Síntese de moléculas contendo a unidade de piperazina como potenciais agentes antipsicóticos

Tese de Doutoramente em Ciências (área de especialização em Química).

Enzymatic phosphorylation of hair keratin enhances fast adsorption of cationic moieties

The current study describes the in vitro phosphorylation of a human hair keratin, using protein kinase for the first time. Phosphorylation of keratin was demonstrated by 31P NMR (Nuclear Magnetic Resonance) and Diffuse Reflectance Infrared Fourier Transform (DRIFT) techniques. Phosphorylation induced a 2.5 fold increase of adsorption capacity in the first 10 minutes for cationic moiety like Methylene Blue (MB). Thorough description of MB adsorption process was performed by several isothermal models. Reconstructed fluorescent microscopy images depict distinct amounts of dye bound to the differently treated hair. The results of this work suggest that the enzymatic phosphorylation of keratins might have significant implications in hair shampooing and conditioning, where short application times of cationic components are of prime importance.

Nickel(II) complexes of bidentate N-N′ ligands containing mixed pyrazole, pyrimidine and pyridine aromatic rings as catalysts for ethylene polymerisation

This work describes the synthesis and characterisation of Ni(II) complexes of the following neutral bidentate nitrogen ligands containing pyrazole (pz), pyrimidine (pm) and pyridine (py) aromatic rings: 2-pyrazol-1-yl-pyrimidine (pzpm), 2-(4-methyl-pyrazol-1-yl)-pyrimidine (4-Mepzpm), 2-(4-bromo-pyrazol-1-yl)-pyrimidine (4-Brpzpm), 2-(3,5-dimethyl-pyrazol-1-yl)-pyrimidine (pz*pm), 2-pyrazol-1-yl-pyridine (pzpy) and bis(3,5-dimethylpyrazol-1-yl)phenylmethane (bpz*mph). The complexes [NiBr2(pzpm)] (1), [NiBr2(4-Mepzpm)] (2), [NiBr2(4-Brpzpm)] (3), [NiBr2(pz*pm)] (4), [NiBr2(pzpy)] (5) and [NiBr2(bpz*mph)] (6) were tested as catalysts for ethylene polymerisation, in the presence of the cocatalysts methylaluminoxane (MAO) or diethylaluminium chloride (AlEt2Cl), the catalyst systems 1-3/MAO showing moderate to high activities up to the temperature of 20 °C only in the presence of MAO, whereas 4-6/MAO revealed to be inactive. Other related Pd(II) complexes, already reported in previous works, such as [PdClMe(pzpm)], [PdClMe(pz*pm)], [PdClMe(pzpy)] and [PdClMe(bpz*mph)], also showed to be inactive in the polymerisation of ethylene, when activated by MAO or AlEt2Cl. Selected samples of polyethylene products were characterised by GPC/SEC, 1H and 13C NMR and DSC, showing to be low molecular weight polymers with Mn values ranging from ca. 550 to 1500 g mol−1 and unusually low dispersities of 1.2–1.7, with total branching degrees generally varying between 2 and 12%, melting temperatures from 40 to 120 °C and crystallinities from 40 to 70%.

Aromatic amines sources, environmental impact and remediation

Aromatic amines are widely used industrial chemicals as their major sources in the environment include several chemical industry sectors such as oil refining, synthetic polymers, dyes, adhesives, rubbers, perfume, pharmaceuticals, pesticides and explosives. They result also from diesel exhaust, combustion of wood chips and rubber and tobacco smoke. Some types of aromatic amines are generated during cooking, special grilled meat and fish, as well. The intensive use and production of these compounds explains its occurrence in the environment such as in air, water and soil, thereby creating a potential for human exposure. Since aromatic amines are potential carcinogenic and toxic agents, they constitute an important class of environmental pollutants of enormous concern, which efficient removal is a crucial task for researchers, so several methods have been investigated and applied. In this chapter the types and general properties of aromatic amine compounds are reviewed. As aromatic amines are continuously entering the environment from various sources and have been designated as high priority pollutants, their presence in the environment must be monitored at concentration levels lower than 30 mg L1, compatible with the limits allowed by the regulations. Consequently, most relevant analytical methods to detect the aromatic amines composition in environmental matrices, and for monitoring their degradation, are essential and will be presented. Those include Spectroscopy, namely UV/visible and Fourier Transform Infrared Spectroscopy (FTIR); Chromatography, in particular Thin Layer (TLC), High Performance Liquid (HPLC) and Gas chromatography (GC); Capillary electrophoresis (CE); Mass spectrometry (MS) and combination of different methods including GC-MS, HPLC-MS and CE-MS. Choosing the best methods depend on their availability, costs, detection limit and sample concentration, which sometimes need to be concentrate or pretreated. However, combined methods may give more complete results based on the complementary information. The environmental impact, toxicity and carcinogenicity of many aromatic amines have been reported and are emphasized in this chapter too. Lately, the conventional aromatic amines degradation and the alternative biodegradation processes are highlighted. Parameters affecting biodegradation, role of different electron acceptors in aerobic and anaerobic biodegradation and kinetics are discussed. Conventional processes including extraction, adsorption onto activated carbon, chemical oxidation, advanced oxidation, electrochemical techniques and irradiation suffer from drawbacks including high costs, formation of hazardous by-products and low efficiency. Biological processes, taking advantage of the naturally processes occurring in environment, have been developed and tested, proved as an economic, energy efficient and environmentally feasible alternative. Aerobic biodegradation is one of the most promising techniques for aromatic amines remediation, but has the drawback of aromatic amines autooxidation once they are exposed to oxygen, instead of their degradation. Higher costs, especially due to power consumption for aeration, can also limit its application. Anaerobic degradation technology is the novel path for treatment of a wide variety of aromatic amines, including industrial wastewater, and will be discussed. However, some are difficult to degrade under anaerobic conditions and, thus, other electron acceptors such as nitrate, iron, sulphate, manganese and carbonate have, alternatively, been tested.

Aproximación al manejo de Nemátodos Fitófagos de los genéros nacobbus y meloidogyne en la Provincia de Córdoba: cultivos hortícolas y aromáticos. Approach to the management of phytophagous nematodes of de genera nacobbus and meloidogyne in the province of Córdoba: horticultural and aromatic crops.

El proyecto que se propone es continuación de investigaciones relacionadas con dos géneros de nematodos fitófagos de interés para cultivos hortícolas (Nacobbus y Meloidogyne) y plantas aromáticas (Meloidogyne) en la Provincia de Córdoba. Hasta el momento, los conocimientos sobre estos parásitos son limitados, de allí la justificación del tema propuesto. Se realizarán muestreos de los citados cultivos, desarrollados a campo y bajo cubierta, para identificar poblaciones de esos nematodos perjudiciales. Se caracterizará morfológicamente poblaciones de N. aberrans con el objeto de consolidar criterios que permitan su reconocimiento. Se llevarán a cabo estudios de relaciones nematodo-hospedador poniendo énfasis en aspectos para los cuales no existen antecedentes en el país. Por un lado, se evaluará el grado de susceptibilidad de diferentes cultivares de pimiento a la agresión de poblaciones de N. aberrans de reconocida patogenicidad para el cultivo. Por otro, se estudiará el efecto de la colonización de micorrizas arbusculares en raíces de tomate y pimiento, como agente que brindaría resistencia al ataque de N. aberrans. Cabe hacer notar que con respecto a la interacción del complejo representado por N. aberrans-planta-micorrizas no ha sido publicada información alguna a nivel mundial. La importancia del proyecto reside en la posibilidad de obtener datos originales, transferibles al sector de producción agrícola. Al mismo tiempo, contribuirá a definir estrategias alternativas de manejo de problemas ocasionados por nematodos fitófagos, basadas en aspectos biológicos y no en la utilización sistemática de agroquímicos contaminantes.

Inducción sistémica de la biosintesis de metabolitos secundarios en plantas aromáticas mediada por rizobacterias. Systemic induction of secondary metablolites biosynthesis in aromatic plants by rhizobacteria.

El uso de microorganismos como inoculantes para incrementar la disponibilidad y toma de nutrientes por parte de los cultivos, es una nueva tecnología que ha dado buenos resultados, observándose un incremento en la emergencia, vigor, mayor desarrollo en la parte aérea y de raíces, registrándose aumentos considerables de los rendimientos en cultivos de interés comercial. Esto es debido a que los microorganismos PGPR (Plant Growth promoting rhizobacteria) sintetizan ciertas sustancias reguladoras del crecimiento como giberelinas, citoquininas y auxinas; las cuales estimulan la densidad y longitud de los pelos radicales, aumentando así la cantidad y longitud de las raíces de los vegetales. Así, se incrementa la capacidad de absorción de agua y nutrientes, haciendo que las plantas sean más vigorosas, productivas y tolerantes a condiciones climáticas adversas, como sequías o heladas. Otro factor benéfico es que ciertos microorganismos solubilizan nutrientes poco móviles en el suelo como el caso del fósforo, segundo nutriente, después del nitrógeno en importancia para el crecimiento de los cultivos. Estos microorganismos también tienen una función muy importante en el control natural de agentes patógenos, a través de la inducción del sistema de defensa en las plantas, aumentando su resistencia a enfermedades, a través de la producción de compuestos bacterianos como antibióticos y sideróforos. Los variados mecanismos mediante los cuales la acción PGPR se lleva a cabo no son plenamente conocidos y, por lo tanto, es necesario determinar con precisión su efecto particular en la biología de la planta beneficiada. Las plantas aromáticas y medicinales inoculadas con microorganismos (rizobacterias) registran un incremento en varios parámetros de crecimiento vegetal (peso fresco parte aérea, peso seco de raíz, número de hojas, etc) y en el rendimiento de aceite esencial (AE). El aumento de la síntesis, y la variación de los porcentajes relativos de los componentes principales de AE en plantas aromáticas, como efecto de la inoculación, podría considerarse como una respuesta defensiva de la planta frente a la colonización de microorganismos dado que varios AE poseen propiedades antimicrobianas. El incremento de estos metabolitos también se ha registrado como respuesta frente a la herbivoría. En el presente proyecto se propone dilucidar la existencia de una relación entre las defensas inducidas por rizobacterias con la producción de metabolitos secundarios en plantas aromaticas y medicinales. The use of microorganisms as inoculants to increase the availability and nutrient uptake by crops, is a new technology that has been successfully applied, with an increase in the emergence, vigor, greater development in the shoot and roots, recording significant increases in yields of crop with commercial interest. This is because microorganisms PGPR (Plant Growth Promoting rhizobacteria) synthesize certain growth regulating substances such as gibberellins, cytokinins and auxins, which stimulate the density and length of root hairs, increasing the number and length of roots. Thus, increase the capacity of absorbing water and nutrients, make the plants more vigorous, productive and tolerant to adverse climatic conditions such as drought or frost.Another beneficial factor is that some microorganisms solubilize nutrients mobile in the soil as the case of phosphorus, second nutrient after nitrogen important for plant growth. These organisms also have an important role in the natural control of pathogens through the induction of the plants defense system, increasing their resistance to disease through the production of compounds such as antibiotics and bacterial siderophores. The various mechanisms by which PGPR action takes place are not fully known and therefore it is necessary to accurately determine its particular effect on the biology of the specific plant benefit. Aromatic and medicinal plants inoculated with microorganisms (rhizobacteria) recorded an increase in several parameters of plant growth (shoot fresh weight, root dry weight, leaf number, etc) and essential oil yield (AE). The increase in the biosynthesis, and changes in the relative percentages of the main components of AE in aromatic plants inoculated with rizobacterias, could be regarded as a plant defense response against microbial colonization, since several AE have antimicrobial properties. The increase of these metabolites have also been recorded as a response to herbivory.

The use of forensic polycyclic aromatic hydrocarbon signatures and Compound Ratio Analysis Techniques (CORAT) for the source characterisation of petrogenic/pyrogenic environmental releases.

This study utilised recent developments in forensic aromatic hydrocarbon fingerprint analysis to characterise and identify specific biogenic, pyrogenic and petrogenic contamination. The fingerprinting and data interpretation techniques discussed include the recognition of: The distribution patterns of hydrocarbons (alkylated naphthalene, phenanthrene, dibenzothiophene, fluorene, chrysene and phenol isomers), • Analysis of “source-specific marker” compounds (individual saturated hydrocarbons, including n-alkanes (n-C5 through 0-C40) • Selected benzene, toluene, ethylbenzene and xylene isomers (BTEX), • The recalcitrant isoprenoids; pristane and phytane and • The determination of diagnostic ratios of specific petroleum / non-petroleum constituents, and the application of various statistical and numerical analysis tools. An unknown sample from the Irish Environmental Protection Agency (EPA) for origin characterisation was subjected to analysis by gas chromatography utilising both flame ionisation and mass spectral detection techniques in comparison to known reference materials. The percentage of the individual Polycyclic Aromatic Hydrocarbons (PAIIs) and biomarker concentrations in the unknown sample were normalised to the sum of the analytes and the results were compared with the corresponding results with a range of reference materials. In addition, to the determination of conventional diagnostic PAH and biomarker ratios, a number of “source-specific markers” isomeric PAHs within the same alkylation levels were determined, and their relative abundance ratios were computed in order to definitively identify and differentiate the various sources. Statistical logarithmic star plots were generated from both sets of data to give a pictorial representation of the comparison between the unknown sample and reference products. The study successfully characterised the unknown sample as being contaminated with a “coal tar” and clearly demonstrates the future role of compound ratio analysis (CORAT) in the identification of possible source contaminants.

Burkholderia sartisoli sp. nov., isolated from a polycyclic aromatic hydrocarbon-contaminated soil.

A Gram-negative, rod-shaped, aerobic bacterium, designated strain RP007(T), was isolated from a polycyclic aromatic hydrocarbon-contaminated soil in New Zealand. Two additional strains were recovered from a compost heap in Belgium (LMG 18808) and from the rhizosphere of maize in the Netherlands (LMG 24204). The three strains had virtually identical 16S rRNA gene sequences and whole-cell protein profiles, and they were identified as members of the genus Burkholderia, with Burkholderia phenazinium as their closest relative. Strain RP007(T) had a DNA G+C content of 63.5 mol% and could be distinguished from B. phenazinium based on a range of biochemical characteristics. Strain RP007(T) showed levels of DNA-DNA relatedness towards the type strain of B. phenazinium and those of other recognized Burkholderia species of less than 30 %. The results of 16S rRNA gene sequence analysis, DNA-DNA hybridization experiments and physiological and biochemical tests allowed the differentiation of strain RP007(T) from all recognized species of the genus Burkholderia. Strains RP007(T), LMG 18808 and LMG 24204 are therefore considered to represent a single novel species of the genus Burkholderia, for which the name Burkholderia sartisoli sp. nov. is proposed. The type strain is RP007(T) (=LMG 24000(T) =CCUG 53604(T) =ICMP 13529(T)).

Physiology and transcriptome of the polycyclic aromatic hydrocarbon-degrading Sphingomonas sp. LH128 after long-term starvation.

The survival, physiology and gene expression profile of the phenanthrene-degrading Sphingomonas sp. LH128 was examined after an extended period of complete nutrient starvation and compared with a non-starved population that had been harvested in exponential phase. After 6 months of starvation in an isotonic solution, only 5 % of the initial population formed culturable cells. Microscopic observation of GFP fluorescent cells, however, suggested that a larger fraction of cells (up to 80 %) were still alive and apparently had entered a viable but non-culturable (VBNC) state. The strain displayed several cellular and genetic adaptive strategies to survive long-term starvation. Flow cytometry, microscopic observation and fatty acid methyl ester (FAME) analysis showed a reduction in cell size, a change in cell shape and an increase in the degree of membrane fatty acid saturation. Transcriptome analysis showed decreased expression of genes involved in ribosomal protein biosynthesis, chromosomal replication, cell division and aromatic catabolism, increased expression of genes involved in regulation of gene expression and efflux systems, genetic translocations, and degradation of rRNA and fatty acids. Those phenotypic and transcriptomic changes were not observed after 4 h of starvation. Despite the starvation situation, the polycyclic aromatic hydrocarbon (PAH) catabolic activity was immediate upon exposure to phenanthrene. We conclude that a large fraction of cells maintain viability after an extended period of starvation apparently due to tuning the expression of a wide variety of cellular processes. Due to these survival attributes, bacteria of the genus Sphingomonas, like strain LH128, could be considered as suitable targets for use in remediation of nutrient-poor PAH-contaminated environments.

Generation of polycyclic aromatic hydrocarbons (PAHs) during woodworking operations.

Occupational exposures to wood dust have been associated with an elevated risk of sinonasal cancer (SNC). Wood dust is recognized as a human carcinogen but the specific cancer causative agent remains unknown. One possible explanation is a co-exposure to; wood dust and polycyclic aromatic hydrocarbons (PAHs). PAHs could be generated during incomplete combustion of wood due to heat created by use of power tools. To determine if PAHs are generated from wood during common wood working operations, PAH concentrations in wood dust samples collected in an experimental chamber operated under controlled conditions were analyzed. In addition, personal air samples from workers exposed to wood dust (n = 30) were collected. Wood dust was generated using three different power tools: vibrating sander, belt sander, and saw; and six wood materials: fir, Medium Density Fiberboard (MDF), beech, mahogany, oak and wood melamine. Monitoring of wood workers was carried out by means of personal sampler device during wood working operations. We measured 21 PAH concentrations in wood dust samples by capillary gas chromatography-ion trap mass spectrometry (GC-MS). Total PAH concentrations in wood dust varied greatly (0.24-7.95 ppm) with the lowest being in MDF dust and the highest in wood melamine dust. Personal PAH exposures were between 37.5-119.8 ng m(-3) during wood working operations. Our results suggest that PAH exposures are present during woodworking operations and hence could play a role in the mechanism of cancer induction related to wood dust exposure.

Occupational exposure of truck drivers to dust and polynuclear aromatic hydrocarbons: a pilot study in Geneva, Switzerland.

The exposure to dust and polynuclear aromatic hydrocarbons (PAH) of 15 truck drivers from Geneva, Switzerland, was measured. The drivers were divided between "long-distance" drivers and "local" drivers and between smokers and nonsmokers and were compared with a control group of 6 office workers who were also divided into smokers and nonsmokers. Dust was measured on 1 workday both by a direct-reading instrument and by sampling. The local drivers showed higher exposure to dust (0.3 mg/m3) and PAH than the long-distance drivers (0.1 mg/m3), who showed no difference with the control group. This observation may be due to the fact that the local drivers spend more time in more polluted areas, such as streets with heavy traffic and construction sites, than do the long-distance drivers. Smoking does not influence exposure to dust and PAH of professional truck drivers, as measured in this study, probably because the ventilation rate of the truck cabins is relatively high even during cold days (11-15 r/h). The distribution of dust concentrations was shown in some cases to be quite different from the expected log-normal distribution. The contribution of diesel exhaust to these exposures could not be estimated since no specific tracer was used. However, the relatively low level of dust exposure dose not support the hypothesis that present day levels of diesel exhaust particulates play a significant role in the excess occurrence of lung cancer observed in professional truck drivers.