PREPARAÇÃO DE PONTOS DE CARBONO E SUA CARACTERIZAÇÃO ÓPTICA: UM EXPERIMENTO PARA INTRODUZIR NANOCIÊNCIA NA GRADUAÇÃO

Carbon dots (CDs) constitute a new class of carbon-based nanomaterials that measure less than 10 nm and display attractive physical and chemical features such as fluorescence. CDs have been considered the new “power” carbon nanomaterials since their accidental discovery in 2004. This study reports a simple, easy, and accessible experiment for undergraduate courses. The experiment involves the preparation of CDs by pyrolysis using commercial gelatin as a low cost precursor as well as CD purification and optical characterization. The optical properties of CDs such as absorption and emission properties make them a promising material for teaching the basic concepts and techniques used for characterization of nanomaterials. Also, the reactants and final product are suitable for undergraduate courses since they are non-toxic materials. The prepared CDs can be used in such applications as bioimaging, solar cells, and photocatalysis.

Alusöljyvahingon seurauksena rantautuvan öljyn lajitteluohjeiston muodostaminen

Työn tärkeimpänä päämääränä oli muodostaa öljyvahinkojätejakeille yksityiskohtaiset ja käytännön olosuhteissa mahdollisimman hyvin toimivat lajitteluohjeet. Lähtökohtana oli se, että edeltävien lajitteluohjeiden soveltuvuutta haluttiin tarkastella useista eri näkökulmista, kuten muodostuvien kustannusten kannalta. Työn muut tavoitteet olivat: jäteastioiden määrän ja laadun selvitys sekä lainsäädännön asettamien rajoitteiden selvittäminen. Riskijäte rajattiin työn ulkopuolelle. Tutkimus toteutettiin pääasiassa kirjallisiin lähteisiin, sähköpostikyselyihin ja puhelinhaastatteluihin perustuvien tietojen avulla. Tärkeimmäksi selvitettäväksi seikaksi osoittautui lajittelusta aiheutuvien kustannusten määrittäminen. Etenkin käsittelykustannuksista saatiin viitteitä optimaalisesta lajitteluvaihtoehdosta. Taloudellisessa tarkastelussa käytiin läpi öljyvahinkojätteiden kulkeutuminen rannalta käsittelyyn saakka, jolloin eri vaihtoehtojen eroavaisuudet saatiin selville. Taloudellisen tarkastelun perusteella paras vaihtoehto oli lajitellun jätteen käsittely siirrettävällä termodesorptiolaitoksella yhdistettynä Kotkan hyötyvoimalaitokseen. Tämän perusteella voidaan päätellä, että öljyinen maa-aines ja öljyinen sekajäte kannattaa käsitellä erillisinä jakeina. Tällöin öljyinen maa-aines ja öljyinen sekajäte kannattaa myös lajitella omiin jakeisiinsa. Keräysastioista on vaikeaa antaa suosituksia ilman riittävän kattavia kenttäkokeita. Taloudellisessa tarkastelussa muoviastiat osoittautuivat edullisimmaksi vaihtoehdoksi. Monissa selvityksissä on öljyvahinkojätteiden käsittelyvaihtoehdoksi valittu Riihimäen Ekokem Oy Ab. Se tuli kuitenkin huomattavasti kalliimmaksi kuin siirrettävän termodesorptiolaitoksen sisältävät laskuesimerkit, joten myös muita vaihtoehtoja kannattaisi harkita. Muita kuin muovisia keräysastioita tulisi vielä testata käytännön öljyntorjuntaharjoituksissa, jotta niiden lopullinen käyttökelpoisuus varmistuu. Harjoitukset tulisi suorittaa mahdollisimman vaihtelevissa sää- ja maasto-olosuhteissa, jotta saadaan tarpeeksi kattavaa tutkimustietoa astioiden soveltuvuudesta.

Alusöljyvahingossa kuolleiden eläinten turvallinen käsittely

Diplomityö on osa Kymenlaakson ammattikorkeakoulun SÖKÖ II-hanketta, missä tehdään toimintamalli suuren öljyntorjuntaoperaation koordinointiin rannikon öljytorjunnasta vastaaville viranomaisille. Työn tavoitteena oli selvittää alusöljyvahingossa kuolleiden eläinten sekä eläinten keräilystä ja hoidosta syntyvien jätteiden käsittelymenetelmät ja soveltuvat käsittelylaitokset. Lisäksi tavoitteena oli selvittää kuolleiden eläinten välivarastointialueen tekniset vaatimukset ja öljyiselle eläinjätteelle soveltuva kuljetuskalusto. Työn teoriaosuudet toteutettiin kirjallisten lähteiden ja asiantuntijahaastatteluiden avulla. Tärkeäksi osaksi työtä muodostui kustannuslaskelma, missä laskettiin kuolleiden eläinten keräilystä aina loppukäsittelyyn asti muodostuvat kustannukset. Työn teoriaosuudet loivat pohjan kustannuslaskelmalle. Kuolleiden eläinten loppukäsittelyyn valittiin kolme eri laitosta ja kolme eri käsittelytekniikkaa, mitkä ovat: rumpu-uuni, siirrettävä termodesorptio ja renderöinti. Kuolleet eläimet kannattaa keskitetysti kerätä ja kuljettaa yhdelle alueelle, missä suoritetaan kuolleiden eläinten välivarastointi kylmäkonteissa ja tilastointi sekä elävien eläinten pesu ja hoito. Kustannuslaskelman perusteella siirrettävä termodesorptiolaitos on halvin käsittelyvaihtoehto, mutta kustannuserot muihin laitoksiin ovat pienet. Koko kuolleiden eläinten turvallisen käsittelyn kustannukset ovat alle promillen koko suuren öljyvahingon kustannuksista. Loppukäsittelylaitosta valittaessa on käsittelykustannuksia tärkeämpää selvittää laitoksen sen hetkinen jätteenkäsittelytilanne ja vapaa käsittelykapasiteetti kuolleille eläimille.

Thermoanalytical study and characterization of native starches of Paraná pine seeds (Araucaria angustiofolia, Bert O. Ktze) and European chestnut seeds (Castanea sativa, Mill)

Starch is the most important carbohydrate storage in plants. It is a raw material with diverse botanical origins, and is used by the food, paper, chemical, pharmaceutical, textile and other industries. In this work, native starches of Paraná pine seeds (pinhão) (Araucária angustiofolia, Bert O. Ktze) and european chestnut seeds (Castanea sativa, Mill) were studied by thermoanalytical techniques: thermo-gravimetry (TG), differential thermal analysis (DTA) and differential scanning calorimetry (DSC), as well as X-ray powder patterns diffractometry. Apparent and total amylose content was also determined.

Lanthanum based high surface area perovskite-type oxide and application in CO and propane combustion

The perovskite-type oxides using transition metals present a promising potential as catalysts in total oxidation reaction. The present work investigates the effect of synthesis by oxidant co-precipitation on the catalytic activity of perovskite-type oxides LaBO3 (B= Co, Ni, Mn) in total oxidation of propane and CO. The perovskite-type oxides were characterized by means of X-ray diffraction, nitrogen adsorption (BET method), thermo gravimetric and differential thermal analysis (ATG-DTA) and X-ray photoelectron spectroscopy (XPS). Through a method involving the oxidant co-precipitation it's possible to obtain catalysts with different BET surface areas, of 33-44 m²/g, according the salts of metal used. The characterization results proved that catalysts have a perovskite phase as well as lanthanum oxide, except LaMnO3, that presents a cationic vacancies and generation for known oxygen excess. The results of catalytic test showed that all oxides have a specific catalytic activity for total oxidation of CO and propane even though the temperatures for total conversion change for each transition metal and substance to be oxidized.

Quantification of Proteins and Cells: Luminometric Nonspecific Particle-Based Methods

New luminometric particle-based methods were developed to quantify protein and to count cells. The developed methods rely on the interaction of the sample with nano- or microparticles and different principles of detection. In fluorescence quenching, timeresolved luminescence resonance energy transfer (TR-LRET), and two-photon excitation fluorescence (TPX) methods, the sample prevents the adsorption of labeled protein to the particles. Depending on the system, the addition of the analyte increases or decreases the luminescence. In the dissociation method, the adsorbed protein protects the Eu(III) chelate on the surface of the particles from dissociation at a low pH. The experimental setups are user-friendly and rapid and do not require hazardous test compounds and elevated temperatures. The sensitivity of the quantification of protein (from 40 to 500 pg bovine serum albumin in a sample) was 20-500-fold better than in most sensitive commercial methods. The quenching method exhibited low protein-to-protein variability and the dissociation method insensitivity to the assay contaminants commonly found in biological samples. Less than ten eukaryotic cells were detected and quantified with all the developed methods under optimized assay conditions. Furthermore, two applications, the method for detection of the aggregation of protein and the cell viability test, were developed by utilizing the TR-LRET method. The detection of the aggregation of protein was allowed at a more than 10,000 times lower concentration, 30 μg/L, compared to the known methods of UV240 absorbance and dynamic light scattering. The TR-LRET method was combined with a nucleic acid assay with cell-impermeable dye to measure the percentage of dead cells in a single tube test with cell counts below 1000 cells/tube.

Influence of Heat Treatment on Strength Properties of Finger-jointed Components

The purpose for the thesis was to study the thermo treatment of finger-jointed wood. The thesis concentrated on examining the tensile and bending strength of finger-jointed and thermo treated wood. The aim was to find out how different treatment temperature levels and adhesives influence the strength of wood that has been finger-jointed before heat treatment. Secondary objectives were to examine the influence of the treatment time at one temperature, determine differences in the strength between the joints in heartwood and sapwood, examine the visual appearance of the finger joints after the treatment and establish possibilities to reach a characteristic strength level corresponding to C14. Only minor differences in strength properties were measured between the finger-jointed wood treatments II and III. A greater difference was shown between these two treatment temperatures I, which lead to reduced strength. The average strength of joints glued with adhesive 2 was higher after treatments II and III compared to those glued with the adhesive 1. At the treatment temperature I, the adhesive 1 strength properties were at the same level compared to the adhesive 2 or better. There were not any significant differences.

Uuteaineet BCTMP-prosessissa

Diplomityö tehtiin M-real Joutsenon BCTMP-tehtaalle. Työn aiheena oli tehdä uuteaine- ja metallikartoitus eri prosessivaiheiden massoista ja suodoksista BCTMP-tehtaassa. BCTMPmassaa käytetään muun muassa elintarvikekartongin keskikerroksessa. Pakattaviin kartonkeihin suoraan kosketuksissa olevat elintarvikkeet ovat alttiita haju- ja makuvivahteille. Hajut ja maut voivat saada alkunsa kartongin komponenteista, kuten BCTMP-massasta, kartongin päällystysaineista ja kartongin jatkojalostuksessa käytetyistä painoväreistä. Diplomityön teoriaosuudessa keskityttiin puun lipofiilisiin uuteaineisiin ja erityisesti komponentteihin, jotka aiheuttavat haju- ja makuongelmia. Teoriaosuudessa käsiteltiin myös haju- ja makuominaisuuksien muodostumiseen vaikuttavia tekijöitä sekä kerrottiin mahdollisista uuteaineiden poistomenetelmistä. Teoriaosuudessa esiteltiin analysointimenetelmiä, joita käytettiin kokeellisessa osassa. Kokeellisessa osassa esiteltiin valmistettavan massan laatuspesifikaatiot ja toteutuneet laatutrendit. Samassa tutkittiin massan analysointimenetelmien tuloksien korrelaatioita kokonaisuuteaineiden toteamiseksi. Seuraavaksi käsiteltiin syksyllä 2010 ja talvella 2011 tehtyjä prosessin uuteainekartoituksia, jossa muun muassa tutkittiin eri prosessivaiheiden massojen ja suodoksien uuteainemääriä ja –komponentteja. Kartoituksien yhteydessä laskettiin prosessin uuteainetaseita ja prosessin pesutehokkuuksia. Samalla tutkittiin siirtymämetallimääriä, jotka osaltaan vaikuttavat haju- ja makuominaisuuksiin. Kokeellisen osan korrelaatiovertailuissa huomattiin, että merkittävin korrelaatio kokonaisuuteaineisiin on viimeisen massan pesuvaiheen suodoksen sameudella, liki 80 %. Heksanaalipotentiaalin ja uuteaineen korrelaatio oli myös noin 80 %. Valmiista massasta tehdyn kemiallisen hapen kulutuksen eli COD:n korrelaatio uuteaineisiin oli vain 38 %. Prosessikartoituksen tuloksista laskettujen pesuvaiheiden erotustehokkuudet olivat hyvät.

Upconverting Phosphor Technology: Exceptional Photoluminescent Properties Light Up Homogeneous Bioanalytical Assays

The aim of the present study was to demonstrate the wide applicability of the novel photoluminescent labels called upconverting phosphors (UCPs) in proximity-based bioanalytical assays. The exceptional features of the lanthanide-doped inorganic UCP compounds stem from their capability for photon upconversion resulting in anti-Stokes photoluminescence at visible wavelengths under near-infrared (NIR) excitation. Major limitations related to conventional photoluminescent labels are avoided, rendering the UCPs a competitive next-generation label technology. First, the background luminescence is minimized due to total elimination of autofluorescence. Consequently, improvements in detectability are expected. Second, at the long wavelengths (>600 nm) used for exciting and detecting the UCPs, the transmittance of sample matrixes is significantly greater in comparison with shorter wavelengths. Colored samples are no longer an obstacle to the luminescence measurement, and more flexibility is allowed even in homogeneous assay concepts, where the sample matrix remains present during the entire analysis procedure, including label detection. To transform a UCP particle into a biocompatible label suitable for bioanalytical assays, it must be colloidal in an aqueous environment and covered with biomolecules capable of recognizing the analyte molecule. At the beginning of this study, only UCP bulk material was available, and it was necessary to process the material to submicrometer-sized particles prior to use. Later, the ground UCPs, with irregular shape, wide size-distribution and heterogeneous luminescence properties, were substituted by a smaller-sized spherical UCP material. The surface functionalization of the UCPs was realized by producing a thin hydrophilic coating. Polymer adsorption on the UCP surface is a simple way to introduce functional groups for bioconjugation purposes, but possible stability issues encouraged us to optimize an optional silica-encapsulation method which produces a coating that is not detached in storage or assay conditions. An extremely thin monolayer around the UCPs was pursued due to their intended use as short-distance energy donors, and much attention was paid to controlling the thickness of the coating. The performance of the UCP technology was evaluated in three different homogeneous resonance energy transfer-based bioanalytical assays: a competitive ligand binding assay, a hybridization assay for nucleic acid detection and an enzyme activity assay. To complete the list, a competitive immunoassay has been published previously. Our systematic investigation showed that a nonradiative energy transfer mechanism is indeed involved, when a UCP and an acceptor fluorophore are brought into close proximity in aqueous suspension. This process is the basis for the above-mentioned homogeneous assays, in which the distance between the fluorescent species depends on a specific biomolecular binding event. According to the studies, the submicrometer-sized UCP labels allow versatile proximity-based bioanalysis with low detection limits (a low-nanomolar concentration for biotin, 0.01 U for benzonase enzyme, 0.35 nM for target DNA sequence).

Substrate with Organosuper® for cucumber seedlings formation in protected environments and polystyrene trays

Culture environments, trays and doses of organic compost were evaluated in the formation of cucumber seedlings (Cucumis sativus L.). Five environmental conditions were tested: (A1) a greenhouse with height of 2.5 m, covered with polyethylene film, (A2) nursery with height of 2.5 m, monofilament fabric, 50% shading, (A3) nursery with height of 2.5 m, heat-reflective screen, 50% shading, (A4) nursery with a height of 1.8 m, covered with coconut tree straw and (A5) greenhouse with height of 4.0 m, covered with polyethylene film, with zenith opening and thermo-reflective cloth under the plastic. Trays of 72 (R1) and 128 (R2) cells were filled with 93% soil and 7% organic compound (S1), 86% soil and 14% organic compound (S2) and 79% soil and 21% organic compound (S3). It was used a randomized design in split-split-plot scheme, with five replicates (environments x trays x substrates). The greenhouses provide the best environments for the formation of cucumber seedlings. A tray of 72 cells is the best container, promoting more vigorous seedlings in substrate with soil and 7 or 14% organic compound.

Production of baruzeiro seedling in different protected environments and substrates

The high seedlings quality is essential for deployment of homogeneous orchards. This study evaluated the baruzeiro (Dipteryx alata Vog) seedlings formation on different substrates within protected environments. It was used substrates with100% of cattle manure; 100% of cassava stems; 100% of vermiculite; 50% of cattle manure + 50% of cassava stems; 50% of cattle manure + 50% of vermiculite; 50% of cassava stems + 50% of vermiculite; and + ⅓ of cattle manure + ⅓ of cassava stems + ⅓ of vermiculite. These substrates were tested in protected areas: greenhouse; black shade net of 50% shading; and aluminized thermo-reflective screen of 50% shading. A completely randomized experimental design with five replicates of four plants was adopted. Initially, data were submitted to analysis of individual variance of the substrates, in each environment of cultivation, then performing the evaluation of the residual mean square and the analysis of these environments together for comparison. The best substrate for baruzeiro seedlings was pure vermiculite. The substrates with 100% of manure and the substrate with 33.33% of the mixed studied materials can be used for seedlings formation. The environment with screen can be indicated for the production of baruzeiro seedlings, since it gave vigor to the seedlings.

Seedlings of Acrocomia aculeata in diferent substrates and protected environments

The seedling production stage is the key to achieve uniformity in tree breeding stage. This study evaluated "bocaiúva" (Acrocomia aculeata) seedling formation, with pre-germinated seeds in different substrates and protected environments, in the University of Mato Grosso do Sul State, Aquidauana, MS. As substrates, we used 100% cattle manure (M), 100% cassava branches (CB), 100% vermiculite (V), 50% cattle manure + 50% cassava branches, 50 % cattle manure + 50% vermiculite, 50% cassava branches + 50% vermiculite and ⅓ cattle manure + ⅓ cassava branches + ⅓ vermiculite. These substrates were tested in a greenhouse covered with 150 µm low density polyethylene (LDPE) film under thermo-reflective screen with 50% shading under film; black screen with 50% shading on the sides; black monofilament screen with 50% shading set on roof and sides; and aluminized thermo- reflective screen with 50% shading set on roof and sides. The completely randomized experimental design with 5 replications of 5 plants each was adopted. Initially, data were submitted to analysis of substrate individual variance in each growing environment, then performing the waste mean square evaluation and their environment joint analysis for comparison. The best growing environment is the thermo-reflective screen compared to LDPE greenhouse and black screen set. All substrates containing manure are recommended for bocaiúva seedlings formation. The pure cassava branch is not indicated for seedling, even using chemical fertilizer.

Soursop seedlings: emergence and development under different cultivation environments and substrates – Part I

ABSTRACT Seedling quality is crucial to obtain vigorous plants in the field. This study aims to evaluate the emergence and development of soursop seedlings in different substrates in protected environments. The experiment was conducted at the Mato Grosso do Sul State University and carried out using five protected environments: greenhouse, greenhouse with thermo-reflective screen, nursery with monofilament screen, nursery with thermo-reflective screen, and nursery with palm thatch. The substrates (S) consisted of cattle manure (M), humus (H), cassava branches (C), and vermiculite (V) as in the following ratios: S1 = H + V (1:3), S2 = H + V (1:1), S3 = H + V (3:1), S4 = H + C (1:3), S5 = H + C (1:1), S6 = H + C (3:1), S7 = M + V (1:3), S8 = M + V (1:1), S9 = M + V (3:1), S10 = M + C (1:3), S11 = M + C (1:1), S12 = M + C (3:1), S13 = H + M + V (1:1:1), S14 = H + M + C (1:1:1), and S15 = H + M + V + C (1:1:1:1). For the statistical analysis, each of those environments was considered as an experiment in which was used the completely randomized design; subsequently, it was performed a combined analysis of them. In summary, the greenhouse with thermo-reflective screen and combined substrates with “M + V” promote greater development of the seedlings. High concentrations of “V” or “C” cause no beneficial effect on soursop seedlings.

Soursop seedlings: biomasses and biometric relations in different farming environments and substrates – Part II

ABSTRACT The quality of seedling is critical to obtain vigorous plants in the field. The present study aimed to assess biomasses and biometric relations of soursop seedlings. We used different substrates in protected environments. The experiment was performed at the Universidade Estadual do Mato Grosso do Sul (UFMS) (State University of Mato Grosso do Sul). Five farming environments were developed in greenhouses: one covered with low-density polyethylene film (LDPE), another with with polyethylene and heat-reflective cloth under film under 50% shading in aluminized color, monofilament cloth under 50% shading in black, thermo-reflective cloth under 50% shading in aluminized color, and an environment covered with bacuri coconut straw. Substrates were made of manure, humus, cassava branches and vermiculite at different proportions. Each of them varying from 25%, 33.3%, 50% and 75% in mixture combination. Each environment was considered an experiment. A completely randomized design was adopted and later a joint analysis of them. Agricultural greenhouse covered with LDPE and thermo-reflective cloths under 50% of shading, proportionated seedlings with greater biomass. Substrates containing manure are the most suitable for soursop seedlings. High percentages of earthworm humus produce low quality soursop seedlings. Soursop seedlings had a Dickson’s quality index around 0.335. The greenhouse covered only with LDPE film did not produce high quality seedlings.

Luminescent Lanthanide Reporters: New Concepts for Use in Bioanalytical Applications

Lanthanides represent the chemical elements from lanthanum to lutetium. They intrinsically exhibit some very exciting photophysical properties, which can be further enhanced by incorporating the lanthanide ion into organic or inorganic sensitizing structures. A very popular approach is to conjugate the lanthanide ion to an organic chromophore structure forming lanthanide chelates. Another approach, which has quickly gained interest, is to incorporate the lanthanide ions into nanoparticle structures, thus attaining improved specific activity and binding capacity. The lanthanide-based reporters usually express strong luminescence emission, multiple narrow emission lines covering a wide wavelength range, and exceptionally long excited state lifetimes enabling timeresolved detection. Because of these properties, the lanthanide-based reporters have found widespread applications in various fields of life. This study focuses on the field of bioanalytical applications. The aim of the study was to demonstrate the utility of different lanthanide-based reporters in homogeneous Förster resonance energy transfer (FRET)-based bioaffinity assays. Several different model assays were constructed. One was a competitive bioaffinity assay that utilized energy transfer from lanthanide chelate donors to fluorescent protein acceptors. In addition to the conventional FRET phenomenon, a recently discovered non-overlapping FRET (nFRET) phenomenon was demonstrated for the first time for fluorescent proteins. The lack of spectral overlap in the nFRET mechanism provides sensitivity and versatility to energy transfer-based assays. The distance and temperature dependence of these phenomena were further studied in a DNA-hybridization assay. The distance dependence of nFRET deviated from that of FRET, and unlike FRET, nFRET demonstrated clear temperature dependence. Based on these results, a possible excitation mechanism operating in nFRET was proposed. In the study, two enzyme activity assays for caspase-3 were also constructed. One of these was a fluorescence quenching-based enzyme activity assay that utilized novel inorganic particulate reporters called upconverting phosphors (UCPs) as donors. The use of UCPs enabled the construction of a simple, rather inexpensive, and easily automated assay format that had a high throughput rate. The other enzyme activity assay took advantage of another novel reporter class, the lanthanidebinding peptides (LBPs). In this assay, energy was transferred from a LBP to a green fluorescent protein (GFP). Using the LBPs it was possible to avoid the rather laborious, often poorly repeatable, and randomly positioned chemical labeling. In most of the constructed assays, time-resolved detection was used to eliminate the interfering background signal caused by autofluorescence. The improved signal-to-background ratios resulted in increased assay sensitivity, often unobtainable in homogeneous assay formats using conventional organic fluorophores. The anti-Stokes luminescence of the UCPs, however, enabled the elimination of autofluorescence even without time-gating, thus simplifying the instrument setup. Together, the studied reporters and assay formats pave the way for increasingly sensitive, simple, and easily automated bioanalytical applications.