Phylogenetic Analysis of Phaeosphaeria Species Using Mating Type Genes and Distribution of Mating Types in Iran

Phaeosphaeria species are pathogenic on wheat, barley and a wide range of wild grasses. To analyze mating type loci of the Phaeosphaeria species and investigate mating type distribution in Iran, we sequenced mating type loci of 273 Phaeosphaeria isolates including 67 isolates obtained from symptomatic leaves and ears of wheat, barley, and wild grasses from two wheat-growing region in Iran as well as 206 isolates from our collection from other regions in Iran which were isolated in our previous studies. Mating type genes phylogeny was successfully used to determine the species identity and relationships among isolates within the Phaeosphaeria spp. complex. In this study, we reported seven new host records for Phaeosphaeria species and the Phaeosphaeria avenaria f. sp. tritici 3 group was first reported from Iran in this study. Mating type distribution among Phaeosphaeria species was determined. Both mating types were present in all sampling regions from Iran. We observed skewed distribution of mating types in one region (Kohgiluyeh va Boyer-Ahmad) and equal distribution in the other region (Bushehr). However, when considering our entire dataset of 273 Iranian Phaeosphaeria isolates, the ratio of mating types was not deviated significantly from 1:1 suggesting possibilities for isolates of opposite mating type to interact and reproduce sexually, although the sexual cycle may infrequently occur in some regions especially when the climatic conditions are unfavorable for teleomorph development.     

Cytogenetic and crossability studies in hulled wheat collected from Central Zagros in Iran

Underutilized hulled wheat species may be valuable sources of genes for wheat improvement. Hulled wheats of mountainous Central Zagros area of Iran are poorly studied with unclear classification status. In this study, the region was extensively searched and hulled wheats were sampled from local farmers’ fields. Cytogenetic studies of several samples revealed that these wheats are tetraploid with a chromosome number of 2n = 28. Considerable level of karyotypic resemblance was detected between these hulled wheats and durum wheat genotypes, suggesting their genomic similarity. To compare the degree of cross compatibility, F1 progenies were produced using three durum accessions and a bread wheat cultivar as female parent in crosses with two of the collected hulled wheat samples, namely Zarneh and Jonghan. The highest crossability rate was observed when Aria, a durum wheat cultivar, was used as female parent. In general, Jonghan showed higher crossability rate with both bread and durum cultivars as compared to Zarneh. The meiotic behavior of F1 genotypes was analyzed to determine genomic characteristics of hulled wheat samples. Microscopic examination of pollen mother cells at meiotic metaphase strongly suggested the AABB type genome for the hulled wheat samples. Meiotic behaviors of hybrids derived from cross with Zarneh as male parent showed more abnormalities than the other male parent. Meiotic restitution, chromosomes laggards and chromosomal bridges were noticed and the extent varied for each cross. It may be concluded that the hulled wheats of Central Zagros that are currently cultivated are mainly tetraploids emmer containing AABB genome and are crossable with durum wheat, producing hybrids mostly with normal meiotic behaviors.     

Mutational analysis of uroporphyrinogen III cosynthase gene in Iranian families with congenital erythropoietic porphyria

Porphyrias are rare metabolic hereditary diseases originating from defects in specific enzymes involved in the heme biosynthesis pathway. Congenital erythropoietic porphyria (CEP) is the rarest autosomal recessive porphyria resulting from a deficiency of uroporphyrinogen III cosynthase (UROS), the fourth enzyme in heme biosynthesis. CEP leads to an excessive production and accumulation of type Ι porphyrins in bone marrow, skin and several other tissues. Clinical manifestations are presented in childhood with severe cutaneous photosensitivity, blistering, scarring and deformation of the hands and the loss of eyebrows and eyelashes. Less than 200 cases of CEP have been reported to date. Four CEP patients and their family members were studied for the first time in Iran. A missense mutation in the UROS gene was identified in this family. A, T to C change at nucleotide 34313, leading to a substitution of Leucine by Proline at codon 237, was observed in the homozygous state in these 4 patients and heterozygous state in their parents. Our data from the Iranian population emphasizes the importance of codon 237 alone, given the rarity of this disease. This fact can be taken into consideration in the mutational analysis of UROS. This work emphasizes the advantages of molecular genetic techniques as diagnostic tools for the detection of clinically asymptomatic heterozygous mutation carriers as well as CEP within families.     

Acute 17β-Estradiol Pretreatment Protects Against Abdominal Aortic Occlusion Induced Spinal Cord Ischemic-Reperfusion Injury

Postoperative neurologic deficit due to spinal cord ischemia-reperfusion (I/R) injury is the most devastating complication following thoracoabdominal aortic aneurysm repairs. The protective potential for 17β-Estradiol has not been yet studied in such injury. In this study, ischemia induction for 18 min in male New Zealand White rabbits resulted in the highest percentage (80%) of biphasic paraplegic outcome assessed by Tarlov’s score. Acute Estradiol pretreatment (1 mg/kg, i.p., 30 min before I/R induction) altered this outcome and significantly prevented the worsening pattern of neurologic deficits over 48 h of observation. Histopathologic and oxidative stress evaluations of lumbar spinal cords taken in delayed permanent paraplegic phase (48 h after ischemia induction), further confirmed protective efficacy of Estradiol in such context. In western blot analysis, the expression of cleaved caspase-3 and heat shock protein 70 declined in Estradiol pretreated group compared to ischemic control group. TUNEL assay also showed the efficacy of Estradiol to abate motor neuron apoptosis. Interestingly, Estradiol respectively increased and decreased the expression of Cyclooxygenase (COX)-1 and COX-2, to a significant extent. Estradiol, exerting its protection through affecting one or a combination of involved biochemical factors can constitute a potential candidate to protect against thoracoabdominal aortic aneurysm repairs induced spinal cord I/R injury.     

Intraguild predation among Scolothrips longicornis (Thysanoptera: Thripidae), Neoseiulus californicus and Typhlodromus bagdasarjani (Acari: Phytoseiidae) under laboratory conditions

This study was carried out on the ability of predatory thrips Scolothrips longicornis Priesner to feed on 2 phytoseiid species and vice versa. Also the effect of predation of Neoseiulus californicus (McGregor) on Typhlodromus bagdasarjani Wainstein and Arutunjan and vice versa was evaluated. The larvae, prepupae, and pupae of thrips and the eggs, larvae, and protonymphs of phytoseiids were selected as intraguild prey. The intraguild predation (IGP) among S. longicornis and 2 phytoseiid species was unidirectional and in favor of phytoseiids, i.e., S. longicornis was not able to feed on larval stages of 2 phytoseiids. However, N. californicus and T. bagdasarjani fed on the 1st instar larvae (1.39 and 0.80 per day), 2nd instar larvae (0.87 and 0.55 per day), prepupae (0.51 and 0.48 per day), and pupae of thrips (0.51 and 0.49 per day, respectively). Both phytoseiids fed on eggs, larvae, and protonymphal stages of each other. Females of N. californicus consumed more phytoseiid larvae (2.49 per day) than T. bagdasarjani, which consumed 1.08 N. californicus larvae per day. When Tetranychus urticae was presented as an extraguild prey, intensity of IGP between 2 species of phytoseiids and on larval stages of S. longicornis reduced significantly. Therefore, it is concluded that (i) IGP existed among the 3 examined species and lack of feeding of S. longicornis on 2 phytoseiid species can be justified by its feeding type (monophagy), (ii) N. californicus was much more prone to IGP than was T. bagdasarjani.     

A Metabolic Shift Favoring Sphingosine 1-Phosphate at the Expense of Ceramide Controls Glioblastoma Angiogenesis

Studies in cell culture and mouse models of cancer have indicated that the soluble sphingolipid metabolite sphingosine 1-phosphate (S1P) promotes cancer cell proliferation, survival, invasiveness, and tumor angiogenesis. In contrast, its metabolic precursor ceramide is prodifferentiative and proapoptotic. To determine whether sphingolipid balance plays a significant role in glioma malignancy, we undertook a comprehensive analysis of sphingolipid metabolites in human glioma and normal gray matter tissue specimens. We demonstrate, for the first time, a systematic shift in sphingolipid metabolism favoring S1P over ceramide, which increases with increasing cancer grade. S1P content was, on average, 9-fold higher in glioblastoma tissues compared with normal gray matter, whereas the most abundant form of ceramide in the brain, C18 ceramide, was on average 5-fold lower. Increased S1P content in the tumors was significantly correlated with increased sphingosine kinase 1 (SPHK1) and decreased sphingosine phosphate phosphatase 2 (SGPP2) expression. Inhibition of S1P production by cultured glioblastoma cells, using a highly potent and selective SPHK1 inhibitor, blocked angiogenesis in cocultured endothelial cells without affecting VEGF secretion. Our findings validate the hypothesis that an altered ceramide/S1P balance is an important feature of human cancers and support the development of SPHK1 inhibitors as antiangiogenic agents for cancer therapy.     

Relative motion of transmembrane segments S0 and S4 during voltage sensor activation in the human BK(Ca) channel

Large-conductance voltage- and Ca(2+)-activated K(+) (BK(Ca)) channel α subunits possess a unique transmembrane helix referred to as S0 at their N terminus, which is absent in other members of the voltage-gated channel superfamily. Recently, S0 was found to pack close to transmembrane segments S3 and S4, which are important components of the BK(Ca) voltage-sensing apparatus. To assess the role of S0 in voltage sensitivity, we optically tracked protein conformational rearrangements from its extracellular flank by site-specific labeling with an environment-sensitive fluorophore, tetramethylrhodamine maleimide (TMRM). The structural transitions resolved from the S0 region exhibited voltage dependence similar to that of charge-bearing transmembrane domains S2 and S4. The molecular determinant of the fluorescence changes was identified in W203 at the extracellular tip of S4: at hyperpolarized potential, W203 quenches the fluorescence of TMRM labeling positions at the N-terminal flank of S0. We provide evidence that upon depolarization, W203 (in S4) moves away from the extracellular region of S0, lifting its quenching effect on TMRM fluorescence. We suggest that S0 acts as a pivot component against which the voltage-sensitive S4 moves upon depolarization to facilitate channel activation.     

Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method

Accurate morphological differentiation between the liver fluke species Fasciola hepatica and Fasciola gigantica is difficult. We evaluated PCR-restriction enzyme profiles of internal transcribed spacer 1 (ITS1) that could aid in their identification. Fifty F. hepatica and 30 F. gigantica specimens were collected from different hosts in three provinces of Iran. For DNA extraction, we crushed fragments of the worms between two glass slides as a new method to break down the cells. DNA from the crushed materials was then extracted with a conventional phenol-chloroform method and with the newly developed technique, commercial FTA cards. A primer pair was selected to amplify a 463-bp region of the ITS1 sequence. After sequencing 14 samples and in silico analysis, cutting sites of all known enzymes were predicted and TasI was selected as the enzyme that yielded the most informative profile. Crushing produced enough DNA for PCR amplification with both the phenol-chloroform and commercial FTA card method. The DNA extracted from all samples was successfully amplified and yielded a single sharp band of the expected size. Digestion of PCR products with TasI allowed us to distinguish the two species. In all samples, molecular identification was consistent with morphological identification. Our PCR-restriction enzyme profile is a simple, rapid and reliable method for differentiating F. hepatica and F. gigantica, and can be used for diagnostic and epidemiological purposes.