status. This continual interaction between gut microbiota and endocrine organs sometimes can be considered as the etiology of diseases such as type 2 diabetes mellitus (T2DM), obesity, osteoporosis, polycystic ovary syndrome (PCOS), and thyroid diseases. Microbiota is introduced for a total collection of microbial organisms in our bodies and microbiome referred for their genome and their collective functions. Near 100 trillion microorganisms live in our body and almost all of them occupy the human gut gastrointestinal tract. Precision medicine can play a crucial role in health maintenance by affecting gut microbiota composition in every individual. It can also develop special treatments specifically for every individual. In this review, we addressed any correlation between gut microbiota and endocrine disorders including T2DM, obesity, PCOS, thyroid disorders and osteoporosis.

Low temperature (LT) and Ascochyta blight are two major stresses in chickpea (Cicer arietinum L.) cultivation. After exposure to LT treatments (acclimation, LT, acclimation + LT and control conditions), we evaluated induced responses by pathogen infection in LT-sensitive (ILC533) and -tolerant (Sel96Th11439) chickpea genotypes. In relation to genotype-specific defense reactions, tolerant plants responded more effective to the infection compared to sensitive plants, particularly after an acclimation treatment. LT treatments, and in particular an acclimation treatment, induced a significant decrease in disease symptoms 7 and 14 days after inoculation. Histopathological studies showed more infection throughout the cells of the sensitive genotype under control conditions. The minimal spread of the pathogen was observed in the tolerant genotype after acclimation treatment, and the infected plants did not contain pycnidia or hyphae of the pathogen. The electrolyte leakage index (ELI) and amount of malondialdehyde (MDA) after LT treatments were less than the values compared to post pathogen infection, indicating a difference in the degree of oxidative stress. Lower levels of pathogen-induced oxidative stress (ELI and MDA) and better photosynthetic efficiency (measured as Fv/Fm values) show that the tolerant genotype exposed to LT treatments acclimate better to the stress conditions compared to the sensitive genotype. Higher transcript levels for the superoxide dismutase, catalase and ascorbate peroxidase were detected during pathogen infection, particularly during the acclimation treatment, which might confirm lower oxidative stress in tolerant plants compared to sensitive plants. It is concluded that LT increased the capacity for tolerance to Ascochyta blight in chickpea plants and this was correlated with the stimulation of mechanisms involved in defense responses.

Amyotrophic lateral sclerosis is a fatal, devastating, rapidly progressive, adult onset neurodegenerative disease, which is involved in the formation of proteinaceous virulent aggregates from superoxide dismutase 1 (SOD1) as a Cu/Zn metalloenzyme in motor neurons. However, the particular mechanism of mutant SOD1 toxicity has remained elusive. Here, we use complementary biochemical and biophysical methods to investigate thermodynamic parameters of wild-type (WT), E100K and D125H mutants in both metal-free (apo) and metalized (holo) forms of SOD1, to find out if there is any relationship between the structural/thermodynamic stability with aggregation propensity in apo and holo forms of WT/mutant SOD1 variants. Also, in this study, we compared the aggregation propensity of WT SOD1 and the mutants (E100K, D125H) under reducing conditions; the aggregation process and protein surface hydrophobicity were monitored by Thioflavin T (ThT) and ANS fluorescence, respectively. To determine molecular sizes, and intramolecular secondary structures, dynamic light scattering (DLS) and X-ray diffraction analyses were also applied, respectively. In addition to thermodynamic parameters measurements of both apo- and holo-proteins, melting temperature (Tm) of apo proteins was assessed. Furthermore, for comparative evaluation of toxicity of SOD1 aggregates, LDH cytotoxicity as well as hemolysis assays were performed. The hydrodynamic diameters of all SOD1 variants were the same. ThT/ANS results, however, confirmed that DTT-mediated reduction in intramolecular S–S bonds induces gross conformational changes followed by the protein assembly. Thermodynamic studies showed that neither metal deficiency/mutation nor intramolecular S–S reduction, alone, can significantly reduce stability of SOD1 proteins, and demetallation (reduced stability) and intramolecular S–S reduction, together, are required for initiation of SOD1 misfolding/aggregation. CR binding, CD and XRD results, also together, confirmed the non-amyloid character of relatively toxic SOD1 aggregates. It appears that reduced thermodynamic stability as prerequisite for aggregation of SOD1 mutants must be accompanied by intramolecular disulfides reduction.

There are five described Rhynchosporium species, Rhynchosporium commune, R. secalis, R. agropyri, R. orthosporum and R. lolii, that cause scald diseases on Poaceae. This study used morphological (conidial shape and size) and phylogenetic analyses of two loci (the internal transcribed spacer region (ITS) and β-tubulin (TUBB)) to identify Rhynchosporium species and their host ranges in Iran. Despite the large variation observed for Rhynchosporium conidial dimensions, the phylogenetic analyses of the ITS region and concatenated ITS and TUBB loci revealed that all isolates from wild grasses in Iran belong to R. commune. R. commune was isolated from Hordeum murinum ssp. glaucum, Hordeum vulgare ssp. spontaneum, Lolium multiflorum and Avena sativa in Iran. A. sativa has only been reported from Iran as a host for R. commune. After cross inoculation, A. sativa was considered as the most resistant host showing the lowest susceptibility to R. commune isolates. Of the grass hosts tested, H. vulgare ssp. spontaneum was the most susceptible. The most aggressive isolate across all tested hosts was isolated from Hordeum murinum ssp. glaucum. Cross-infection of the R. commune isolates from all hosts onto uncultivated grasses and cultivated barley suggests the potential of the uncultivated grasses as inoculum sources for cultivated barley epidemics and pathogen evolution. Thus, management of uncultivated grasses in the vicinity of barley fields should assist in managing the disease on cultivated barley.

Colletotrichum lindemuthianum, causal agent of bean anthracnose is one of the most destructive fungal bean diseases that potentially have an enormous economic impact on bean cultivation worldwide. To obtain novel insights into how bean regulates its defense pathways to react efficiently against this invading pathogen, cvs Khomein and Naz representing contrasting interactions were investigated by histochemical and real-time quantitative RT-PCR (RT-qPCR) approaches. Our histochemical analysis conducted at 24, 48, 72 and 96 h post inoculation (hpi) demonstrated that hydrogen peroxide (H2O2) may have a bifunctional role in establishing the incompatibility and compatibility. This free radical (H2O2) can, therefore, be elevated at an early stage (24 hpi) of the incompatible context to arrest fungal growth, whereas H2O2 was promoted at the late stage (72 hpi) of the compatible context to facilitate the infection process. Additionally, we demonstrated that O2− probably plays an essential function in launching the resistance response since O2− increased significantly at 36 hpi in incompatible interaction compared with that of the compatible interface. Our expression analysis showed that the expression of six defense-related genes are differentially regulated regarding the types of interactions, shedding light on how the beans adaptively regulate defense pathways. This study contributes to a better understanding of the kinetics of reactions triggered following C. lindemuthianum infection and may be applied to develop novel strategies to manage bean anthracnose effectively.

Background: Pseudomonas aeruginosa and Acinetobacter baumannii are widely ubiquitous in nature. In addition, they are opportunistic pathogens for humans and the common cause of nosocomial infections. Objective: Due to the increased antibiotic resistance in the treatment of nosocomial infections, this study aimed to evaluate the antibiotic susceptibility pattern of P. aeruginosa and A. baumannii in the pediatrics intensive care unit (PICU). Materials and Methods: Totally, 280 clinical samples from PICU patients were evaluated in this study. The samples were examined for P. aeruginosa and A. baumannii using standard microbiological methods. Finally, the Epsilometer test method was performed to investigate the antibiotic susceptibility pattern of these bacteria. Results: The results revealed a total of 21 isolates (7.5%) of P. aeruginosa and 11 isolates (3.9%) of A. baumannii. P. aeruginosa isolates showed the highest susceptibility to colistin (85.7%) and gentamicin (66.7%) while A. baumannii isolates were more susceptible to colistin (100%), ceftazidime (54.5%), and amikacin (45.5%), respectively. Conclusion: Due to the antibiotic susceptibility patterns of bacterial isolates in the recent study, colistin and gentamicin are recommended for the treatment of P. aeruginosa infections and colistin, ceftazidime, and amikacin are suggested for A. baumannii infections.

Because it is important to develop new sustainable sources of edible protein, insects have been recommended as a new protein source. This study applied Life Cycle Assessment (LCA) to investigate the environmental impact of small-scale edible insect production unit in South Korea. IMPACT 2002 + was applied as the baseline impact assessment (IA) methodology. The CML-IA baseline, EDIP 2003, EDP 2013, ILCD 2011 Midpoint, and ReCiPe midpoint IA methodologies were also used for LCIA methodology sensitivity analysis. The protein, fat contents, and fatty acid profile of the investigated insect (Protaetia brevitarsis seulensis larvae) were analyzed to determine its potential food application. The results revealed that the studied edible insect production system has beneficial environmental effects on various impact categories (ICs), i.e., land occupation, mineral extraction, aquatic and terrestrial ecotoxicity, due to utilization of bio-waste to feed insects. This food production system can mitigate the negative environmental effects of those ICs, but has negative environmental impact on some other ICs such as global warming potential. By managing the consumption of various inputs, edible insects can become an environmentally efficient food production system for human nutrition. This article has been corrected. Link to the correction: [https://cherry.chem.bg.ac.rs/handle/123456789/4586] Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4587]

The main purposes of this study were to examine the effects of protoplast fusion on chitinase production, antifungal activity and major antifungal compounds in parental strains and fusants and to i...

In the continuation of studies on Alternaria species from wheat plants with black head mold symptoms, 10 isolates with typical characteristics of the section Pseudoalternaria were isolated and studied by a combined approach based on morphological characteristics and phylogenetic analyses using sequence data sets of the internal transcribed spacer of ribosomal DNA (ITS-rDNA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and plasma membrane adenosine triphosphatase (ATPase) genes. These isolates represented a new and previously undescribed species in this section which is illustrated and described here as Alternaria ershadii sp. nov. Also based on our phylogenetic analyses, it was revealed that Alternaria brassicifolii, which was recently described from Brassica rapa subsp. pekinensis in Korea and placed in this section, formed a well separated monotypic lineage distinct from the other members in the section Pseudoalternaria. Comparisons of three-dimensional sporulation patterns and conidial characteristics supported the molecular results. So, at present, Alternaria section Pseudoalternaria contains only seven species viz. Alternaria altcampina, A. arrhenatheri, A. ershadii, A. inflata, A. kordkuyana, A. parvicaespitosa and A. rosae. Morphological comparisons of Alternaria ershadii with other species in the section Pseudoalternaria and with Alternaria brassicifolii is also provided.

During a survey of sedge plants in northern Iran, three specimens of Pseudopyricularia cyperi were isolated from Cyperus sp. Their taxonomical identity was established by their spore and conidiophore morphology and ITS rDNA sequence analysis. The specimens are described and illustrated. Pseudopyricularia cyperi is a new record for Iran.

The objectives of the present study were to identify composition of fungal communities associated with healthy and declined oak trees and seedlings in Ilam Province, Iran, and to evaluate their role in occurrence of the oak decline. Fungal isolates were obtained from branches of healthy and declining Persian oak trees and seedlings in Ilam province during summer and autumn 2014–2015. Fungal species were identified according to both morphological and molecular characteristics obtained from ITS, 28S and 18S regions of ribosomal DNA. Some fungal species such as Neoscytalidium dimidiatum and Obolarina persica were obtained only from branches of Persian oak trees with decline symptoms. The Acremonium sp., Coniochaeta sp., Cytospora ribis, Fusarium tricinctum, Microsphaeriopsis olivacea and Neoetophoma samarorum species were found only in healthy trees as endophytic species. While, B. mediterrana, Didymella glomerata, Fusarium solani and Tricothecium roseum were isolated from both healthy and declined trees. The F. tricinctum and T. roseum species were found in healthy seedlings. However, D. glomerata was isolated from both healthy and dried seedlings. The species B. mediterrana, D. glomerata, N. dimidiatum and O. persica showed pathogenicity on the Persian oak seedlings in the greenhouse conditions. Finally, it could be concluded that for the first time two species, D. glomerata and N. dimidiatum, were recorded as pathogens associated with Persian oak. In addition, Acremonium sp., Coniochaeta sp., C. ribis, F. solani, F. tricinctum, N. samarorum and T. roseum were recorded for the first time as endophytic fungi on Persian oak trees.

Upgrading of pyrolysis bio-oil by a novel catalytic hydrotreating process, including hydrodeoxygenation (HDO) and hydrodenitrogenation (HDN) was found as an effective technical method for the improvement of biofuel characteristics. In this study, for the first time, the performance of a novel meso-microporous composite material, HMS-ZSM-5, as a support on the catalytic activity of NiMo-based catalysts in the bio-oil hydrotreating was evaluated. The experiments were carried out in a flow fixed-bed reactor at the temperature range of 300–360 °C, 30 bar pressure, and LHSV = 4 h-1. Also, the results were and compared with those of HMS, ZSM-5, and γ-Al2O3 supports. For all catalysts, the increase in temperature resulted in the enhancement of HDO and HDN reactions efficiency. NiMo/HMS-ZSM-5 possessed a high acid property which contributed to the removal of oxygen and nitrogen from bio-oil, with the conversion of 84.10% and 69.60%, respectively. Therefore, the novel catalyst of this study represented much superior upgrading performances compared with those of stand-alone NiMo/HMS and NiMo/ZSM-5 catalysts and also the conventional catalyst of NiMo/γ-Al2O3.

Despite the seasonal nature of reproduction in goats, creating a capacity that enables getting goats pregnant throughout the year is of economical importance in commercial settings. The objective was to compare natural and artificial (hormonal) methods of estrus synchronization on pregnancy rate of Murciano-Granadina goats under extremely hot climate of southern Iran. To synchronize estrus, experimental goats received one of three treatments. The first group (n = 123) was treated with CIDR (intravaginal progesterone release) on day-0 and which was later removed on day-19 plus eCG injection followed by introduction to bucks on day-21. The second group (n = 157) was treated with CIDR on day-0 and with eCG injection on day-17, then CIDR removed on day-19 followed by introduction to bucks on day-21. The third group (n = 257) did not receive any hormonal treatments and were only synchronized naturally via introduction to bucks (natural mating). Findings revealed that natural synchronization (male effect) resulted in significantly greater pregnancy rates compared to the second group (69 vs. 53%, P < 0.05). The first group tended to have greater pregnancy than did the first group (63 vs. 53%, P < 0.10). Improved reproductive performance of dairy goats under hot stressful climate signifies the economical importance and practicality of natural mating as an effective method of estrus synchronization in commercial goat production.

Current techniques for plant virus detection, such as RT- PCR and ELISA, require multistep procedures and rely on sophisticated equipment. Due to the global spread of plant viruses, the development of simpler, faster and cheaper assay methods is inevitable. Gold nanoparticles (AuNPs) had raised much interest during recent years due to their novel optical properties or diagnostic purposes. The localized surface plasmon resonance (LSPR1) of AuNPs had been used in the development of novel colorimetric nano-biosensing systems. The frequency and intensity of the LSPR peak generally depend on the shape, size and the surrounding medium of the AuNPs. In this study, unmodified AuNPs had been used to detect the unamplified Tomato yellow leaf curl virus (TYLCV) genome in infected plants. A specific DNA probe complementary to the coat protein region of virus genome was designed. The extracted total DNA of uninfected and infected plants was mixed with hybridization buffer and the designed probe. The mixture was denatured, annealed and then cooled to room temperature and was followed by AuNPs addition. The color changes in the samples indicating the presence of target virus infections were assessed visually after the addition of salt and confirmed by UV-Vis spectroscopy. The results showed that this strategy allowed for fast and sensitive detection of TYLCV genome and eliminated the need for PCR amplification and detection equipment.

The population structure and genetic diversity of Pyrenophora tritici-repentis from hexaploid wheat were examined using 51 simple sequence repeat (SSR) markers designed to represent each chromosome...

Scrophularia subaphylla ( S. subaphylla ) L., a medicinal plant from the Scrophulariaceae family, has been reported to possess potential profits in the treatment and prophylaxis of different diseases. Some phenolic compounds in this genus have been displayed decent effects on different types of cancer via multiple mechanisms. The current study aimed to bioassay guided isolation of cytotoxic constituents from the aerial parts of S. subaphylla against breast (MCF-7) and colon (HT-29) cancer cell lines as well as normal cells (L929). Different extracts of S. subaphylla were acquired by Soxhlet apparatus and then subjected to brine shrimp lethality test and MTT assay for assessing their cytotoxic characteristics. Cytotoxic extract subjected to further phytochemical fractionation using solid phase extraction, reversed-phase high pressure liquid chromatography (RP-HPLC), and one dimensional nuclear magnetic resonance (1D-NMR) spectroscopy. The biological activity of the isolated pure components, verbascoside and 3 ' O rhamnosyl -4' O para coumaryl 7- hydroxyl salidroside , was assessed using MTT assay against MCF-7 and HT-29 carcinoma cells. Two known phenylpropanoid compounds were isolated from this species. Their structures were elucidated by spectroscopic data (using 1 H-NMR and 13 C-NMR) and compared with the previous literature. Both pure compounds in comparison with control group demonstrated significant antiproliferative activity against cancerous cells ( P < 0.001). In our study, verbascoside and its derivative could inhibit proliferation of cancerous cells without any side effects on normal cells.

The salinity tolerance of 17 breeding wheat genotypes along with three local varieties was evaluated under control and salinity stress (160 mM NaCl) conditions. At the seedling stage, shoot and root dry weights, relative water content (RWC), membrane stability index (MSI), relative chlorophyll content (SPAD index), root and shoot Na+ (RN and SN), root and shoot K+ (RK and SK), root and shoot K+/Na+ ratios (RKN and SKN), root-to-shoot Na+ translocation (RTSN), root-to-shoot K+ translocation (RTSK), stomatal conductance (GS), transpiration rate (TE), and photosynthesis rate (PN) were measured. Moreover, the investigated genotypes were assessed in terms of grain yield across four saline regions during the 2018–2019 cropping seasons. Salinity stress caused a significant reduction in the RDW, SDW, PN, GS, TE, SK, RKN, SKN, RTSN, and RTSK, but resulted in increased RN, RK, and SN. The results of AMMI analysis of variance also indicated significant differences among test locations, genotypes, and their interaction effects. The PCA-based biplot revealed that grain yield strongly correlated with RKN and RK. Furthermore, the correlation among PN, GS, and TE traits was strong and positive and had a positive correlation with RWC, MSI, RDW, and SPAD index. Considering our results, RK and RKN were identified as useful physiological tools to screen salt tolerance at the early-growth stage. According to the ranking patterns obtained by the average sum of ranks method (ASR) and grain yield, we observed that genotype number G5 had considerable physiological potential at the early-growth stage and also responded well to soil salinity at the farm; thus this genotype can be promoted for commercial production.

Fusarium verticillioides is one of the most important fungal pathogens of maize since it causes severe yield losses and produces the mycotoxins fumonisins that represent a major concern for human and animal health. Information about genetic diversity and population structure of fungal pathogens is essential for developing disease management strategies. The aim of this research was to investigate the genetic structure of F. verticillioides isolated from different provinces of Iran through determination of mating type idiomorphs, phylogenetic analyses based on translation elongation factor-1 alpha (EF-1α), RNA Polymerase II Subunit (RPB2), beta-tubulin (tub2) and Calmodulin (cmdA) genes and genetic diversity analyses based on 6 simple-sequence repeats (SSRs). Both mating types were detected in Iranian populations of F. verticillioides, particularly in Qazvin and Khuzestan, with equal frequency, which highlighted that sexual reproduction is favorable under field conditions. However, the linkage disequilibrium indices did not support the hypothesis of random mating in Khuzestan and Fars. Although assessment of nucleotide diversity based on housekeeping genes showed low level of variation among strains, genotype diversity based on SSRs revealed a high level of genetic diversity within Iranian populations. AMOVA analysis highlighted that the genetic variation of F. verticillioides in Iran was mainly distributed within population of a single area (97%), while a small proportion of genetic variation (3%) resided among populations. These patterns of variation are likely explained by the continuous gene flow among populations isolated from different areas. On the other hand, principal coordinate analysis indicated that the distribution of genetic variation among populations could be explained by the geographical distances. Consequently, to reduce pathogen gene flow among regions, the quarantine processes in Iran should be intensified.

Salinity stress is one of the major limiting abiotic stresses that decrease crop production worldwide. To recommend genotypes for cultivation under saline stress conditions, comprehensive understanding of genetic basis and plant responses to this stress is need. In the present study, a total of 20 barley genotypes were investigated to isolate potential salt-tolerant genotypes at the early growth stage using hydroponic system, and adult plant under field conditions. Different growth and physiological traits including root fresh and dry weights (RFW and RDW), shoot fresh and dry weights (SFW and SDW), relative water content (RWC), membrane stability index (MSI), relative chlorophyll content (SPAD index), root and shoot Na+ (RN and SN), root and shoot K+ (RK and SK), root and shoot K+:Na+ ratios (RKN and SKN), root-to-shoot Na+ translocation (RTSN), root-to-shoot K+ translocation (RTSK), stomatal conductance (GS), transpiration rate (TE), and photosynthesis rate (PN) were measured. Barley seedling were treated with two salinity levels (0 mM NaCl (as control conditions) and 200 mM NaCl (as stress conditions)) for 30 days. Moreover, the yield performance and stability of investigated barley genotypes were evaluated across five environments during the 2018–2020 cropping seasons. Salinity stress significantly decreased growth and physiological traits in all seedling plants; however, some salt-tolerant genotypes showed the lowest reduction in measured traits. Multivariate analysis grouped measured traits and tested genotypes into different clusters. The multi-trait genotype–ideotype distance index (MGIDI) selected genotypes G12, G14, G6, G7, and G16 as the salt-tolerant barley genotypes. Considering the results of the AMMI analysis showed that grain yields of tested barley genotypes were influenced by environment (E), genotype (G) and GE interaction effects. Based on the weighted average of absolute scores of the genotype index (WAASB) and other stability statistics, G7, G8, G14, and G16 were selected as superior genotypes. Considering the outputs of MGIDI and WAASB indices revealed that three genotypes G7, G14 and G16 can be recommended as new genetic resources for improving and stabilizing grain yield in barley programs for the moderate climate and saline regions of Iran. In conclusion, our results suggest that the using MGIDI index in the early growth stage can accelerate screening nurseries in barley breeding programs.

Acne vulgaris is one of the most common dermatologic disorders affects people of all races and ethnicities and has many adverse effects on the quality of life. The increased bacterial resistance to antibiotics has reduced the effectiveness of treatment with these agents. There is an increasing focus on the involvement of oxidative stress in the pathophysiology of acne. This study investigates the effect of N-acetylcysteine (NAC) as an antioxidant in the treatment of acne vulgaris. This systematic review was conducted through a search in databases such as Science Direct, PubMed, Scielo, and Medline using keywords including acne vulgaris, anti and NAC, and all the keywords associated with each of the subtitles. The factors affecting the occurrence and expansion of acne include increased sebum synthesis, hyperkeratinization of pilosebaceous units, colonization with Propionibacterium acnes, and increased release of inflammatory mediators and ROS. Studies have shown that glutathione stimulation following the administration of NAC increases glutathione levels for the detoxification of oxygen-free radicals. Moreover, NAC prevents the synthesis and release of inflammatory cytokines such as TNF-α, IL-8, IL-6, MP9, and IL-1β and has shown antibacterial activities against important bacteria including E. coli, S. epidermidis, Pseudomonas, and Klebsiella. This medication has anti-proliferative effects and is also used for excoriation and PCOD. The results of the present study showed the beneficial effects of using NAC in patients with acne vulgaris in terms of the disease complications and comorbidities. Given its diverse functional mechanisms, this medication can be used to treat acne and its consequences.

Cancer has now been established as one of the most common chronic diseases due to high mortality rate. The early stage of non-invasive tumors can now be successfully treated leading to have high survival rates; however, the late stage invasive and metastatic tumors still suffer from poor treatment outcomes. Among multiple contributing factors, the role of tumor microenvironment and its complexities has been well recognized in cancer progression. Stromal cells including cancer-associated fibroblasts (CAFs), endothelial cells, adipocytes, immune cells as well as extracellular matrix (ECM) continuously interact with malignant cells and regulate various hallmarks of cancer including tumor growth, invasion, and intravasation. To better understand the role of the interaction between tumor cells and their surrounding microenvironment, numerous model systems ranging from two-dimensional (2D) assays to 3D hydrogels and in vivo murine xenografts have been utilized. While each one of these model systems exhibit certain advantages in studying biological facets of tumor progression, they are often limited to perform well-controlled mechanistic studies due to various factors including lack of tumor-stroma organotypic organization and presence of confounding biochemical and biophysical factors within the tumor microenvironment. In this regard, in the past few years, 3D in vitro microengineered model systems are becoming instrumental to precisely mimic the complexities of the native tumor microenvironment to conduct fundamental and well-designed studies for multiple purposes ranging from biological discovery to therapeutic screening. These model systems include microfluidics, micro-patterned features, and 3D organoids. In this chapter, we will outline the fabrication strategy of our microengineered 3D co-culture tumor-stromal model which comprises high-density array of tumor seeded microwells surrounded by stromal cells, such as CAFs encapsulated within collagen-based hydrogel. The developed platform provides excellent spatial organization of tumor and stromal entities with designated initial architecture and cellular positioning, therefore enabling to study the specific role of cell-cell and cell-ECM interaction on tumor proliferation/expansion, cancer cell migration as well as stromal activation. The developed platform is compatible with standard biological assays enabling gene and protein expression analyses across different types of cancer and co-culture of tumor and stromal cells.

α-Synuclein (αS) aggregates plays a pivotal role in the pathogenesis of synucleinopathies including Parkinson's Disease. The toxicity of αS aggregates has been broadly studied and variant defects have been reported through which these aggregates lead in cell death. Although cell death through apoptosis pathway has been proposed in many studies, the molecular details underlying in this pathway have not been uncovered. To shed a light on the relationships between αS aggregates and apoptotic cell death, changes in levels and behavior of molecular indicators of the intrinsic apoptotic pathway was investigated in HEK-293T cells overexpressing wild-type α-synuclein and A53T-α-synuclein. Overexpression of both WT-αS and A53T-αS resulted in the increase of caspase-9 activity, and rise in Cytochrome c (Cyt c) and PARC content, concurrently. We assume that rising in PARC level may result in Cyt c degradation, and consequently suppressing/attenuating intrinsic apoptosis pathway. Besides, increasing of Casp-9 activity can be related to αS aggregates and subsequent degradation of Cyt c. To understand the mechanisms behind this using theoretical model, molecular dynamic simulation was also applied to investigate the possible interaction of Casp-9 with α-synuclein aggregates. The results showed that the interaction between Casp-9 with αS aggregates could activate Casp-9 by changing the conformation of some crucial residues.

Saccharomyces cerevisiae is being used for long as a rich source of proteins, sugars, nucleotides, vitamins and minerals. Autolyzed and hydrolyzed yeast biomass has found numerous applications in the health food industry as well as livestock feeds. Here, we have compared three lysis methods for production of yeast lysates using autolysis, plasmolysis (ethyl acetate 1.5%), and enzymatic hydrolysis (Alcalase 0.2%). The efficiency of each process was compared according to soluble solid and protein contents, cell lysis monitoring, and release of intracellular materials, cell viability and microscopic analysis. Results showed that plasmolysis by ethyl acetate was found to be more efficient compared to autolysis, with a higher recovery of yeast extract (YE) content. In comparison, the content of released solids and proteins were higher during the enzymatic hydrolysis using Alcalase compared to autolysis and plasmolysis treatments. The highest decrease in optical density of 600 nm was monitored for the hydrolyzed cells. Besides, we defined "Degree of Leakage (DL)" as a new index of the lysis process, referring to the percentage of total released proteins from the cells and it was estimated to about 65.8%, which represents an appropriate indicator of the cell lysis. The biochemical and biophysical properties of the hydrolyzed yeast product as well as its biological activity (free radical scavenging activity and bacterial binding capacity) suggest that Alcalase could be used to accelerate the lysis of yeast cells and release the valuable intracellular components used for foodstuffs, feed and fermentation media applications. Production of baker's yeast lysates using autolysis, plasmolysis, and enzymatic hydrolysis methods.

Background: Adenotonsillar hypertrophy (ATH) is a common problem in children that causes the obstruction of airways and other relevant complications. Helicobacter pylori (H. pylori) is considered one of the microbial causes of ATH development. Objectives: The study aimed to determine the H. pylori colonization in patients with adenotonsillar hypertrophy and assess the prevalence and clinical characteristics of colonized patients and possible association with complications. Methods: This study was carried out on 114 children with ATH referring to the Bouali Hospital of Sari from December 2017 to December 2018. Adenotonsillar samples were prepared from the right and left tonsils and adenoid of each patient. The Rapid Urease test (RUT) and nested-PCR were performed on the samples. Data analysis was performed with SPSS version 20 software. Results: The participants included 52 females and 62 males with a mean age of 7.19 ± 3.08 years. The RUT was positive for 59 participants: 26 in right tonsil, 31 in left tonsil, and two in adenoid samples. Helicobacter pylori was detected in the adenotonsillar tissue of 95 patients using nested-PCR. No significant association was found between the PCR results and gender (P = 0.123). Conclusions: This study approved the presence of H. pylori in the adenotonsillar tissue of children with ATH and highlighted the concept that the pharynx could be an extra gastric reservoir of H. pylori. However, we failed to prove an association between H. pylori adenotonsillar colonization with ATH and acute otitis media.

The center of origin of the globally distributed wheat pathogenParastagnospora nodorumhas remained uncertain because only a small number of isolates from the Fertile Crescent, a region in the Middle East where wheat was domesticated from wild grasses, were included in earlier population genetic and phylogeographic studies. We isolated and genetically analyzed 193P. nodorumstrains from three naturally infected wheat fields distributed across Iran, a country located within the Fertile Crescent, using eleven neutral microsatellite loci. Compared to previous studies that included populations from North America, Europe, Africa, Australia and China, the populations from Iran had the highest genetic diversity globally and also exhibited greater population structure over smaller spatial scales, patterns typically associated with a species’ center of origin. Genes encoding the necrotrophic effectorsSnToxA, SnTox1andSnTox3were found at a high frequency in the Iranian population. By sequencing 96 randomly chosen Iranian strains, we detected new alleles for all three effector genes. Analyses of allele diversity showed that all three effector genes had higher diversity in Iran than in any population included in previous studies, with Iran acting as a hub for the effector diversity that was found in other global populations. Taken together, these findings support the hypothesis thatP. nodorumoriginated either within or nearby the Fertile Crescent with a genome that already encoded all three necrotrophic effectors during its emergence as a pathogen on wheat. Our findings also suggest thatP. nodorumwas the original source of theToxAgenes discovered in the wheat pathogensPhaeosphaeria avenariaf. sp.tritici1,Pyrenophora tritici-repentisandBipolaris sorokiniana.

The apoptotic protease-activating factor 1 (Apaf-1) split luciferase biosensor has been used as a biological tool for the detection of early stage of apoptosis. The effect of doxorubicin in a cell-based assay and the addition of cytochrome c and ATP in a cell-free system have been used to test the functionality of the reporter for the detection of apoptosome formation. Here, our data established a drug- and cytochrome c/ATP-independent way of apoptosis induction relying on the expression of the biosensor itself to induce formation of apoptosome. Overexpression of Apaf-1 constructs led to increased split luciferase activity and caspase-3 activity in the absence of any drug treatment. Caspase-3 activity was significantly inhibited when caspase-9DN was co-overexpressed, while the activity of the Apaf1 biosensor was significantly increased. Our results show that the Apaf-1 biosensor does not detect etoposide-induced apoptosis.

Despite significant efforts in the study of cardiovascular diseases (CVDs), they persist as the leading cause of mortality worldwide. Considerable research into human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) has highlighted their immense potential in the development of in vitro human cardiac tissues for broad mechanistic, therapeutic, and patient-specific disease modeling studies in the pursuit of CVD research. However, the relatively immature state of hPSC-CMs remains an obstacle in enhancing clinical relevance ofengineered cardiac tissue models. In this study, we describe development of a microfluidic platform for 3D modeling of cardiac tissues, derived from both rat cells and hPSC-CMs, to better recapitulate the native myocardium through co-culture with interstitial cells (specifically cardiac fibroblasts), biomimetic collagen hydrogel encapsulation, and induction of highly anisotropic tissue architecture. The presented platform is precisely engineered through incorporation of surface topography in the form of staggered microposts to enable long-term culture and maturation of cardiac cells, resulting in formation of physiologically relevant cardiac tissues with anisotropy that mimics native myocardium. After two weeks of culture, hPSC-derived cardiac tissues exhibited well-defined sarcomeric striations, highly synchronous contractions, and upregulation of several maturation genes, including HCN1, KCNQ1, CAV1.2, CAV3.1, PLN, and RYR2. These findings demonstrate the ability of the proposed engineered platform to mature animal- as well as human stem cell-derived cardiac tissues over an extended period of culture, providing a novel microfluidic chip with the capability for cardiac disease modeling and therapeutic testing.

Some Stemphylium isolates reported from Malaysia as well as other regions of the world have been identified by morphology and blast analysis of the ITS region only, and eventually deposited in NCBI GenBank. To our knowledge, some of these isolates have been misidentified. The re-examination of identity of all Stemphylium isolates reported from Malaysia using morphological and phylogenetic analyses of the ITS and gpd, as well as phylogenetic analyses of all isolates of S. lycopersici and S. solani available in NCBI GenBank from other regions of the world indicated that a remarkable number of isolates deposited in NCBI GenBank under S. lycopersici and S. solani species were misidentified.

Background Accurate diagnosis of mucormycosis, a life-threatening fungal infection, remains a challenge for physicians. Objectives To identify the causative Mucorales in fresh clinical samples and formalin-fixed paraffin-embedded (FFPE) samples of patients with proven mucormycosis by molecular method. Patients/methods Fresh clinical samples of patients with proven mucormycosis according to the EORTC/MSG criteria admitted between 2015 and 2017 and histopathologically proven FFPE archives collected during 2004-2007 and 2015-2017 from Mazandaran University-affiliated hospitals of northern Iran were included. Seminested PCR targeting the 18S rDNA of Mucorales and ITS region was performed, and PCR products were then sequenced. Results While culture was positive only in 5 of 9 (56%) of fresh specimen cases, PCR was positive in all 9 (100%) histologically proven mucormycosis. Ten of 18 (56%) FFPE samples were PCR-positive. Overall, Mucorales PCR was positive in 19 of 27 (70%) samples. Mucorales species were Rhizopus arrhizus in 16 (84%) cases, R. arrhizus/Amylomyces rouxii in 2 (10.5%) cases and Rhizopus stolonifer in one case (5.5%). Among 27 mucormycosis cases, 25 (93%) cases were rhinocerebral, and 2 (7%) cases were disseminated. Diabetes mellitus (74%) and neutropaenia (63%) were the main risk factors. Conclusions Seminested PCR targeting 18S rDNA region of Mucorales is useful for identification of the causative agents of mucormycosis.

Mucormycosis is a rare but fulminant opportunistic fungal infection, which occurs most often in diabetic and immunocompromised patients. Dental extractions may create a portal of entry for the fungal infection. The mucormycosis may be the original cause of the pain and can be misdiagnosed as dental pain. In this paper, two cases of mucormycosis are reported after dental extractions and successfully treated with amphotericin B (case #1) and combined with posaconazole (case #2). The two cases we describe exemplify the fulminant mucormycosis of maxillary sinuses after dental extraction inpatients with uncontrolled diabetic support the findings that this predisposing condition created a suitable environment for the Mucorales growth. These case reports emphasize early recognition and urgent treatment of mucormycosis is necessary to prevent the spread of infection Therefore, dental surgeons and healthcare practitioners should become familiar with mucormycosis.

Background: Coronary artery disease is a leading cause of death worldwide. The present study has been designed to investigate efficacy of methanolic extract of Scrophularia subuphylla (S. subuphylla) on ischemia and reperfusion-induced myocardial injury in isolated rat heart. Methods: The isolated male Wistar rat hearts (n= 5) were perfused by Krebs-Henseleit solution enriched with the extract (0, 1, 5, and 10 μg/ml), using the langendorff method. After 15 minutes stabilization, the hearts were subjected to 30 minutes regional ischemia and then 120 minutes reperfusion. Results: Administration of the extract did not improve any of cardiac markers of flow rate, heart rate and developed pressure. Number, percentage and duration of arrhythmias were not affected by any concentrations of the extract. However, the concentration of 1 and 5 µg/ml increased the VT duration compared to control group (P

Background: Transplantation of fetal mesencephalic tissue is a well-established concept for functional reinnervation of the dopamine-depleted rat striatum. However, there is no extensive description of the glial response of the host brain following this procedure.Aims: The present study aimed to quantitatively and qualitatively analyse astrogliosis surrounding intrastriatal grafts and compare it to the reaction to mechanical injury with the transplantation instrument only.Study Design: Animal experimentation.Methods: The standard 6-hydroxydopamine-induced unilateral model of Parkinson’s disease was used. The experimental animals received transplantation of a single-cell suspension of E14 ventral mesencephalic tissue. Control animals (sham-transplanted) were subjected to injury by the transplantation cannula, without injection of a cell suspension. Histological analyses were carried out 7 and 28 days following the procedure by immunohistochemistry assays for tyrosine hydroxylase and glial fibrillary acidic protein. To evaluate astrogliosis, the cell density and immunopositive area were measured in distinct zones within and surrounding the grafts or the cannula tract.Results: Statistical analysis revealed that astrogliosis in the grafted striatum increased from day 7 to day 28, as shown by a significant change in both cell density and the immunopositive area. The cell density increased from 816.7±370.6 to 1403±272.1 cells/mm2 (p