Betulin silver nanoparticles qualify as efficient antimelanoma agents in in vitro and in vivo studies.

The current study was purported to assess the: (i) in vitro toxicity of betulin silver nanoparticles (AgNPs-B), bare and capped with polyethylene glycol (PEG), on two murine melanoma cell lines (B164A5 and B16Ova) and on healthy cell lines (keratinocytes and melanocytes), and (ii) in vivo antitumor efficacy of PEGylated AgNPs-B in an experimental melanoma model. Bare and PEG-capped AgNPs-B were synthesized by a chemical reduction method resulting in stable and non-aggregated spherical AgNPs-B and PEG-AgNPs-B, of narrow size distributions and mean hydrodynamic diameters of 25 nm and 75 nm, respectively. In vitro assessments were achieved by MTT and Annexin V-FITC assays and in vivo evaluation involved non-invasive techniques for the surveillance of the physiological skin parameters changes and histopathological examination of the harvested organs. The in vitro results revealed selective cytotoxicity against melanoma cells, at low doses that are nontoxic to normal cells; higher doses were associated with the loss of selectivity and toxicity for healthy cells. PEGylated formulation of betulin exerted a dose-dependent pro-apoptotic effect, more obvious in the case of B164A5 cells. Histopathological analysis suggested that PEGylated AgNPs-B developed relevant in vivo effects as antimelanoma agents by decreasing the tumor volume and inhibiting the development of secondary tumors.

Short-term effects of very low dose cadmium feeding on copper, manganese and iron homeostasis: A gastropod perspective.

The available information on the interplay between low-dose cadmium intake and copper, manganese, and iron homeostasis in invertebrates is limited. We have currently studied the accumulation of these trace metals in the hepatopancreas of adult snails, Cantareus aspersus, following 14 and 28 days of exposure to low doses of dietary cadmium, up to 1 mg/kg dw (dry weight). The cadmium dose, but not the duration of exposure, had a significant effect on hepatopancreas copper deposition, the values being significantly elevated compared to controls. A significant peak in manganese levels at 14 days was found in snails administered the lowest cadmium dose. These increases occurred even in the absence of cadmium increase in the hepatopancreas. Our data suggest that low dose cadmium feeding can produce a transient disturbance in hepatopancreas copper and manganese homeostasis. Such responses may serve as early biomarkers of physiological changes occurring during the initial stages of cadmium intoxication.

High-level dietary cadmium exposure is associated with global DNA hypermethylation in the gastropod hepatopancreas.

5-methylcytosine (5mC) is a key epigenetic mark which influences gene expression and phenotype. In vertebrates, this epigenetic mark is sensitive to Cd exposure, but there is no information linking such an event with changes in global 5mC levels in terrestrial gastropods despite their importance as excellentecotoxicological bioindicators of metal contamination. Therefore, we first evaluated total 5mC content in DNA of the hepatopancreas of adult Cantareus aspersus with the aim to determine whether this epigenetic mark is responsive to Cd exposure. The experiment was conducted under laboratory conditions and involved a continuous exposure, multiple dose- and time-point (14, 28, and 56 days) study design. Hepatopancreas cadmium levels were measured using Flame Atomic Absorption Spectrometry and the percentage of 5-mC in samples using an ELISA-based colorimetric assay. Snail death rates were also assessed. Our results, for the first time, reveal the presence of 5mC in C. aspersus and provide evidence for Cd-induced changes in global 5mC levels in DNA of gastropods and mollusks. Although less sensitive than tissue accumulation, DNA methylation levels responded in a dose- and time-dependent manner to dietary cadmium, with exposure dose having a much stronger effect than exposure duration. An obvious trend of increasing 5mC levels was observed starting at 28 days of exposure to the second highest dose and this trend persisted at the two highest treatments for close to one month, when the experiment was terminated after 56 days. Moreover, a strong association was identified between Cd concentrations in the hepatopancreas and DNA methylation levels in this organ. These data indicate an overall trend towards DNA hypermethylation with elevated Cd exposure. No consistent lethal effect was observed, irrespective of time point and Cd-dosage. Overall, our findings suggest that the total 5mC content in DNA of the hepatopancreas of land snails is responsive to sublethal Cd exposure and give new insights into invertebrate environmental epigenetics.

Design, synthesis and pharmaco-toxicological assessment of 5-mercapto-1,2,4-triazole derivatives with antibacterial and antiproliferative activity.

The extensive biochemical research of multiple types of cancer has revealed important enzymatic signaling pathways responsible for tumor occurrence and progression, thus compelling the need for the discovery of new means with which to block these signaling cascades. The phosphoinositide 3-kinase/ protein kinase B (PI3K/AKT) pathway, which plays an important role in maintaining relevant cellular functions, exhibits various alterations in common human cancers, thus representing a suitable target in cancer treatment. Molecules bearing the 1,2,4-triazole moiety are known to possess multiple biological activities, including anticancer activity. The current study used molecular docking in the design of 5-mercapto-1,2,4-triazole derivatives with antiproliferative activity targeting the PI3K/AKT pathway. Three structures emerged as the result of this method, which indicated for these a highly favorable accommodation within the active binding site of PI3K protein, thus acting as potential PI3K inhibitors, and hence interfering with the above-mentioned pathway. The molecules were synthesized and their chemical structure was confirmed. The antiproliferative activity of these compounds was tested on 4 cancer cell lines (A375, B164A5, MDA-MB-231 and A549) and on normal human keratinocytes (HaCaT) by in vitro alamarBlue assay. The 3 compounds revealed antitumor activity against the breast cancer cell line (MDA-MB-231) and reduced toxicity on the normal cell line. The antibacterial activity of the compounds was also tested in vitro on Gram-positive and Gram-negative bacterial strains, revealing moderate activity.

Mistletoe and Garlic Extracts as Polyurethane Carriers - A Possible Remedy for Choroidal Melanoma.

BACKGROUND: Melanoma is known as the most dangerous form of skin cancer; whereas the malignant choroidal melanoma is an orphan disease known as the most common primary intraocular malignancy in adults. Literature suggests that the consumption of garlic and mistletoe leads to a reduced risk of developing cancer. OBJECTIVE: The aim of this study was the obtaining and the characterization of polymer structures containing mistletoe or garlic extract. METHODS: The structures were obtained in a polyaddition process combined with a spontaneous emulsification; they were characterized by pH, size, Zeta potential and DSC measurements, evaluation of encapsulation efficacy, penetrability through membranes and in vitro cytotoxicity tests. RESULTS: The microstructures present sizes between 1.05 and 2.60 µm and Zeta potentials between -7 and +36 mV. A good encapsulation was observed on different evaluations (88-92%). It was determined that approx. 30% of polymer microstructures containing vegetal extracts pass through an artificial membrane in 4 days. An in vitro cytotoxicity test revealed that these products are safe for administration. The analysis of antitumor efficacy indicates that garlic extracts have important effects after 48 and 72 hours on A375 cells; however, polymer microstructures with herbal extracts did not reveal antiproliferative activities on A375 cells because these polymer structures present a slow degradation. CONCLUSION: Sterile eye drops solutions based on polymer microstructures containing garlic or mistletoe extracts were obtained; the sample based on garlic extracts may be used in the pharmaceutical field as drug carrier with an antiproliferative effect which occurs after a prolong period.

Effects of Quercetin on Blood Pressure: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: Quercetin, the most abundant dietary flavonol, has antioxidant effects in cardiovascular disease, but the evidence regarding its effects on blood pressure (BP) has not been conclusive. We assessed the impact of quercetin on BP through a systematic review and meta-analysis of available randomized controlled trials. METHODS AND RESULTS: We searched PUBMED, Cochrane Library, Scopus, and EMBASE up to January 31, 2015 to identify placebo-controlled randomized controlled trials investigating the effect of quercetin on BP. Meta-analysis was performed using either a fixed-effects or random-effect model according to I(2) statistic. Effect size was expressed as weighted mean difference (WMD) and 95% CI. Overall, the impact of quercetin on BP was reported in 7 trials comprising 9 treatment arms (587 patients). The results of the meta-analysis showed significant reductions both in systolic BP (WMD: -3.04 mm Hg, 95% CI: -5.75, -0.33, P=0.028) and diastolic BP (WMD: -2.63 mm Hg, 95% CI: -3.26, -2.01, P<0.001) following supplementation with quercetin. When the studies were categorized according to the quercetin dose, there was a significant systolic BP and diastolic BP-reducing effect in randomized controlled trials with doses ≥500 mg/day (WMD: -4.45 mm Hg, 95% CI: -7.70, -1.21, P=0.007 and -2.98 mm Hg, 95% CI: -3.64, -2.31, P<0.001, respectively), and lack of a significant effect for doses <500 mg/day (WMD: -1.59 mm Hg, 95% CI: -4.44, 1.25, P=0.273 and -0.24 mm Hg, 95% CI: -2.00, 1.52, P=0.788, respectively), but indirect comparison tests failed to significant differences between doses. CONCLUSIONS: The results of the meta-analysis showed a statistically significant effect of quercetin supplementation in the reduction of BP, possibly limited to, or greater with dosages of >500 mg/day. Further studies are necessary to investigate the clinical relevance of these results and the possibility of quercetin application as an add-on to antihypertensive therapy.

Cardiotoxicity of anthracycline therapy: current perspectives.

Anthracyclines, especially doxorubicin and daunorubicin, are the drugs of first choice in the treatment of patients with hematologic malignancies, soft-tissue sarcomas, and solid tumors. Unfortunately, the use of anthracyclines is limited by their dose-dependent and cumulative cardiotoxicity. The molecular mechanism responsible for anthracycline-induced cardiotoxicity remains poorly understood, although experimental and clinical studies have shown that oxidative stress plays the main role. Hence, antioxidant agents, especially dexrazoxane, and also other drug classes (statins, ß-blockers) proved to have a beneficial effect in protecting against anthracycline-induced cardiotoxicity. According to previous clinical trials, the major high-risk factors for anthracycline-induced cardiotoxicity are age, body weight, female gender, radiotherapy, and other diseases such as Down syndrome, familial dilated cardiomyopathy, diabetes and hypertension. Consequently, further studies are needed to elucidate the molecular pathogenesis of anthracycline-induced cardiotoxicity and also to discover new cardioprotective agents against anthracycline-induced cardiotoxicity.

Effects of supplementation with pomegranate juice on plasma C-reactive protein concentrations: A systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Pomegranate juice (PJ) has a high content of antioxidants and bioactive polyphenols, being widely used for its antioxidant, anti-inflammatory and chemopreventive effects. PURPOSE: The objective of this meta-analysis consisted in investigating the impact of PJ on plasma C-reactive protein (CRP) concentrations. METHODS: The search included SCOPUS, Medline and two Iranian bibliographic databases namely MagIran and Scientific Information Database (from inception to December 09, 2014) to identify prospective trials for investigating the impact of pomegranate preparations on serum concentrations of CRP. Two independent reviewers extracted data on study characteristics, methods and outcomes. RESULTS: Among 427 participants in the selected studies, 216 were allocated to PJ groups, and 211 to control group. Meta-analysis of data from 5 eligible randomized controlled trials (RCTs) arms did not provide compelling evidence as to a significant CRP-lowering effect of supplementation with pomegranate juice (WMD: -0.22 mg/l, 95% CI: -0.45, 0.01, p = 0.061). The impact of pomegranate juice on plasma CRP levels was found to be independent of duration of supplementation (slope: 0.003; 95% CI: -0.005, 0.011; p = 0.444). CONCLUSION: In conclusion, this meta-analysis of data from 5 prospective trials did not indicate a significant effect of PJ on plasma CRP levels, and this effect was independent of duration of supplementation.

Impact of L-carnitine on plasma lipoprotein(a) concentrations: A systematic review and meta-analysis of randomized controlled trials.

We aimed to assess the impact of L-carnitine on plasma Lp(a) concentrations through systematic review and meta-analysis of available RCTs. The literature search included selected databases up to 31(st) January 2015. Meta-analysis was performed using fixed-effects or random-effect model according to I(2) statistic. Effect sizes were expressed as weighted mean difference (WMD) and 95% confidence interval (CI). The meta-analysis showed a significant reduction of Lp(a) levels following L-carnitine supplementation (WMD: -8.82 mg/dL, 95% CI: -10.09, -7.55, p < 0.001). When the studies were categorized according to the route of administration, a significant reduction in plasma Lp(a) concentration was observed with oral (WMD: -9.00 mg/dL, 95% CI: -10.29, -7.72, p < 0.001) but not intravenous L-carnitine (WMD: -2.91 mg/dL, 95% CI: -10.22, 4.41, p = 0.436). The results of the meta-regression analysis showed that the pooled estimate is independent of L-carnitine dose (slope: -0.30; 95% CI: -4.19, 3.59; p = 0.878) and duration of therapy (slope: 0.18; 95% CI: -0.22, 0.59; p = 0.374). In conclusion, the meta-analysis suggests a significant Lp(a) lowering by oral L-carnitine supplementation. Taking into account the limited number of available Lp(a)-targeted drugs, L-carnitine might be an effective alternative to effectively reduce Lp(a). Prospective outcome trials will be required to fully elucidate the clinical value and safety of oral L-carnitine supplementation.

A systematic review and meta-analysis of the impact of Spirulina supplementation on plasma lipid concentrations.

BACKGROUND & AIMS: The impact of Spirulina supplementation on plasma lipid concentrations has not been conclusively studied. Therefore the aim of the meta-analysis was to assess the effect of Spirulina supplementation on plasma lipid concentrations. METHODS: We searched PubMed and Scopus (up to July 03, 2015) to identify randomized controlled trials (RCTs) that investigate the effect Spirulina supplementation on plasma lipid concentrations. Meta-analysis and meta-regression were performed using random-effects models. RESULTS: Random-effect meta-analysis of data from 7 RCTs showed a significant effect of supplementation with spirulina in reducing plasma concentrations of total cholesterol (WMD: -46.76 mg/dL, 95% CI: -67.31 to -26.22, p < 0.001), LDL-C (WMD: -41.32 mg/dL, 95% CI: -60.62 to -22.03, p < 0.001) and triglycerides (WMD: -44.23 mg/dL, 95% CI: -50.22 to -38.24, p < 0.001), and elevating those of HDL-C (WMD: 6.06 mg/dL, 95% CI: 2.37-9.76, p = 0.001). The impact of spirulina on plasma concentrations of total cholesterol (slope: -1.32; 95% CI: -8.58 to 5.93; p = 0.720), LDL-C (slope: -1.01; 95% CI: -8.03 to 6.02; p = 0.778), triglycerides (slope: -1.39; 95% CI: -4.26 to 1.48; p = 0.342) and HDL-C (slope: 1.79, 95% CI: -0.48 to 4.05; p = 0.122) was independent of administered dose. Regarding duration of supplementation with Spirulina, significant associations were found with changes in plasma concentrations of total cholesterol (slope: -1.77; 95% CI: -3.48 to -0.07; p = 0.042), LDL-C (slope: -1.73; 95% CI: -3.40 to -0.06; p = 0.042) HDL-C (slope: 0.91; 95% CI: 0.68-1.14; p < 0.001) and triglycerides (slope: -1.39; 95% CI: -2.28 to -0.50; p = 0.002). CONCLUSIONS: This meta-analysis showed a significant effect of supplementation with Spirulina in reducing plasma concentrations of total cholesterol, LDL-C, triglycerides and elevating those of HDL-C.

Effects of flaxseed supplements on blood pressure: A systematic review and meta-analysis of controlled clinical trial.

BACKGROUND & AIMS: Many experimental and clinical trials suggested that flaxseed might be a potent antihypertensive, but the evidences concerning the effects of flaxseed supplements on blood pressure (BP) has not been fully conclusive. We aimed to assess the impact of the effects of flaxseed supplements on blood pressure through systematic review of literature and meta-analysis of available randomized controlled trials (RCTs). METHODS: The literature search included PUBMED, Cochrane Library, Scopus, and EMBASE up to February 2015 to identify RCTs investigating the effect of flaxseed supplements on plasma blood pressure. Effect size was expressed as weighed mean difference (WMD) and 95% confidence interval (CI). RESULTS: 15 trials (comprising 19 treatment arms) with 1302 participants were included in this meta-analysis. Random-effects meta-analysis suggested significant reductions in both systolic BP (SBP) (WMD: -2.85 mmHg, 95%CI: -5.37 to -0.33, p = 0.027) and diastolic BP (DBP) (WMD: -2.39 mmHg, 95%CI: -3.78 to -0.99, p = 0.001) following supplementation with flaxseed products. When the studies were stratified according to their duration, there was a greater effect on both SBP and DBP in the subset of trials with ≥12 weeks of duration (WMD: -3.10 mmHg, 95%CI: -6.46 to 0.27, p = 0.072 and -2.62 mmHg, 95%CI: -4.39 to -0.86, p = 0.003, respectively) vs the subset lasting <12 weeks (WMD: -1.60 mmHg, 95%CI: -5.44 to 2.24, p = 0.413, and -1.74 mmHg, 95%CI: -4.41 to 0.93, p = 0.202, respectively). Another subgroup analysis was performed to assess the impact of flaxseed supplement type on BP. Reduction of SBP was significant with flaxseed powder (WMD: -1.81 mmHg, 95% CI: -2.03 to -1.59, p < 0.001) but not oil (WMD: -4.62 mmHg, 95%CI: -11.86 to 2.62, p = 0.211) and lignan extract (WMD: 0.28 mmHg, 95% CI: -3.49 to 4.04, p = 0.885). However, DBP was significantly reduced with powder and oil preparations (WMD: -1.28 mmHg, 95% CI: -2.44 to -0.11, p = 0.031, and -4.10 mmHg, 95%CI: -6.81 to -1.39, p = 0.003, respectively), but not with lignan extract (WMD: -1.78 mmHg, 95% CI: -4.28 to 0.72, p = 0.162). CONCLUSIONS: This meta-analysis of RCTs showed significant reductions in both SBP and DBP following supplementation with various flaxseed products.

Tibolone decreases Lipoprotein(a) levels in postmenopausal women: A systematic review and meta-analysis of 12 studies with 1009 patients.

INTRODUCTION: Circulating lipoprotein (a) (Lp(a)) is a recognized risk factor for cardiovascular disease (CVD). Tibolone, a synthetic steroid, may lower Lp(a) levels; however, evidence of the effects of tibolone on Lp(a) still remain to be defined. Therefore, we investigated the effects of tibolone treatment on circulating Lp(a) levels in postmenopausal women. METHODS: The search included PUBMED, Web of Science, Scopus, and Google Scholar (up to January 31st, 2015) to identify controlled clinical studies investigating the effects of oral tibolone treatment on Lp(a) levels in postmenopausal women. Random-effects meta-regression was performed using unrestricted maximum likelihood method for the association between calculated weighted mean difference (WMD) and potential moderators. RESULTS: Meta-analysis of data from 12 trials (16 treatment arms) suggested a significant reduction of Lp(a) levels following tibolone treatment (WMD: -25.28%, 95% confidence interval [CI]: -36.50, -14.06; p < 0.001). This result was robust in the sensitivity analysis and its significance was not influenced after omitting each of the included studies from the meta-analysis. When the studies were categorized according to the tibolone dose, there were consistent significant reductions of Lp(a) in the subsets of studies with doses <2.5 mg/day (WMD: -17.00%, 95%CI: -30.22, -3.77; p < 0.012) and 2.5 mg/day (WMD: -29.18%, 95%CI: -45.02, -13.33; p < 0.001). Likewise, there were similar reductions in the subsets of trials with follow-up either <24 months (WMD: -26.79%, 95%CI: -38.40, -15.17; p < 0.001) or ≥24 months (WMD: -23.10%, 95%CI: -40.17, -6.03; p = 0.008). CONCLUSIONS: This meta-analysis shows that oral tibolone treatment significantly lowers circulating Lp(a) levels in postmenopausal women. Further studies are warranted to explore the mechanism of this effect and the potential value and place of tibolone or its analogues in the treatment of elevated Lp(a) in individuals at risk of CVD.

Effect of sour tea (Hibiscus sabdariffa L.) on arterial hypertension: a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Hibiscus sabdariffa L. is a tropical wild plant rich in organic acids, polyphenols, anthocyanins, polysaccharides, and volatile constituents that are beneficial for the cardiovascular system. Hibiscus sabdariffa beverages are commonly consumed to treat arterial hypertension, yet the evidence from randomized controlled trials (RCTs) has not been fully conclusive. Therefore, we aimed to assess the potential antihypertensive effects of H. sabdariffa through systematic review of literature and meta-analysis of available RCTs. METHODS: The search included PUBMED, Cochrane Library, Scopus, and EMBASE (up to July 2014) to identify RCTs investigating the efficacy of H. sabdariffa supplementation on SBP and DBP values. Two independent reviewers extracted data on the study characteristics, methods, and outcomes. Quantitative data synthesis and meta-regression were performed using a fixed-effect model, and sensitivity analysis using leave-one-out method. Five RCTs (comprising seven treatment arms) were selected for the meta-analysis. In total, 390 participants were randomized, of whom 225 were allocated to the H. sabdariffa supplementation group and 165 to the control group in the selected studies. RESULTS: Fixed-effect meta-regression indicated a significant effect of H. sabdariffa supplementation in lowering both SBP (weighed mean difference -7.58 mmHg, 95% confidence interval -9.69 to -5.46, P < 0.00001) and DBP (weighed mean difference -3.53 mmHg, 95% confidence interval -5.16 to -1.89, P < 0.0001). These effects were inversely associated with baseline BP values, and were robust in sensitivity analyses. CONCLUSION: This meta-analysis of RCTs showed a significant effect of H. sabdariffa in lowering both SBP and DBP. Further well designed trials are necessary to validate these results.