Chimeric small subunits influence catalysis without causing global conformational changes in the crystal structure of ribulose-1,5-bisphosphate carboxylase/oxygenase

Comparison of subunit sequences and X-ray crystal structures of ribulose-1,5-bisphosphate carboxylase/oxygenase indicates that the loop between beta-strands A and B of the small subunit is one of the most variable regions of the holoenzyme. In prokaryotes and nongreen algae, the loop contains 10 residues. In land plants and green algae, the loop is comprised of approximately 22 and 28 residues, respectively. Previous studies indicated that the longer betaA-betaB loop was required for the assembly of cyanobacterial small subunits with plant large subunits in isolated chloroplasts. In the present study, chimeric small subunits were constructed by replacing the loop of the green alga Chlamydomonas reinhardtii with the sequences of Synechococcus or spinach. When these engineered genes were transformed into a Chlamydomonas mutant that lacks small-subunit genes, photosynthesis-competent colonies were recovered, indicating that loop size is not essential for holoenzyme assembly. Whereas the Synechococcus loop causes decreases in carboxylation V(max), K(m)(O(2)), and CO(2)/O(2) specificity, the spinach loop causes complementary decreases in carboxylation V(max), K(m)(O(2)), and K(m)(CO(2)) without a change in specificity. X-ray crystal structures of the engineered proteins reveal remarkable similarity between the introduced betaA-betaB loops and the respective loops in the Synechococcus and spinach enzymes. The side chains of several large-subunit residues are altered in regions previously shown by directed mutagenesis to influence CO(2)/O(2) specificity. Differences in the catalytic properties of divergent Rubisco enzymes may arise from differences in the small-subunit betaA-betaB loop. This loop may be a worthwhile target for genetic engineering aimed at improving photosynthetic CO(2) fixation.     

Sublethal concentration of H<Subscript>2</Subscript>O<Subscript>2</Subscript> enhances the protective effect of mesenchymal stem cells in rat model of spinal cord injury

Objective

To investigate the effect of H₂O₂ on the migration and antioxidant defense of mesenchymal stem cells (MSCs) and the neurotrophic effects of H₂O₂-treated MSCs on spinal cord injury (SCI).

Results

Sublethal concentrations of H₂O₂ decreased cell migration and expression of CXCR4 and CCR2 as well as Nrf2 expression in MSCs. In the second phase, transplantation of treated and untreated MSCs to SCI caused minor changes in locomotor dysfunction. There was a significantly difference between cell-treated and spinal cord injury groups in expression of BDNF (brain-derived neurotrophic factor). Transplantation of H₂O₂-treated cells caused an increase in BDNF expression compared to non-treated cells.

Conclusion

Transplantation of H₂O₂-treated stem cells may have protective effects against SCI through by increasing neurotrophic factors.

     

Impact of the erineum strain of <Emphasis Type="Italic">Colomerus vitis</Emphasis> (Acari: Eriophyidae) on the development of plants of grapevine cultivars of Iran

The present experiment was aimed at determining the influence of the grape erineum strain of Colomerus vitis (GEM) (Acari: Eriophyidae) on responses of local grapevine cultivars. GEM was applied at five density levels to each of five cultivars, i.e. Shahani, Sahebi Uroomie, Khalili Bovanat, Rishbaba and Sezdang Ghalat (listed from early to late grape ripening). The experiment was performed in a full factorial design (12 replicates each) and effects of the mite on the relative content of leaf chlorophyll, internode and cane length, leaf area and weight, number and size of the erinea, and percentage of leaves with erinea were investigated. Also mite density on leaves and in buds was assessed. Data were analyzed with a two-way ANOVA followed by Tukey’s test to separate means among treatment levels and cultivars. The relative content of chlorophyll (expressed in Spad units) in infested leaves was reduced along with an increase in mite density and it was shown to be highly significant at the two higher mite density levels for Khalili Bovanat, Rishbaba and Sezdang Ghalat; Shahani and Sahebi Uroomie leaves appeared to be less affected by mite infestation. The highest mite density treatment displayed a strong correlation with weight (positive correlation) and size (negative correlation) of the leaves of four cultivars; leaves of Sahebi Uroomie appeared to be less affected. The reduced internode length was weak in infested plants. Most infested plants produced shorter canes and their lengths appeared to have a strong negative correlation with the highest mite density in four cultivars; canes of Sahebi Uroomie did not appear affected. At the highest mite density, canes of Khalili Bovanat and Sahebi Uroomie displayed the most and the least shortening effects, respectively. The percentage of leaves with erinea, as well as the number of erinea per leaves and the diameter of erinea increased along with the mite population density. The mite densities in buds (April 2014) and on leaves with erinea (in November 2013) were higher at the highest treatment level in the medium-late (Rishbaba) and late ripening (Sezdang Ghalat) cultivars, than in the early and early-medium ripening ones. Almost all data collected in the current experiment allowed the conclusion that Sahebi Uroomie and Shahani were less affected than the other cultivars (Khalili Bovanat, Rishbaba and Sezdang Ghalat).     

The role of antioxidants in sperm freezing: a review

Cryopreservation of spermatozoa is becoming more important because of new clinical requirements and current clinical practice. Despite the success of sperm cryopreservation this routinely used procedure induces serious detrimental changes in sperm function. Some researchers believe that cryopreservation is associated with DNA fragmentation and DNA single strand breaks in sperm. Mechanisms of cryodamage to human spermatozoa are thought to be multifactorial including: cold shock, osmotic stress, intracellular ice crystal formation, oxidative stress, and combinations of these conditions. Additives showing antioxidative properties reported to reduce the impact of ROS-induced and cold shock damages. Many studies exist as regards the effects of antioxidants on the cryopreservation aimed at improving the quality of post-thaw semen. Hence, this review will clarify results of recent applications of various antioxidants used in numerous research efforts to improve cryopreservation of spermatozoa. This review is to increase the understanding of the roles of these antioxidants concerning mechanisms which enhance resistance to cryodamage of spermatozoa.     

Experimental and theoretical investigations of Dy(III) complex with 2,2′-bipyridine ligand: DNA and BSA interactions and antimicrobial activity study

Abstract

In this study, the interactions of a novel metal complex [Dy(bpy)₂Cl₃.OH₂] (bpy is 2,2'-bipyridine) with fish salmon DNA (FS-DNA) and bovine serum albumin (BSA) were investigated by experimental and theoretical methods. All results suggested significant binding between the Dy(III) complex with FS-DNA and BSA. The binding constants (K_b), Stern-Volmer quenching constants (K_SV) of Dy(III)-complex with FS-DNA and BSA at various temperatures as well as thermodynamic parameters using Van’t Hoff equation were obtained. The experimental results from absorption, ionic strength, iodide ion quenching, ethidium bromide (EtBr) quenching studies and positive ΔH? and ΔS? suggested that hydrophobic groove-binding mode played a predominant role in the binding of Dy(III)-complex with FS-DNA. Indeed, the molecular docking results for DNA-binding were in agreement with experimental data. Besides, the results found from experimental and molecular modeling indicated that the Dy(III)-complex bound to BSA via Van der Waals interactions. Moreover, the results of competitive tests by phenylbutazone, ibuprofen, and hemin (as a site-I, site-II and site-III markers, respectively) considered that the site-III of BSA is the most possible binding site for Dy(III)-complex. In addition, Dy(III) complex was concurrently screened for its antimicrobial activities. The presented data provide a promising platform for the development of novel metal complexes that target nucleic acids and proteins with antimicrobial activity.

Communicated by Ramaswamy H. Sarma