Electrochemical sensors, MTT and immunofluorescence assays for monitoring the proliferation effects of cissus populnea extracts on Sertoli cells

<p>Abstract</p> <p>Background</p> <p>We describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells.</p> <p>Methods</p> <p>The proliferation activities of the extracts on Sertoli cells were studied using a high-throughput electrochemical sensor array (DOX-96) and the analytical sensor characteristics were compared with conventional colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and fluorescence spectroscopy.</p> <p>Results</p> <p>This work shows that there is a definite positive trend in the proliferation effect of the extract of <it>Cissus populnea </it>on the TM4 Sertoli cells. All of the three techniques confirmed that the most effective concentration for the proliferation is 10 ppm. At this concentration, the proliferation effect was established around 120% for both DOX-96 and MTT techniques, whereas fluorescence assays showed a higher level (120-150%). DOX-96 showed a lower limit of detection (1.25 × 10(4) cells/ml); whereas the LOD recorded for both MTT and fluorescence techniques was 2.5 × 10(4) cells/ml. Visual examination of the cells by means of confocal fluorescence microscopy confirmed the proliferation of Sertoli cells as was determined using the MTT assay. This investigation provides a confident interpretation of the results and proved that the most effective concentration for the proliferation using <it>Cissus populnea </it>plant extract is 10 ppm.</p> <p>Conclusions</p> <p>Overall, the DOX results compared well with the conventional methods of checking proliferation of cells. The fascinating feature of the sensor array is the ability to provide continuous proliferation experiments with no additional reagents including 96 simultaneous electrochemical experiments. The use of the DOX-96 could reduce a typical bioassay time by 20-fold. Thus the DOX-96 can be used as both a research tool and for practical cell culture monitoring.</p

A framework of criteria for the sustainability assessment of nanoproducts

AbstractNanotechnology applications (nanoproducts) have entered the market or are expected to do so in the near future. Robust and science-based criteria are required to appraise and manage their sustainability. This paper describes the approach used to develop a comprehensive and reliable framework of criteria, which was missing until now, for evaluating the sustainability of nanoproducts. A literature review of the frameworks and tools employed to assess nanoproducts sustainability implications was firstly performed to select an initial set of criteria. A survey of experts in the sustainable nanotechnology domain was then conducted to elicit their knowledge in terms of completeness, reliability and validity of the criteria set. Ranking and correlation analyses completed the research by identifying the parameters of major interest as well as the links and dependencies between them. A total of 54 and 65 experts replied to the pilot and main survey, respectively. The reliability and validity of the criteria was assessed with the responses from both questionnaires, whereas the answers from the main survey were used to calculate the relative index of the criteria as well as their correlations. This research resulted in a framework composed of 68 criteria, which are structured into six main areas: (i) economic performance; (ii) environmental impacts, (iii) environmental risk assessment; (iv) human health risk assessment; (v) social implications and (vi) technical performance. This study helps to broaden the understanding on the identification of criteria for sustainability assessments. It also provides those interested in evaluating nanotechnology implications with the basis for real case studies, possibly by integrating available information with the stakeholders using tools that support decision-making

Co-exposure risks of pesticides residues and bacterial contamination in fresh fruits and vegetables under smallholder horticultural production systems in Tanzania.

This research article published by PLOS ONE,2020This study was carried out to investigate the risks of simultaneous exposure to pesticide residues and bacteria contaminants in locally produced fresh vegetables and vegetables in Tanzania. A total of 613 samples were analyzed for pesticide residues, out of which 250 were also analyzed for bacterial contamination. Overall, 47.5% had pesticide residues, 74.2% exceeded Maximum Residue Levels (MRLs). Organophosphorus (95.2%), organochlorines (24.0%), pyrethroids (17.3%), and carbamates (9.2%) residues dominated. MRL values were mostly exceeded in tomatoes, onions, watermelons, cucumbers, Chinese cabbage, and sweet paper. Tetramethrin (0.0329-1.3733 mg/kg), pirimiphos-methyl (0.0003-1.4093 mg/kg), permethrin (0.0009-2.4537 mg/kg), endosulfan (beta) (0.0008-2.3416 mg/kg), carbaryl (0.0215-1.5068 mg/kg), profenofos (0.0176-2.1377 mg/kg), chlorpyrifos (0.0004-1.2549 mg/kg) and dieldrin (0.0011-0.5271 mg/kg) exceeded MRLs. The prevalence of bacteria contamination was high (63.2%). Enterobacter (55.6%) Pseudomonas aeruginosa (32.4%), E. coli (28.2%), Citrobacter (26.8%), Klebsiella oxytoca (14.8%), and Salmonella (7.7%) were isolated. Furthermore, 46.4% tested positive for both pesticide residues and bacterial contaminants. Vegetables from farms (60.7%) contained more dual contaminants than market-based vegetables (41.8%). This may have resulted from excessive pesticide use and unhygienic handling of fresh fruits and vegetables at production level. Binary logistic regression showed that fresh fruits and vegetables with pesticide residues were 2.231 times more likely to have bacteria contaminants (OR: 2.231; 95% CI: 0.501, 8.802). The contamination levels of pesticide residues and bacterial contaminants could be perceived as a serious problem as most fresh fruits and vegetables recorded values of pesticide residues far above the MRLs with pathogenic bacteria isolated in higher proportions. MRLs was higher in most vegetables consumed raw or semi-cooked such as watermelons, carrots, cucumber, tomatoes, onion and sweet paper. There is an urgent need to develop pesticide monitoring and surveillance systems at farmer level, educating farmers and promoting the use of greener pesticides to mitigate the health effects of pesticides and bacterial contaminants