Non-esterified fatty acid-associated ability of follicular fluid to support porcine oocyte maturation and development.

PURPOSE: The effect of supplementing maturation medium with follicular fluid (FF) was examined according to its non-esterified fatty acid (NEFA) content or with a fatty acid mixture (FA-Mix) on the developmental competence of oocytes, as well as the mitochondrial quality and quantity in the oocytes and cumulus cells. METHOD: Porcine oocytes from a slaughterhouse were used. RESULTS: The FF or FA-Mix in maturation medium increased the lipid content in both the oocytes and the cumulus cells, but the adenosine triphosphate content was differentially affected. The FF supplementation increased the mitochondrial DNA copy number, survival of cumulus cells, and rate of oocyte development to the blastocyst stage, whereas the FA-Mix supplementation did not show these effects. The expression levels of GPC4,PFKP,PRDX3, and TFAM in the cumulus cells increased after FF supplementation, but the expression of GJA1 decreased, compared with the cells that were cultured without FF. CONCLUSION: Adding FF and FA-Mix to the maturation medium increased the lipid content in the oocytes and cumulus cells. The effects of FF on the cumulus cells and oocytes were not observed after FA-Mix supplementation, indicating that the concentration of the NEFAs in the FF are closely associated with an ability to support oocyte maturation and the metabolism of cumulus cells and oocytes.

Cooperative inhibitory effects of uremic toxins and other serum components on OATP1B1-mediated transport of SN-38.

PURPOSE: Half-life of SN-38, an active metabolite of irinotecan, remarkably increases in patients with end-stage kidney disease (ESKD), even though SN-38 is excreted in bile. Uremic toxins (UTs), which accumulate in the serum of ESKD patients, were reported to inhibit organic anion-transporting polypeptide (OATP) 1B1-mediated uptake of SN-38; however, the relevance of this finding in a clinical setting is unknown. This study focused on cooperative effects of serum components and UTs on OATP1B1-mediated transport of SN-38. METHODS: Uptake of SN-38 by OATP1B1 was evaluated using cells stably expressing OATP1B1. Serum was obtained from > 400 ESKD patients undergoing hemodialysis. Deproteinized serum was combined with human serum albumin (HSA) to explore the effects of albumin-bound and unbound serum compounds. RESULTS: Uptake clearance of SN-38 in OATP1B1 cells decreased by 40% in the presence of uremic serum residue with albumin compared to that in the presence of normal serum residue. Additional UTs (3-carboxy-4-methyl-5-propyl-2-furanpropionic acid, hippuric acid, indole-3-acetic acid, and 3-indoxyl sulfate) combined with normal serum residue in HSA decreased OATP1B1-mediated SN-38 transport by 32.1% compared to that in the presence of normal serum residue. The inhibitory effect of albumin-unbound fraction with UTs and normal serum residue was comparable to that of uremic serum residue, with an uptake decrease of 17.2% compared to that reported in the presence of normal serum residue. CONCLUSIONS: Hepatic uptake of SN-38 via OATP1B1 decreases in ESKD patients through cooperative inhibitory effects of UTs and serum components.

FOXO3a potentiates hTERT gene expression by activating c-MYC and extends the replicative life-span of human fibroblast.

In our previous studies, we reported that SIRT1 prevents cellular senescence in human fibroblast, and that SIRT1-induced inhibition of cellular senescence is due to enhanced hTERT gene expression. In this study, we investigate the molecular mechanisms behind SIRT1-induced potentiation of hTERT transcription and show that FOXO3a functions downstream of SIRT1 and prevents the induction of cellular senescence by enhancing hTERT gene expression. Furthermore, we found that FOXO3a-induced potentiation of hTERT gene expression is regulated in a c-MYC/E-box dependent manner. In addition, we found that FOXO3a binds to the novel binding element in the c-MYC promoter, and this interaction activates the transcription of the c-MYC gene. The resulting increase in c-MYC leads to higher levels of c-MYC recruited to the hTERT promoter and, in turn, activates hTERT gene expression. Taken together, this pathway might constitute the molecular basis for the anti-senescence effects of SIRT1 and FOXO3a.

[The investigation of understanding and comfort of patients taking Divigel(®) 1 mg].

Hormone replacement therapy (HRT) given by injection or administered orally or topically can improve the QOL of patients with menopausal symptoms. Because patient comfort is influenced largely by the dosage form, pharmacists should understand the properties of each dosage form and provide appropriate information to individual patients. In this study, we investigated the understanding of medicines and diseases of patients receiving HRT and discuss the approaches pharmacists can take to improve patients' adherence. Thirty-seven patients (mean age 51.7±3.6 years) taking estradiol gel (Divigel(®) 1 mg) completed a questionnaire asked by their pharmacist. Responses indicated 70% of patients failed to use the gel as prescribed, and they had poor knowledge of both the sites where the gel shouldn't be applied and appropriate measures to take if having forgotten to apply the gel (43% and 11% correct understanding, respectively). Since the duration of HRT treatment for menopausal symptom is 2-5 years, patients should be administered the minimum effective dose in the shortest amount of time. Hence it is important to maintain patients' adherence particularly in this limited administration period. HRT guidelines define HRT outcome as not only improvement of menopausal symptoms but also suppression of bone resorption, improvement of glucose and lipid metabolism, and reduced prevalence of Alzheimer's disease. Accordingly, pharmacists should facilitate proper adherence to HRT to improve and maintain women's QOL in the perimenopausal period, necessitating they actively provide pharmaceutical care such as preparing useful instructions patients can repeatedly use and periodically checking patients' understanding of their HRT medications.

Lipidomics analysis of mesenteric lymph after trauma and hemorrhagic shock.

BACKGROUND: After trauma and hemorrhagic shock (T/HS), a variety of inflammatory mediators enter the systemic circulation through mesenteric lymph ducts, leading to acute lung injury and multiple-organ dysfunction syndrome. Recent studies have demonstrated that post-HS mesenteric lymph (PHSML) activates polymorphonuclear leukocytes (PMNs) and causes vascular endothelial cell and red blood cell dysfunction. Furthermore, PHSML contains proinflammatory mediators, such as biologically active lipids. The purpose of this study was to identify the lipid mediators in PHSML and plasma by liquid chromatography/electrospray ionization mass spectrometry and then estimate the biologic activities of the identified lipids on PMNs. METHODS: PHSML was collected from male Sprague-Dawley rats undergoing trauma (laparotomy) plus HS (40 mm Hg, 30 minutes) or sham shock (SS). The lipids in PHSML and plasma were extracted using the methods of Bligh and Dyer, and liquid chromatography/electrospray ionization mass spectrometry was performed. The biologic activities (superoxide production and elastase release) of identified lipids on human PMNs were tested. RESULTS: Phosphatidylcholine, lysophosphatidylcholine (LPC), phosphatidylethanolamine, lysophosphatidylethanolamine (LPE), and sphingomyelin were detected in the PHSML. Furthermore, linoleoyl, arachidonoyl, and docosahexaenoyl LPCs and LPEs significantly increased in the PHSML of the T/HS group as compared with those of the T/SS group. In the plasma, arachidonoyl and docosahexaenoyl LPCs of the T/HS group also significantly increased in comparison with that of the T/SS group. Linoleoyl and arachidonoyl LPCs and LPEs showed the priming activity on N-formyl-methionyl-leucyl-phenylalanine-activated PMNs. The elastase release was also induced by linoleoyl and arachidonoyl LPCs. CONCLUSION: Mesenteric lymph after T/HS contains biologically active lipids, such as LPCs and LPEs with polyunsaturated fatty acids, which may be involved in the pathogenesis of acute lung injury/multiple-organ dysfunction syndrome.

SIRT1 prevents replicative senescence of normal human umbilical cord fibroblast through potentiating the transcription of human telomerase reverse transcriptase gene.

SIRT1, the mammalian homolog of sirtuins, has emerged as a mediator of the beneficial effects of calorie restriction. Among them, we focused on the SIRT1-induced prevention of cellular senescence, and tried to reveal the molecular mechanisms that define the effects of SIRT1. Firstly in this study, we observed that overexpression of SIRT1 resulted in the prevention of cellular senescence of normal human umbilical cord fibroblast HUC-F2 cells. Here, we focused on the human telomerase reverse transcriptase (hTERT) gene as a target of the SIRT1-induced prevention of cellular senescence. Results showed that SIRT1, SIRT1 activator, resveratrol, and SIRT1 activating condition, starved condition, increased the transcription of hTERT in HUC-F2 cells. Next, we found that SIRT1 increased hTERT transcription in a c-MYC-dependent manner, triggered the transcription of the c-MYC gene and increased the amount of c-MYC recruited to the hTERT promoter. Further, SIRT1 increased the transcriptional activation ability of c-MYC and correspondingly increased the amount of acetylated H4 histone at the hTERT promoter. All of these results indicated that SIRT1 activates hTERT transcription through the involvement of c-MYC, and suggested that this SIRT1-induced augmentation of hTERT transcription resulted in the extension of the cellular life span of HUC-F2 cells.

Identification and physiological evaluation of the components from citrus fruits as potential drugs for anti-corpulence and anticancer.

On the basis of monitoring the prevention of accumulation of lipid droplets in mouse 3T3-L1 preadipocyte cells and inhibition of the proliferation of human colon cancer HT-29 cells, effective anti-corpulence and anticancer compounds were isolated from the peel of Citrus fruits. These bioactive components were identified as polymethoxyflavones and coumarin derivatives by spectroscopic analyses. 5-Hydroxy-6,7,8,3',4'-pentamethoxyflavone had the greatest anti-corpulence effects and 3,5,6,7,8,3',5'-heptamethoxyflavone had the greatest anticancer effects. Furthermore, distributions of those bioactive components in the peel of 10 species of Citrus fruits were demonstrated by HPLC analyses.