Metabolomic Response of Calotropis procera Growing in the Desert to Changes in Water Availability

Water availability is a major limitation for agricultural productivity. Plants growing in severe arid climates such as deserts provide tools for studying plant growth and performance under extreme drought conditions. The perennial species Calotropis procera used in this study is a shrub growing in many arid areas which has an exceptional ability to adapt and be productive in severe arid conditions. We describe the results of studying the metabolomic response of wild C procera plants growing in the desert to a one time water supply. Leaves of C. procera plants were taken at three time points before and 1 hour, 6 hours and 12 hours after watering and subjected to a metabolomics and lipidomics analysis. Analysis of the data reveals that within one hour after watering C. procera has already responded on the metabolic level to the sudden water availability as evidenced by major changes such as increased levels of most amino acids, a decrease in sucrose, raffinose and maltitol, a decrease in storage lipids (triacylglycerols) and an increase in membrane lipids including photosynthetic membranes. These changes still prevail at the 6 hour time point after watering however 12 hours after watering the metabolomics data are essentially indistinguishable from the prewatering state thus demonstrating not only a rapid response to water availability but also a rapid response to loss of water. Taken together these data suggest that the ability of C. procera to survive under the very harsh drought conditions prevailing in the desert might be associated with its rapid adjustments to water availability and losses

Gene expression and histopathology alterations during rat mammary carcinogenesis induced by 7,12- dimethylbenz[a]anthracene and the protective role of Neem (Azadirachta indica) leaf extract

Abstract: The present study was investigated to evaluate the protective role of ethanolic neem leaf extract (ENLE) against 7,12-dimethylbenz[a]anthracene (DMBA)-induced expression alterations of the Bcl-2, CK8, CK19, p53, p21, p27 and PCNA genes and histopathological lesions in the mammary tissues of female rats. Eighty Swiss albino female rats were divided into eight groups. Group 1 supplemented with corn oil as control. Group 2 females received DMBA. Groups 3, 4 and 5 females received 100, 200 and 400 mg/kg of ENLE alternate to the DMBA application from the beginning of the experimental period, respectively. Groups 6, 7, and 8 females treated similar to groups 3, 4 and 5 plus DMBA. All the animals were sacrificed after an experimental period of 12 weeks. The expression of Bcl-2, CK8, CK19, p53, p21, p27 and PCNA genes was investigated using reverse transcription polymerase chain reaction (RT-PCR). In addition, histopathology analysis of the mammary tissues was confirmed. The results revealed that DMBA treatment induced expression alterations of genes related to cancer. Also histopathological lesions were found in mammary tissues of female rats. These alterations of the gene expression as well as the histopathological lesions were markedly suppressed when female rats were treated with ENLE combined with DMBA. Conclusion: These findings suggest that ENLE exerts its anticancer properties by inhibiting alterations in the gene expression and the histopathological lesions in the mammary tissues of female rats exposed to DMBA

Production and characterization of thermostable xylanase from Bacillus subtilis XP10 isolated from marine water

This study aims is to characterize an efficient bacterium, capable of producing thermostable xylanases. Different bacterial isolates were obtained on medium containing xylan as the sole carbon source from samples collected from Jeddah, Saudi Arabia. Out of 15 xylanase producing isolates, the best xylanase producer was XP10 which was selected for the enzyme production. It was identified based on morphological and some biochemical characters as a species belonging to the genus Bacillus and identified as Bacillus subtilis XP10. Identification was confirmed by 16S rDNA studies and restriction fragment length polymorphism. Factors affecting enzymes production were studied. Maximum xylanase production was 2.82 U/ml obtained at pH 8 for 4 days at 40°C and 120 rpm. The molecular weight of the purified enzyme was 23 kDa. Fivefold increasing in xylanase production was obtained by construction of recombinant Escherichia coli 107 harboring recombinant vector pJET 1.2/blunt/xynA. The purified thermostable alkalotolerant xylanase can be used in the treatment of agricultural wastes as well as in the bioremediation of xylan-containing materials.Keywords: Xylanase, polymerase chain reaction based restriction fragment length polymorphism (PCR-based RFLP), 16S rDNA, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), xynA geneAfrican Journal of Biotechnology Vol. 12(8), pp. 780-79

<i>Actinidia deliciosa</i> Extract as a Promising Supplemental Agent for Hepatic and Renal Complication-Associated Type 2 Diabetes (In Vivo and In Silico-Based Studies)

Type 2 diabetes (T2D) is a chronic metabolic condition associated with obesity, oxidative stress-mediated inflammation, apoptosis, and impaired insulin signaling. The utilization of phytochemical therapy generated from plants has emerged as a promising approach for the treatment of diabetes and its complications. Kiwifruit is recognized for its substantial content of antioxidative phenolics. Therefore, this work aimed to examine the effect of Actinidia deliciosa (kiwi fruit) on hepatorenal damage in a high-fat diet (HFD) and streptozotocin (STZ)-induced T2D in rats using in vivo and in silico analyses. An increase in hepatic and renal lipid peroxidation was observed in diabetic rats accompanied by a decrease in antioxidant status. Furthermore, it is important to highlight that there were observable inflammatory and apoptotic responses in the hepatic and renal organs of rats with diabetes, along with a dysregulation of the phosphorylation levels of mammalian target of rapamycin (mTOR), protein kinase B (Akt), and phosphoinositide 3-kinase (PI3K) signaling proteins. However, the administration of kiwi extract to diabetic rats alleviated hepatorenal dysfunction, inflammatory processes, oxidative injury, and apoptotic events with activation of the insulin signaling pathway. Furthermore, molecular docking and dynamic simulation studies revealed quercetin, chlorogenic acid, and melezitose as components of kiwi extract that docked well with potential as effective natural products for activating the silent information regulator 1(SIRT-1) pathway. Furthermore, phenolic acids in kiwi extract, especially syringic acid, P-coumaric acid, caffeic acid, and ferulic acid, have the ability to inhibit the phosphatase and tensin homolog (PTEN) active site. In conclusion, it can be argued that kiwi extract may present a potentially beneficial adjunctive therapy approach for the treatment of diabetic hepatorenal complications

Experimental Setup and plants chosen.

<p>(a) Representative photo of the plants chosen for this experiment growing in its natural habitat in Saudi Arabia near to Jeddah. For this study representative species of similar size and performance were chosen. (b) Experimental set-up. At day 1 (control) leaves of three independent plants were harvested 1 h post-dawn, at midday and 1 h pre-dusk. One day later (Day 2), plants were watered at dawn and leaves were harvested 1 h post dawn, at midday and 1 h pre-dusk. Harvested leaves were frozen immediately in liquid –N and processes as described in Experimental procedures.</p

Clustered heatmap visualization of different lipid classes.

<p>Shown is the average abundance of several complex lipids visualized in a false-color heatmap at the three time points before and after watering ordered according to their presence in photosynthetic membranes, in cellular membranes or representing storage lipids.</p

Relative water content of leaves of <i>C. procera</i> plants taken at dawn, midday and one hour pre-dusk one day before watering and up to three days after watering.

<p>Relative water content of leaves of <i>C. procera</i> plants taken at dawn, midday and one hour pre-dusk one day before watering and up to three days after watering.</p

Principal Component Analysis (PCA) and ANOVA for metabolomic analysis of leaf samples from control and watered plants.

<p>(a) PCA (upper part) and ANOVA (lower part) for primary metabolites. (b) PCA (upper part) and ANOVA (lower part) for complex lipids. (c) PCA (upper part) and ANOVA (lower part) for secondary metabolites. Shown are always three independent samples per time point (dawn (1 hour post dawn/after watering), midday (6 hours after dawn/after watering) and pre-dusk (12 hours after dawn/after watering). Watered samples are shown in blue, non-watered in red. The lower part shows the results of a Bonferroni corrected ANOVA displaying the influence of treatment (watering) for all samples and of harvesting time for primary and secondary metabolites.</p

Boxplots of representative species of photosynthetic, structural and storage lipids.

<p>Boxplot-visualizations for a subset of complex lipids as determined for the three independent samples for the different time points and treatments as indicated on the x-axis of three replicates that were harvested at 1 h post-dawn, midday and 1 h pre-dusk for control and rehydrated plants.</p