Tunisian Clematis flammula Essential Oil Enhances Wound Healing: GC-MS Analysis, Biochemical and Histological Assessment.

The aerial part of Clematis flammula (Ranunculaceae) has been traditionally used in the treatment of skin diseases including mycotic infection in the Tunisian traditional medicine. The study was undertaken to extract and determine the essential oil chemical composition of Clematis flammula aerial parts and to assess the potential of anemonin in wound healing on mechanically wounded wistar rats. The essential oil was obtained by hydrodistillation and analyzed by GC-MS. Anemonin was isolated and then incorporated as active in a cream for which the cytotoxicity was evaluated by methyl thiazolyl tetrazolium (MTT)-based colorimetric assay. Then, its potential in wound healing on mechanically wounded wistar rats was assessed. The GC-MS analysis showed that the major compound was protoanemonin (86.74%) which spontaneously dimerised in part to form the anemonin. The wound healing activity of anemonin cream exhibited a non toxic potential of anemonin at a concentration of 25 µg/mL with a cell migration efficiency that reaches more than 80% after 48 hours of treatment. Wound healing efficiency was evaluated by monitoring morphological and skin histological analyses. Comparable wound surface reduction of the group treated by anemonin cream (p ≥ 0.05) when compared to the reference treated group. The skin histological analysis showed the completely wound closure. Antioxidant activity was assessed by the malondialdehyde (MDA) rates and antioxidant enzymes (glutathione peroxidase (GPx) and catalase) determination. The results provided strong support for the effective wound healing activity of anemonin cream, making it a promising candidate as a therapeutic agent in tissue repairing processes.

Low dose assessment of the carcinogenicity of furan in male F344/N Nctr rats in a 2-year gavage study.

Furan is a volatile organic chemical that is a contaminant in many common foods. Furan is hepatocarcinogenic in mice and rats; however, the risk to humans from dietary exposure to furan cannot be estimated accurately because the lowest tested dose of furan in a 2-year bioassay in rats gave nearly a 100% incidence of cholangiocarcinoma. To provide bioassay data that can be used in preparing risk assessments, the carcinogenicity of furan was determined in male F344/N Nctr rats administered 0, 0.02, 0.044, 0.092, 0.2, 0.44, 0.92, and 2 mg furan/kg body weight (BW) by gavage 5 days/week for 2 years. Exposure to furan was associated with the development of malignant mesothelioma on membranes surrounding the epididymis and on the testicular tunics, with the increase being significant at 2 mg furan/kg BW. There was also a dose-related increase in the incidence of mononuclear cell leukemia, with the increase in incidence being significant at 0.092, 0.2, 0.92, and 2 mg furan/kg BW. Dose-related non-neoplastic liver lesions included cholangiofibrosis, mixed cell foci, basophilic foci, biliary tract hyperplasia, oval cell hyperplasia, regenerative hyperplasia, and cytoplasmic vacuolization. The most sensitive non-neoplastic lesion was cholangiofibrosis, the frequency of which increased significantly at 0.2 mg furan/kg BW.

Preliminary assessment of the risk linked to furan ingestion by babies consuming only ready-to-eat food.

The risk linked to furan ingestion has been assessed in previous papers for Belgian adults and children. The present paper focuses on infants consuming only ready-to-eat baby food. As there is no Belgian baby dietary database, the furan exposure assessment was carried out by using an Italian infant consumption database and Belgian contamination data. The estimated daily intake (EDI) was calculated according to a deterministic methodology. It involved 42 commercially available ready-to-eat baby food and 36 baby consumption records. The mean EDI was 1460 ng*(kg(bw)*day)⁻¹ which is 3.8 times higher than the 381 ng*(kg(bw)*day)⁻¹ reported for Belgian adults, and 3.5 times higher than the 419 ng*(kg(bw)*day)⁻¹ measured for Belgian children. To assess and characterise the risk for babies' exposure, the margin of exposure (MoE) was calculated. It highlighted that 74% of infants have a MoE < 1000, with a minimum of 140. However, these are only preliminary results as they were calculated from a very small dataset and the infant cytochrome P450 activity is significantly different compared with the adult's. Therefore, the risk linked to furan ingestion by babies should be assessed in a different manner. To this end, additional data regarding a baby diet as well as a better understanding of furan toxicity for babies are needed to characterise more accurately the risk for infants.

The European medicines agency review of eribulin for the treatment of patients with locally advanced or metastatic breast cancer: summary of the scientific assessment of the committee for medicinal products for human use.

The European Commission issued on March 17, 2011, a marketing authorization valid throughout the European Union (EU) for eribulin (Halaven; Eisai Limited). The decision was based on the favorable opinion of the Committee for Medicinal Products for Human Use recommending a marketing authorization for the treatment of patients with locally advanced or metastatic breast cancer who have progressed after at least 2 chemotherapeutic regimens for advanced disease. Eribulin mesylate is a structurally simplified synthetic analogue of halichondrin B, which is a natural product isolated from the marine sponge Halichondria okadai (ATC code L01XX41). Eribulin is a nontaxane, microtubule dynamics inhibitor belonging to the halichondrin class of antineoplastic agents. Eribulin inhibits the growth phase of microtubules without affecting the shortening phase and sequesters tubulin into nonproductive aggregates leading to G(2)-M cell-cycle block, disruption of mitotic spindles, and, ultimately, apoptotic cell death after prolonged mitotic blockage. The recommended dose of eribulin is 1.23 mg/m(2) (equivalent to 1.4 mg/m(2) eribulin mesylate) to be administered intravenously over 2 to 5 min on days 1 and 8 of a 3-week cycle. In the pivotal trial, eribulin was associated with increased overall survival in patients with locally advanced or metastatic breast cancer who received at least 2 prior chemotherapy lines for advanced disease (median overall survival was 13.2 months in the eribulin arm vs. 10.6 months in the control arm; HR = 0.805; 95% confidence interval, 0.677-0.958; P = 0.014). The most common side effects are asthenia or fatigue and neutropenia. The objective of this article is to summarize the scientific review of the application leading to approval in the EU. The detailed scientific assessment report and product information, including the summary report and product information, including product characteristics, are available on the European Medicines Agency website.

Risk assessment for furan contamination through the food chain in Belgian children.

Young, old, pregnant and immuno-compromised persons are of great concern for risk assessors as they represent the sub-populations most at risk. The present paper focuses on risk assessment linked to furan exposure in children. Only the Belgian population was considered because individual contamination and consumption data that are required for accurate risk assessment were available for Belgian children only. Two risk assessment approaches, the so-called deterministic and probabilistic, were applied and the results were compared for the estimation of daily intake. A significant difference between the average Estimated Daily Intake (EDI) was underlined between the deterministic (419 ng kg⁻¹ body weight (bw) day⁻¹) and the probabilistic (583 ng kg⁻¹ bw day⁻¹) approaches, which results from the mathematical treatment of the null consumption and contamination data. The risk was characterised by two ways: (1) the classical approach by comparison of the EDI to a reference dose (RfD(chronic-oral)) and (2) the most recent approach, namely the Margin of Exposure (MoE) approach. Both reached similar conclusions: the risk level is not of a major concern, but is neither negligible. In the first approach, only 2.7 or 6.6% (respectively in the deterministic and in the probabilistic way) of the studied population presented an EDI above the RfD(chronic-oral). In the second approach, the percentage of children displaying a MoE above 10,000 and below 100 is 3-0% and 20-0.01% in the deterministic and probabilistic modes, respectively. In addition, children were compared to adults and significant differences between the contamination patterns were highlighted. While major contamination was linked to coffee consumption in adults (55%), no item predominantly contributed to the contamination in children. The most important were soups (19%), dairy products (17%), pasta and rice (11%), fruit and potatoes (9% each).

Risk assessment of Belgian adults for furan contamination through the food chain.

Risk assessment is an interdisciplinary process used to quantify the risk linked to a hazard. In the present paper it is applied to quantify the risk linked to furan ingestion through the food chain for the Belgian adult population. Two approaches, deterministic and probabilistic, were carried out in parallel. The deterministic method relied on a case study, whereas the probabilistic approach involved statistical distributions of contamination and consumption data to calculate a statistical distribution of the daily intake. First, the deterministic method revealed a low estimated daily intake (EDI) for the average population (380 ng*(kg(bw)*day)⁻¹) and a huge contribution of coffee consumption to the EDI (55%). Increasing or decreasing the daily coffee consumption by one cup can affect the EDI by about 22%. Afterwards, the probabilistic approach showed that the average population has a low EDI (494 ng*(kg(bw)*day)⁻¹), and that high contamination levels were only registered in a small proportion of the population. Finally, a comparison of the RfD(chronic oral) showed that less than 10% of the Belgian population had an EDI above the reference dose proposed by the USEPA; the majority of the population had an EDI 20% below the reference dose. The margin of exposure (MoE) approach indicated that the level of risk related to furan intake through ingestion is low, with a MoE > 10,000 for more than 10% of the population and no result < 100.

Detailed exposure assessment of dietary furan for infants consuming commercially jarred complementary food based on data from the DONALD study.

Furan is a possible human carcinogen regularly occurring in commercially jarred complementary foods. This paper will provide a detailed exposure assessment for babies consuming these foods considering different intake scenarios. The occurrence data on furan in complementary foods were based on our own headspace-gas chromatography/mass spectrometry (HS-GC/MS) analytical results (n = 286). The average furan content in meals and menus was between 20 and 30 µg kg(-1), which is in excellent agreement with results from other European countries. Using measured food consumption data from the Dortmund Nutritional and Anthropometric Longitudinally Designed (DONALD) study, the average exposures for consumers of commercially jarred foods ranged between 182 and 688 ng kg(-1) bw day(-1), with a worst case scenario for P95 consumers ranging between 351 and 1066 ng kg(-1) bw day(-1). The exposure data were then used to characterize risk using the margin of exposure method based on a benchmark dose lower confidence limit for a 10% response (BMDL10) of 1.28 mg kg(-1) bw day(-1) for hepatocellular tumours in rats. The margin of exposures (MOEs) were below the threshold of 10 000, which is often used to define public health risks, in all scenarios, ranging between 7022 and 1861 for average consumers and between 3642 and 1200 for the P95 consumers. Mitigative measures to avoid furan in complementary foods should be of high priority for risk management.

Eribulin mesylate (Halaven) for breast cancer.

Eribulin mesylate (Halaven-Eisai) has been approved by the FDA for treatment of patients with metastatic breast cancer who have previously received at least 2 chemotherapy regimens for metastatic cancer. Prior therapy should have included an anthracycline and a taxane in either an adjuvant or metastatic setting. Other drugs used to treat anthracycline- and taxane-refractory metastatic breast cancer include capecitabine (Xeloda), gemcitabine (Gemzar, and others) and vinorelbine (Navelbine, and others).

Assessment of seasonality in exposure to dioxins, furans and dioxin-like PCBs by using long-term food-consumption data.

According to the European Food Safety Authority (EFSA) guidance related to uncertainties in dietary exposure assessment, exposure assessment based on short-term food-consumption surveys, such as 24-h recalls or 2-day records, tend to overestimate long-term exposure because of the assumption that the dietary pattern will be similar day after day over a lifetime. The aim of this study was to make an assessment of dietary exposure to polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), also called 'dioxins' and 'dioxin-like PCBs', using long-term household purchase and consumption survey data collected by TNS-Secodip. Weekly purchases of the major dioxins and dl-PCB vector products of these contaminants were collected for 328 single-person households, who participated at TNS-Secodip consumption surveys from 2003 to 2005 and who were single-person households in order to estimate better their consumption. These data were combined with average contamination levels of food products. Weekly gross average exposure was estimated at 10.2 pg toxic equivalent (WHO TEQ) kg(-1) bw week(-1) (95% confidence interval [9.6, 10.9]). According to the typical shape of the distribution of individual weekly exposures, it is sensible to fit an exponential law to these data. The mean was therefore 12.1 pg WHO TEQ kg(-1) bw week(-1). This value is higher than the arithmetic mean because it better takes into account inter-individual variability. It was estimated that about 20% of persons in this sample were exceeding the current health-based guidance value mainly due to high consumption of seafood and/or dairy products. Thanks to long survey duration (3 years) and the weekly recording of food consumption, it was possible to demonstrate the actual seasonality of dietary exposure to dioxins and dl-PCBs with a maximum between March and September; similar seasonality is observable for fish consumption. Autoregressive integrated moving average (ARIMA) models were adjusted to the time series and it was demonstrated that the number of times the upper limit of confidence intervals exceeds the provisional tolerable weekly intake (PTWI) is about 15 weeks per year on average. Finally, compared with the results obtained from data collected in the short-term surveys (1 week), this study does not suggest that short-term consumption surveys tend to overestimate the long-term exposure.

An age-adjusted bootstrap-based Poly-k test.

The assumption of an asymptotic normal distribution of some test statistics may be invalid in certain dose-response trend tests. For instance, the survival-adjusted Cochran-Armitage test, known as the Poly-k test, is asymptotically standard normal under the null hypothesis. However, the asymptotic normality is not valid if there is a deviation from the tumour onset distribution that is assumed in this test or if the competing risks survival rates differ across groups. We develop an age-adjusted bootstrap-based method to assess the significance of assumed asymptotic normal tests for animal carcinogenicity data. The proposed method differs from conventional bootstrap methods in the aspect of preserving the mortality rate in each dose group under the null hypothesis of equal tumour incidence rates among the groups. We investigate an empirical distribution of the Poly-3 (P3) trend test statistic using the proposed age-adjusted bootstrap-based method and compare it with the P3 test statistic referenced to the assumed standard normal distribution. A simulation study is conducted to evaluate the robustness of these tests to various Weibull-family tumour onset distributions. The proposed method is applied to National Toxicology Program data sets to evaluate a dose-related trend of a test substance on the incidence of neoplasms.

Isolation of pectenotoxin-2 from Dinophysis acuta and its conversion to pectenotoxin-2 seco acid, and preliminary assessment of their acute toxicities.

We have developed a simple and effective method for isolating pectenotoxin-2 (PTX-2) from Dinophysis cells collected from a natural bloom. A two-step extraction procedure followed by two column chromatography steps produced PTX-2 in high purity suitable for use as an analytical standard and for toxicological studies. Incubation of purified PTX-2 with the supernatant from ultracentrifuged blue (Mytilus edulis) or Greenshell (Perna canaliculus) mussel hepatopancreas homogenate caused rapid conversion to pectenotoxin-2 seco acid (PTX-2 SA). Purification of PTX-2 SA was achieved by solvent extraction followed by column chromatography. PTX-2 and PTX-2 SA were fully characterized by LC-MS and NMR, and full (1)H and (13)C NMR assignments were obtained. Okadaic acid C(8)-diol ester was isolated during the purification of PTX-2, and its identity confirmed by NMR and LC-MS analyses. Pectenotoxin-2 seco acid methyl ester, identified by LC-MS, was also produced during the hydrolytic procedure due to the presence of methanol. PTX-2 was acutely toxic to mice by i.p. injection (LD(50)=219 microg/kg) but no effects were seen with PTX-2 SA at 5000 microg/kg. Neither PTX-2 nor PTX-2 SA was overtly toxic to mice by the oral route at doses up to 5000 microg/kg. No diarrhea was observed in mice dosed with either compound, suggesting that pectenotoxins do not belong in the diarrhetic shellfish poison group.

Exposure assessment of dioxins/furans consumed in dairy foods and fish.

Dioxins/furans are ubiquitous environmental contaminants whose primary route of human exposure occurs via the consumption of fatty foods of animal origin. The US FDA conducted a market basket survey of dairy products and commercial fish and shellfish to obtain data on levels of 17 dioxin/furan congeners (2, 3, 7, 8-congeners) in the US. The dairy products sampled included various cheeses (American, cheddar, Swiss, cottage), ice cream, yogurt, butter, and milk. The finfish and shellfish (molluscs and crustacea) sampled are those marine species consumed in the greatest amounts and include canned tuna, shrimp, cod, blue crab, and oysters. Catfish was sampled because it is the dominant aquaculture species. Samples were collected in 1995/96 and analysis for 17 dioxin/furan congeners was performed by high-resolution gas chromatography following extraction and clean-up. Limits of detection (LOD) and quantitation (LOQ) for each congener in each food were reported. Point estimates of exposure were calculated using a 3-day (1-day diary plus 2-day recall) food consumption survey for eaters-only and for the general population (USDA/CSFII, 1989-92). Toxicity equivalency factors (TEFs) developed by the World Health Organization (1997) were used to derive overall dioxin/furan toxicity equivalents (TEQ) for each sample food. Mean estimates of TEQ exposure for each food were derived using five values for non-detects (ND = 0; ND = 1/2 LOD or LOQ, ND = LOD or LOQ) on both a total sample and eaters-only basis. Using zero and the LOD provide lower and upper bounds on the range of estimated exposure, respectively. The bounds on mean dioxin intakes (pg/person/day) calculated for consumers of specific foods were estimated as follows (using zero or LOD for non-detects): butter (0.5-11), cheese (1.6-3.2), ice cream (4-19), yogurt (0.8-28), catfish (148-150), fish (other than catfish) (0.03-9), crustacea (32-35), mollusks (16.1-16.6), and shrimp (0.09-4.5). Exposure estimates derived by the five ND-methods are strongly dependent on the LOD and LOQ and represent upper bound estimates of exposure. Uncertainty in the exposure estimates is reduced with refinements in the analytical method.

[Assessment by transcutaneous PO2 measurement of the treatment of venous ulcers with naftidrofuryl]. / Appréciation par la mesure de la PO2 transcutanée d'un traitement des ulcéres veineux par le naftidrofuryl.

A double-blind therapeutic trial was undertaken in 40 subjects with large venous ulcers and distributed on a homogeneous basis. They were treated with naftidrofuryl by intravenous infusions for one week followed by oral treatment for three months, in comparison with a placebo (infusions of saline and lactose tablets). All the patients received the same local treatment. Results were assessed not only clinically (changes in ulcer) but also objectively by the measurement of TcPO2. TcPO2 by the measurement of transcutaneous tissue oxygenation is an indicator of the state of the microcirculation and hence the trophic condition of the tissues. Study of the results (Student t) showed that in the treated group and at the ulcer site there was a significant (p less than 0.05) difference from D7 onwards, which increased further during treatment (p less than 0.02 at 3 months). For the placebo group, significance occurred only at D90 (p less than 0.05). The intergroup course difference was itself significant at D45 and D90. The tolerance of naftidrofuryl in both of its forms was unremarkable. Clinical study showed a good correlation between improvement in PO2 and ulcer healing. This trial confirmed the efficacy of naftidrofuryl in the treatment of venous ulcers.