Gallic acid modulates purine metabolism and oxidative stress induced by ethanol exposure in zebrafish brain

Gallic acid (GA) is a secondary metabolite found in plants. It has the ability to cross the blood-brain barrier and, through scavenging properties, has a protective effect in a brain insult model. Alcohol metabolism generates reactive oxygen species (ROS); thus, alcohol abuse has a deleterious effect on the brain. The zebrafish is a vertebrate often used for screening toxic substances and in acute ethanol exposure models. The aim of this study was to evaluate whether GA pretreatment (24 h) prevents the changes induced by acute ethanol exposure (1 h) in the purinergic signaling pathway in the zebrafish brain via degradation of extracellular nucleotides and oxidative stress. The nucleotide cascade promoted by the nucleoside triphosphate diphosphohydrolase (NTPDase) and 5′-nucleotidase was assessed by quantifying nucleotide metabolism. The effect of GA alone at 5 and 10 mg L^?1 did not change the nucleotide levels. Pretreatment with 10 mg L^?1 GA prevented an ethanol-induced increase in ATP and ADP levels. No significant difference was found between the AMP levels of the two pretreatment groups. Pretreatment with 10 mg L^?1 GA prevented ethanol-enhanced lipid peroxidation and dichlorodihydrofluorescein (DCFH) levels. The higher GA concentration was also shown to positively modulate against ethanol-induced effects on superoxide dismutase (SOD), but not on catalase (CAT). This study demonstrated that GA prevents the inhibitory effect of ethanol on NTPDase activity and oxidative stress parameters, thus consequently modulating nucleotide levels that may contribute to the possible protective effects induced by alcohol and purinergic signaling.

     

Host suitability and feeding preference of the African citrus triozid Trioza erytreae Del Guercio (Hemiptera: Triozidae), natural vector of “Candidatus Liberibacter africanus”

African citrus greening (ACGD) and huanglongbing (HLB) diseases are the most damaging diseases of citrus worldwide. Currently, the disease has no cure and has been attributed to the collapse of the citrus industry in several countries. In Africa, the causative agent “Candidatus” Liberibacter africanus is vectored by African citrus triozid (ACT) Trioza erytreae Del Guercio (Hemiptera: Triozidae). African citrus triozid is native to Africa but has been recently reported in Asia and Europe. Apart from citrus, Murraya koenigii (L.) and Clausena anisata (Willd) Hook. F. ex Benth. are also considered as preferred host plants. At present, there is scant information on host plant suitability and preference of T. erytreae. Also, there are contradictory reports on its reproduction and survival on rutaceous and non‐rutaceous host plants. In the present study, we tested the suitability and preference of rutaceous and non‐rutaceous trees and shrubs as potential ACT host plants in choice and no‐choice bioassays. The development from egg to the adult stage was longest on Calodendrum capense (Wright & Arn.) Engl. Host plants of superior quality accordingly to several ACT's biological parameters measured also revealed significantly higher morphometric characteristics. Our findings on the host status of the five rutaceous plants imply that these plants can greatly influence the population dynamics of ACT as well as the epidemiology of ACGD, and these can be a useful guide in the area‐wide management of the pest in Kenya.     

S6 kinase deletion suppresses muscle growth adaptations to nutrient availability by activating AMP kinase

S6 kinase (S6K) deletion in metazoans causes small cell size, insulin hypersensitivity, and metabolic adaptations; however, the underlying molecular mechanisms are unclear. Here we show that S6K-deficient skeletal muscle cells have increased AMP and inorganic phosphate levels relative to ATP and phosphocreatine, causing AMP-activated protein kinase (AMPK) upregulation. Energy stress and muscle cell atrophy are specifically triggered by the S6K1 deletion, independent of S6K2 activity. Two known AMPK-dependent functions, mitochondrial biogenesis and fatty acid β-oxidation, are upregulated in S6K-deficient muscle cells, leading to a sharp depletion of lipid content, while glycogen stores are spared. Strikingly, AMPK inhibition in S6K-deficient cells restores cell growth and sensitivity to nutrient signals. These data indicate that S6K1 controls the energy state of the cell and the AMPK-dependent metabolic program, providing a mechanism for cell mass accumulation under high-calorie diet.     

Topological remodeling of cultured endothelial cells by characterized cyclic strains

Evaluation of mechanical environment on cellular function is a major field of study in cellular engineering. Endothelial cells lining the entire vascular lumen are subjected to pulsatile blood pressure and flow. Mechanical stresses caused by such forces determine function of arteries and their remodeling. Critical values of mechanical stresses contribute to endothelial damage, plaque formation and atherosclerosis. A device to impose cyclic strain on cultured cells inside an incubator was designed and manufactured operating with different load amplitudes, frequencies, numbers of cycles and ratios of extension to relaxation. Endothelial cells cultured on collagen coated silicon scaffolds were subjected to cyclic loading. Effects of mechanical loading on cell morphology were quantified using image processing methods. Results showed change in cell orientation from a randomly oriented before the test up to 80 degrees alignment from load axis after loading. Endothelial cells were elongated with shape index reductions up to 47% after cyclic stretch. By increase of strain amplitude, loading frequency and number of cycles, significant decrease in shape index and significant increase in orientation angle were observed. Change of load waveform similar to arterial pulse pressure waveform resulted in alteration of cell alignment with 9.7% decrease in shape index, and 10.8% increase in orientation angle. Results of cyclic loading tests in a disturbed environment with elevated PH showed lack of remodeling. It was concluded that tensile loading of endothelial cells influences cell morphology and alignment, a mechanism for structural regulation, functional adaptation and remodeling. Disturbed environment results in endothelial dysfunction and injury.     

Comparative analysis of development and survival of two Natal fruit fly Ceratitis rosa Karsch (Diptera,Tephritidae) populations from Kenya and South Africa

Comparative analysis of development and survivorship of two geographically divergent populations of the Natal fruit fly Ceratitis rosa Karsch designated as Ceratitis rosa R1 and Ceratitis rosa R2 from Kenya and South Africa were studied at seven constant temperatures (10, 15, 20, 25, 30, 33, 35 °C). Temperature range for development and survival of both populations was 15–35 °C. The developmental duration was found to significantly decrease with increasing temperature for Ceratitis rosa R1 and Ceratitis rosa R2 from both countries. Survivorship of all the immature stages of Ceratitis rosa R1 and Ceratitis rosa R2 from Kenya was highest over the range of 20–30 °C (87–95%) and lowest at 15 and 35 °C (61–76%). Survivorship of larvae of Ceratitis rosa R1 and Ceratitis rosa R2 from South Africa was lowest at 35 °C (22%) and 33 °C (0.33%), respectively. Results from temperature summation models showed that Ceratitis rosa R2 (egg, larva and pupa) from both countries were better adapted to low temperatures than R1, based on lower developmental threshold. Minimum larval temperature threshold for Kenyan populations were 11.27 °C and 6.34 °C (R1 and R2, respectively) compared to 8.99 °C and 7.74 °C (R1 and R2, respectively) for the South African populations. Total degree-day (DD) accumulation for the Kenyan populations were estimated at 302.75 (Ceratitis rosa R1) and 413.53 (Ceratitis rosa R2) compared to 287.35 (Ceratitis rosa R1) and 344.3 (Ceratitis rosa R2) for the South African populations. These results demonstrate that Ceratitis rosa R1 and Ceratitis rosa R2 from both countries were physiologically distinct in their response to different temperature regimes and support the existence of two genetically distinct populations of Ceratitis rosa. It also suggests the need for taxonomic revision of Ceratitis rosa, however, additional information on morphological characterization of Ceratitis rosa R1 and Ceratitis rosa R2 is needed.     

A structural basis for the biosynthesis of the major chlorogenic acids found in coffee

Chlorogenic acids (CGAs) are a group of phenolic secondary metabolites produced by certain plant species and an important component of coffee (Coffea spp.). The CGAs have been implicated in biotic and abiotic stress responses, while the related shikimate esters are key intermediates for lignin biosynthesis. Here, two hydroxycinnamoyl-coenzyme A shikimate/quinate hydroxycinnamoyl transferases (HCT/HQT) from coffee were biochemically characterized. We show, to our knowledge for the first time, that in vitro, HCT is capable of synthesizing the 3,5-O-dicaffeoylquinic acid diester, a major constituent of the immature coffee grain. In order to further understand the substrate specificity and catalytic mechanism of the HCT/HQT, we performed structural and mutagenesis studies of HCT. The three-dimensional structure of a native HCT and a proteolytically stable lysine mutant enabled the identification of important residues involved in substrate specificity and catalysis. Site-directed mutagenesis confirmed the role of residues leucine-400 and phenylalanine-402 in substrate specificity and of histidine-153 and the valine-31 to proline-37 loop in catalysis. In addition, the histidine-154-asparagine mutant was observed to produce 4-fold more dichlorogenic acids compared with the native protein. These data provide, to our knowledge, the first structural characterization of a HCT and, in conjunction with the biochemical and mutagenesis studies presented here, delineate the underlying molecular-level determinants for substrate specificity and catalysis. This work has potential applications in fine-tuning the levels of shikimate and quinate esters (CGAs including dichlorogenic acids) in different plant species in order to generate reduced or elevated levels of the desired target compounds.     

Comparison of therapeutic effect of aqueous extract of garlic and nystatin mouthwash in denture stomatitis

doi: 10.1111/j.1741‐2358.2011.00544.x Comparison of therapeutic effect of aqueous extract of garlic and nystatin mouthwash in denture stomatitis Introduction: Denture stomatitis (DS) is the most common form of chronic oral candidiasis. The standard treatment for DS is nystatin, which is accompanied with complications such as a bitter taste. The aim of this study was to compare the effect of garlic with nystatin in DS. Material and Methods: This randomised clinical trial study was performed on 40 patients with DS. After obtaining written consent, patients were divided into two groups while members of each group were given either nystatin or garlic extract for 4 weeks. The length and width of erythema area was measured at the end of the first, second, third, and the fourth weeks using a calliper. Data were analysed by SPSS and statistical tests including variance analysis with anova repeated measures, chi‐square, and least square differences. Results: The changes in the length and width of erythema at different times according to the type of treatment were found to be significant while an accelerated recovery was demonstrated for nystatin (p < 0.001). Both regimens resulted in significant recovery (p < 0.0001). Greater satisfaction with the use of garlic rather than nystatin was mentioned (p < 0.0001). Conclusion: Considering the efficacy of garlic and lack of side effects for this compound and also regarding the nystatin‐associated complications, garlic extract can be introduced as a substitution for standard treatment in DS.     

Deoxycholic acid induces intracellular signaling through membrane perturbations

Secondary bile acids have long been postulated to be tumor promoters in the colon; however, their mechanism of action remains unclear. In this study, we examined the actions of bile acids at the cell membrane and found that they can perturb membrane structure by alteration of membrane microdomains. Depletion of membrane cholesterol by treating with methyl-beta-cyclodextrin suppressed deoxycholic acid (DCA)-induced apoptosis, and staining for cholesterol with filipin showed that DCA caused a marked rearrangement of this lipid in the membrane. Likewise, DCA was found to affect membrane distribution of caveolin-1, a marker protein that is enriched in caveolae membrane microdomains. Additionally, fluorescence anisotropy revealed that DCA causes a decrease in membrane fluidity consistent with the increase in membrane cholesterol content observed after 4 h of DCA treatment of HCT116 cells. Significantly, by using radiolabeled bile acids, we found that bile acids are able to interact with and localize to microdomains differently depending on their physicochemical properties. DCA was also found to induce tyrosine phosphorylation and activate the receptor tyrosine kinase epidermal growth factor receptor in a ligand-independent manner. In contrast, ursodeoxycholic acid did not exhibit any of these effects even though it interacted significantly with the microdomains. Collectively, these data suggest that bile acid-induced signaling is initiated through alterations of the plasma membrane structure and the redistribution of cholesterol.