Determinação turbidimétrica em fluxo de cloridrato de fluoxetina em formulações farmacêuticas

A simple, accurate and precise flow-injection turbidimetric procedure for the determination of fluoxetine hydrochloride in pharmaceutical formulations is reported. The procedure is based on the precipitation of chloride of fluoxetine hydrochloride with silver nitrate solution and the yielded insoluble AgCl(s) was monitored at 420 nm. The analytical curve was linear in the fluoxetine hydrochloride concentration range 3.0 x 10-5 - 5.0 x 10-4 mol L-1 with a detection limit of 10 µmol L-1 and, a sample throughout of 60 h-1.

Determinação de citrato de sildenafila e de tadalafila por cromatografia líquida de ultraeficiência com detecção por arranjo de diodos (CLUE-DAD)

A simple ultra-performance liquid chromatographic method for the simultaneous determination of sildenafil citrate and tadalafil was developed and validated. Sample preparation was dissolution in methanol, followed by centrifugation and dilution (1:10) with methanol. Analysis was performed in an Acquity® UPLC system with Acquity® BEH C18 column (2.1 x 50 mm, with 1.7 μm particles). The elution was isocratic with phosphate buffer pH 2.3 and acetonitrile (65:35, v/v) at a flow rate of 0.7 mL/min. The method presented adequate specificity, linearity, precision and accuracy and allowed the determination of the drugs in seized forensic samples.

Determinação de bussulfano em plasma através de cromatografia líquida de alta eficiência com detector de arranjo de diodos e derivatização com dietilditiocarbamato de sódio

A high performance liquid chromatographic-diode array detection method for the determination of busulfan in plasma was developed and validated. Sample preparation consisted of protein precipitation followed by derivatization with sodium diethyldithiocarbamate and liquid-liquid extraction with methyl-tert-butyl ether. Chromatograms were monitored at 277 nm. Separation was carried out on a Lichrospher RP 18 column (5 µm, 250 x 4 mm). The mobile phase consisted of water and acetonitrile (20:80, v/v). The method presented adequate specificity, linearity, precision and accuracy and allowed reliable determination of busulfan in clinical plasma samples, being applied to three patients submitted to bone marrow transplantation.

Análise exploratória rápida e não destrutiva (screening) da presença de elementos químicos tóxicos em material escolar por fluorescência de raios x

Compulsory in Brazil, toy certification aims to prevent possible risks in toy use. The Brazilian National Institute of Metrology (INMETRO) establishes the maximum concentration of toxic elements (TE) that may be present in toys. This study evaluates the presence of TE in different crayons, gouache and modeling clays using X-ray fluorescence. This technique is fast, has low operating cost and minimum sample pretreatment, resulting in a clean procedure without reagent consumption and waste generation. Bromine (in gouache) and Barium (in crayons) were the only potentially TE identified in the samples studied.

VALIDAÇÃO INTRALABORATORIAL DA DETERMINAÇÃO DE METILTESTOSTERONA EM ÁGUAS NATURAIS POR VOLTAMETRIA USANDO ELETRODO DE GOTA PENDENTE DE MERCÚRIO

17α-methyltestosterone (17α-MT) is a hormone used in medicine and aquaculture which can be found in natural waters. It is considered an emerging organic pollutant and its accurate and precise analysis is required. Therefore, this study presents the in-house validation of a new voltammetric methodology for 17α-MT analysis in natural waters. A hanging mercury drop electrode and NaOH 5.0×10-3 mol L-1 solution as the electrolyte were employed in the voltammetric determinations. To confirm the presence of 17α-MT in water samples quantitative analysis was performed by ESI-MS/MS. The methodology exhibited good selectivity, linearity, accuracy (recovery of between 100.4 and 108.8%) and precision (RSDs for repeatability of 2.85% and for intermediate precision of 14.10%, with n = 5, respectively). LOD and LOQ were 3.07 µg L-1 and 10.78 µg L-1, respectively. The methodology was suitable for determination of 17α-MT in natural waters due to low-cost, good performance and no sample pre-treatment required.

Direct determination of calcium in milk by atomic absorption spectrometry using flow-injection analysis

A flow-injection system with sample and reagent addition by the synchronous merging zones approach for calcium determination in milk by flame AAS is proposed. Main parameters were optimized using a factorial design with central point. The optimum conditions were 2.5% (m/v) for La concentration, 8 mL min-1 for the carrier flow-rate, 20 cm for coiled reactor and 250 ìL for sample volume. Different sample preparation procedures were evaluated such as dilution in water or acid and microwave-assisted decomposition using concentrated or diluted acids. The optimized flow system was applied to determine Ca in eleven commercial milk samples and two standard reference materials diluted in water. Similar calcium levels were encountered comparing the results obtained by the proposed method (dilution in water) with those obtained using microwave-oven digestion. Results obtained in two standard reference materials were in agreement at 95% confidence level with those certified. Recoveries of spiked samples were in the 93% - 116% range. Relative standard deviation (n = 12) was < 5.4% and the sample throughput was 150 measurements per hour, corresponding to a consumption of 250 µL of sample and 6.25 mg La per determination.

Transcriptional Profiling of Organ‐Specific Autoimmunity in Human

Our understanding of the pathogenesis of organ‐specific autoinflammation has been restricted by limited access to the target organs. Peripheral blood, however, as a preferred transportation route for immune cells, provides a window to assess the entire immune system throughout the body. Transcriptional profiling with RNA stabilizing blood collection tubes reflects in vivo expression profiles at the time the blood is drawn, allowing detection of the disease activity in different samples or within the same sample over time. The main objective of this Ph.D. study was to apply gene‐expression microarrays in the characterization of peripheral blood transcriptional profiles in patients with autoimmune diseases. To achieve this goal a custom cDNA microarray targeted for gene‐expression profiling of human immune system was designed and produced. Sample collection and preparation was then optimized to allow gene‐expression profiling from whole‐blood samples. To overcome challenges resulting from minute amounts of sample material, RNA amplification was successfully applied to study pregnancy related immunosuppression in patients with multiple sclerosis (MS). Furthermore, similar sample preparation was applied to characterize longitudinal genome‐wide expression profiles in children with type 1 diabetes (T1D) associated autoantibodies and eventually clinical T1D. Blood transcriptome analyses, using both the ImmunoChip cDNA microarray with targeted probe selection and genome‐wide Affymetrix U133 Plus 2.0 oligonucleotide array, enabled monitoring of autoimmune activity. Novel disease related genes and general autoimmune signatures were identified. Notably, down‐regulation of the HLA class Ib molecules in peripheral blood was associated with disease activity in both MS and T1D. Taken together, these studies demonstrate the potential of peripheral blood transcriptional profiling in biomedical research and diagnostics. Imbalances in peripheral blood transcriptional activity may reveal dynamic changes that are relevant for the disease but might be completely missed in conventional cross‐sectional studies.

Reflexive Space. A Constructionist Model of the Russian Reflexive Marker

This study examines the structure of the Russian Reflexive Marker ( ся/-сь) and offers a usage-based model building on Construction Grammar and a probabilistic view of linguistic structure. Traditionally, reflexive verbs are accounted for relative to non-reflexive verbs. These accounts assume that linguistic structures emerge as pairs. Furthermore, these accounts assume directionality where the semantics and structure of a reflexive verb can be derived from the non-reflexive verb. However, this directionality does not necessarily hold diachronically. Additionally, the semantics and the patterns associated with a particular reflexive verb are not always shared with the non-reflexive verb. Thus, a model is proposed that can accommodate the traditional pairs as well as for the possible deviations without postulating different systems. A random sample of 2000 instances marked with the Reflexive Marker was extracted from the Russian National Corpus and the sample used in this study contains 819 unique reflexive verbs. This study moves away from the traditional pair account and introduces the concept of Neighbor Verb. A neighbor verb exists for a reflexive verb if they share the same phonological form excluding the Reflexive Marker. It is claimed here that the Reflexive Marker constitutes a system in Russian and the relation between the reflexive and neighbor verbs constitutes a cross-paradigmatic relation. Furthermore, the relation between the reflexive and the neighbor verb is argued to be of symbolic connectivity rather than directionality. Effectively, the relation holding between particular instantiations can vary. The theoretical basis of the present study builds on this assumption. Several new variables are examined in order to systematically model variability of this symbolic connectivity, specifically the degree and strength of connectivity between items. In usage-based models, the lexicon does not constitute an unstructured list of items. Instead, items are assumed to be interconnected in a network. This interconnectedness is defined as Neighborhood in this study. Additionally, each verb carves its own niche within the Neighborhood and this interconnectedness is modeled through rhyme verbs constituting the degree of connectivity of a particular verb in the lexicon. The second component of the degree of connectivity concerns the status of a particular verb relative to its rhyme verbs. The connectivity within the neighborhood of a particular verb varies and this variability is quantified by using the Levenshtein distance. The second property of the lexical network is the strength of connectivity between items. Frequency of use has been one of the primary variables in functional linguistics used to probe this. In addition, a new variable called Constructional Entropy is introduced in this study building on information theory. It is a quantification of the amount of information carried by a particular reflexive verb in one or more argument constructions. The results of the lexical connectivity indicate that the reflexive verbs have statistically greater neighborhood distances than the neighbor verbs. This distributional property can be used to motivate the traditional observation that the reflexive verbs tend to have idiosyncratic properties. A set of argument constructions, generalizations over usage patterns, are proposed for the reflexive verbs in this study. In addition to the variables associated with the lexical connectivity, a number of variables proposed in the literature are explored and used as predictors in the model. The second part of this study introduces the use of a machine learning algorithm called Random Forests. The performance of the model indicates that it is capable, up to a degree, of disambiguating the proposed argument construction types of the Russian Reflexive Marker. Additionally, a global ranking of the predictors used in the model is offered. Finally, most construction grammars assume that argument construction form a network structure. A new method is proposed that establishes generalization over the argument constructions referred to as Linking Construction. In sum, this study explores the structural properties of the Russian Reflexive Marker and a new model is set forth that can accommodate both the traditional pairs and potential deviations from it in a principled manner.

Comparison of model-based above ground biomass predictions from LiDAR

In order to reduce greenhouse emissions from forest degradation and deforestation the international programme REDD (Reducing Emissions from Deforestation and forest Degradation) was established in 2005 by the United Nations Framework Convention on Climate Change (UNFCCC). This programme is aimed to financially reward to developing countries for any emissions reductions. Under this programm the project of setting up the payment system in Nepal was established. This project is aimed to engage local communities in forest monitoring. The major objective of this thesis is to compare and verify data obtained from di erect sources - remotely sensed data, namely LiDAR and field sample measurements made by two groups of researchers using two regression models - Sparse Bayesian Regression and Bayesian Regression with Orthogonal Variables.

Molecular systematics of Thorea (Rhodophyta, Thoreales) species in Brazil

This study aimed to evaluate species level taxonomy and phylogenetic relationship among Thorea species in Brazil and other regions of the world using two molecular markers - RUBISCO large subunit plastid gene (rbcL) and nuclear small-subunit ribosomal DNA (SSU rDNA). Three samples of Thorea from Brazil (states of Mato Grosso do Sul and São Paulo) and one sample from Dominican Republic (DR) were sequenced. Analyses based on partial sequences of rbcL (1,282 bp) and complete sequences of SSU (1,752 bp) were essentially congruent and revealed that Thoreales formed a distinct monophyletic clade, which had two major branches with high support, representing the genera Thorea and Nemalionopsis. Thorea clade had four main branches with high support for all analyses, each one representing the species: 1) T. gaudichaudii C. Agardh from Asia (Japan and Philippines) - this clade occurred only in the rbcL analyses; 2) T. violacea Bory from Asia (Japan) and North America (U.S.A. and DR); 3) T. hispida (Thore) Desvaux from Europe (England) and Asia (Japan); 4) a distinct group with the three Brazilian samples (sequence identity: rbcL 97.2%, 1,246 bp; SSU 96.0-98.1%, 1,699-1,720 bp). The Brazilian samples clearly formed a monophyletic clade based on both molecular markers and was interpreted as a separate species, for which we resurrected the name T. bachmannii Pujals. Morphological and molecular evidences indicate that the Thoreales is well-resolved at ordinal and generic levels. In contrast, Thorea species recognized by molecular data require additional characters (e.g. reproductive and chromosome numbers) to allow consistent and reliable taxonomic circumscription aiming at a world revision based on molecular and morphological evidences.

Multivariate analysis of pollen frequency of the native species Escallonia pulverulenta (Saxifragaceae) in Chilean honeys

The aim of this work was the identification of geographic zones suitable for the production of honeys in which pollen grains of Escallonia pulverulenta (Ruiz & Pav.) Pers. (Saxifragaceae) can be detected. The analysis of botanical origin of 240 honey samples produced between La Serena and Puerto Mont (the IV and X Administrative Regions of Chile), allowed the detection of pollen grains of E. pulverulenta in 46 Chilean honeys. The geographic distribution of the honeys studied is presented together with their affinities, through factor analysis and frequency tables. The study was based on the presence of E. pulverulenta pollen. Escallonia pulverulenta pollen percentages oscillated between 0.24% and 78.5%. Seventeen of the studied samples were designated as unifloral - i.e. samples showing more than 45% pollen of a determined plant species. Two of these corresponded to E. pulverulenta (corontillo, madroño or barraco) honeys. The remaining unifloral honeys correspond to 8 samples of Lotus uliginosus Schkuhr (birdsfoot trefoil), 2 samples of Aristotelia chilensis (Molina) Stuntz (maqui) and 1 sample of Escallonia rubra (Ruiz & Pav.) Pers. (siete camisas), Eucryphia cordifolia Cav. (ulmo or muemo), Weinmannia trichosperma Cav. (tineo), Rubus ulmifolius Schott (blackberry) and Brassica rapa L. (turnip). Honeys with different percentages of E. pulverulenta pollen - statistically analyzed through correspondence analysis - could be associated and assigned to one of three geographic types, defined on the basis of this analysis. The geographical type areas defined were the Northern Mediterranean Zone (samples from the IV Region), Central Mediterranean Zone (samples from the V to the VIII regions including two samples of unifloral Escallonia pulverulenta honey), and Southern Mediterranean Zone (samples from the IX Region).

Human group C rotavirus in children with diarrhea in the Federal District, Brazil

Group C rotaviruses are fastidious in their in vitro cell culture requirements. Recent serosurveys indicate that antibody to group C rotavirus is present in 3-45% of the human population in certain geographic locations, suggesting that rotavirus group C infection is more prevalent than previously believed and that the low rate of detection of these agents is probably due to the lack of sensitive diagnostic assays. From March to December 1994, 406 fecal specimens were collected from children under five years of age who were outpatients at the emergency services of nine public hospitals in Brasília, Federal District, Brazil. In addition to the samples from children, one public outpatient unit requested virological investigation of a stool sample from an HIV-seropositive adult male with diarrhea of sudden onset. All samples were analyzed by enzyme immunoassay for group A rotavirus and adenovirus (EIARA) and by polyacrylamide gel electrophoresis (PAGE). One hundred and seven (26%) were positive for group A rotavirus. Four samples from children and the sample from the HIV-seropositive patient, although negative by EIARA, showed a group C rotavirus profile by PAGE and were positive for rotavirus by electron microscopy. Using specific VP6 and VP7 primers for group C rotavirus, a reverse transcriptase-polymerase chain reaction (RT-PCR) was performed and products were detected by agarose gel electrophoresis and ethidium bromide staining. These products were confirmed to be specific for group C rotavirus by using digoxigenin-oligonucleotide probes, Southern hybridization and chemiluminescent detection. The five positive group C rotavirus samples were detected in August (3 samples) and September (2 samples). To the best of our knowledge, this is the first report of group C rotavirus detected in the Federal District, Brazil and in an HIV-seropositive patient with acute gastroenteritis.

Omistuksen tavoitteellisuus perheyrityksissä - kartoitus omistamisen tavoitteista ja tavoitteiden määrittelystä

Suomi tarvitsee menestyviä yrittäjiä, uusia yrityksiä, kasvuyrityksiä ja kansainvälistymistä. Yrittä-jyydelle asetetaankin suuria odotuksia. Suurin huomio yrittäjyyden tarkastelussa on kuitenkin keskittynyt liiketoimintojen kehittämiseen – sen sijaan omistajuuden ja yrittäjyyden yhteyttä ei ole juurikaan huomioitu. Tämän seurauksena myös omistajuuden vaikutusta yrityksen kehittymi-seen ja menestykseen on analysoitu vähän. Edistääkseen yrityksen kehittymistä, tulee omistuksen olla tavoitteellista. Omistamisen tavoittei-den tarkastelu on kuitenkin harvinaista ja parhaimmillaankin yksiulotteista: omistamisen tavoit-teiden nähdään liittyvän lähes yksinomaan taloudellisiin tavoitteisiin, ja tavoitteiden moninaisuus unohdetaan. Tässä tutkimuksessa tarkastellaan suomalaisten perheyrittäjien omistamisen tavoit-teellisuutta, tavoitteiden määrittelyä sekä omistajaohjauksen keinoja, lähinnä hallituksen roolia, omistukseen liittyvien ratkaisujen tekijänä. Tutkimushankkeen aineisto kerättiin EK:n ja Perheyritysten liiton jäsenyrityksistä. Aineisto kerät-tiin joulukuussa 2013 ja tammikuussa 2014 web-pohjaisella kyselyllä ja siihen vastasi määräajan kuluessa 332 vastaajaa. Vastaajista perheyrityksiä oli 241 kpl. Tarkemman analyysin kohteeksi jäi 233 perheyritystä. Tutkimuksen tulokset osoittavat, että suomalaisten yrittäjien omistajuuteen liittyvät tavoitteet ovat monipuolisempia kuin yleisesti oletetaan. Puhtaasti taloudellisiin motiiveihin perustuvan yri-tysten omistamisen ohella liiketoimintaan kohdistuva mielenkiinto ja halu toimia yrittäjänä ovat merkittäviä omistamisen vaikuttimia. Suuri joukko yritysten omistajista ei kuitenkaan tietoisesti pohdi omistamisen tavoitteita vaan omistuksen tavoitteet määrittyvät liiketoiminnan tavoitteiden tai totuttujen tapojen ja perinteiden kautta. Osalla yritysten omistajista omistus muuttuu tavoit-teellisemmaksi yrityksen kehittyessä ja omistajajoukon kasvaessa. Ensimmäisen sukupolven yrittäjät näyttävät toimivan ennen kaikkea yrittäjäideologian mukaisesti pyrkien kasvattamaan yritystä ja painottaen liiketaloudellisia arvoja päätöksenteossa. Yritystoi-minnan kehittyessä ja sukupolvien lisääntyessä yrityksen omistamiseen liittyvät tavoitteet moni-puolistuvat ja monimutkaistuvat, eivätkä enää perustu pelkästään liiketoiminnallisiin menes-tysodotuksiin. Hallitustoiminnan aktivointi aktivoi myös omistuksen tavoitteellisuuden kehittymistä. Omistus-pohjan hajaantuminen lisää tarvetta erilaisten omistajaohjauksen mekanismien hyödyntämiselle. Hallituksella näyttää olevan rooli omistusratkaisuiden arvioinnissa sekä yrityksen operatiiviseen toimintaan liittyvissä omistusratkaisuissa, mutta myös läheisemmin omistamiseen ja omistajapoh-jaan liittyvissä omistusratkaisuissa. Perheomistukseen liittyvien tavoitteiden huomioiminen on tärkeää perheyrityksen hallituksen toiminnassa myös yrityksen toiminnan vakiintuessa. Sen sijaan, että perheeseen liittyvät omista-misen tavoitteet katoaisivat yrityksestä, muuttuu niiden määrittely tietoisemmaksi ja osaksi yri-tyksen hallintoa. Määräysvallan merkitys tavoitteena pienenee, kun omistajan etäisyys operatiivi-sesta toiminnasta, tai sen johtamisesta kasvaa. Tutkimuksen tulokset kertovat omistamisen tavoitteista ja tavoitteiden määrittelystä. Selvityksen lopussa luotiin määritelmä päämäärätietoiselle omistajalle: Päämäärätietoinen omistaja on omista-ja, joka tietoisesti pohtii tavoitteita omistamiselleen sekä käyttää aktiivisesti omistajavaltaansa valitsemiensa päämäärien saavuttamiseksi.

Karyotype of cryopreserved bone marrow cells

The analysis of chromosomal abnormalities is important for the study of hematological neoplastic disorders since it facilitates classification of the disease. The ability to perform chromosome analysis of cryopreserved malignant marrow or peripheral blast cells is important for retrospective studies. In the present study, we compared the karyotype of fresh bone marrow cells (20 metaphases) to that of cells stored with a simplified cryopreservation method, evaluated the effect of the use of granulocyte-macrophage colony-stimulating factor (GM-CSF) as an in vitro mitotic index stimulator, and compared the cell viability and chromosome morphology of fresh and cryopreserved cells whenever possible (sufficient metaphases for analysis). Twenty-five bone marrow samples from 24 patients with hematological disorders such as acute myeloid leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myeloid leukemia, megaloblastic anemia and lymphoma (8, 3, 3, 8, 1, and 1 patients, respectively) were selected at diagnosis, at relapse or during routine follow-up and one sample was obtained from a bone marrow donor after informed consent. Average cell viability before and after freezing was 98.8 and 78.5%, respectively (P < 0.05). Cytogenetic analysis was successful in 76% of fresh cell cultures, as opposed to 52% of cryopreserved samples (P < 0.05). GM-CSF had no proliferative effect before or after freezing. The morphological aspects of the chromosomes in fresh and cryopreserved cells were subjectively the same. The present study shows that cytogenetic analysis of cryopreserved bone marrow cells can be a reliable alternative when fresh cell analysis cannot be done, notwithstanding the reduced viability and lower percent of successful analysis that are associated with freezing.

Sensory descriptors of cocoa beans from cultivated trees of Soconusco, Chiapas, Mexico

The odor and taste profile of cocoa bean samples obtained from trees cultivated in southern Mexico were evaluated by trained panelists. Seven representative samples (groups) of a total of 45 were analyzed. Four attributes of taste (sweetness, bitterness, acidity and astringency), and nine of odor (chocolate, nutty, hazelnut, sweet, acidity, roasted, spicy, musty and off-odor) were evaluated. A sample (G7) with higher scores in sweet taste and sweet and nutty odors was detected, as well as a high association between these descriptors and the sample, analyzed through principal component analysis (PCA). Similarly, samples that showed high scores for non-desired odors in cocoas such as off-odor and musty were identified and related by PCA to roasted odor and astringent taste (G2 and G4). Based on this scores, the samples were listed in descending order by their sensory quality as G7> G5> G6> G3> G1> G4> G2.