Prospecting for candidate molecules from Conus virgo toxins to develop new biopharmaceuticals

Background: A combination of pharmacological and biomedical assays was applied in this study to examine the bioactivity of Conus virgo crude venom in order to determine the potential pharmacological benefit of this venom, and its in vivo mechanism of action. Methods: Two doses (1/5 and 1/10 of LC50, 9.14 and 4.57 mg/kg) of the venom were used in pharmacological assays (central and peripheral analgesic, anti-inflammatory and antipyretic), while 1/2 of LC50 (22.85 mg/kg) was used in cytotoxic assays on experimental animals at different time intervals, and then compared with control and reference drug groups. Results: The tail immersion time was significantly increased in venom-treated mice compared with the control group. Also, a significant reduction in writhing movement was recorded after injection of both venom doses compared with the control group. In addition, only the high venom concentration has a mild anti-inflammatory effect at the late inflammation stage. The induced pyrexia was also decreased significantly after treatment with both venom doses. On the other hand, significant increases were observed in lipid peroxidation (after 4 hours) and reduced glutathione contents and glutathione peroxidase activity, while contents of lipid peroxidation and nitric oxide (after 24 hours) and catalase activity were depleted significantly after venom administration. Conclusion: These results indicated that the crude venom of Conus virgo probably contain bioactive components that have pharmacological activities with low cytotoxic effects. Therefore, it may comprise a potential lead compound for the development of drugs that would control pain and pyrexia.(AU)

Ocular surface and tear film changes in workers exposed to organic solvents used in the dry-cleaning industry.

Workers in the dry-cleaning industry are exposed to organic solvents that may cause eye irritation and tear film changes. Objective To quantify changes in the ocular surface and tear film in dry cleaners exposed to organic solvents and associate these changes with ocular irritation as reported in a symptom questionnaire for dry eye diagnosis. Methods This was a case and control study in which the characteristics and eye-irritation symptoms were compared between two groups of 62 participants that were either exposed or not exposed to organic solvents. A general optometric examination and the following test were performed: lipid interferometry, Lissamine Green Stain, tear breakup time, Schirmer I, conjunctival impression cytology and the Donate dry eye symptoms questionnaire. Results Sixty-five percent of exposed workers obtained a higher score than 13 on the Donate dry eye symptoms questionnaire which indicated the presence of more irritation symptoms than those in the non- exposed group. A Chi-square analysis indicated the exposed group reported significantly higher incidences (P <0.005) for eye irritation symptoms of sandy sensation; tearing eyes sensation; foreign body sensation; tearing; dry eye; dryness; eyestrain and heavy eyelids. A Mann Whitney-U indicated greater severity only for symptoms relating to dry eye; sandy sensation; foreign body sensation, tearing; tearing eyes and dryness. There was a statistically significant difference (P <0.05) for Schirmer I; tear break up time; and the ocular surface assessed with Lissamine green staining and conjunctival impression cytology between groups. A reduction in the thickness of the lipid layer in the exposed group compared to the non-exposed group was observed. Surprisingly, clinical test outcomes were not significantly correlated with dry eye symptoms nor years of exposure. Conclusion Workers in the dry-cleaning industry exposed to organic solvents are associated with changes in ocular surface and tear film generating irritation symptoms commonly present in evaporative dry eye.

Effect of platform subway depth on the presence of Airborne PM2.5, metals, and toxic organic species.

PM2.5 that have been related to public health risks, were collected during two seasons with High-Vol samplers in platforms of a Mexican subway station, which interconnects through transfers three lines having different depths. The objective was to study the influence of depth on the PM2.5 concentrations and their species. PM2.5 concentrations in cold-dry and warm-dry seasons presented statistical differences, being in average 57 and 66 µgm-3 respectively, in the shallower line 9; 90 µgm-3 and 111 µgm-3 in line 1; and 104 and 122 µgm-3in the deepest line 7. During the cold-dry season and warm-dry season PM2.5concentrations in the subway environment were respectively up to 3.5 times and up to 5 times greater than in the ambient air. Like PM2.5, metals analyzed with an OES-ICP presented higher concentrations in deeper lines as well as PAHs quantified with CG-MS, which ranged from 4.5 to 11.7 ngm-3. High PM2.5, metals and organic toxic concentrations found in deeper lines of the subway environment represent a risk for commuters endorsing the need for ventilation systems to reduce them. Zn, Pb, V and Ni in subway particles presented the highest solubility in artificial lysosomal fluid suggesting high bioavailability in the lung fluids.

DNA damage in different wildlife species exposed to persistent organic pollutants (POPs) from the delta of the Coatzacoalcos river, Mexico.

The delta of the Coatzacoalcos river is a priority region for the biological conservation in the Gulf of Mexico. Environmental studies in the area have detected a complex mixture of contaminants where the presence of Persistent organic compounds (POPs) is highlighted. Deoxyribonucleic acid (DNA) integrity of biological populations are global concerns due to their ecological implications. The purpose of this study was to measure the exposure to POPs and DNA damage in nine species residing in the Coatzacoalcos river classified by taxonomic group, type of habitat and feeding habits. Total POPs concentrations (minimum and maximum) detected for all species were from 22.7 to 24,662.1 ng/g l.w; and the values of DNA damage (minimum and maximum) varied from 0.7 to 20.5 and from 6.5 to 56.8 µm (Olive tail moment and tail length respectively). Broadly speaking, reptiles, species residing in the wetland and the ones with a carnivorous diet showed higher levels of POPs and DNA damage. This study provides us with a baseline of the state of POPs contamination and shows the degree of environmental stress to which the different components of the ecosystem of the Coatzacoalcos river delta are subject to.

Predictive ecotoxicity of MoA 1 of organic chemicals using in silico approaches.

Persistent organic products are compounds used for various purposes, such as personal care products, surfactants, colorants, industrial additives, food, pesticides and pharmaceuticals. These substances are constantly introduced into the environment and many of these pollutants are difficult to degrade. Toxic compounds classified as MoA 1 (Mode of Action 1) are low toxicity compounds that comprise nonreactive chemicals. In silico methods such as Quantitative Structure-Activity Relationships (QSARs) have been used to develop important models for prediction in several areas of science, as well as aquatic toxicity studies. The aim of the present study was to build a QSAR model-based set of theoretical Volsurf molecular descriptors using the fish acute toxicity values of compounds defined as MoA 1 to identify the molecular properties related to this mechanism. The selected Partial Least Squares (PLS) results based on the values of cross-validation coefficients of determination (Qcv2) show the following values: Qcv2 = 0.793, coefficient of determination (R2) = 0.823, explained variance in external prediction (Qext2) = 0.87. From the selected descriptors, not only the hydrophobicity is related to the toxicity as already mentioned in previously published studies but other physicochemical properties combined contribute to the activity of these compounds. The symmetric distribution of the hydrophobic moieties in the structure of the compounds as well as the shape, as branched chains, are important features that are related to the toxicity. This information from the model can be useful in predicting so as to minimize the toxicity of organic compounds.

Toxicity and genotoxicity assessment in sediments from the Matanza-Riachuelo river basin (Argentina) under the influence of heavy metals and organic contaminants.

The aim of this study was to investigate the parameters of chemical extraction associated with the detection of toxicity and genotoxicity in sediment sample extracts. Quantitative analysis of metals and polycyclic aromatic hydrocarbons (PAHs), together with a battery of four bioassays, was performed in order to evaluate the extraction efficiency of inorganic and organic toxicants. The extracts were carried out using two inorganic solvents, two organic solvents and two extraction methodologies, making a total of five extracts. Two toxicity tests, the algal growth inhibition of Pseudokirchneriella subcapitata and the root elongation inhibition of Lactuca sativa, and two genotoxicity tests, the analysis of revertants of Salmonella typhimurium and the analysis of micronuclei and chromosomal aberrations in Allium cepa, were performed. According to the chemical analysis, the acidic solution extracted more heavy metal concentrations than distilled water, and dichloromethane extracted more but fewer concentrations of PAH compounds than methanol. Shaker extracts with distilled water were non-toxic to P. subcapitata, but were toxic to L. sativa. The acidic extracts were more toxic to P. subcapitata than to L. sativa. The methanolic organic extracts were more toxic to the alga than those obtained with dichloromethane. None of these extracts resulted toxic to L. sativa. Mutagenic effects were only detected in the organic dichloromethane extracts in the presence of metabolic activation. All the inorganic and organic extracts were genotoxic to A. cepa. This study showed that the implementation of different extraction methods together with a battery of bioassays could be suitable tools for detecting toxicity and genotoxicity in sediment samples.

Toxicidade equivalente da atmosfera por dioxinas, furanos e bifenilas policloradas,com uso de duas técnicas de coleta, passiva e ativa / Equivalent toxicity of the atmosphere for dioxins, furans and polychlorinated biphenyls, using two air sampling techniques, passive and active

Introdução: Dioxinas, furanos e bifenilas policloradas são poluentes tóxicos para a saúde humana incluindo riscos de incidência de cânceres, efeitos de neurodesenvolvimento, lesões dérmicas, cloroacne. Estes compostos são poluentes orgânicos persistentes (POPs) que podem ser transportados de longas distâncias da fonte de emissão e se bioacumular em ecossistemas. A atmosfera poluída foi recentemente classificada como carcinogênica para os seres humanos pela Organização Mundial da Saúde, mostrando a importância de sua caracterização, principalmente para compostos tóxicos. Entretanto, técnica de coleta ativa tem custo elevado para POPs, e existem poucos estudos de calibração que validem a substituição. Objetivos: Avaliar a toxicidade equivalente da atmosfera por dioxinas, furanos e bifenilas cloradas, utilizando técnicas de coleta ativa e passiva, e verificar gradiente de concentração nos ambientes urbano, urbano/industrial e de background. Método: Amostras de ar foram coletadas, utilizando coletores ativos e passivos, durante dois períodos consecutivos de quatro meses: de setembro a dezembro de 2014 (período 1) e de maio a agosto de 2015 (período 2) em três cidades de São Paulo, SP, em ambientes urbano, urbano/industrial e de background. Todas as amostras foram extraídas com solução de tolueno:acetona (9:1) em Soxhlet por 24 h e padrões marcados (13C12-PCDD/Fs e 13C12-PCBs) foram adicionados em cada amostra antes do processo de extração. Os extratos foram purificados em coluna de sílica mista (40 por cento H2SO4 e 10 por cento AgNO3) seguida por coluna de alumina. O procedimento analítico foi realizado utilizando HRGC/HRMS (High Resolution Gas Chromatograph/High Resolution Mass Spectrometer) operando em ionização de impacto de elétrons com energia de 35 eV no modo SIM (Select Ion Monitoring) e resolução de 10.000. Resultados mostraram que: (1) existe variação sazonal para concentrações de PCDD/Fs no ar entre os períodos 1 e 2 (p=0,03), enquanto as concentrações de dl-PCBs não foram estatisticamente diferentes nestes períodos (p=0,52); (2) existe gradiente de concentração de PCDD/Fs e dl- PCBs que aumenta na seguinte ordem: background

Introduction: Dioxins, furans and polychlorinated biphenyls are toxic pollutants for human health including risks of cancer incidence, neurodevelopmental effects, dermal lesions, chloracne. These compounds are persistent organic pollutants (POPs) that can be transported to long distances from the emission source and they are bioaccumulated in ecosystems. Recently, the outdoor air pollution were classified as carcinogenic to humans by the World Health Organization, showing the importance of its characterization for toxic compounds. However, active air monitoring has a high cost for POPs, and there is a few calibration studies which support that substitution. Objective: To assess the equivalent toxicity of the atmosphere regarding the measurement of dioxins, furans and polychlorinated biphenyls, using active and passive air samplers, and to evatuate the contrasting concentrations at urban, urban/industrial and background sites. Method: Air samples were collected, using active and passive samplers, over two consecutive periods of four months: from September to December 2014 (period 1) and from May to August 2015 (period 2) at three cities in São Paulo, SP, covering urban, urban/industrial and background sites. All samples were extracted with toluene:acetone (9:1) in a Soxhlet apparatus for 24 hours and surrogate standards (13C12-PCDD/F and 13C12-PCBs) were spiked on each sample media prior to extraction procedure. The extracts were purified on an silica column (40 per cent H2SO4 and 10 per cent AgNO3) followed by an alumina column. The analytical procedure was carried out using HRGC/HRMS (High Resolution Gas Chromatograph/High Resolution Mass Spectrometer) operating in electron impact ionization with an energy of 35 eV in SIM (selected ion monitoring) mode and 10.000 resolution power. Results show that (1) there are seasonal variations for PCDD/F concentrations in air between period 1 and 2 (p=0.03), whereas dl-PCB levels were not statistically different (p=0.52) in those periods. (2) PCDD/F and dl-PCB air levels are in the following order: background

DNA damage and cytotoxicity in pathology laboratory technicians exposed to organic solvents.

The aim of this study was to evaluate potential DNA damage and cytotoxicity in pathology laboratory technicians exposed to organic solvents, mainly xylene. Peripheral blood and buccal cells samples were collected from 18 technicians occupationally exposed to organic solvents and 11 non-exposed individuals. The technicians were sampled at two moments: Monday and Friday. DNA damage and cytotoxicity were evaluated using the Comet Assay and the Buccal Micronucleus Cytome assay. Fifteen subjects (83.5%) of the exposed group to solvents complained about some symptom probably related to contact with vapours of organic solvents. DNA damage in the exposed group to solvents was nearly 2-fold higher on Friday than on Monday, and in both moments the individuals of this group showed higher levels of DNA damage in relation to controls. No statistical difference was detected in buccal cell micronucleus frequency between the laboratory technicians and the control group. However, in the analysis performed on Friday, technicians presented higher frequency (about 3-fold) of karyolytic and apoptotic-like cells (karyorrhectic and pyknotic) in relation to control group. Considering the damage frequency and the working time, a positive correlation was found in the exposed group to solvents (r=0.468; p=0.05). The results suggest that pathology laboratory workers inappropriately exposed to organic solvents have increased levels of DNA damage.

Response of Rhodococcus erythropolis strain IBBPo1 to toxic organic solvents.

Recently, there has been a lot of interest in the utilization of rhodococci in the bioremediation of petroleum contaminated environments. This study investigates the response of Rhodococcus erythropolis IBBPo1 cells to 1% organic solvents (alkanes, aromatics). A combination of microbiology, biochemical, and molecular approaches were used to examine cell adaptation mechanisms likely to be pursued by this strain after 1% organic solvent exposure. R. erythropolis IBBPo1 was found to utilize 1% alkanes (cyclohexane, n-hexane, n-decane) and aromatics (toluene, styrene, ethylbenzene) as the sole carbon source. Modifications in cell viability, cell morphology, membrane permeability, lipid profile, carotenoid pigments profile and 16S rRNA gene were revealed in R. erythropolis IBBPo1 cells grown 1 and 24 h on minimal medium in the presence of 1% alkanes (cyclohexane, n-hexane, n-decane) and aromatics (toluene, styrene, ethylbenzene). Due to its environmental origin and its metabolic potential, R. erythropolis IBBPo1 is an excellent candidate for the bioremediation of soils contaminated with crude oils and other toxic compounds. Moreover, the carotenoid pigments produced by this nonpathogenic Gram-positive bacterium have a variety of other potential applications.

Response of Rhodococcus erythropolis strain IBBPo1 to toxic organic solvents

Abstract Recently, there has been a lot of interest in the utilization of rhodococci in the bioremediation of petroleum contaminated environments. This study investigates the response of Rhodococcus erythropolis IBBPo1 cells to 1% organic solvents (alkanes, aromatics). A combination of microbiology, biochemical, and molecular approaches were used to examine cell adaptation mechanisms likely to be pursued by this strain after 1% organic solvent exposure. R. erythropolis IBBPo1 was found to utilize 1% alkanes (cyclohexane, n-hexane, n-decane) and aromatics (toluene, styrene, ethylbenzene) as the sole carbon source. Modifications in cell viability, cell morphology, membrane permeability, lipid profile, carotenoid pigments profile and 16S rRNA gene were revealed in R. erythropolis IBBPo1 cells grown 1 and 24 h on minimal medium in the presence of 1% alkanes (cyclohexane, n-hexane, n-decane) and aromatics (toluene, styrene, ethylbenzene). Due to its environmental origin and its metabolic potential, R. erythropolis IBBPo1 is an excellent candidate for the bioremediation of soils contaminated with crude oils and other toxic compounds. Moreover, the carotenoid pigments produced by this nonpathogenic Gram-positive bacterium have a variety of other potential applications.

Exposure to organic solvents used in dry cleaning reduces low and high level visual function.

PURPOSE: To investigate whether exposure to occupational levels of organic solvents in the dry cleaning industry is associated with neurotoxic symptoms and visual deficits in the perception of basic visual features such as luminance contrast and colour, higher level processing of global motion and form (Experiment 1), and cognitive function as measured in a visual search task (Experiment 2). METHODS: The Q16 neurotoxic questionnaire, a commonly used measure of neurotoxicity (by the World Health Organization), was administered to assess the neurotoxic status of a group of 33 dry cleaners exposed to occupational levels of organic solvents (OS) and 35 age-matched non dry-cleaners who had never worked in the dry cleaning industry. In Experiment 1, to assess visual function, contrast sensitivity, colour/hue discrimination (Munsell Hue 100 test), global motion and form thresholds were assessed using computerised psychophysical tests. Sensitivity to global motion or form structure was quantified by varying the pattern coherence of global dot motion (GDM) and Glass pattern (oriented dot pairs) respectively (i.e., the percentage of dots/dot pairs that contribute to the perception of global structure). In Experiment 2, a letter visual-search task was used to measure reaction times (as a function of the number of elements: 4, 8, 16, 32, 64 and 100) in both parallel and serial search conditions. RESULTS: Dry cleaners exposed to organic solvents had significantly higher scores on the Q16 compared to non dry-cleaners indicating that dry cleaners experienced more neurotoxic symptoms on average. The contrast sensitivity function for dry cleaners was significantly lower at all spatial frequencies relative to non dry-cleaners, which is consistent with previous studies. Poorer colour discrimination performance was also noted in dry cleaners than non dry-cleaners, particularly along the blue/yellow axis. In a new finding, we report that global form and motion thresholds for dry cleaners were also significantly higher and almost double than that obtained from non dry-cleaners. However, reaction time performance on both parallel and serial visual search was not different between dry cleaners and non dry-cleaners. CONCLUSIONS: Exposure to occupational levels of organic solvents is associated with neurotoxicity which is in turn associated with both low level deficits (such as the perception of contrast and discrimination of colour) and high level visual deficits such as the perception of global form and motion, but not visual search performance. The latter finding indicates that the deficits in visual function are unlikely to be due to changes in general cognitive performance.

New environmentally friendly MSPD solid support based on golden mussel shell: characterization and application for extraction of organic contaminants from mussel tissue.

The use of golden mussel shells as a solid support in vortex-assisted matrix solid-phase dispersion (MSPD) was evaluated for the first time for extraction of residues of 11 pesticides and nine pharmaceutical and personal care products from mussel tissue samples. After they had been washed, dried, and milled, the mussel shells were characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, infrared spectroscopy, and Brunauer-Emmett-Teller analysis. The MSPD procedure with analysis by liquid chromatography-tandem mass spectrometry allowed the determination of target analytes at trace concentrations (nanograms per gram), with mean recoveries ranging from 61 to 107 % and relative standard deviations lower than 18 %. The optimized method consisted of dispersion of 0.5 g of mussel tissue, 0.5 g of NaSO4, and 0.5 g of golden mussel shell for 5 min, and subsequent extraction with 5 mL of ethyl acetate. The matrix effect was evaluated, and a low effect was found for all compounds. The results showed that mussel shell is an effective material and a less expensive material than materials that have traditionally been used, i.e., it may be used in the MSPD dispersion step during the extraction of pesticides and pharmaceutical and personal care products from golden mussel tissues. Graphical Abstract Vortex-assited matrix solid-phase dispersion for extraction of 11 pesticides and 9 PPCPs care products from mussel tissue samples.

Use, development and improvements in the protocol of whole-sediment toxicity identification evaluation using benthic copepods.

The whole-sediment Toxicity Identification Evaluation (TIE) approach is a useful technique that allows for the identification of the contaminants responsible for the toxicity of complex sediment samples. This study aimed to compare the effectiveness of this technique in identifying the causes of toxicity when the test organism used in the toxicity test is capable of ingesting sediment particles. Two forms of exposure were compared: whole-sediment (WS), which integrates dermic and dietary exposures; and sediment-water interface (SWI), which involves dermic exposure only. The combined analysis of the TIE experiments revealed that metals, ammonia and, at one station, organic compounds, were responsible for sediment toxicity. The integrated use of WS and SWI TIE manipulations provided a more complete overview of the causes of toxicity, and thus enabled a better comprehension of complex contamination situations and, consequently, a better ecological assessment.

[Economic assessment in health and environment from control of persistent organic pollutants in Colombia]. / Valoración económica en salud y medio ambiente del control de contaminantes orgánicos persistentes en Colombia.

Objective To estimate the economic benefits related to environment and health in the context of the implementation of the Stockholm Convention for the control of Persistent Organic Pollutants in the country. METHOD: The estimation was conducted based on two scenarios: non-compliance with the agreement and compliance with the Convention. Gross profit was derived from the difference in present value between the health and environmental costs that are assumed in each scenario. Results Gross profit by decreasing health costs arising from the implementation of the Convention was estimated at USD $ 511 and USD $ 501 million. By introducing variables such as management costs and agreement on potential benefits for access to international markets, the benefits to the country were estimated at between USD $1 631 and USD $ 3 118 million. Discussion Despite the economic benefits generated by lower expenditure on health for the Convention implementation, the costs associated with reducing pollutant emissions generated a negative balance, compensated only by the expectation of higher revenues for international market access. We consider this initial economic assessment an important contribution, but it should be reviewed to include valuation methodologies involving other social profitability variables and different scenarios for emerging technologies, new scientific knowledge about these pollutants, changes in legislation and / or changes in trade agreement conditions, among others.

Long-term occupational exposure to organic solvents affects color vision, contrast sensitivity and visual fields.

The purpose of this study was to evaluate the visual outcome of chronic occupational exposure to a mixture of organic solvents by measuring color discrimination, achromatic contrast sensitivity and visual fields in a group of gas station workers. We tested 25 workers (20 males) and 25 controls with no history of chronic exposure to solvents (10 males). All participants had normal ophthalmologic exams. Subjects had worked in gas stations on an average of 9.6 ± 6.2 years. Color vision was evaluated with the Lanthony D15d and Cambridge Colour Test (CCT). Visual field assessment consisted of white-on-white 24-2 automatic perimetry (Humphrey II-750i). Contrast sensitivity was measured for sinusoidal gratings of 0.2, 0.5, 1.0, 2.0, 5.0, 10.0 and 20.0 cycles per degree (cpd). Results from both groups were compared using the Mann-Whitney U test. The number of errors in the D15d was higher for workers relative to controls (p<0.01). Their CCT color discrimination thresholds were elevated compared to the control group along the protan, deutan and tritan confusion axes (p<0.01), and their ellipse area and ellipticity were higher (p<0.01). Genetic analysis of subjects with very elevated color discrimination thresholds excluded congenital causes for the visual losses. Automated perimetry thresholds showed elevation in the 9°, 15° and 21° of eccentricity (p<0.01) and in MD and PSD indexes (p<0.01). Contrast sensitivity losses were found for all spatial frequencies measured (p<0.01) except for 0.5 cpd. Significant correlation was found between previous working years and deutan axis thresholds (rho = 0.59; p<0.05), indexes of the Lanthony D15d (rho=0.52; p<0.05), perimetry results in the fovea (rho= -0.51; p<0.05) and at 3, 9 and 15 degrees of eccentricity (rho= -0.46; p<0.05). Extensive and diffuse visual changes were found, suggesting that specific occupational limits should be created.

Psychophysical evaluation of achromatic and chromatic vision of workers chronically exposed to organic solvents.

The purpose of this paper was to evaluate achromatic and chromatic vision of workers chronically exposed to organic solvents through psychophysical methods. Thirty-one gas station workers (31.5 ± 8.4 years old) were evaluated. Psychophysical tests were achromatic tests (Snellen chart, spatial and temporal contrast sensitivity, and visual perimetry) and chromatic tests (Ishihara's test, color discrimination ellipses, and Farnsworth-Munsell 100 hue test--FM100). Spatial contrast sensitivities of exposed workers were lower than the control at spatial frequencies of 20 and 30 cpd whilst the temporal contrast sensitivity was preserved. Visual field losses were found in 10-30 degrees of eccentricity in the solvent exposed workers. The exposed workers group had higher error values of FM100 and wider color discrimination ellipses area compared to the controls. Workers occupationally exposed to organic solvents had abnormal visual functions, mainly color vision losses and visual field constriction.

Green chemistry and the evolution of flow analysis. A review.

Flow analysis has achieved its majority as a well-established tool to solve analytical problems. Evolution of flow-based approaches has been analyzed by diverse points of view, including historical aspects, the commutation concept and the impact on analytical methodologies. In this overview, the evolution of flow analysis towards green analytical chemistry is demonstrated by comparing classical procedures implemented with different flow approaches. The potential to minimize reagent consumption and waste generation and the ability to implement processes unreliable in batch to replace toxic chemicals are also emphasized. Successful applications of greener approaches in flow analysis are also discussed, focusing on the last 10 years.

Partially ordered sets: ranking and prediction of substances' properties.

There are at least two significant applications of partial order theory in chemistry: Ranking methods and substances' properties prediction. In both cases, a set of objects is endowed with a partial order relation e.g. "more polluting than", "can be obtained from", "more reactive than" etc. The couple of set and partial order relation is known in mathematics as a partially ordered set (poset). Ranking methods, such as the Hasse diagram technique, lead to a partial order where several incomparabilities (lack of order) appear between pairs of objects. This phenomenon is quite common in ranking studies, and it often is circumvented by a combination of object features leading to a total order. However, such a combination introduces subjectivities and bias in the ranking process. Here a step-by-step procedure is shown to turn incomparabilities into comparabilities taking into account all the possible bias by a linear combination of features. In such a manner, it is possible to predict how probable it is to obtain a particular total order from a given poset. Similarly, it is possible to calculate the needed bias over certain attributes to obtain a particular total order. An example application is shown where substances are ranked according to their bioconcentration factor and biodegradation potential. Another application of partial order theory to chemistry has to do with the prediction of properties for a set of substances related in a (preferably systematic) chemical fashion. A customary relation is "can be obtained from"; if such a relation is set up for a given molecular structure e.g. benzene, and all its substituted derivatives (say chlorinated ones) are considered, then the set of benzene and its chlorinated derivatives are partially ordered. Taking advantage of the poset generated, different methods can be applied to predict properties of the substances considered in the poset. Such methods include the poset-average, cluster expansion, and splinoid methods. In this paper we discuss each one of these methods, its advantages and disadvantages and we outline its applicability to estimate cooperative free energies of hemoglobins with different degree of oxygenation.

Cys mutants in functional regions of Sticholysin I clarify the participation of these residues in pore formation.

Experimental evidence shows that the mechanism of pore formation by actinoporins is a multistep process, involving binding of the water-soluble monomer to the membrane and subsequent oligomerization on the membrane surface, leading to the formation of a functional pore. However, as for other eukaryotic pore-forming toxins, the molecular details of the mechanism of membrane insertion and oligomerization are not clear. In order to obtain further insight with regard to the structure-function relationship in sticholysins, we designed and produced three cysteine mutants of recombinant sticholysin I (rStI) in relevant functional regions for membrane interaction: StI E2C and StI F15C (in the N-terminal region) and StI R52C (in the membrane binding site). The conformational characterization derived from fluorescence and CD spectroscopic studies of StI E2C, StI F15C and StI R52C suggests that replacement of these residues by Cys in rStI did not noticeably change the conformation of the protein. The substitution by Cys of Arg5² in the phosphocholine-binding site, provoked noticeable changes in rStI permeabilizing activity; however, the substitutions in the N-terminal region (Glu², Phe¹5) did not modify the toxin's permeabilizing ability. The presence of a dimerized population stabilized by a disulfide bond in the StI E2C mutant showed higher pore-forming activity than when the protein is in the monomeric state, suggesting that sticholysins pre-ensembled at the N-terminal region could facilitate pore formation.

DNA damage in earthworms (Eisenia spp.) as an indicator of environmental stress in the industrial zone of Coatzacoalcos, Veracruz, Mexico.

Coatzacoalcos, Veracruz is one of the major industrial areas of Mexico. Presently, the Coatzacoalcos River and the areas surrounding the industrial complex are considered by various authors to be some of most polluted sites in Mexico. The objective of this study was to determine if earthworms could be used as indicators of environmental stress in the Coatzacoalcos industrial zone. Often, detritivores and decomposers such as earthworms are the first to be affected when the soil is contaminated. We collected soil samples to be used for persistent organic pollutants (POPs) quantification by gas chromatography. Concentrations of hexachlorobenzene, lindane and total polychlorinated biphenyls (PCBs) in the soil were above the maximum permissible limits of the Canadian Environmental Quality Guidelines (CEQG). Comet assay was conducted in coelomocytes of wild earthworms collected in Coatzacoalcos and compared with the control earthworms. We found DNA damage in earthworms from Coatzacoalcos that was significantly higher (P < 0.05) in comparison to laboratory earthworms. Earthworms are an appropriate organism to use as an indicator of environmental impact in contaminated sites. DNA damage recorded in the earthworms provides clear evidence of environmental impacts by the chemical industry on the wildlife of this region.