DETECTION AND QUANTIFICATION OF PHYTOCHEMICAL MARKERS OF Ilex paraguariensis BY LIQUID CHROMATOGRAPHY

Ilex paraguariensis (yerba-mate) is used as a beverage, and its extract requires adequate quality control methods in order to guarantee quality and safe use. Strategies to develop and optimize a chromatographic method to quantify theobromine, caffeine, and chlorogenic acid in I. paraguariensis extracts were evaluated by applying a quality by design (QbD) model and ultra high-performance liquid chromatography (UHPLC). The presence of these three phytochemical markers in the extracts was evaluated using UHPLC-MS and was confirmed by the chromatographic bands in the total ion current traces (m/z of 181.1 [M+H]+, 195.0 [M+H]+, and 353.0 [M−H]−, respectively). The developed method was then transferred to a high-performance liquid chromatography (HPLC) platform, and the three phytochemical markers were used as external standards in the validation of a method for analyses of these compounds in extracts using a diode array detector (DAD). The validated method was applied to quantify the chlorogenic acid, caffeine, and theobromine in the samples. HPLC-DAD chromatographic fingerprinting was also used in a multivariate approach to process the entire data and to separate the I. paraguariensis extracts into two groups. The developed method is very useful for qualifying and quantifying I. paraguariensis extracts.

TREATMENT OF WASTE FROM ATOMIC EMISSION SPECTROMETRIC TECHNIQUES AND REUSE IN UNDERGRADUATE LAB CLASSES FOR QUALITATIVE ANALYSIS

Flame atomic absorption spectrometry (FAAS) and inductively coupled plasma optical emission spectrometry (ICP OES) are widely used in academic institutions and laboratories for quality control to analyze inorganic elements in samples. However, these techniques have been observed to underperform in sample nebulization processes. Most of the samples processed through nebulization system are discarded, producing large volumes of waste. This study reports the treatment and reuse of the waste produced from ICP OES technique in a laboratory of analytical research at the Universidade Federal do Ceará, Brazil. The treatment of the waste was performed by the precipitation of elements using (NH4)2CO3. Subsequently, the supernatant solution can be discarded in accordance with CONAMA 430/2011. The precipitate produced from the treatment of residues can be reused as a potential sample in undergraduate qualitative analytical chemistry lab classes, providing students the opportunity to test a real sample.

DESENVOLVIMENTO DE ENSAIO PARA ANÁLISE QUALITATIVA DE BIODIESEL EM MISTURAS DIESEL-BIODIESEL PARA APLICAÇÃO EM POSTOS REVENDEDORES DE COMBUSTÍVEIS

In this study, we developed a method for the visual detection of biodiesel in petrodiesel-biodiesel (BX) blends through the aminolysis of the methyl or ethyl esters of fatty acids that are found in biodiesel and that are absent from diesel and vegetable oils. This method is based on three reactional stages, which produce a dark red and easily visualized complex in the presence of biodiesel. In the absence of biodiesel, there is no dark red coloring, whereas in the presence of diesel or vegetable oil, there is a light red to yellow coloring. This simple, practical, inexpensive, and effective procedure may be applied by petrol stations to guarantee to consumers and resellers the presence of biodiesel in diesel blends, regardless of the BX blend's initial coloring or of the sulfur found in the diesel. In short, it ensures a safe fuel tank fill-up with BX blend.

Imaging and Characterisation of Dirt Particles in Pulp and Paper

Dirt counting and dirt particle characterisation of pulp samples is an important part of quality control in pulp and paper production. The need for an automatic image analysis system to consider dirt particle characterisation in various pulp samples is also very critical. However, existent image analysis systems utilise a single threshold to segment the dirt particles in different pulp samples. This limits their precision. Based on evidence, designing an automatic image analysis system that could overcome this deficiency is very useful. In this study, the developed Niblack thresholding method is proposed. The method defines the threshold based on the number of segmented particles. In addition, the Kittler thresholding is utilised. Both of these thresholding methods can determine the dirt count of the different pulp samples accurately as compared to visual inspection and the Digital Optical Measuring and Analysis System (DOMAS). In addition, the minimum resolution needed for acquiring a scanner image is defined. By considering the variation in dirt particle features, the curl shows acceptable difference to discriminate the bark and the fibre bundles in different pulp samples. Three classifiers, called k-Nearest Neighbour, Linear Discriminant Analysis and Multi-layer Perceptron are utilised to categorize the dirt particles. Linear Discriminant Analysis and Multi-layer Perceptron are the most accurate in classifying the segmented dirt particles by the Kittler thresholding with morphological processing. The result shows that the dirt particles are successfully categorized for bark and for fibre bundles.

Asiakkuuksien johtaminen kuljetusalan pk-yrityksessä

The profitability on transportation field has been low for a long time. The re-cession that started in the fall of 2008 has caused even more trouble to transportation companies as the number of transportations has gone down and the competition has gotten tougher since the time before the recession. Because the low profitability level does not allow competing with prices, transportation companies must come up with other means to differ from their competitors. The main goal of the research was to find out how a small and medium-sized enterprise (SME) on transportation field should manage its customer relationships. Due to limited resources, companies should direct their resources to serving their most valuable customers. Companies should first make a determination of their customer base. This works as a base for establishing customer portfolios, in other words making customer groups based on e.g. profitability, buying volumes and loyalty. Companies can gain competitive advantage also by improving their service quality. Important information regarding the selection criteria of a transporta-tion company, transaction methods, service time and accessibility of person-nel, customers’ expectations on transportation company’s personnel, utiliza-tion of IT-solutions as a part of transportation service, the image of a transportation company and necessity of quality control meetings was gathered for this research by interviewing customers. By utilizing this information, transportation company can adapt its service offering to equal the customer needs and expectations better. A service process example was created to improve the service quality of the transportation company. It shows the whole process from the customer sending the transportation order to the point when the customer receives an invoice from the transportation company. The example process shows the most fragile stages of the service process: the interaction points. An example of this is when the driver and the customer meet at the unloading place. The results show that even a SME can have the opportunity to succeed in competitive markets by managing CRM. Expensive software are usually out of reach of a SME, but having a CRM software does not automatically guar-antee success in customer relationship management. Small and medium-sized firms can commit their current customers and recruit new customers by utilizing customer knowledge, offering top quality service, being innovative and offering functional services.

Hitsauksen laadunhallinta offshore-teollisuuden alihankintakonepajassa

Diplomityön tavoitteena oli kuvata hitsauksen laadunhallintaa sekä hitsauksen laadunhallintajärjestelmän rakentamista offshore-teollisuuden konepajaan standardin ISO 3834-2 mukaisesti. Diplomityö koostuu teoriaosasta sekä käytännön osasta. Teoriaosa käsittelee laadunhallintaa yleisesti, minkä jälkeen se tarkentuu käsittelemään hitsauksen laadunhallintaa. Esimerkkinä on esitetty standardin NORSOK M-101 mukaisia laatuvaatimuksia. Laadunhallintaan liittyy oleellisesti myös hitsausliitoksen NDT-tarkastaminen, josta on käsitelty silmämääräinen tarkastus ja ultraäänitarkastus. Teoriaosan lopussa on käsitelty hitsauksen työturvallisuutta sekä hitsauksen laadunvalvontaa online-seurantajärjestelmällä. Käytännön osassa kerrotaan hitsauksen laatujärjestelmän rakentamisesta Meteco Oy:n Kuhmoisten tehtaalla. Yrityksen nykytilanne kartoitettiin, minkä jälkeen ryhdyttiin rakentamaan hitsauksen laadunhallintajärjestelmää. Tuloksena syntyi hitsauksen laatukäsikirja, joka pohjautuu standardiin ISO 3834-2. Käytännön osuudessa on kerrottu myös online-seurantajärjestelmän soveltuvuudesta alihankintakonepajan käyttöön. Hitsauksen laatujärjestelmän rakentamiseen kuului henkilöstön koulutustilaisuudet, joissa informoitiin edellä mainitun laatukäsikirjaan kuuluvista asioista. Tästä syntyi yritykseen myös koulutusohjelmasuunnitelma, joka antaa valmiudet yrityksen henkilökunnalle kehittyä työssään ja näin tehostaa toimintaa.

Spectrophotometric determination of metronidazole through Schiff's base system using vanillin and PDAB reagents in pharmaceutical preparations

Two simple sensitive and reproducible spectrophotometric methods have been developed for the determination of metronidazole either in pure form or in their tablets. The proposed methods are based on the reduction of the nitro group to amino group of the drug. The reduction of metronidazole was carried out with zinc powder and 5 N hydrochloric acid at room temperature in methanol. The resulting amine was then subjected to a condensation reaction with aromatic aldehyde namely, vanillin and p-dimethyl amino benzaldehyde (PDAB) to yield yellow colored Schiff's bases. The formed Schiff's bases are quantified spectrophotometrically at their absorption maxima at 422 nm for vanillin and 494 nm for PDAB. Beer's law was obeyed in the concentration ranges 10 to 65 µg mL-1 and 5 to 40 µg mL-1 with a limit of detection (LOD) of 0.080 µg mL-1 and 0.090 µg mL-1 for vanillin and PDAB, respectively. The mean percentage recoveries were found to be 100.05 ± 0.37 and 99.01 ± 0.76 for the two methods respectively. The proposed methods were successfully applied to determine the metronidazole in their tablet formulations and the results compared favorably to that of reference methods. The proposed methods are recommended for quality control and routine analysis.

Supersaturation-controlled crystallization

Identification of product requirements and quality, together with the management of production are key issues in chemical engineering. Quality control of crystalline products is part of the quality of many industrially manufactured products like paper, paintings, medicines and fertilizers. In most crystallization cases, quality is described with the size, polymorph, shape and purity of the crystal. The chemical composition, hydrodynamics and driving force, together with the operating temperature are in a key position when the properties of a crystalline product are controlled with the crystallization process. This study concentrates on managing the identified properties of a crystalline product with the control of a driving force. The controlling of the driving force can be based on the change of solubility or the change of concentration. Solubility can be changed with temperature, pressure and an antisolvent. The concentration of crystallizing compound, the solute can be changed with the evaporation of the solvent and with the addition of a reagent. The present study focuses on reagent addition and temperature change as methods of changing the level of the driving force. Three control structures for direct control of supersaturation are built, one for cooling crystallization and two for reactive crystallization. Closed loop feedback control structures are based on the measurement of the solute concentration with attenuated total reflection - Fourier transform infrared spectrometer. The details of the reagent feed are analyzed with experimental studies and with results of computational fluid dynamic simulations of the inert particle pulse in the premixer and inert particle injection to the mixing tank. Nucleation in conditions of controlled reactive crystallization is analyzed with Nielsen’s equation of homogeneous nucleation. The resulting control systems, based on regulation of supersaturation, can be used to produce the desired polymorph of an organic product. The polymorph composition of product crystals is controlled repeatably with the decision of a set value of supersaturation level.

Role and regulatory mechanisms of heat shock factor 2

Protein homeostasis is essential for cells to prosper and survive. Various forms of stress, such as elevated temperatures, oxidative stress, heavy metals or bacterial infections cause protein damage, which might lead to improper folding and formation of toxic protein aggregates. Protein aggregation is associated with serious pathological conditions such as Alzheimer’s and Huntington’s disease. The heat shock response is a defense mechanism that protects the cell against protein-damaging stress. Its ancient origin and high conservation among eukaryotes suggest that the response is crucial for survival. The main regulator of the heat shock response is the transcription factor heat shock factor 1 (HSF1), which induces transcription of genes encoding protective molecular chaperones. In vertebrates, a family of four HSFs exists (HSF1-4), with versatile functions not only in coping with acute stress, but also in development, longevity and cancer. Thus, knowledge of the HSFs will aid in our understanding on how cells survive suboptimal circumstances, but will also provide insights into normal physiological processes as well as diseaseassociated conditions. In this study, the function and regulation of HSF2 have been investigated. Earlier gene inactivation experiments in mice have revealed roles for HSF2 in development, particularly in corticogenesis and spermatogenesis. Here, we demonstrate that HSF2 holds a role also in the heat shock response and influences stress-induced expression of heat shock proteins. Intriguingly, DNA-binding activity of HSF2 upon stress was dependent on the presence of intact HSF1, suggesting functional interplay between HSF1 and HSF2. The underlying mechanism for this phenomenon could be configuration of heterotrimers between the two factors, a possibility that was experimentally verified. By changing the levels of HSF2, the expression of HSF1-HSF2 heterotrimer target genes was altered, implementing HSF2 as a modulator of HSF-mediated transcription. The results further indicate that HSF2 activity is dependent on its concentration, which led us to ask the question of how accurate HSF2 levels are achieved. Using mouse spermatogenesis as a model system, HSF2 was found to be under direct control of miR-18, a miRNA belonging to the miR-17~92 cluster/Oncomir-1 and whose physiological function had remained unclear. Investigations on spermatogenesis are severely hampered by the lack of cell systems that would mimic the complex differentiation processes that constitute male germ cell development. Therefore, to verify that HSF2 is regulated by miR-18 in spermatogenesis, a novel method named T-GIST (Transfection of Germ cells in Intact Seminiferous Tubules) was developed. Employing this method, the functional consequences of miR-18-mediated regulation in vivo were demonstrated; inhibition of miR- 18 led to increased expression of HSF2 and altered the expression of HSF2 target genes Ssty2 and Speer4a. Consequently, the results link miR-18 to HSF2-mediated processes such as germ cell maturation and quality control and provide miR-18 with a physiological role in gene expression during spermatogenesis.Taken together, this study presents compelling evidence that HSF2 is a transcriptional regulator in the heat shock response and establishes the concept of physical interplay between HSF2 and HSF1 and functional consequences thereof. This is also the first study describing miRNA-mediated regulation of an HSF.

Vuokamallisten kartonkipakkausten laadunvarmistuslaitteiston kehittäminen

Diplomityön tavoitteena oli kehittää vuokamallisille kartonkipakkauksille laadunvarmistuslaitteisto. Kirjallisen osan alussa esiteltiin vuokamallisten kartonkipakkausten valmistusprosessia. Tästä siirryttiin laatuasioihin, jossa tärkeimmät asiat olivat kartonkivuokien valmistuksessa esiintyvät laatupoikkeamat ja konenäkö. Tutkimusosan alussa esitellään Lappeenrannan teknillisessä yliopistossa kehitetty kartonkivuokien valmistuslinjasto. Tämän jälkeen vaatimuslistan pohjalta suunnitellaan kyseiseen linjastoon sopiva automaattinen laadunvalvontalaite, johon sisältyy myös kartonkivuokien siirtolaite. Suunnitteluprosessi aloitettiin koekuvaamalla kartonkivuokia erilaisilla kameroilla ja valaistusmenetelmillä. Koekuvausten perusteella valittiin konenäkölaitteisto. Tämän jälkeen toiminnoista luotiin periaatepiirroksia, joista kehitettiin varsinainen suunnitelma. Työn tuloksena saatiin suunnitelma konenäköön perustuvan automaattisen laadunvalvontalaitteen rakentamiselle.

Characterisation of Human Neutral and Lysosomal alpha-Mannosidases

Neutral alpha-mannosidase and lysosomal MAN2B1 alpha-mannosidase belong to glycoside hydrolase family 38, which contains essential enzymes required for the modification and catabolism of asparagine-linked glycans on proteins. MAN2B1 catalyses lysosomal glycan degradation, while neutral α-mannosidase is most likely involved in the catabolism of cytosolic free oligosaccharides. These mannose containing saccharides are generated during glycosylation or released from misfolded glycoproteins, which are detected by quality control in the endoplasmic reticulum. To characterise the biological function of human neutral α-mannosidase, I cloned the alpha-mannosidase cDNA and recombinantly expressed the enzyme. The purified enzyme trimmed the putative natural substrate Man9GlcNAc to Man5GlcNAc, whereas the reducing end GlcNAc2 limited trimming to Man8GlcNAc2. Neutral α-mannosidase showed highest enzyme activity at neutral pH and was activated by the cations Fe₂+, Co2+ and Mn₂+, Cu2+ in turn had a strong inhibitory effect on alpha-mannosidase activity. Analysis of its intracellular localisation revealed that neutral alpha-mannosidase is cytosolic and colocalises with proteasomes. Further work showed that the overexpression of neutral alpha-mannosidase affected the cytosolic free oligosaccharide content and led to enhanced endoplasmic reticulum associated degradation and underglycosylation of secreted proteins. The second part of the study focused on MAN2B1 and the inherited lysosomal storage disorder α-mannosidosis. In this disorder, deficient MAN2B1 activity is associated with mutations in the MAN2B1 gene. The thesis reports the molecular consequences of 35 alpha-mannosidosis associated mutations, including 29 novel missense mutations. According to experimental analyses, the mutations fall into four groups: Mutations, which prevent transport to lysosomes are accompanied with a lack of proteolytic processing of the enzyme (groups 1 and 3). Although the rest of the mutations (groups 2 and 4) allow transport to lysosomes, the mutated proteins are less efficiently processed to their mature form than is wild type MAN2B1. Analysis of the effect of the mutations on the model structure of human lysosomal alpha-mannosidase provides insights on their structural consequences. Mutations, which affect amino acids important for folding (prolines, glycines, cysteines) or domain interface interactions (arginines), arrest the enzyme in the endoplasmic reticulum. Surface mutations and changes, which do not drastically alter residue volume, are tolerated better. Descriptions of the mutations and clinical data are compiled in an α-mannosidosis database, which will be available for the scientific community. This thesis provides a detailed insight into two ubiquitous human alpha-mannosidases. It demonstrates that neutral alpha-mannosidase is involved in the degradation of cytosolic oligosaccharides and suggests that the regulation of this α-mannosidase is important for maintaining the cellular homeostasis of N-glycosylation and glycan degradation. The study on alpha-mannosidosis associated mutations identifies multiple mechanisms for how these mutations are detrimental for MAN2B1 activity. The α-mannosidosis database will benefit both clinicians and scientific research on lysosomal alpha‑mannosidosis.

Kinetic behavior of nitrification in the post-treatment of poultry wastewater in a sequential batch reactor

A sequential batch reactor with suspended biomass and useful volume of 5 L was used in the removal of nutrients and organic matter in workbench scale under optimal conditions obtained by central composite rotational design (CCRD), with cycle time (CT) of 16 h (10.15 h, aerobic phase, and 4.35 h, anoxic phase) and carbon: nitrogen ratio (COD/NO2--N+NO3--N) equal to 6. Complete cycles (20), nitrification followed by denitrification, were evaluated to investigate the kinetic behavior of degradation of organic (COD) and nitrogenated (NH4+-N, NO2--N and NO3--N) matter present in the effluent from a bird slaughterhouse and industrial processing facility, as well as to evaluate the stability of the reactor using Shewhart control charts of individual measures. The results indicate means total inorganic nitrogen (NH4+-N+NO2- -N+NO3--N) removal of 84.32±1.59% and organic matter (COD) of 53.65±8.48% in the complete process (nitrification-denitrification) with the process under statistical control. The nitrifying activity during the aerobic phase estimated from the determination of the kinetic parameters had mean K1 and K2 values of 0.00381±0.00043 min-1 and 0.00381±0.00043 min-1, respectively. The evaluation of the kinetic behavior of the conversion of nitrogen indicated a possible reduction of CT in the anoxic phase, since removals of NO2--N and NO3--N higher than 90% were obtained with only 1 h of denitrification.

The use of infrared thermography in the identification of pododermatitis in broilers

ABSTRACT International trade in broiler’ feet, mainly to Asian markets, has demanded better quality control. The objective of this research was to study the suitability of using chicken footpad surface temperature to determine early lesions of pododermatitis. The project was conducted in two houses A1 and A2) in a commercial farm during one production flock. A1 had reused litter of wood shavings and rice hulls, and A2 had a new litter of sawdust. Both houses had positive pressure ventilation. The inner area of the poultry was virtually divided into three quadrants. The footpads were checked for the feet quality, and a degree of pododermatitis was awarded. Thermal images were made to test the surface temperature of the foot and identify inflammation in a total of 30 birds per house, at ages 5, 19, 29, 28 and 40 days of grow-out. Conditions of the rearing environment as well as the surface temperature of the litter, litter moisture, and degree of compression, were recorded. The environment within the houses did not differ. The surface temperatures of the footpad did not differ between the groups. The minimum footpad surface temperatures within the scores were similar, except for the score 3, which did not occur in A1. There was a prevalence of severe injury in the house with a new litter.

Analyysityökalun kehittäminen C#-kielisten lähdekoodien tarkastamiseen

Katselmoinnit ja tarkastusmenettelyt ovat osa ohjelmistotuotantoprosessin laadunvarmistusta. Staattisella tarkastamisella tarkoitetaan ohjelmistotuotteen visuaalista tarkastamista ohjelmistovirheiden havaitsemiseksi ja korjaamiseksi. Ohjelmiston lähdekoodin tarkastaminen voidaan suorittaa automaattisesti tarkoitukseen sopivalla ohjelmistolla l. analyysityökalulla. Tässä työssä toteutettiin analyysityökalu C#-kielisten lähdekoodien tarkastamiseen. Työkalulla suoritetussa kenttätestauksessa havaittiin tarkastettavissa ohjelmistoissa ohjelmiston ylläpitoon vaikuttavia puutteita. Lisäksi työssä tarkasteltiin katselmointeja osana ohjelmistotuotantoprosessin laadunvarmistusta sekä erilaisia ohjelmistovirheitä ja niiden lähteitä.