Genomic damages in peripheral blood lymphocytes and association with polymorphisms of three glutathione S-transferases in workers exposed to formaldehyde

DNA and chromosome damages in peripheral blood lymphocytes were evaluated in 151 workers occupationally exposed to formaldehyde (FA) and 112 non-FA exposed controls. The effects of polymorphisms in three glutathione-S-transferase (GSTs) genes on the DNA and chromosome damages were assessed as well. Alkaline comet assay and cytokinesis-block micronucleus (CBMN) assay were used to determine DNA and chromosome damages, respectively. The genotypes of GSTP1 (Ile105Val), GSTT1, and GSTM1 were assayed. The mean 8-h time-weighted average (TWA) concentrations of FA in two plywood factories were 0.83 ppm (range: 0.08–6.30 ppm). FA-exposed workers had higher olive tail moment (TM) and CBMN frequency compared with controls (Olive TM, 3.54, 95%CI = 3.19–3.93 vs. 0.93, 95%CI = 0.78–1.10, P < 0.01; CBMN frequency, 5.51 ± 3.37 vs. 2.67 ± 1.32, P < 0.01). Olive TM and the CBMN frequency also had a dose-dependent relation with the personal FA exposure. Significant association between FA exposure history and olive TM and CBMN frequency were also identified. The level of olive TM was slightly higher in FA-exposed workers with GSTM1 null genotype than those with non-null genotype (3.86, 95%CI = 3.31–4.50 vs. 3.27, 95%CI = 2.83–3.78, P = 0.07) with adjustment of covariates. We also found that FA-exposed workers carrying GSTP1 Val allele had a slightly higher CBMN frequency compared with workers carrying only the wild-type allele (6.32 ± 3.78 vs. 5.01 ± 2.98, P = 0.05). Our results suggest that the FA exposure in this occupational population increased DNA and chromosome damages and polymorphisms in GSTs genes may modulate the genotoxic effects of FA exposure.     

A novel fluorescence-based cellular permeability assay

Vascular permeability is a pathologic process in many disease states ranging from metastatic progression of malignancies to ischemia–reperfusion injury. In order to more precisely study tissue, and more specifically cell layer permeability, our goal was to create a fluorescence-based assay which could quantify permeability without radioactivity or electrical impedance measurements. Human aortic endothelial cells were grown in monolayer culture on Costar®-Transwell® clear polyester membrane 6-well cell culture inserts. After monolayer integrity was confirmed, vascular endothelial growth factor (VEGF₁₆₅) at varying concentrations with a fixed concentration of yellow-green fluorescent 0.04 μm carboxylate-modified FluoSpheres® microspheres were placed in the luminal chamber and incubated for 24 h. When stimulated with VEGF₁₆₅ at 20, 40, 80, and 100 ng/ml, this assay system was able to detect increases in trans-layer flux of 8.2 ± 2.4%, 16.0 ± 3.7%, 41.5 ± 4.9%, and 58.6 ± 10.1% for each concentration, respectively. This represents the first fluorescence-based permeability assay with the sensitivity to detect changes in the permeability of a cell layer to fluid flux independent of protein flux; as well as being simpler and safer than previous radioactive-and impedance-based permeability assays. With the application of this in vitro assay to a variety of pathologic conditions, both the dynamics and physiology relating to cellular permeability can be more fully investigated.     

Valoración de la musculatura isquiosural en personas mayores

Objectiveto evaluate hamstring flexibility in older adults.Materials and methodsa total of 177 subjects (13 men and 164 women) aged between 43 and 80 years old (mean age = 63.4 ± 6.7 years) who attended fitness classes were evaluated. The mean height was 161 ± 10 cm and the mean weight was 74 ± 5.6 kg. Hamstring flexibility was evaluated using the straight leg raise test.Resultsthe mean flexion in the right hip was 72° ± 13.2° and the mean flexion of the left hip was 72.4° ± 13.8°. No significant differences were found in the values obtained from the straight leg raise test in the left and right legs. For the right leg, 48.02% of the subjects’ values were within the normal range, while 28.81% showed grade I shortness, and 23.61% showed grade II shortness. For the left leg, 49.94% showed normal values, 29.94% showed grade I shortness, and 22.03% showed grade II shortness.Conclusionshalf of the adults and elderly individuals that took part in the present study had hamstring shortness, which increased in frequency with age.     

The preparation and antihyperlipidaemic assay of piperlonguminine in vivo

The natural product piperlonguminine (GBN) which is extracted from Piper longum Linn., has high antihyperlipidaemic activity and low toxicity. However, the content of natural GBN in P. longum (0.20–0.25%) is low, and it is not easy to prepare enough sample of natural GBN for further studies, such as large-scale animal experiments. Therefore, in the present study, we tested and confirmed the antihyperlipidaemic activity of chemically synthesised GBN in rats for the first time. The results of the antihyperlipidaemic assay in vivo showed that synthetic GBN had significant lipid-lowering activities. Synthetic GBN not only inhibited body weight gain (66.3 ± 22.50 g vs. 83.9 ± 19.95 g) but also significantly decreased total cholesterol (TC, 9.67 ± 3.32 mmol/L vs. 22.26 ± 5.84 mmol/L), total glycerol (TG, 1.47 ± 0.08 mmol/L vs. 2.86 ± 0.75 mmol/L), and low-density lipoprotein cholesterol (LDL-C, 3.57 ± 1.15 mmol/L vs. 5.44 ± 1.42 mmol/L) while increasing high-density lipoprotein cholesterol (HDL-C, 1.31 ± 0.56 mmol/L vs. 0.68 ± 0.20 mmol/L) in the serum of rats fed a high-fat diet.     

Correlation between serum trace elements and risk of preeclampsia: A case controlled study in Riyadh,Saudi Arabia

Preeclampsia is a serious medical complication during pregnancy. In response to an increasing number of preeclamptic cases and scarcity of data concerning the interrelation between trace element levels and preeclampsia, we carried out a hospital based case–control study in Riyadh, Saudi Arabia to study the correlation between levels of serum trace elements and risk of preeclampsia. One hundred and twenty pregnant women were enrolled in this study and divided into three groups of 40 each—Control group, HR group (women at high risk of preeclampsia) and PET group (Preeclampsia group). Serum trace element levels were estimated by inductively coupled plasma optical emission spectrophotometer. The analysis found that mean values of Ca, Mg and Zn were 90.08 ± 6.38, 19.33 ± 3.32 and 1.30 ± 0.83 mg/L respectively in normotensive control and 77.85 ± 4.47, 15.44 ± 1.43 and 0.98 ± 0.63 mg/L respectively in the HR group. The mean values of Ca, Mg and Zn in the preeclamptic group were 70.37 ± 4.66, 13.58 ± 1.98 and 0.67 ± 0.59 mg/L, respectively. Interelement analysis reflected a negative correlation between Ca and Mg and between Mg and Zn whereas positive correlation between Ca and Zn in preeclamptic women. However the correlation was not statistically significant. In conclusion, our study suggests that decreased levels of these trace elements in serum may act as predisposing factors in pathogenesis of Preeclampsia.     

A FRET-based assay for the discovery of West Nile Virus NS2B-NS3 protease inhibitors

The West Nile Virus (WNV) has been a worldwide epidemic since the early 1990s. Currently there are no therapeutic treatments for WNV infections. One particular avenue of treatment is inhibition of the NS2B-NS3 protease, an enzyme that is crucial for WNV replication. In our effort to increase the number of NS2B-NS3 protease inhibitors, we report a novel FRET-based high throughput assay for the discovery of WNV NS2B-NS3 protease inhibitors. For this assay, a FRET-based peptide substrate was synthesized and kinetically characterized with the NS2B-NS3 protease. The new substrate exhibits a K_m of 3.35 ± 0.31 μM, a k_cat of 0.0717 ± 0.0016 s^?1 and a k_cat/K_m of 21,400 ± 2000 M^?1 s^?1.     

Biodosimetry using micronucleus assay in acute partial body therapeutic irradiation

Biological dosimetry provides information on the absorbed dose and its distribution in the body for an early assessment of irradiation consequences in exposed individuals. In this study, an effort has been made to see the applicability of biological dosimetry using micronuclei assay for dose estimation in therapeutic irradiation of cancer patients in acute high dose partial body irradiation. Dose estimation in partial body irradiations was done on the basis of Dolphin's contaminated Poisson method, using the in vitro dose response calibration curve. The equivalent whole body dose and the dose to the irradiated part of the body were estimated to be 1.8 ± 0.1 Gy and 6.4 ± 0.3 Gy, respectively. The estimated percentage of irradiated blood and the fraction of cells exposed were 41.5 ± 1.6% and 10.4 ± 0.8%, respectively. The estimated fraction of irradiated cells was comparable with the actual volume of irradiation.     

A colorimetric assay method to measure acetyl-CoA synthetase activity: Application to woodchuck model of hepatitis virus-induced hepatocellular carcinoma

A new spectrophotometric method for quantitation of acetyl-CoA synthetase (ACAS) activity is developed. It has been applied for ACAS assay in the liver tissues of a woodchuck model of hepatitis virus-induced hepatocellular carcinoma (HCC). The assay is based on the established pyrophosphate (PPi) detection system. ACAS activity is indexed by the amount of PPi, the product of ACAS reaction system of activated form of acetate (acetyl-CoA) with ACAS catalysis. PPi is determined quantitatively as the amount of chromophore formed with molybdate reagent, 1-amino-2-naphthol-4-sulfonic acid in bisulfite and 2-mercaptoethanol. PPi reacts with molybdate reagent to produce phosphomolybdate and PPi-molybdate complexes. 2-mercaptoethanol is responsible for color formation which has the peak absorbance at 580 nm. This method was sensitive from 1 to 20 nmol of PPi in a 380-μl sample (1-cm cuvette). A ten-fold excess of Pi did not interfere with the determination of PPi. To study the major metabolic pathways of imaging tracer [1-¹¹C]-acetate in tumors for detection of HCC by Positron Emission Tomography (PET), the activity of one of the key enzymes involved in acetate or [1-¹¹C]-acetate metabolism, ACAS was assayed by this newly developed assay in the tissue samples of woodchuck HCCs. A significant increase of ACAS activity was observed in the liver tissues of woodchuck HCCs as compared with neighboring regions surrounding the tumors (P < 0.05). The respective ACAS activities in the subcellular locations were also significantly higher in HCCs than in the surrounding tissues (P < 0.05) (total soluble fraction: 876.61 ± 34.64 vs. 361.62 ± 49.97 mU/g tissue; cytoplasmic fraction: 1122.02 ± 112.39 vs. 732.32 ± 84.44 mU/g tissue; organelle content: 815.79 ± 100.77 vs. 547.91 ± 97.05 mU/ g tissue; sedimentable fragment: 251.92 ± 51.56 vs. 90.94 ± 18.98 mU/ g tissue). The finding suggests an increase in ACAS activity in the liver cancer of woodchuck models of HCC as compared to that in the normal woodchuck liver. The developed assay is rapid, simple and accurate and is suitable for the investigation of ACAS activity under physiologic and pathophysiologic conditions.     

Nanosecond electric pulses induce DNA breaks in cisplatin-sensitive and -resistant human ovarian cancer cells

Human ovarian cancer cells COC1 and COC1/DDP (cisplatin-resistant subline) were exposed to 6 kV/cm nanosecond electric pulses (nsEP) with a pulse length of 8, 16 or 24 ns. The potential in a subcellular unit was calculated using a multilayer dielectric spherical model, and area under the voltage–time curves (AUC) integrated with a lower limit of 0.2 V. Cell viability was determined, and double-stand and total DNA breaks detected with the neutral and alkaline comet assays. nsEP evoked a higher voltage and AUC in nucleoplasm, and the levels in COC1 cells was just above those in COC1/DDP cells. Comets only appeared in the alkaline assay demonstrating single-stand DNA break. Fewer DNA break (16.51% vs. 35.13% at 24 ns, p = 0.0150) and more survival (22.42% vs. 13.19% at 24 ns, p = 0.0015) occurred in COC1/DDP cells despite an equal electric energy and almost equal cell sizes. 24-ns EP led to higher rates of cell-death and comet. The comet rate correlated with cell-death fraction in either cell line (r = 0.5701, p = 0.0135; r = 0.5110, p = 0.0302). There was no a correlation between the tail length, tail moment or Olive tail moment and cell-death rate. The data showed that response of chemosensitive cells differed from that of chemoresistant cells and DNA damage contributed to percent of cell death.     

The mycolyltransferase 85A,a putative drug target of Mycobacterium tuberculosis: Development of a novel assay and quantification of glycolipid-status of the mycobacterial cell wall

The enzymes of the antigen 85 complex (Ag85A, B, and C) possess mycolyltransferase activity and catalyze the synthesis of the most abundant glycolipid of the mycobacterial cell wall, the cord factor. The cord factor (trehalose 6,6′-dimycolate, TDM) is essential for the integrity of the mycobacterial cell wall and pathogenesis of the bacillus. Thus, TDM biosynthesis is regarded as a potential drug target for control of Mycobacterium tuberculosis infections. Trehalose 6,6′-dimycolate (TDM) is synthesized from two molecules of trehalose-6′-monomycolate (TMM) by antigen 85A. We report here a novel enzyme assay using the natural substrate TMM. The novel colorimetric assay is based on the quantification of glucose from the degradation of trehalose, which is the product from catalytic activity of antigen 85A. Using the new assay, K_m and K_cat were determined with values of 129.6 ± 8.1 µM and 65.4 ± 4.1 min^− 1, respectively. This novel assay is also suitable for robust high-throughput screening (HTS) for compound library screening against mycolyltransferase (antigen 85A). The assay is significantly faster and more convenient to use than all assays currently in use. The assay has a very low coefficient of variance (0.04) in 96-well plates and shows a Z′ factor of 0.67–0.73, indicating the robustness of the assay. In addition, this new assay is highly suitable for the quantification of total TMM of the mycobacterial cell envelope.     

A high pressure liquid chromatography-based assay for glutathione-S-transferase class distinction assay

In order to expedite the process of classification of the members of the family of glutathione-S-transferases (GSTs) high performance liquid chromatography with photodiode array detection (HPLC-PDA) was used as a means for measuring enzymatic activity. The GST chosen for the development of the HPLC-PDA technique was from equine liver (E-GST). The characterizing substrates, ethacrynic acid (EA) and bromosulfophthalein (BSP), along with previously gathered characterization data allowed for the distinction of α, μ or π-class enzymes. In an initial characterization of the previously unclassified E-GST it was determined that the enzyme was of the π-class with specific activities of 0.062, ± 0.0015 μmol min^− 1 mg^− 1 and 0.0019, ± 0.00064 μmol min^− 1 mg^− 1 for EA and BSP, respectively. Finally, the activity of the E-GST with the EA and BSP substrates, was measured by HPLC-PDA, and was found to be 0.027, ± 0.003 μmol min^− 1 mg^− 1 and 0.002, ± 0.0005 μmol min^− 1 mg^− 1, respectively. While the HPLC-PDA data do not mirror the spectrophotometric results quantitatively the overall response by the E-GST was the same. In general, the E-GSTs were shown to belong to the π-class when characterized by HPLC-PDA due to an EA specific activity greater than 0.01 μmol min^− 1 mg^− 1 and a negligible BSP activity (≤ 0.002 μmol min^− 1 mg^− 1).     

Inhalation of formaldehyde does not induce genotoxic effects in broncho-alveolar lavage (BAL) cells of rats

Male Fischer-344 rats were exposed to formaldehyde (FA) by inhalation for 4 weeks (6 h/day, 5 days/week). Groups of six rats each were exposed to the target concentrations of 0, 0.5, 1, 2, 6, 10 and 15 ppm. Potential genotoxic effects in the lung were investigated as part of a comprehensive study on local and systemic toxic and genotoxic effects. Broncho-alveolar lavage (BAL) cells were obtained by lung lavage with physiological saline and counted. From one half of the cells, slides for the micronucleus test (MNT) were prepared by cytocentrifugation; with the other half, the comet assay was performed. DNA migration in the comet assay was measured both directly and after irradiation of the cells with 2 Gy gamma-radiation. The latter modification of the comet assay was included to increase its sensitivity for the detection of DNA-protein cross-links (DPX). For the comet assay, four slides were analysed from each cell sample, two without and two with irradiation. From each slide, 50 randomly selected cells were measured by image analysis and tail intensity (% tail DNA) and tail moment were evaluated. The frequency of micronucleated BAL cells was determined in acridine orange-stained slides by analysing 2000 cells per animal. FA did not induce any significant effect in any of the genotoxicity tests performed. It can be concluded that inhalation of FA in a 28 days study with FA concentrations up to 15 ppm does not lead to genotoxic effects in BAL cells of rats. Because detection of DPX by the comet assay is a very sensitive biomarker of FA exposure of cells, our results suggest that there is no genetically relevant exposure of the lung after FA inhalation. The results of our inhalation study, which was performed under GLP conditions, call into question the biological significance of previously reported genotoxic effects in the lung of rats after FA inhalation.     

A convenient synthesis and molecular modeling study of novel purine and pyrimidine derivatives as CDK2/cyclin A3 inhibitors

A series of novel purine and pyrimidine derivatives were prepared and biologically evaluated for their in vitro anti-CDK2/cyclin A3 and antitumor activities in Ehrlich ascites carcinoma (EAC) cell based assay. The novel purine derivatives 13a,b demonstrated potent inhibitor activities with IC₅₀ values of 14 ± 9 and 13 ± 9 μM, respectively. Additionally, compound 15a showed the highest potency (IC₅₀ = 10 ± 6 μM) in EAC cell based assay. Molecular modeling study, including fitting to a 3D-pharmacophore model and their docking into cyclin dependant kinase2 (CDK2) active site showed high fit values and docking scores.     

Increased Atherogenic Low-Density Lipoprotein Cholesterol in Untreated Subclinical Hypothyroidism

ObjectiveTo evaluate the effects of physiologic doses of levothyroxine replacement on the lipoprotein profile in patients with subclinical hypothyroidism (SCH).MethodsIn a prospective, double-blind, placebo- controlled study, we enrolled 120 patients—mostly, but not exclusively, premenopausal women—with SCH. Patients were randomly assigned to either a levothyroxine- treated group (n = 60) or a placebo (control) group (n = 60). Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TG) were measured before and 52 weeks after assignment to either group.ResultsIn the levothyroxine-treated group, the lipoprotein mean values before and after the 52-week study were as follows: TC, 5.05 ± 0.98 mmol/L versus 4.74 ± 0.87 mmol/L (P < .0001); LDL-C, 3.30 ± 0.90 mmol/L versus 2.89 ± 0.59 mmol/L (P < .01); TG, 1.18 ± 0.71 mmol/L versus 0.95 ± 0.53 mmol/L (P < .002); and HDL-C, 1.20 ± 0.33 mmol/L versus 1.19 ± 0.32 mmol/L (P = .29). In the control group, TC, HDL-C, and TG values remained unchanged after 52 weeks in comparison with baseline, but LDL-C mean values increased from 2.79 ± 0.60 mmol/L to 3.11 ± 0.77 mmol/L, a change that was statistically significant (P < .001). At the end of the study, the lipid profile changes between levothyroxine- treated and control groups were compared. Total cholesterol and LDL-C were significantly lower in the levothyroxine-receiving group (P < .029 and P < .0001, respectively) in comparison with the control group. The difference did not reach statistical significance for TG and HDL-C values.ConclusionIn premenopausal women, SCH has a negative effect on the lipoprotein profile and may translate into a sizable cardiovascular risk if left untreated. (Endocr Pract. 2008;14:570-575)     

Individualising the exposure of −110 °C whole body cryotherapy: The effects of sex and body composition

The purpose of this study was to investigate the effects of whole body cryotherapy (WBC) on a range of thermoregulatory measures. We also sought to examine the influence of sex and body composition. A convenience sample of 18 healthy participants (10 males and 8 females) (27±6 yr) volunteered for this study. Temperature (core, tympanic, skin and mean body), heart rate, blood pressure, and thermal comfort and sensation were recorded pre- and post- (immediately and every 5 min until 35 min post) exposure to a single bout of WBC (30 s at −60 °C, 150 s at 110 °C). Anthropometric data (height, weight, body surface area, body mass index, fat mass and fat free mass) were also recorded. No significant differences in temperature (core, tympanic, skin and mean body), heart rate, blood pressure, or thermal comfort / sensation were observed between male and females at baseline. Immediately post WBC mean body (male:31.9±0.8 °C; female:31.0±0.9 °C; ∆ mean body temperature:0.9±0.1 °C; P≤0.05, d=0.64) and mean skin (male:22.1±2.2 °C; female:19.6±2.8 °C; ∆ mean skin temperature:−2.5±0.6 °C; d=0.99, P≤0.05) temperature was significantly different between sexes. Sex differences were also observed in regional skin temperature (male thigh, 20.8±1.1 °C; female thigh, 16.7±1.1 °C, ∆ mean thigh skin temperature:−4.1 °C; d=3.72; male calf, 20.5±1.1 °C; female calf, 18.2±1 °C, ∆ mean calf skin temperature:−2.3±0.1 °C; d=3.61; male arm, 21.7±1 °C; female arm, 19±0.4 °C, ∆ mean arm skin temperature: −2.7±0.3 °C; d=3.54; P≤0.05). Mean arterial pressure was significantly different over time (P≤0.001) and between sexes (male 0 mins:94±10 mmHg; female 0 mins:85±7 mmHg; male 35 mins:88±7 mmHg; female 35 mins:80±6 mmHg; P≤0.05). Combined data set indicated a strong negative relationship between skin temperature and body fat percentage 35 min’ post WBC (r=−0.749, P≤0.001) and for core temperature and body mass index in males only (r=0.726, P≤0.05) immediately after WBC. There were no significant differences between sexes in any other variables (heart rate, tympanic and perceptual variables). We observed sex differences in mean skin and mean body temperature following exposure to whole body cryotherapy. In an attempt to optimise treatment, these differences should be taken into account if whole body cryotherapy is prescribed.     

Intra- and interday reliability of voluntary and electrically stimulated isometric contractions of the quadriceps femoris

The reliability of voluntary and electrically stimulated isometric contractions of m. quadriceps femoris of male participants (n = 10; age 30 ± 8 years; height 1.79 ± 0.05 m; body mass 79.4 ± 8.3 kg) was investigated using ratio limits of agreement (LoA) on a time scale common to examine recovery from muscle damaging exercise. No systematic changes in reliability occurred over time (baseline versus 2, 24, 48, and 72 h). Maximal voluntary contraction (MVC) and interpolated twitch technique (ITT) showed no mean bias (P > 0.05) with 95% LoA of ±12.7 and ±5.4, respectively. Resting twitch and potentiated doublet peak force showed no mean bias (P > 0.05). However, 95% LoA were smaller for the doublet (±13.9) than the twitch (±32.0). Twitch and doublet rates showed similar trends. Ratio of low (20 Hz) to high (50 Hz) frequency forces showed no mean bias (P > 0.05) and 95% LoA of (±9.2). However, there was significant mean bias (P < 0.05) and wider 95% LoA for peak force, contraction and relaxation parameters of the low and high frequency forces. In conclusion, MVC, ITT, potentiated doublet and the ratio of low to high frequency forces are recommended to most reliably examine functional muscle recovery between 2 and 72 h after damaging exercise.     

Soil quality assessment of coastal wetlands in the Yellow River Delta of China based on the minimum data set

Little information is available to assess the dynamic changes in wetland soil quality in coastal regions, though it is essential for wetland conservation and management. Soil samples were collected in Suaeda salsa wetlands (SWs), Tamarix chinensis wetlands (TWs), Suaeda salsa–Tamarix chinensis wetlands (STWs), freshwater Phragmites australis wetlands (FPWs) and saltwater Phragmites australis wetlands (SPWs) in three sampling periods (i.e., summer and autumn of 2007 and spring of 2008). According to the flooding characteristics of these wetlands, the study area could be grouped into three sub-regions: short-term flooding region (STFR), seasonal flooding region (SFR) and tidal flooding region (TFR). Soil quality was evaluated using the soil quality index (SQI), which was calculated using the selected minimum data set (MDS) based on principal components analysis (PCA). Our results showed that soil salt content (SSC), total carbon (TC), magnesium (Mg), nitrate nitrogen (NO₃⁻-N) and total sulfur (TS) consisted of a MDS among 13 soil properties. The SQI values varied from 0.18 to 0.66 for all soil samples, of which the highest and lowest SQI values were observed in TFR. The average SQI values were significantly higher in summer (0.50 ± 0.13) than in spring (0.37 ± 0.13) and autumn (0.36 ± 0.11) in the whole study area (p < 0.05). The average SQI values followed the order STFR (0.44 ± 0.12) > TFR (0.41 ± 0.15) > SFR (0.35 ± 0.09) although no significant differences were observed among the three regions (p > 0.05). SPWs and SWs soils showed higher SQI values (0.50 ± 0.10 and 0.47 ± 0.15, respectively) than TWs (0.30 ± 0.08) soils (p < 0.05). The SSC was the dominant factor of soil quality with its proportion of 34.1% contributing to the SQI values, followed by TC (24.5%) and Mg (24.1%). Correlation analysis also showed that SQI values were significantly negatively correlated with SSC. SSC might be a characteristic indicator of wetland soil quality assessment in coastal regions. The findings of this study showed that the SQI based on MDS is a powerful tool for wetland soil quality assessment.     

Evaluation of genotoxicity, cytotoxicity and cytostasis in human lymphocytes exposed to patulin by using the cytokinesis-block micronucleus cytome (CBMN cyt) assay

Patulin (PAT) is a fungal secondary metabolite commonly present in apples and apple products. In the present study, PAT was evaluated for its genotoxic, cytotoxic and cytostatic effects to human peripheral blood lymphocytes by using the cytokinesis-block micronucleus cytome (CBMN Cyt) assay. Lymphocyte cultures were treated with PAT at the following concentrations, 0.1, 0.3, 0.5, 1.0, 2.5, 5.0, and 7.5 μM, as well as 0.5 μM mitomycin c (MMC) as a positive control and dimethyl sulfoxide (DMSO) as a vehicle control. PAT was found to induce nucleoplasmic bridges (NPBs) at 5.0 and 7.5 μM concentrations (P?<?0.05), apoptotic cells at 0.1, 1.0, 5.0 μM (P?<?0.05), 7.5 μM concentrations (P?<?0.01) and necrotic cells at 0.3 and 2.5 μM (P?<?0.05), 5.0 and 7.5 μM (P?<?0.01) concentrations in human lymphocytes. The 2.5, 5.0, and 7.5 μM PAT concentrations also led to a clear decrease in the nuclear division index (NDI) (P?<?0.05). PAT caused a significant dose-dependent increase in the number cells of NPBs, in the frequency of apoptotic and necrotic cells, and a significant dose-dependent decrease in the NDI values in lymphocytes. These results indicate that PAT at high concentrations is genotoxic, cytotoxic and cytostatic in cultured human lymphocytes.     

Assessing the ecological stress in a Garonne River stretch,southwest France

In order to assess the level of ecological stress caused by the pollution from local disturbances in a stretch of the Garonne River, France, we applied the Abundance-Biomass Comparison (ABC) index, using fish assemblages. Data were collected in a 10-year span (1992–2002) in a reference site and in two pollution-exposed sites. The ABC index mean value in the reference site (S1) was 0.03 ± 0.002 (95% Confidence Interval – CI); for the polluted sites (S2 and S3), the values were −0.09 ± 0.002 (95% CI) and −0.12 ± 0.002 (95% CI), respectively. The ABC index showed that, besides flow variations, both downstream sites are statistically different (p < 0.05) from the reference site, but all three seem to be under moderate stress. Furthermore, we related our ABC scores to water quality and flow regime variables in the reference site and one of the polluted sites by means of a cluster analysis. The results showed that, in the reference site, the ABC scores are closely related to the flow regime, while in the polluted site, downstream a urban area, ABC is related to water quality variables such as phosphates and total phosphorous. We argue that ecological indicators can help decisions on environmental damage liability.     

Antioxidant prenylated phenols of Artocarpus plants attenuate ultraviolet radiation-induced damage on human keratinocytes and fibroblasts

Two novel prenylated phenols, cyclocomunoindenol (1) and cyclocomunohexanol (2) were isolated from the cortex of roots of Artocarpus communis. Their structures were determined by spectroscopic methods. Compounds 2 and 6 revealed significant DPPH-scavenging activity with an IC₅₀ values of 435.48 ± 0.93 and 53.55 ± 8.73 μM, respectively. Compounds 1, 2, and 6 displayed significant ABTS⁺ scavenging activity with an IC₅₀ values of 164.26 ± 2.44, 227.01 ± 3.64, and 44.09 ± 0.88 μM, respectively. Compound 6 exhibited an inhibitory effect on XO activity with an IC₅₀ value of 10.91 ± 1.77 μM and the relative oxygen radical absorbance capacity (ORAC) values of 1–6, using ORAC-pyrogallol red (PGR) assay, were determined to be 0.28 ± 0.06, 0.31 ± 0.02, 0.55 ± 0.25, 0.84 ± 0.36, 0.35 ± 0.15, and 0.70 ± 0.09, respectively. Antioxidants 5 and 6 significantly attenuate UVA radiation-induced damage on human HaCaT keratinocytes and Hs68 fibroblasts. These finding showed that 1–6 may be used as antioxidants and 5 and 6 may protect skin against the adverse effects of UV radiation.