Antioxidant and cytotoxic activities of 'açaí' (Euterpe precatoria Mart.)

Decoction of roots of the Amazonian palm açaí (Euterpe precatoria Mart.) is widely used by Brazilian and Peruvian people as an anti-inflammatory, to heal kidney and liver diseases and against snake bites. In this study, the antioxidant activity of extracts and flavonoids (quercetin, catechin, epicatechin, rutin and astilbin) isolated from roots and leaf stalks of E. precatoria was investigated using β-carotene in TLC plates and DPPH radical scavenging in a spectrophotometric bioassay. All extracts and flavonoids showed activity. Also, the cytotoxic activity of these extracts was evaluated by the brine shrimp (Artemia salina) larvicide bioassay and was lower than that of lapachol, used as control. The presence of flavonoids and sitosterol-3-O-β-D-glucopyranoside in the extracts can justify the use of the plant in traditional medicine.

Toxicidade e atividade antioxidante de flavonoides das cascas das raízes de Lonchocarpus filipes

The phytochemical investigation of dichloromethane extract from root bark of Lonchocarpus filipes Benth (Leguminosae) afforded four flavonoids including three dibenzoylmethane derivatives rarely found in nature. The structures were established based on their spectral data (¹H and 13C NMR, 2D-NMR) as being: lanceolatin B (1), pongamol (2), (E)-7-O-methylpongamol (3) and (E)-9-O-methylpongamol (4). Compound (4) is described herein for the first time as a natural product. The extracts and the isolated compounds (1), (2) and (3) displayed high toxicity in the brine shrimp lethality assay. Only compound (2) showed antioxidant activity using a DPPH radical scavenging assay. This is the first report on the phytochemical study of Lonchocarpus filipes.

Purificação e caracterização da quitinase de uva (Vitis vinífera L. cv Red Globe) para a produção de quitosana a partir de quitina de camarão

Chitinase is produced by a wide variety of plants as a defense against peste attacks. In this study, grape chitinases were purified 16 times by fractionation in 80% ammonium sulfate followed by dialysis and filtration. Purified chitinases exhibited enzymatic activity toward chitin azure. The yield of purified chitinase was 229 mg/L with chitinase activity of 563 U/g. Chitinases had molecular masses of 24 and 30 kDa, as evaluated by SDS-PAGE 12.5%. Two pH optima were determined 3.0 and 6.0. The optimal temperature was 42 °C. Pre hydrolysis of crystalline shrimp chitin by chitinases caused in an increase in the deacetylation ratio triggered by chitin deacetylase producing chitooligosaccharides with DA (degree acetylation) of 58.8%.

Otimização e validação de método multirresíduo para determinação de sulfonamidas em camarão cultivado por cromatografia líquida de alta eficiência com detecção por UV

This work reports the optimization and method validation for sulfonamides (sulfamethazine, sulfaquinoxaline, sulfadimethoxine and sulfathiazole) in shrimp muscle using HPLC-UV. The sulfonamides were extracted with acetonitrile and acetic acid, and the extract cleaned up with a Strata SCX SPE cartridge prior to analysis. The method presented linearity in the range of 20-120 µg kg-1, good linear correlation (r > 0.99), and limits of quantification in the range of 4.7-20.2 µg kg-1. The recovery for shrimp muscles spiked with 50-150 µg kg-1 ranged from 63.2-108.0%. Precision and accuracy analysis showed acceptable relative standard deviation. Commercial shrimps were analyzed and sulfonamides don't were found above of the method limit of quantification.

Synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides: evaluation of antioxidant and antimicrobial activities

A facile one-step synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides and TMSA was described. This paper determines their antimicrobial activity against nine human bacterial pathogens by the broth microdilution method; antioxidant activity by DPPH• inactivation and a ferric-reducing power assay; and toxicity by a brine shrimp, Artemia salina, assay. The 1H-benzoxazine-2,4-dione yields were in the range of 57 to 98%. The novel compound 1H-pyrazino[2,3-][1,3]oxazine-2,4-dione 4c showed the highest antioxidant capacity (DPPH 35.4% and FRAP 0.063 µmol TEs/µmol).

Hydroksihappojen erotus mustalipeästä nanosuodatuksella

Hydroksihapoille on olemassa useita käyttömahdollisuuksia teollisuudessa, joten niiden hyödyntäminen sellunvalmistuksen sivuvirrasta eli mustalipeästä on suuren kiinnostuksen kohteena. Tässä työssä selvitettiin, onko hydroksihappojen erotus ja puhdistus mustalipeästä mahdollista nanosuodatuksella. Kokeellisessa osassa suodatettiin emäksisen mustalipeän lisäksi hapotettua ja jäähdytyskiteytettyä mustalipeää, johon oli lisätty liuotinta. Mustalipeäsuodatuksissa käytettiin viittä erilaista nanosuodatusmembraania (Microdyn Nadir® NP010 ja NP030, Dow Chemical Company NF-90, Woongjin Chemical NE-70 sekä Ge-Osmonics Desal 5 DK). Kirjallisuusosassa käsiteltiin puun sisältämien yhdisteiden kemiallista koostumusta, sellun valmistuksen pääperiaatteita, mustalipeän ja hydroksihappojen ominaisuuksia sekä hydroksihappojen käyttömahdollisuuksia. Lisäksi tarkasteltiin erilaisia hydroksihappojen erotusmenetelmiä, nanosuodatuksen teoriaa ja prosessiin sopivan membraanin valintakriteerejä. Työn kokeellisessa osassa tutkittiin emäksisen mustalipeän monivaiheisen nanosuodatuksen tehokkuutta hydroksihappojen erotuksessa. Hapotetun ja jäähdytyskiteytetyn mustalipeän suodatuskokeissa tutkittiin erityyppisten membraanien erotuskykyä sekä syötön liuotinlisäyksen vaikutusta hydroksihappojen erottumiseen. Lisäksi tarkasteltiin membraanien kestävyyttä ja foulaantumista suodatusolosuhteissa. Työn tulokset osoittivat, että hydroksihappoja voidaan fraktioida mustalipeästä nanosuodatuksella. Hydroksihappojen fraktiointiin vaikuttaa merkittävästi mustalipeässä käytetyn liuottimen läsnäolo sekä suodatuspaine. Lisäksi koetulosten perusteella havaittiin, että monivaiheisella nanosuodatuksella hydroksihapot läpäisevät membraanin ja permeaattiin saavutetaan puhtaampi happojae.

Hitsauksen tehostaminen robotisoinnin käyttöönotolla

Kansainvälisen kilpailun kiristyessä yritysten on tehostettava tuotantoaan kustannusten alentamiseksi. Eräs tehostamisen keino on tuotannon automatisointi. Tässä diplomityössä käsitellään hitsauksen tehostamista robottiaseman käyttöönotolla GaV Group Oy:ssä. Työn tarkoituksena on tuoda esille oleellisimmat uuden valmistusteknologian tehokkaaseen käyttöönottoon kuuluvat asiat. Teoriaosassa käsitellään robottisolun investoinnin jälkeisiä toimenpiteitä esivalmistelusta, käyttöönotosta ja tuotannon stabiloinnista. Pääpaino teoriaosuudessa on esivalmistelulla, jolla on suurin vaikutus uuden teknologian käyttöönotossa. Lisäksi teoriaosuudessa käydään läpi laadunvarmistuksen pääperiaatteet, turvallisuusnäkökulmat ja muutoksen mittareiden määrittäminen. Käytännön osiossa on esitelty robottisolun käyttöönotto kohdeyrityksessä. Työn alussa valittiin robotisoidusti hitsattavat kappaleet, parannettiin mahdollisuuksien mukaan konstruktioita paremmin robotisoituun hitsaukseen soveltuviksi ja rakennettiin niille hitsauskiinnittimet. Robotilla tuotteille etsittiin sopivat parametrit ja tehtiin hitsausohjelmat tämän työn puitteissa. Työssä esitellään myös tuotannon käynnistyminen laatuongelmineen ja tuotannon tunnusluvut. Hitsauksen lisäksi robottiin liitettiin esivalmistuksen laadun takaamiseksi hienosädeplasma, jolla aukotetaan robotilla hitsattavia tuotteita. Hienosädeplasmalla saadaan entistä tarkemmat ja railopinnoiltaan siistit reiät yhteille, joka on välttämätöntä robotisoidussa hitsauksessa.

Rational use of water in a poultry slaughterhouse in the state of Paraná, Brazil: a case study

Agroindustries are major consumers of water. However, to adapt to environmental trends and be competitive in the market, they have sought rational use of water through water management in their activities. Cleaner Production can result in economic, environmental and social benefits, and in actions that promote reduction in water consumption. This case study was conducted in a slaughterhouse and poultry cold storage processing plant and aimed to identify points of excessive water consumption, and to propose alternatives for managing water resources by reducing consumption. Consumption data are presented in relation to the processing stages with alternatives proposed for the rational use of water, such as closure of mains water during shift changes. Following the implementation of recommendations, a reduction in water consumption of approximately 11,137 m³ per month was obtained, which equates to a savings of US$ 99,672 per year. From this study, it was concluded that the company under review could develop various improvement actions and make an important contribution to the preservation of water resources in the region where it operates.

Coverage mobilization by different no-tillage in-line handling mechanisms

Vegetation cover on soil acts positively in maintaining temperature and soil moisture, yet, it has been imposing specific operational conditions on seeders. The objective of this study was to evaluate performance of different mechanisms regarding straw mobilization, employed in a no-till seeder. The experimental area was conducted on clayey soil under no-tillage with a large quantity of sorghum residue. The experiment was established in a randomized block design, as the treatments consisted of a combination of two mechanisms at front of the furrow opener composed of cutting disc and row cleaners, and three mechanisms behind the seed furrower, covering discs prototype model M1, Spider and commercial model, with the combination of cutting disc and Spider model not being evaluated. We assessed the coverage permanence on soil index, vegetation mass on surface and inside the line. The treatment containing the row cleaner mechanism efficiently removed straw from the surface of sowing line as well as the return one acted on straw replacement. It was identified that use of the cutting disc at the front of seeder contributed to the increase of straw installation inside the line, three times more than in the row cleaner system when operating individually. Covering mechanism with row cleaners reduced straw inside the line and kept line covering similar to treatment of cutting disc operating alone.

An Overview of the Cleantech Sector - Evaluating Environmental Indicators

The purpose of the thesis is to give an overview of the cleantech sector and to give an answer how cleantech companies can evaluate the environmental sustainability of their business by utilizing various indicators and measures. The thesis is a literature study and it is based on a secondary data. Thesis presents the definitions to cleantech as well as its history and the main industries. Cleantech market overview in Finland and worldwide is also introduced. Furthermore, various indicators are presented in order to evaluate the environmental sustainability of companies' business. In the end, indicators used in cleantech sector are evaluated. As a result, the thesis presents the following methodologies that can be used in evaluating the environmental sustainability in the cleantech sector: Sustainability assessment framework, Environmental value analysis, COMPLIMENT - Environmental performance index for industries and Environmental assessment for cleaner production. More tools are still needed to evaluate environmental sustainability in the cleantech sector.

Optimization and validation of the solid-liquid extraction technique for determination of picloram in soils by high performance liquid chromatography

The objective of this study was to optimize and validate the solid-liquid extraction (ESL) technique for determination of picloram residues in soil samples. At the optimization stage, the optimal conditions for extraction of soil samples were determined using univariate analysis. Ratio soil/solution extraction, type and time of agitation, ionic strength and pH of extraction solution were evaluated. Based on the optimized parameters, the following method of extraction and analysis of picloram was developed: weigh 2.00 g of soil dried and sieved through a sieve mesh of 2.0 mm pore, add 20.0 mL of KCl concentration of 0.5 mol L-1, shake the bottle in the vortex for 10 seconds to form suspension and adjust to pH 7.00, with alkaline KOH 0.1 mol L-1. Homogenate the system in a shaker system for 60 minutes and then let it stand for 10 minutes. The bottles are centrifuged for 10 minutes at 3,500 rpm. After the settlement of the soil particles and cleaning of the supernatant extract, an aliquot is withdrawn and analyzed by high performance liquid chromatography. The optimized method was validated by determining the selectivity, linearity, detection and quantification limits, precision and accuracy. The ESL methodology was efficient for analysis of residues of the pesticides studied, with percentages of recovery above 90%. The limits of detection and quantification were 20.0 and 66.0 mg kg-1 soil for the PVA, and 40.0 and 132.0 mg kg-1 soil for the VLA. The coefficients of variation (CV) were equal to 2.32 and 2.69 for PVA and TH soils, respectively. The methodology resulted in low organic solvent consumption and cleaner extracts, as well as no purification steps for chromatographic analysis were required. The parameters evaluated in the validation process indicated that the ESL methodology is efficient for the extraction of picloram residues in soils, with low limits of detection and quantification.

Toxic, antimicrobial and hemagglutinating activities of the purple fluid of the sea hare Aplysia dactylomela Rang, 1828

The antimicrobial, hemagglutinating and toxic activities of the purple fluid of the sea hare Aplysia dactylomela are described. Intact or dialyzed purple fluid inhibited the growth of species of Gram-positive and Gram-negative bacteria and the action was not bactericidal but bacteriostatic. The active factor or factors were heat labile and sensitive to extreme pH values. The fluid preferentially agglutinated rabbit erythrocytes and, to a lesser extent, human blood cells, and this activity was inhibited by the glycoprotein fetuin, a fact suggesting the presence of a lectin. The fluid was also toxic to brine shrimp nauplii (LD50 141.25 µg protein/ml) and to mice injected intraperitoneally (LD50 201.8 ± 8.6 mg protein/kg), in a dose-dependent fashion. These toxic activities were abolished when the fluid was heated. Taken together, the data suggest that the activities of the purple fluid are due primarily to substance(s) of a protein nature which may be involved in the chemical defense mechanism of this sea hare.

P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships

Resistance to chemotherapy in cancer cells is mainly mediated by overexpression of P-glycoprotein (Pgp), a plasma membrane ATP-binding cassette (ABC) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Pgp consists of two homologous halves each containing a transmembrane domain and a cytosolic nucleotide-binding domain (NBD) which contains two consensus Walker motifs, A and B, involved in ATP binding and hydrolysis. The protein also contains an S signature characteristic of ABC transporters. The molecular mechanism of Pgp-mediated drug transport is not known. Since the transporter has an extraordinarily broad substrate specificity, its cellular function has been described as a "hydrophobic vacuum cleaner". The limited knowledge about the mechanism of Pgp, partly due to the lack of a high-resolution structure, is well reflected in the failure to efficiently inhibit its activity in cancer cells and thus to reverse multidrug resistance (MDR). In contrast to the difficulties encountered when studying the full-length Pgp, the recombinant NBDs can be obtained in large amounts as soluble proteins. The biochemical and biophysical characterization of recombinant NBDs is shown here to provide a suitable alternative route to establish structure-function relationships. NBDs were shown to bind ATP and analogues as well as potent modulators of MDR, such as hydrophobic steroids, at a region close to the ATP site. Interestingly, flavonoids also bind to NBDs with high affinity. Their binding site partly overlaps both the ATP-binding site and the steroid-interacting region. Therefore flavonoids constitute a new promising class of bifunctional modulators of Pgp.

Infrequent p53 gene alterations in ulcerative colitis

The purpose of this study was to determine whether point mutations and loss of the p53 gene take place in ulcerative colitis which is histologically negative for dysplasia. DNA was extracted from 13 frozen rectal or colon biopsies and blood samples. Ulcerative colitis was classified histologically as active (10 cases) and inactive (3 cases). Exons 5-8 were amplified by PCR, treated with exonuclease and shrimp alkaline phosphatase and sequenced by the dideoxy chain termination method with the Sequenase Version 2.0 DNA sequencing kit. PCR products of intron 6 and exon 4 were digested with MspI and AccII, respectively, for RFLP analysis. No p53 gene mutation was detected in these cases. The number of informative patients for loss of heterozygosity (LOH) at the p53 intron 6 was high, 11 out of 12 (92%), whereas no LOH was observed. LOH affecting p53 exon 4 was not detected in lesions from 5 of 12 patients (42%). In ulcerative colitis, tumor progression is similar to that in sporadic colon cancer, and other oncogenes and tumor suppressor genes are likely to be mutated before the p53 gene.

The Role of Oxidative Stress in Environmental Responses of Fennoscandian Animals

All aerobic organisms have to deal with the toxicity of oxygen. Oxygen enables more efficient energy production compared to anaerobic respiration or fermentation, but at the same time reactive oxygen species (ROS) are being formed. ROS can also be produced by external factors such as UV-radiation and contamination. ROS can cause damage to biomolecules such as DNA, lipids and proteins and organisms try to keep the damage as small as possible by repairing biomolecules and metabolizing ROS. All ROS are not harmful, because they are used as signaling molecules. To cope against ROS organism have an antioxidant (AOX) system which consists both enzymatic and non-enzymatic AOX defense. Some AOX are produced by the organism itself and some are gained via diet. In this thesis I studied environmentally caused changes in the redox regulation of different wild vertebrate animals to gain knowledge on the temporal, spatial and pollution-derived-effects on the AOX systems. As study species I used barn swallow, ringed seal and the Baltic salmon. For the barn swallow the main interest was the seasonal fluctuation in the redox regulation and its connection to migration and breeding. The more contaminated ringed seals of the Baltic Sea were compared to seals from cleaner Svalbard to investigate whether they suffered from contaminant induced oxidative stress. The regional and temporal variation in redox regulation and regional variation in mRNA and protein expressions of Baltic salmon were studied to gain knowledge if the salmon from different areas are equally stressed. As a comparative aspect the redox responses of these different species were investigated to see which parts of the AOX system are substantial in which species. Certain parts of AOX system were connected to breeding and others to migration in barn swallows, there was also differences in biotransformation between birds caught from Africa and Finland. The Baltic ringed seal did not differ much from the seals from Svalbard, despite the difference in contaminant load. A possible explanation to this could be the enhanced AOX mechanisms against dive-associated oxidative stress in diving air-breathing animals, which also helps to cope with ROS derived from other sourses. The Baltic salmon from Gulf of Finland (GoF) showed higher activities in their AOX defense enzymes and more oxidative damage than fish from other areas. Also on mRNA and proteomic level, stress related metabolic changes were most profound in in the fish from GoF. Mainly my findings on species related differences followed the pattern of mammals showing highest activities and least damage and birds showing lower activities and most damage, fish being intermediate. In general, the glutathione recycling-related enzymes and the ratio of oxidized and reduced glutathione seemed to be the most affected parameters in all of the species.