LC/ESI-MS method applied to characterization of flavonoids glycosides in B. forficata subsp. pruinosa

Bauhinia forficata is used in folk medicine for its hypoglycemiant effect. In the south of Brazil, the subspecies pruinosa is found in a region with the characteristic flora, pampa biome. This species has been consumed by the local population as a tea for diabetes treatment. We studied the chemical composition of hydroethanolic extracts using LC/ESI-MS. The leaf extracts were prepared by percolation with 50% (v/v) ethanol. The chromatographic analyses were performed using a reverse-phase system, gradient elution with acetonitrile:phosphoric acid 0.05%, and ESI-MS in the positive ion mode. The chemical profile of the flavonoids was suggested to involve four quercetin and kaempferol glycosides.

Avaliação da labilidade de alumínio em infusões de erva-mate empregando voltametria adsortiva de redissolução catódica

Infusions of yerba mate obtained at different stages of industrialization were evaluated to determine the bioavailable fraction of Al. Adsorptive Cathodic Stripping Voltammetry using DASA (complexing agent) was applied to determine the labile fraction of Al at pH 5.0 and pH 8.0 for the total fraction of dissolved Al. The results indicate that on average 60% of Al is complexed with organic compounds, minimizing their bioavailability; however, the labile fraction exceeds by up to 4 times the maximum weekly intake recommended by the World Health Organization.

USO DA RMN DE BAIXA RESOLUÇÃO NA AVALIAÇÃO DA DINÂMICA MOLECULAR DO Origanum vulgare

In this work, proton NMR relaxometry was used to measure the behavior of spin-lattice relaxation time with T1H as the time constant, and also of spin-spin relaxation time with the time constant T2H. These relaxometry parameters were determined to better understand the changes in the main structures present in commercial and in nature forms of origanum. The T1H relaxation data showed that the structures which had higher molecular mass were more sensitive to degradation with increased temperature treatment. According to the values of the T2H parameter, up to 150 degrees no significant change in the mobility and organization of water was observed. These data infer that the ideal cooking temperature and tea preparation mode for this herb should be around 100 degrees for the sample not to lose its characteristics. Also, it is not advisable to cook this herb at higher than 150 degrees but better to consume it at room temperature, especially give commercial herb has already been dehydrated.

Sosiaalisen sukupuolen rakentuminen, suurkaupunki ja kammottavan kokemus Katherine Mansfieldin novelleissa

Tutkielmani tarkastelee englantilaisen modernistin Katherine Mansfieldin (1888–1923) novelleja The Woman at the Store, The Little Governess, Bliss, Pictures, Miss Brill ja A Cup of Tea sosiaalisen sukupuolen rakentumisen näkökulmasta. Novellien kirjoitus- ja julkaisuajankohta ajoittuu vuosille 1911–1924. Uudesta-Seelannista Lontooseen muuttanutta Mansfieldia pidetään yhtenä englantilaisen modernistisen kirjallisuuden uranuurtajista, jonka kehittämän modernin novellimuodon keskeisiä innovaatioita olivat tarinan juonettomuus, tajunnanvirta osana kerrontaa ja psykologisen hetken korostaminen. Mansfieldin tuotannon taustalla vaikuttivat voimakkaasti myös ajankohdalle ominaiset diskurssit naisen asemasta ja mahdollisuuksista. Lähestyn aihettani gender-teoreettisesta viitekehyksestä käsin, jonka keskeisin näkemys on käsitys sukupuolesta sosiaalisesti rakentuneena. Ensimmäisessä käsittelyluvussa pohdin niitä naisen ja miehen representaatioita, joita valitsemissani novelleissa esiintyy. Toisessa käsittelyluvussa syvennyn tarkastelemaan naishahmojen suhdetta suurkaupunkiympäristöön, johon monet novelleista sijoittuvat, ja analysoimaan sitä, millä lailla kaupunki tilana muovaa naissubjekteja. Kolmannessa eli viimeisessä käsittelyluvussa kiinnitän huomioni kammottavan kokemukseen, joka on ominainen kaikille käsittelemilleni novelleille ja joka juontuu sekä niiden kerronnallisista keinoista että novellihenkilöiden konflikteista itsensä ja ulkomaailman kanssa, joilla näen vahvan yhteyden sukupuolitematiikkaan. Sigmund Freud on määritellyt kammottavan joksikin ammoin tutuksi ja torjutuksi, joka on yhtäkkiä murtautunut esiin. Tätä ajatusta kehitellyt Julia Kristeva näkee kammottavan ilmenevän muukalaisena meissä itsessämme. Edellä esitetyin tavoin on mahdollista hahmottaa myös sitä novellien loppuratkaisua, jossa arkinen muuttuu yllättäen uhkaavaksi ja pelottavaksi ja päähenkilöille avautuu jokin uusi tietoisuus itsestään ja ympäröivästä maailmasta. Mansfieldin naiskuva osoittautuu analyysini pohjalta melko monipuoliseksi, mutta miehet esitetään novelleissa lähes poikkeuksetta hegemonisen maskuliinisina. Tällä on yhteytensä novellien kirjoitusajankohtaan ja sen kiivaaseen yhteiskunnalliseen keskusteluun naisen roolista. 1900-luvun alussa naiset alkoivat enenevässä määrin ottaa haltuunsa julkista tilaa. Tarkastelemani novellit ilmentävät monin tavoin sitä ”toisin toistamisen” vaikeutta, minkä totunnaisista sukupuolirooleista poikkeaminen tuottaa. Novellien naishenkilöitä leimaa halu vapautua, mutta rajat tulevat vastaan viimeistään julkisessa tilassa. Käsittelemäni novellit tuovat oivaltavasti esille sukupuolten keskinäisiä valtarakenteita, joita säännöstelee pitkälti vallassa oleva heteronormatiivinen maskuliinisuus. Novellien lopussa henkilöt joutuvat kohtaamaan itsensä uudella tavalla ja määrittelemään uudelleen suhteensa omaan sukupuoleensa ja seksuaalisuuteensa.

Astringent Food Compounds and Their Interactions with Taste Properties

Astringency is traditionally thought to be induced by plant tannins in foods. Because of this current research concerning the mechanism of astringency is focused on tannin‐protein interactions and thus on precipitation, which may be perceived by mechanoreceptors. However, astringency is elicited by a wide range of different phenolic compounds, as well as, some non‐phenolic compounds in various foods. Many ellagitannins or smaller compounds that contribute to astringent properties do not interact with salivary proteins and may be directly perceived through some receptors. Generally, the higher degree of polymerization of proanthocyanidins can be associated with more intense astringency. However, the astringent properties of smaller phenolic compounds may not be directly predicted from the structure of a compound, although glycosylation has a significant role. The astringency of organic acids may be directly linked to the perception of sourness, and this increases along with decreasing pH. Astringency can be divided into different sub‐qualities, including even other qualities than traditional mouth‐drying, puckering or roughing sensations. Astringency is often accompanied by bitter or sour or both taste properties. The different sub‐qualities can be influenced by different astringent compounds. In general, the glycolysation of the phenolic compound results in more velvety and smooth mouthdrying astringency. Flavonol glycosides and other flavonoid compounds and ellagitannins contribute to this velvety mouthdrying astringency. Additionally, they often lack the bitter properties. Proanthocyanidins and phenolic acids elicit more puckering and roughing astringency with some additional bitter properties. Quercetin 3‐O‐rutinoside, along with other quercetin glycosides, is among the key astringent compounds in black tea and red currants. In foods, there are always various other additional attributes that are perceived at the same with astringency. Astringent compounds themselves may have other sensory characteristics, such as bitter or sour properties, or they may enhance or suppress other sensory properties. Components contributing to these other properties, such as sugars, may also have similar effects on astringent sensations. Food components eliciting sweetness or fattiness or some polymeric polysaccharides can be used to mask astringent subqualities. Astringency can generally be referred to as a negative contributor to the liking of various foods. On the other hand, perceptions of astringent properties can vary among individuals. Many genetic factors that influence perceptions of taste properties, such as variations in perceiving a bitter taste or variations in saliva, may also effect the perception of astringency. Individuals who are more sensitive to different sensations may notice the differences between astringent properties more clearly. This may not have effects on the overall perception of astringency. However, in many cases, the liking of astringent foods may need to be learned by repetitive exposure. Astringency is often among the key sensory properties forming the unique overall flavour of certain foods, and therefore it also influences whether or not a food is liked. In many cases, astringency may be an important sub‐property suppressed by other more abundant sensory properties, but it may still have a significant contribution to the overall flavour and thus consumer preferences. The results of the practical work of this thesis show that the astringent phenolic compounds are mostly located in the skin fractions of black currants, crowberries and bilberries (publications I–III). The skin fractions themselves are rather tasteless. However, the astringent phenolic compounds can be efficiently removed from these skin fractions by consecutive ethanol extractions. Berries contain a wide range of different flavonol glycosides, hydroxycinnamic acid derivatives and anthocyanins and some of them strongly contribute to the different astringent and bitterness properties. Sweetness and sourness are located in the juice fractions along with the majority of sugars and fruit acids. The sweet and sour properties of the juice may be used to mask the astringent and bitterness properties of the extracts. Enzymatic treatments increase the astringent properties and fermented flavour of the black currant juice and decrease sweetness and freshness due to the effects on chemical compositions (IV). Sourness and sweetness are positive contributors to the liking of crowberry and bilberry fractions, whereas bitterness is more negative (V). Some astringent properties in berries are clearly negative factors, whereas some may be more positive. The liking of berries is strongly influenced by various consumer background factors, such as motives and health concerns. The liking of berries and berry fractions may also be affected by genetic factors, such as variations in the gene hTAS2R38, which codes bitter taste receptors (V).

ESTUDO DA RELAÇÃO ENTRE CONDIÇÕES DE POLPAÇÃO E PROPRIEDADES FÍSICO-MECÂNICAS DO PAPEL

RESUMOO objetivo deste estudo foi avaliar a influência da condição de polpação nas propriedades físico-mecânicas do papel branqueado de eucalipto. Para este estudo, foram utilizados cavacos de um híbrido clonal de eucalipto (Eucalyptus grandis x Eucalyptus urophylla) de 7 anos de idade. As polpas foram produzidas por duas condições distintas de polpação Lo-SolidsTM, sendo elas: Condição PI – álcali efetivo de 16,5%, sulfidez de 30% e temperatura de cozimento de 155 ºC; e Condição PII – álcali efetivo de 17,5%, sulfidez de 32% e temperatura de cozimento de 147 ºC. Ambos os cozimentos foram realizados buscando-se número kappa de 18,0 ± 0,5. Os branqueamentos foram realizados pela sequência de branqueamento OA(Ze)DP, buscando-se a alvura de 90,0 ± 1,0% ISO. Os refinos foram realizados a 0, 750, 1.500 e 3.000 níveis de revoluções e as correlações, feitas utilizando o índice de tração como variável independente da curva de refino. As propriedades físico-mecânicas do papel foram influenciadas pela condição de polpação, com aumento da drenabilidade, densidade aparente, índice TEA, alongamento e índice de rasgo para a polpa PI, considerando-se um mesmo índice de tração. A polpa PII, entretanto, atingiu o mesmo índice de tração da polpa PI com menor nível de refino e, consequentemente, menor gasto de energia. A condição de polpação PII foi mais adequada para a produção de papel por preservar mais a integridade das fibras, alcançando um mesmo índice de tração com um menor nível de refino.

Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro

A comunicação do estado reprodutivo nos primatas da família Callithrichidae, depende principalmente dos comportamentos sócio-sexuais como um sistema de sinalização primário, uma vez que nestas espécies a ovulação não é percebida pelos machos. Neste trabalho, os padrões de comportamentos sócio-sexuais foram analisados em conjunto com as concentrações de metabólitos fecais dos esteróides sexuais progesterona (MFP), estradiol (MFE) e testosterona (MFT) em casais cativos de Sagüi-de-tufos-pretos (Callithrix penicillata), nas diferentes fases do ciclo ovariano. O grupo estudado era composto por quarto casais adultos, mantidos no Centro de Reabilitação de Animais Selvagens da prefeitura de São Paulo. Os padrões comportamentais foram registrados pelo método de amostragem focal por intervalo de tempo a cada 30 segundos, cinco vezes por semana, totalizando 14.400 registros por animal. A mensuração das concentrações de metabólitos fecais dos esteroides sexuais foram realizados pelo método de enzima imunoensaio (EIE). Os resultados obtidos dessas concentrações possibilitaram a determinação endócrina das fases do ciclo ovariano (folicular e luteal) e de suas respectivas durações, assim como a determinação da fase periovulatória. Foram caracterizados 31 ciclos ovarianos completos, com duração de 24,3±4,1 dias (média ±DP), sendo que a fase folicular compreendeu 13,04±4,8dias e a fase lútea 11,2±4,2 dias. Os comportamentos sócio-sexuais (marcação por cheiro, cheirar genitália, catação e apresentação sexual) e a variável "proximidade" mostraram-se significativamente mais prevalentes na fase periovulatória do que nas demais fases do ciclo. Não houve alteração das concentrações de MFT dos machos ao longo de todo o período estudado. A análise conjunta das concentrações de metabólitos fecais de esteróides sexuais e dos comportamentos sócio-sexuais possibilitou um melhor entendimento das relações endócrino-comportamentais e reprodutivas de C. penicillata.

Työkyvyn johtaminen : Case Pirkanmaan Osuuskauppa

Tämän tutkimuksen tarkoituksena oli tutkia miten Pirkanmaan Osuuskaupan esimiehet ymmärtävät työhyvinvoinnin merkityksen esimiestyössään ja mitä asioita heidän tulisi vielä kehittää työhyvinvointi osaamisessaan. Työkyvyn johtamista verrattiin Elinkeinoelämän keskusliiton luomaan työkyvynjohtamisen malliin. Tutkimuksen pohjalta laadittiin ehdotuksia, jotta esimiesten osaamista, ymmärtämystä ja kykyä hyödyntää työhyvinvoinnin välineitä voitaisiin kehittää ja ylläpitää oikeilla toimenpiteillä. Yrityksissä työhyvinvointiin on alettu kiinnittää entistä enemmän huomiota kiireisen työtahdin, työn vaatimusten sekä työurien mahdollisten pidentymisen vuoksi, jolloin työnantajan lisäksi vaaditaan myös työntekijöiltä panostusta työhyvinvointiin. Tutkimus toteutettiin käyttämällä laadullista menetelmää. Aineiston keruu tehtiin puolistrukturoidun teemahaastattelun avulla. Haastatteluihin valikoitui 4 esimiestä. Haastattelujen avulla kartoitettiin esimiesten tietoja ja näkemyksiä työhyvinvoinnista ja heidän työhyvinvointiosaamisen tasosta. Tutkimuksen avulla selvisi, että esimiesten keskuudessa työhyvinvoinnin ylläpitämiseen ja kehittämiseen panostetaan vähemmän kuin mitä olisi mahdollista. Suurimmiksi esteiksi työhyvinvoinnin toteuttamiselle muodostuivat aika, resurssit ja asenteet. Pirkanmaan Osuuskaupan johtamistyön ja toimintatapojen välillä on eroja verrattuna Elinkeinoelämän keskusliiton työkykyjohtamisen malliin. Esimiestyö on mahdollisuuksia täynnä oleva kilpailuetu, mutta samalla myös huomattava haaste.

Osaamisen johtaminen - käytäntöjä ja kehittämistarpeita henkilöstön näkökulmasta

Tämän pro gradu -tutkimuksen tavoitteena oli tutkia osaamisen johtamisen käytäntöjä ja kehittämistarpeita kohdeorganisaatiossa henkilöstön näkökulmasta. Tutkimuksen tarkoituksena oli selvittää, miten tehtyjä henkilöstötutkimuksia on hyödynnetty kohdeorganisaatiossa ja näkyykö tämä hyödyntäminen käytännön johtamisessa henkilöstön mielestä. Tutkimus suoritettiin laadullisena tapaustutkimuksena ja empiirinen aineisto kerättiin kuudella teemahaastattelulla, jotka analysoitiin sisällönanalyysimenetelmällä. Tutkimuksessa kartoitettiin, millaisia osaamisen johtamisen käytäntöjä kohdeorganisaatiossa on. Tutkimuksen tuloksien perusteella osaamisen johtamisen peruselementit ovat olemassa: säännölliset palaverit koettiin tärkeiksi, mutta koulutuksien ja kehityskeskusteluiden järjestämisessä sekä jälkiseurannassa henkilöstö näki parannettavaa. Myös hiljaisen tiedon siirtäminen ja hyödyntäminen koettiin merkittäväksi tekijäksi organisaation onnistuneen toiminnan ja jatkuvuuden kannalta. Tutkimuksessa todettiin osaamisen johtamisen kehittämisen ja osaamisen kehittämisen menetelmien linkittyvän vahvasti toisiinsa ja tutkimuksen tuloksena esitettiin konkreettisia toimenpide-ehdotuksia osaamisen johtamisen kehittämisestä kohdeorganisaation johdolle. Tutkimuksen mukaan osaamisen johtaminen ja henkilöstöjohtaminen sulautuvat toisiinsa, eikä niitä voida tarkastella erikseen. Johdon ja henkilöstön välisellä vuorovaikutuksella on suuri merkitys onnistuneelle osaamisen johtamiselle. Keskeistä on kytkeä osaamisen johtaminen organisaation käytäntöihin ja varata sen implementointiin riittävästi resursseja.

Connexin hemichannels and cell-cell channels: comparison of properties

Connexin46 (Cx46) forms functional hemichannels in the absence of contact by an apposed hemichannel and we have used these hemichannels to study gating and permeation at the single channel level with high time resolution. Using both cell-attached and -excised patch configurations, we find that single Cx46 hemichannels exhibit some properties expected of half of a gap junction channel, as well as novel properties. Cx46 hemichannels have a large unitary conductance (~300 pS) and a relatively large pore as inferred from permeability to TEA. Both monovalent cations and anions can permeate, but cations are substantially more permeable. The open channel conductance shows marked inward rectification in symmetric salts. We find that the conductance and permeability properties of Cx46 cell-cell channels can be explained by the series addition of two hemichannels. These data suggest that the pore structures of unapposed hemichannels and cell-cell channels are conserved. Also like cell-cell channels, unapposed Cx46 hemichannels are closed by elevated levels of H+ or Ca2+ ions on the cytoplasmic face. Closure occurs in excised patches indicating that the actions of these agents do not require a soluble cytoplasmic factor. Fast (<0.5 ms) application of H+ to either side of the open hemichannel causes an immediate small reduction in unitary conductance followed by complete closure with latencies that are dependent on H+ concentration and side of application; sensitivity is much greater to H+ on the cytoplasmic side. Closure by cytoplasmic H+ does not require that the hemichannel be open. Thus, H+ ions readily permeate Cx46 hemichannels, but at high enough concentration close them by acting at a cytoplasmic site(s) that causes a conformational change resulting in complete closure. Extracellular H+ may permeate to act on the cytoplasmic site or act on a lower affinity extracellular site. Thus, the unapposed hemichannel is a valuable tool in addressing fundamental questions concerning the operation of gap junction channels that are difficult to answer by existing methods. The ability of Cx46, and perhaps other connexins, to form functional unapposed hemichannels that are opened by moderate depolarization may represent an unexplored role of connexins as mediators of transport across the plasma membrane.

The role of calcium, calcium-activated K+ channels, and tyrosine/kinase in psoralen-evoked responses in human melanoma cells

8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 µM). The intracellular Ca2+ chelator BAPTA/AM (1 µM) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 µM protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92% increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.

Participation of ATP-sensitive K+ channels in the peripheral antinociceptive effect of fentanyl in rats

We examined the effect of several K+ channel blockers such as glibenclamide, tolbutamide, charybdotoxin (ChTX), apamin, tetraethylammonium chloride (TEA), 4-aminopyridine (4-AP), and cesium on the ability of fentanyl, a clinically used selective µ-opioid receptor agonist, to promote peripheral antinociception. Antinociception was measured by the paw pressure test in male Wistar rats weighing 180-250 g (N = 5 animals per group). Carrageenan (250 µg/paw) decreased the threshold of responsiveness to noxious pressure (delta = 188.1 ± 5.3 g). This mechanical hyperalgesia was reduced by fentanyl (0.5, 1.5 and 3 µg/paw) in a peripherally mediated and dose-dependent fashion (17.3, 45.3 and 62.6%, respectively). The selective blockers of ATP-sensitive K+ channels glibenclamide (40, 80 and 160 µg/paw) and tolbutamide (80, 160 and 240 µg/paw) dose dependently antagonized the antinociception induced by fentanyl (1.5 µg/paw). In contrast, the effect of fentanyl was unaffected by the large conductance Ca2+-activated K+ channel blocker ChTX (2 µg/paw), the small conductance Ca2+-activated K+ channel blocker apamin (10 µg/paw), or the non-specific K+ channel blocker TEA (150 µg/paw), 4-AP (50 µg/paw), and cesium (250 µg/paw). These results extend previously reported data on the peripheral analgesic effect of morphine and fentanyl, suggesting for the first time that the peripheral µ-opioid receptor-mediated antinociceptive effect of fentanyl depends on activation of ATP-sensitive, but not other, K+ channels.

The methyl ester of rosuvastatin elicited an endothelium-independent and 3-hydroxy-3-methylglutaryl coenzyme A reductase-independent relaxant effect in rat aorta

The relaxant effect of the methyl ester of rosuvastatin was evaluated on aortic rings from male Wistar rats (250-300 g, 6 rats for each experimental group) with and without endothelium precontracted with 1.0 µM phenylephrine. The methyl ester presented a slightly greater potency than rosuvastatin in relaxing aortic rings, with log IC50 values of -6.88 and -6.07 M, respectively. Unlike rosuvastatin, the effect of its methyl ester was endothelium-independent. Pretreatment with 10 µM indomethacin did not inhibit, and pretreatment with 1 mM mevalonate only modestly inhibited the relaxant effect of the methyl ester. Nω-nitro-L-arginine methyl ester (L-NAME, 10 µM), the selective nitric oxide-2 (NO-2) inhibitor 1400 W (10 µM), tetraethylammonium (TEA, 10 mM), and cycloheximide (10 µM) partially inhibited the relaxant effect of the methyl ester on endothelium-denuded aortic rings. However, the combination of TEA plus either L-NAME or cycloheximide completely inhibited the relaxant effect. Inducible NO synthase (NOS-2) was only present in endothelium-denuded aortic rings, as demonstrated by immunoblot with methyl ester-treated rings. In conclusion, whereas rosuvastatin was associated with a relaxant effect dependent on endothelium and hydroxymethylglutaryl coenzyme A reductase in rat aorta, the methyl ester of rosuvastatin exhibited an endothelium-independent and only slightly hydroxymethylglutaryl coenzyme A reductase-dependent relaxant effect. Both NO produced by NOS-2 and K+ channels are involved in the relaxant effect of the methyl ester of rosuvastatin.