Application of PCR for detection and identification of mycoplasma contamination in virus stocks

Summary A nested polymerase chain reaction (PCR) was used to detect and identify mycoplasma contaminants in viral stocks. The results of the PCR assay proved to be a sensitive and accurate indicator of the true status of the stock tested. Those samples positive by agar culture or Hoechst stain were also positive by PCR. Those samples that were inconclusive by Hoechst stain (10.05%) could be clearly determined to be mycoplasma positive or negative by PCR. The PCR assay also detected those fastidious species of mycoplasma that gave false negative results by the direct culture method. In many respects the PCR-based mycoplasma detection method described is superior to the agar culture and Hoechst staining detection methods. In this study, the PCR assay detected substantially more mycoplasma-positive viral stocks than did the agar culture assay. Due to its speed, sensitivity, and reliability, the PCR assay is of particular value in monitoring the process of removing mycoplasma from contaminated stocks. Furthermore, the PCR amplification products can be analyzed by restriction analysis to rapidly identify the species of the mycoplasma contaminating the stock tested.     

Synthesis of soluble coordination polymer nanoparticles using room-temperature ionic liquid

We report a new family of soluble cyano-bridged coordination polymer nanoparticles M₃[Cr(CN)₆]₂/[BMIM][BF₄] (where M²⁺ = Ni, Mn, V^IVO; BMIM = 1-butyl-3-methylimidazolium). These nanoparticles of ca. 6 nm were synthesised in the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate, which acts both as a stabilizing agent and a solvent. The magnetic properties of frozen colloids containing the nanoparticles show that the relaxation of magnetisation is strongly influenced by interparticle interactions leading to the appearance of spin-glass-like dynamics in these systems.     

Empagliflozin inhibits angiotensin II-induced hypertrophy in H9c2 cardiomyoblasts through inhibition of NHE1 expression

Molecular and Cellular Biochemistry - Diabetes mellitus (DM)-induced cardiac morbidities have been the leading cause of death among diabetic patients. Recently, sodium-glucose cotransporter-2...     

Nucleic acid components and their analogs: Design and synthesis of novel cytosine thioglycoside analogs

The synthesis of a new category of novel cytosine 4-thioglycoside analogs has been first accomplished. The main step of this strategy is the synthesis of sodium pyrimidine-4-thiolate through the condensation of 2-cyano-N-arylacetamides with sodium cyanocarbonimidodithioate, followed by coupling with α-bromo-sugars to afford the corresponding cytosine 4-thioglycoside analogs. The free thioglycosides were also prepared. Subsequent studies on the application of this strategy for the preparation of other potent pyrimidine thioglycosides are reported.     

Preliminary screening for water stress tolerance and genetic diversity in wheat (Triticum aestivum L.) cultivars from Sudan

The present study was carried out to screen 12 Sudanese wheat (Triticum aestivum L.) cultivars for their response to water stress at early germination stages and to characterize sources that could be used in breeding programs to develop wheat cultivars with better adaptation to water stress. The effect of osmotic stress on the early growing stages was evaluated, in vitro, using five concentrations of Polyethylene glycol. Genetic diversity was studied using 24 allele specific simple sequence repeats (SSR) markers associated with drought tolerance in wheat. The presence of the drought genes and their chromosomal location was also investigated by isolating and sequencing the dehydration responsive element binding protein (dreb1). Results of the in vitro screening among the cultivars showed significant differences in the root length, shoot length and root/shoot ratio. The 24 drought specific SSR markers used revealed 50 alleles, with an average of 2.0 alleles per locus. Of these, 60% were polymorphic with polymorphism information content (PIC) ranging from 0.16 to 0.89. A dendrogram based on the similarity values generated from the SSR data revealed three major clusters. Of the five specific primers for dreb1 genes, only primer P25F/PR produced amplification products with the expected fragment sizes. Sequencing and BLAST results of the cloned fragments excised from the gels showed 99% homology to the dreb1 gene on chromosome 3A.     

Rosmarinic acid ameliorates the negative effects of salinity in in vitro-regenerated potato explants (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.)

Salinity causes massive loss of economic crops due to the build-up of toxic salts in shoot. Rosmarinic acid (RA) has been known to prevent the inevitable damage to vital biological molecules under stress. There is an increasing interest, therefore, in the use of polyphenols in crop stress physiology studies. An in vitro regeneration protocol was established using explants of the potato cultivar White Desiree to assess tolerance to salinity at a morphological, cellular, and biochemical level. Additionally, we examined the potential ameliorative effects of RA on growth in salt-stressed potato explants. Based on our observations, we propose a model for the ameliorative effects of a caffeic acid conjugate of α-hydroxyhydrocaffeic acid. Explants of White Desiree showed differential responses to salt and RA supplementation. Salt-stressed and control explants grown in RA-supplemented media showed a pronounced expansion of leaf area, suggesting direct effects of RA on cell elongation in potato explants. Root elongation was less affected by salinity in comparison to shoot elongation. Moderate levels of RA resulted in stimulatory effects on growth. The microscopic examination of leaves and stems indicated the presence of unique trichomes at the early stages of explant growth under RA supplementation, which varied considerably in size, shape, and distribution in response to stress. Our data indicated that RA significantly decreased osmolyte content under stress. Salinity resulted in the biosynthesis of high levels of free proline, carbohydrates, and malondialdehyde (MDA). However, the lower contents of MDA in salt-stressed explants treated with RA indicate that RA possesses anti-lipid peroxidation properties. Our observations provide evidence for the stimulatory effects of RA on cellular growth and the protection of membranes from ion toxicity.     

Mosquitocidal and repellent activities of essential oils against Culex pipiens L.

Plants are a prospective source of novel natural insect repellents and botanical insecticides. This study was conducted to investigate the chemical composition of the essential oils of three plants growing in Saudi Arabia, namely Ducrosia anethifolia, Achillea fragrantissima, and Teucrium polium; and to evaluate their potential mosquitocidal and repellent activities against adult female Culex pipiens L. The main components of the three oils were found to be decanal (28.9%) and chrysanthenyl acetate (10.04%), (D. anethifolia); sabinyl acetate (35.79) and artemesia ketone (18.28%) (A. fragrantissima); α‐cadinol (49.53%) and δ‐cadinene (10.23%) (T. polium). The oil of A. fragrantissima was the most toxic (LC₅₀ = 0.11 μL/L air) followed by D. anethifolia and T. polium with LC₅₀ values of 5.22 and 25.98 μL/L air, respectively. T. polium oil was the most repellent (292 min at 2 μL/cm²), followed by D. anethifolia and A. fragrantissima. The results indicate that the essential oils have a potential fumigant insecticidal and repellent activities for mosquito control.     

Trichopsomyia ochrozona (Stackelberg, 1952) (Diptera: Syrphidae) recorded from Iran for the first time with a key to the West Palaearctic Trichopsomyia Williston, 1888 species

The hoverfly Trichopsomyia ochrozona (Stackelberg, 1952) (Diptera: Syrphidae) is recorded for the first time from Iran. An illustrated key to the West Palaearctic species of Trichopsomyia is presented. Illustrations of the puparium of Trichopsomyia joratensis Goeldin, 1997 and the larva and puparium of T. ochrozona are given and their larval and adult habitats are described. A short discussion is given on the known larval habitat of other West Palaearctic Trichopsomyia species. Some of the synonyms have been evaluated and Pipiza melancholica Meigen, 1822 is withdrawn from synonymy of Trichopsomyia flavitarsis (Meigen, 1822) and is considered as a junior synonym of Musca viduata Linnaeus, 1758 syn. nov.