Genetic study of the NOTCH3 gene in CADASIL patients

Background: Cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a monogenic, hereditary, neurological syndrome characterized by small vessel disease (SVD), stroke, vascular cognitive impairment and dementia. It is caused by mutations in the NOTCH3  gene, affecting the number of cysteines in the extracellular domain of the receptor, causing protein misfolding and receptor aggregation. The pathogenic role of cysteine-sparing NOTCH3 missense mutations in patients with typical clinical CADASIL syndrome is unknown. No therapies are available for this condition.Objective: Genetic study of the NOTCH3 gene in CADASIL patients who were referred to the Fazeli-Sanati Genetics Laboratory.Subjects and methods: Peripheral blood samples were collected from 10 CADASIL patients to extract genomic DNA. DNA sequences of exons 2–8, 11–12 and 18–19, where NOTCH3 mutations are typically located; were amplified by using PCR and analyzed by direct sequencing.Results: 11 NOTCH3 exons were analyzed. Homozygous IVS7 + 15A>G mutation were found in five patients, Homozygous IVS7 + 16A>G mutation in one patient, Heterozygous for the Pro109Thr and Pro203His mutations in one patient, which were not reported previously. Heterozygous C395R and R153C mutations were found in two patients. One of the patients has no mutation in 11 analyzed NOTCH3 exons.Conclusion: We found four novel mutations (P109T, P203H, IVS7 + 15A>G and IVS7 + 16A>G) and 2 reported NOTCH3 mutations. Exon 4 and Intron 7 are hotspots in the patients we examined with the NOTCH3 mutations. These findings broaden the mutational spectrum of CADASIL.Keywords:  CADASIL, NOTCH3, Mutation, Exo

Caspian Sea’s Navicula salinicola Hustedt 1939 and effect of the prolonged culture on its fatty acid profile

Diatoms are a potent source of polyunsaturated fatty acids. This study was conducted for screening a Naviculoid diatom strains from the southern Caspian Sea with analyzing its lipid production and accumulation potentials. The isolate was identified as Navicula salinicola strain IBRC-M 5083 based on micro-morphological characterization and analysis of 18S rRNA genomic region. Navicula salinicola were cultured in the f/2 medium under both normal and prolonged culture (21 days) conditions. Total lipid percentages of this strain were found to be 31.83% under normal condition and 43.72±1.4% in prolonged culture respectively on the basis of their dry cell weight (DCW). Also, the oil droplets were detected in 21 days’ cells as shown by Sudan Black B staining experiments. Furthermore, the main fatty acids were found by Gas Chromatography analyses of this strain under prolonged condition to be Eicosapentaenoic acid (25.58%TFA). Such oil accumulation capabilities seem to be promising for performing further studies on this strain as a source of Omega-3 in aquafeed, pharmaceutical and biofuel industries

The isolation and preliminary characterization of native cyanobacterial and microalgal strains from lagoons contaminated with petroleum oil in Khark Island

Introduction: Algae has many applications in terms of ecology, biodiversity, agriculture, medicine, biotechnology, industry, etc. They are potent organisms in bio-active compound production, bioremediation and primary producer. Therefore, it is important to discover local strains with biotechnological and ecological applications. Materials and methods: Soil and water samples were collected from different sites of Khark Island (Persian Gulf). The samples were cultivated and purified using different techniques. Seven different antibiotics together with other physical methods used to purify the isolates. Results: Throughout the project 7 strains including 2 eukaryotic algae and 5 cyanobacteria have been isolated. Imipenem and cycloheximide were the best antibiotics for purification of cultures. Three of isolates were morphologically similar to Arthronema africanum, Pseudanabaena teremula, Anabaenopsis sp. However, they have some different characteristics which according to the present identification keys it is not possible to identify their identity (they have nominated Kh.C.d2, Kh.T.1 and Kh.T.2). Discussion and conclusion: According to the results, isolated strains were identified at the genus level based on morphology characters; therefore the complementary examinations such as molecular identification, ITS, 18s rRNA, 16s rRNA and sequencing can help to approve the strains identity. Upon approval of the new strains account for morphological traits are necessary for their easy identification. The Imipenem antibiotic is the best for eukaryotic algae purification and Cycloheximide is suitable for prokaryotic algae (cyanobacteria) purification

Wickerhamomyces orientalis f.a., sp. nov.: an ascomycetous yeast species belonging to the Wickerhamomyces clade

L

Genome-Wide Analysis of Oceanimonas sp. GK1 Isolated from Gavkhouni Wetland (Iran) Demonstrates Presence of Genes for Virulence and Pathogenicity

Objective: The bacterium Oceanimonas sp. (O. sp.) GK1 is a member of the Aeromonadaceae family and its genome represents several virulence genes involved in fish and human pathogenicity. In this original research study we aimed to identify and characterize the putative virulence factors and pathogenicity of this halotolerant marine bacterium using genome wide analysis. Materials and Methods: The genome data of O. sp. GK1 was obtained from NCBI. Comparative genomic study was done using MetaCyc database. Results: Whole genome data analysis of the O. sp. GK1 revealed that the bacterium possesses some important virulence genes (e.g. ZOT, RTX toxin, thermostable hemolysin, lateral flagella and type IV pili) which have been implicated in adhesion and biofilm formation and infection in some other pathogenic bacteria. Conclusion: This is the first report of the putative pathogenicity of O. sp.GK1. The genome wide analysis of the bacterium demonstrates the presence of virulence genes causing infectious diseases in many warm- and cold-blooded animals

Morphological identification and molecular validation of anchovies (Engraulidae) in the Persian Gulf and Oman Sea

Afrand, Mahboobeh, Sourinejad, Iman, Fazeli, Seyed Abolhassan Shahzadeh, Akbarzadeh, Arash, Yeganeh, Laleh Parsa, Sadeghi, Maryam, Azarbaijani, Reza (2020): Morphological identification and molecular validation of anchovies (Engraulidae) in the Persian Gulf and Oman Sea. Zootaxa 4742 (2): 375-391, DOI: 10.11646/zootaxa.4742.2.1

Graphiola fimbriata: the first species of Graphiolaceae (Exobasidiales, Basidiomycota) described only based on its yeast stage

International audienceThe systematic position of three yeast strains isolated from a plant cell culture, a piece of termite nest, or as a foliar endophyte of Coffea arabica, respectively, is evaluated using morphological, physiological, and phylogenetical characteristics. In culture, all three isolates produced white, pale orange to pink colored colonies of cylindrical cells with monopolar budding and pseudohyphae. Standard phenotypic, biochemical, physiological characterization, and phylogenetic analyses of the combined 26S rRNA gene (D1/D2 domains) and ITS region sequences showed the conspecificity of these isolates and suggest their placement within the Exobasidiales (Ustilaginomycotina) as a sister lineage of the sampled and sequenced Graphiola species. Here, we describe this species as Graphiola fimbriata sp. nov. MycoBank MB 825077 (holotype: PC1T; ex-type cultures: IBRC-M 30158T = CBS 13945T = DSM 104832T). This is the first species described in the genus Graphiola for which only the asexual, saprobic developmental phase is known. The description of the genus Graphiola is therefore emended to allow species known only from a saprobic state

A Rapid and Reproducible Genomic DNA Extraction Protocol for Sequence-Based Identification of Archaea, Bacteria, Cyanobacteria, Diatoms, Fungi, and Green Algae

Background:  Sequence-based identification of various microorganisms including Archaea, Bacteria, Cyanobacteria, Diatoms, Fungi, and green algae necessitates an efficient and reproducible genome extraction procedure though which a pure template DNA is yielded and it can be used in polymerase chain reactions (PCR). Considering the fact that DNA extraction from these microorganisms is time consuming and laborious, we developed and standardized a safe, rapid and inexpensive miniprep protocol. Methods:  According to our results, amplification of various genomic regions including SSU, LSU, ITS, β-tubulin, actin, RPB2, and EF-1 resulted in a reproducible and efficient DNA extraction from a wide range of microorganisms yielding adequate pure genomic material for reproducible PCR-amplifications. Results:   This method relies on a temporary shock of increased concentrations of detergent which can be applied concomitant with multiple freeze-thaws to yield sufficient amount of DNA for PCR amplification of multiple or single fragments(s) of the genome. As an advantage, the recipe seems very flexible, thus, various optional steps can be included depending on the samples used.Conclusion:   Having the needed flexibility in each step, this protocol is applicable on a very wide range of samples. Hence, various steps can be included depending on the desired quantity and quality