Changes in the human peritoneal mesothelial cells during aging

The number of older patients admitted to peritoneal dialysis (PD) programmes is growing. At the same time, there is increasing data about the role of mesothelial cells in determining the functional alteration of the peritoneum during PD. However, little is known about the functional changes accompanying the ageing process in mesothelial cells. We aimed to evaluate whether the aging process is accompanied by changes in some functional characteristic of the human peritoneal mesothelial cells (HPMC), which could account for the poor prognosis observed in old patients with PD. HPMCs were isolated from patients undergoing a nonurgent, nonseptic abdominal surgical procedure, without renal, vascular or inflammatory disease. Cytokine levels (by enzyme-linked immunosorbent assay (ELISA)), nitrates+nitrites, and cyclooxygenase (COX) activity (by a chemiluminescence assay), cytokines, COX, nitric oxide synthase (NOS), and nuclear factor (NF)-κB1, two messenger ribonucleic acid (mRNA) gene expressions (by reverse transcriptase (RT)-Multiplex PCR), COX, and NOS promoter gene activities, and NF-κB-dependent transcription (by transient transfection assays) were determined. Our data show a significant increase in cytokines, COX, and NOS activities, and mRNA expression of cytokines, COX-2, inducible nitric oxide synthase (iNOS) and precursors of NF-κB in HPMCs from old people. This was also the case for COX-2 and iNOS promoter gene activities and NF-κB-dependent transcription. There was a positive correlation between the age of the donor's cell and the proinflammatory profile of the HPMCs. Such age-dependent increase (around two–three times) is partially abolished by different antioxidant or free-radical scavengers. Thus, aging is accompanied by the presence of an inflammatory state in HPMCs, which involves the participation of different reactive oxygen species

HYPOGLYCEMIC AND HYPOLIPIDEMIC EFFECTS OF SPIRULINA PLATENSIS, PHYCOCYANIN, PHYCOCYANOPEPTIDE AND PHYCOCYANOBILIN ON MALE DIABETIC RATS

This study aimed to evaluate hypoglycemic and hypolipidemic activities of Spirulina Platensis and its bioactive components (phycocyanin (PC), phycocyanopeptide (PCP) and phycocyanobilin (PCB)) on male diabetic Rats compared to controls and glibenclamide drug. For this reason, male Albino rats were equally divided into seven groups designated as normal control, diabetic control, diabetic + glibenclamide (Glyburide) drug (600 µg kg−1 body weight), diabetic + Spirulina biomass suspension (50 mg/ml/ kg−1 body weight), diabetic + phycocyanin (50 mg kg−1 body weight), diabetic + phycocyanopeptide (49 mg kg−1 body weight) and diabetic + phycocyanobilin (982 µg kg−1 body weight). The results show a statistically significant reduction (P < 0.05) level of fasting blood glucose, insulin resistance and lipids levels in diabetic animals administration with Spirulina Platensis, phycocyanin, phycocyanopeptide and phycocyanobilin compared with diabetic control.  Also, there were an increase in HDL–cholesterol levels and β-cell function in these treatments. Histopathologically, diabetic rats treated with spirulina, PC, PCP induced a slight improve of pancreatic cells and an obvious recovery of pancreatic cells.  The expression of insulin secretion from cells (β-cells) of diabetic rats was improved in the groups treated with Spirulina, phycocyanin, phycocyanopeptide. While, diabetic rats treated with phycocyanobilin recorded insulin levels lower than them.  From this study it can be concluded that Spirulina Platensis, phycocyanin, phycocyanopeptide and phycocyanobilin possessed hypoglycemic, insulin sensitivity and hypolipidemic effects. Hypoglycemic and hypolipidemic effects of Spirulina Platensis may be attributed to phenolic compounds and phycocyanin. The antidiabetic effect of PC is most likely due to its ability to reduction of insulin resistance, enhance β-cell function and recovery of β-cells. The effect of PC may be attributed to selenium-binding phycocyanopeptide or/ and phycocyanobilin responsible for the antioxidant activity and chromium-binding phycocyanopeptide which activates insulin receptors

Dietary clenbuterol modifies the expression of genes involved in the regulation of lipid metabolism and growth in the liver, skeletal muscle, and adipose tissue of Nile tilapia (Oreochromis niloticus)

The current study aimed to evaluate whether clenbuterol, a β2-adrenergic agonist, supplementation in Nile tilapia (Oreochromis niloticus) diets can influence growth and blood parameters. Besides, assessment of adipogenic genes as fatty acid synthase (FAS) and lipoprotein lipase (LPL) which is a key enzyme in the regulation of the flux of fatty acids in liver, muscle, and adipose tissue as well as muscle growth-regulating genes as myostatin (MYO) in muscle and insulin-like growth factor-1 (IGF-1) in liver. The fish were allocated into three equal groups; control group that fed basal diet only and the other two groups fed a basal diet containing clenbuterol at two doses 5 ppm and 10 ppm/kg diet for 30 consecutive days. Results revealed that clenbuterol supplementation significantly increased body weight, decreased liver, spleen and abdominal fat weights, and decreased total circulatory cholesterol and triacylglycerol levels. Moreover, clenbuterol inhibits lipogenesis by downregulation of FAS gene expression by dose and time-dependent manner in the liver while enhanced lipolysis in both the liver and in the adipose tissue. Moreover, lipolysis was reduced in muscle by dose 10 ppm on day 30. Furthermore, clenbuterol presented higher gene expression of MYO and IGF-1 in muscle and liver respectively by dose 5 ppm at day 15 on the other hand, these findings were reversed by day 30 compared with control. In conclusion, clenbuterol efficacy was apparent in a dose and time response pattern to boost growth and reduce fat deposition rates, indicating for the first time that clenbuterol has a profitable growth impact on Nile tilapia

Exploring the multimodal role of yucca schidigera extract in protection against chronic ammonia exposure targeting: Growth, metabolic, stress and inflammatory responses in nile tilapia (oreochromis niloticus l.)

Ammonia is a critical hazardous nitrogen metabolic product in aquaculture. Despite trials for its control, ammonia intoxication remains one of the most critical issues to overcome. In this study, we explored the modulatory effect and potential mechanism by which Yucca schidigera extract (YSE) can ameliorate ammonia intoxication-induced adverse effects on tilapia health and metabolism. A total number of 120 Nile tilapia were evenly assigned into four groups with three replicates each. The first group served as normal control group; the second group was exposed to ammonia alone from the beginning of the experiment and for four weeks. The third group was supplied with YSE in water at a dose of 8 mg/L and exposed to ammonia. The fourth group was supplied with YSE only in water at a dose of 8 mg/L. YSE supplementation succeeded in improving water quality by reducing pH and ammonia levels. Moreover, YSE supplementation markedly alleviated chronic ammonia-induced adverse impacts on fish growth by increasing the final body weight (FBW), specific growth rate (SGR), feed intake and protein efficiency ratio (PER) while reducing the feed conversion ratio (FCR) via improvements in food intake, elevation of hepatic insulin-like growth factor (ILGF-1) and suppression of myostatin (MSTN) expression levels with the restoration of lipid reserves and the activation of lipogenic potential in adipose tissue as demonstrated by changes in the circulating metabolite levels. In addition, the levels of hepato-renal injury biomarkers were restored, hepatic lipid peroxidation was inhibited and the levels of hepatic antioxidant biomarkers were enhanced. Therefore, the current study suggests that YSE supplementation exerted an ameliorative role against chronic ammonia-induced oxidative stress and toxic effects due to its free radical-scavenging potential, potent antioxidant activities and anti-inflammatory effects

Collagen extract obtained from Nile tilapia (Oreochromis niloticus L.) skin accelerates wound healing in rat model via up regulating VEGF, bFGF, and α-SMA genes expression

Background Collagen is the most abundant structural protein in the mammalian connective tissue and represents approximately 30% of animal protein. The current study evaluated the potential capacity of collagen extract derived from Nile tilapia skin in improving the cutaneous wound healing in rats and investigated the underlying possible mechanisms. A rat model was used, and the experimental design included a control group (CG) and the tilapia collagen treated group (TCG). Full-thickness wounds were conducted on the back of all the rats under general anesthesia, then the tilapia collagen extract was applied topically on the wound area of TCG. Wound areas of the two experimental groups were measured on days 0, 3, 6, 9, 12, and 15 post-wounding. The stages of the wound granulation tissues were detected by histopathologic examination and the expression of vascular endothelial growth factor (VEGF), and transforming growth factor (TGF-ß1) were investigated using immunohistochemistry. Moreover, relative gene expression analysis of transforming growth factor-beta (TGF-ß1), basic fibroblast growth factor (bFGF), and alpha-smooth muscle actin (α-SMA) were quantified by real-time qPCR. Results The histopathological assessment showed noticeable signs of skin healing in TCG compared to CG. Immunohistochemistry results revealed remarkable enhancement in the expression levels of VEGF and TGF-β1 in TCG. Furthermore, TCG exhibited marked upregulation in the VEGF, bFGF, and α-SMA genes expression. These findings suggested that the topical application of Nile tilapia collagen extract can promote the cutaneous wound healing process in rats, which could be attributed to its stimulating effect on recruiting and activating macrophages to produce chemotactic growth factors, fibroblast proliferation, and angiogenesis. Conclusions The collagen extract could, therefore, be a potential biomaterial for cutaneous wound healing therapeutics. Backgroun

Chronic hyperglycemia induces trans-differentiation of human pancreatic stellate cells and enhances the malignant molecular communication with human pancreatic cancer cells

BACKGROUND: Diabetes mellitus is linked to pancreatic cancer. We hypothesized a role for pancreatic stellate cells (PSC) in the hyperglycemia induced deterioration of pancreatic cancer and therefore studied two human cell lines (RLT-PSC, T3M4) in hyperglycemic environment. METHODOLOGY/PRINCIPAL FINDINGS: The effect of chronic hyperglycemia (CHG) on PSCs was studied using mRNA expression array with real-time PCR validation and bioinformatic pathway analysis, and confirmatory protein studies. The stress fiber formation (IC: αSMA) indicated that PSCs tend to transdifferentiate to a myofibroblast-like state after exposure to CHG. The phosphorylation of p38 and ERK1/2 was increased with a consecutive upregulation of CDC25, SP1, cFOS and p21, and with downregulation of PPARγ after PSCs were exposed to chronic hyperglycemia. CXCL12 levels increased significantly in PSC supernatant after CHG exposure independently from TGF-β1 treatment (3.09-fold with a 2.73-fold without TGF-β1, p<0.05). The upregualtion of the SP1 transcription factor in PSCs after CHG exposure may be implicated in the increased CXCL12 and IGFBP2 production. In cancer cells, hyperglycemia induced an increased expression of CXCR4, a CXCL12 receptor that was also induced by PSC's conditioned medium. The receptor-ligand interaction increased the phosphorylation of ERK1/2 and p38 resulting in activation of MAP kinase pathway, one of the most powerful stimuli for cell proliferation. Certainly, conditioned medium of PSC increased pancreatic cancer cell proliferation and this effect could be partially inhibited by a CXCR4 inhibitor. As the PSC conditioned medium (normal glucose concentration) increased the ERK1/2 and p38 phosphorylation, we concluded that PSCs produce other factor(s) that influence(s) pancreatic cancer behaviour. CONCLUSIONS: Hyperglycemia induces increased CXCL12 production by the PSCs, and its receptor, CXCR4 on cancer cells. The ligand-receptor interaction activates MAP kinase signaling that causes increased cancer cell proliferation and migration

Low dose cranial irradiation-induced cerebrovascular damage is reversible in mice

BACKGROUND: High-dose radiation-induced blood-brain barrier breakdown contributes to acute radiation toxicity syndrome and delayed brain injury, but there are few data on the effects of low dose cranial irradiation. Our goal was to measure blood-brain barrier changes after low (0.1 Gy), moderate (2 Gy) and high (10 Gy) dose irradiation under in vivo and in vitro conditions. METHODOLOGY: Cranial irradiation was performed on 10-day-old and 10-week-old mice. Blood-brain barrier permeability for Evans blue, body weight and number of peripheral mononuclear and circulating endothelial progenitor cells were evaluated 1, 4 and 26 weeks postirradiation. Barrier properties of primary mouse brain endothelial cells co-cultured with glial cells were determined by measurement of resistance and permeability for marker molecules and staining for interendothelial junctions. Endothelial senescence was determined by senescence associated β-galactosidase staining. PRINCIPLE FINDINGS: Extravasation of Evans blue increased in cerebrum and cerebellum in adult mice 1 week and in infant mice 4 weeks postirradiation at all treatment doses. Head irradiation with 10 Gy decreased body weight. The number of circulating endothelial progenitor cells in blood was decreased 1 day after irradiation with 0.1 and 2 Gy. Increase in the permeability of cultured brain endothelial monolayers for fluorescein and albumin was time- and radiation dose dependent and accompanied by changes in junctional immunostaining for claudin-5, ZO-1 and β-catenin. The number of cultured brain endothelial and glial cells decreased from third day of postirradiation and senescence in endothelial cells increased at 2 and 10 Gy. CONCLUSION: Not only high but low and moderate doses of cranial irradiation increase permeability of cerebral vessels in mice, but this effect is reversible by 6 months. In-vitro experiments suggest that irradiation changes junctional morphology, decreases cell number and causes senescence in brain endothelial cells

Corrigendum to "European contribution to the study of ROS:A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS)" [Redox Biol. 13 (2017) 94-162]

The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed

Isolation, Identification and Molecular Screening of Pseudomonas sp. Metabolic Pathways NRPs and PKS Associated with the Red Sea Sponge, Hyrtios aff. Erectus, Egypt

Marine coastal areas of the Red sea in Egypt have great biodiversity of sponges which harbors a huge number of endosymbiotic bacteria which are the source of many potential bioactive metabolites and consider as an excellent source. The study focuses on the isolation of Pseudomonas sp associated with marine sponge Hyrtios aff. Erectus from the Red Sea in pure culture, phylogenetically identified and screening of their metabolic pathways polyketide synthases and Nonribosomal peptides (PSK and NRPs). Characterization of the bacterial produced bioactive metabolite was done through antioxidant and cytotoxicity assays using DPPH and cell line. The 16S rRNA gene sequencing of bacterial isolate revealed that the bacterial strain was closely related to Pseudomonas sp., and the active metabolic pathways genes in the Pseudomonas sp was PSK II. The antioxidants and cytotoxicity bioactivity of Pseudomonas sp as is most probably related to the presence of associated bioactive metabolic pathways genes related to polyketides family type II. Furthermore, the antioxidant activity of Pesudomance sp extract, performed by quantification of total phenolic content (TPC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity antioxidant activities, were evaluated. The highest antioxidant activity was exhibited at different concentrations (50 mg-6.25 mg) which showed a potent antioxidant capacity by 100% at 50mg, 25 mg and 12.25 mg. From the cytotoxicity, antioxidant and bioactive metabolic screening pathways this bacterial strain was revealed to be an excellent source of natural antioxidant and cytotoxicity compounds against free radical and cancerous cell line