Root endophytic fungus Serendipita indica modulates barley leaf blade proteome by increasing the abundance of photosynthetic proteins in response to salinity.

AIMS: The present study aimed at analysing the proteome pattern of the leaf blade of barley (Hordeum vulgare L.) in Serendipita indica-colonised plants to decipher the molecular mechanism of S. indica-mediated salt stress. This work is aligned with our previous research on barley leaf sheath to study proteomic pattern variability in leaf blade and sheath of barley plant in response to salinity and S. indica inoculation. METHODS AND RESULTS: The experiment was conducted using a completely randomised factorial design with four replications and two treatments: salinity (0 and 300 mmol l-1 NaCl) and fungus (noninoculated and S. indica-inoculated). The leaf blades of the salt-treated S. indica-colonised and noninoculated plants were harvested after 2 weeks of salt treatment for the physiological and proteomic analyses. After exposure to 300 mmol l-1 NaCl, shoot dry matter production in noninoculated control plants decreased 84% which was about twofold higher than inoculated plants with S. indica. However, the accumulation of sodium in the shoot of S. indica-inoculated plants was significantly lower than the control plants. Analysis of the proteome profile revealed a high number of significantly up-regulated proteins involved in photosynthesis (26 out of 42 identified proteins). CONCLUSIONS: The results demonstrated how the enhanced plant growth and salt stress resistance induced by S. indica was positively associated with the up-regulation of several proteins involved in photosynthesis and carbohydrate metabolism. In fact, S. indica improved photosynthesis in order to reach the best possible performance of the host plant under salt stress. SIGNIFICANCE AND IMPACT OF THE STUDY: Current research provides new insight into the mechanism applied by S. indica in reducing the negative impacts of salt stress in barley at physiological and molecular levels.

Gene Expression Patterns of Royan Human Embryonic Stem Cells Correlate with Their Propensity and Culture Systems.

OBJECTIVE: Human embryonic stem cells (hESCs) have the potential to give rise to all types of cells in the human body when appropriately induced to differentiate. Stem cells can differentiate spontaneously into the three-germ layer derivatives by embryoid bodies (EBs) formation. However, the two-dimensional (2D) adherent culture of hESCs under defined conditions is commonly used for directed differentiation toward a specific type of mature cells. In this study, we aimed to determine the propensity of the Royan hESC lines based on comparison of expression levels of 46 lineage specific markers. MATERIALS AND METHODS: In this experimental study, we have compared the expression of lineage-specific markers in hESC lines during EB versus adherent-based spontaneous differentiation. We used quantitative real-time polymerase chain reaction (qRT-PCR) to assess expressions of 46 lineage-specific markers in 4 hESC lines, Royan H1 (RH1), RH2, RH5, and RH6, during spontaneous differentiation in both EB and adherent cultures at 0, 10, and 30 days after initiation of differentiation. RESULTS: Based on qRT-PCR data analysis, the liver and neuronal markers had higher expression levels in EBs, whereas skin-specific markers expressed at higher levels in the adherent culture. The results showed differential expression patterns of some lineage-specific markers in EBs compared with the adherent cultures. CONCLUSION: According to these results, possibly the spontaneous differentiation technique could be a useful method for optimization of culture conditions to differentiate stem cells into specific cell types such ectoderm, neuron, endoderm and hepatocyte. This approach might prove beneficial for further work on maximizing the efficiency of directed differentiation and development of novel differentiation protocols.

Induction of Sublethal Oxidative Stress on Human Sperm before Cryopreservation: A Time-Dependent Response in Post-Thawed Sperm Parameters.

OBJECTIVE: A recent innovative approach, based on induction of sublethal oxidative stress to enhance sperm cryosurvival, has been applied before sperm cryopreservation. The purpose of this study was to investigate the effects of different induction times of sublethal oxidative stress before cryopreservation on human post-thawed sperm quality. MATERIALS AND METHODS: In this experimental study, we selected semen samples (n=20) from normozoospermic men according to 2010 World Health Organization (WHO) guidelines. After processing the samples by the density gradient method, we divided each sample into 5 experimental groups: fresh, control freezing, and 3 groups exposed to 0.01 µM sodium nitroprusside (SNP) [nitric oxide (NO) donor] for 30 (T30), 60 (T60), or 90 minutes (T90) at 37˚C and 5% CO2 before cryopreservation. Motion characteristics [computer-assisted sperm analyser], viability, apoptosis [annexin V/propidium iodide (PI) assay], DNA fragmentation [sperm chromatin structure assay (SCSA)], and caspase 3 activity (FLICA Caspase Detection Kit) were assessed after thawing. The results were analysed by using one-way ANOVA and Tukey's test. The means were significantly different at P<0.05. RESULTS: Cryopreservation significantly decreased sperm viability and motility parameters, and increased the percentage of apoptosis, caspase 3 activity, and DNA fragmentation (P<0.01) compared to the fresh group. The T60 group had a higher significant percentage of total motility (TM) and progressive motility compared with other cryopreserved groups (P<0.05). We observed a significantly lower percentage of apoptotic rate and caspase 3 activity in the T60 group compared to the other cryopreserved groups (P<0.05). DNA integrity was not significantly affected by this time of sublethal stress induction (P>0.05). CONCLUSION: Our results have demonstrated that the application of sublethal oxidative stress by using 0.01 µM NO for 60 minutes before the freezing process can be a beneficial approach to improve post-thawed human sperm quality.

The suitability of some blood gas and biochemical parameters as diagnostic tools or early indicators of ascites syndrome in broiler sire lines.

In recent few years, there have been some attempts to find a reliable indicator trait as a selection criterion against susceptibility to ascites syndrome (AS). Blood parameters were of great interest as they could be measured in live animals without implementing an ascites-inducing challenge (AIC). In this work, the suitability of some blood parameters was evaluated for diagnosing AS-susceptible chicks in later steps of the disease in trial 1 as well as their early predictive ability in trial 2. In the first trial, one hundred 1-day-old chicks from two pure broiler lines namely S1 and S2 and, in the second trial, 226 1-day-old chicks from line S2 were subjected to AIC. Saline drinking water (1200 mg/l) and lower-than-standard ambient temperatures were the implemented AICs in trials 1 and 2 respectively. The blood parameters including pH, partial pressure of O2 (pO2 ), partial pressure of CO2 (pCO2 ), bicarbonate ion concentration (BIC), percentage of haematocrit (HCT) and saturated haemoglobin (SaO2 ) were measured twice per each bird at days 28 and 35 in trial 1 and once in trial 2 at day 21. The results of the first trial revealed that in line S2 some of the blood parameters differed significantly between the ascitic and non-ascitic groups following exposure to AIC. In this line, the incidence of AS was accompanied by a lower pO2 , SaO2 and BIC, while with higher pCO2 and HCT values. In the second trial, however, although almost all of the parameters showed meaningful differences between the ascitic and non-ascitic broilers, only mean difference of BIC parameter was statistically significant. The general conclusion of this study is that the blood parameters can somewhat have diagnostic ability in the condition in which the AIC is already present, whereas the results did not approve their usefulness as early predictors of AS.