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.

Female novelty and the courtship behavior of male guinea pigs (Cavia porcellus)

In several rodent species, an increase or recovery of sexual behavior can be observed when sexually satiated males are placed in contact with a novel mate. In order to assess the influence of female novelty on the courtship behavior of guinea pigs (Cavia porcellus), four adult males were observed during four daily 15-min sessions while interacting with the same pregnant female (same-female sessions). A new female was presented during the fifth session (switched-female session). The duration of behavioral categories was obtained from videotape records using an observational software. From the first to the second session, all males decreased the time allocated to investigating (sniffing and licking), following, and mounting the female, and that response did not recover by the end of the same-female sessions. No similar decreasing tendencies were detected in the circling or rumba categories. A marked increase of investigating occurred in all males from the last same-female session (8.1, 11.9, 15.1 and 17.3 percent session time) to the switched-female one (16.4, 18.4, 37.1 and 28.9 percent session time, respectively). Increases in following and circling were recorded in three of four males, and full-blown recovery of mounting in one male. No consistent changes in the females' responses to males (following or attacking) were observed throughout testing. These results are consistent with the hypothesis that guinea pig males recognize individual females and that courtship responses may suffer a habituation/recovery process controlled by mate novelty.

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.

Deficiency of sex hormones does not affect 17-ß-estradiol-induced coronary vasodilation in the isolated rat heart

The relaxation of coronary arteries by estrogens in the coronary vascular beds of naive and hypertensive rats has been well described. However, little is known about this action in gonadectomized rats. We investigated the effect of 17-ß-estradiol (E2) in coronary arteries from gonadectomized rats, as well as the contributions of endothelium-derived factors and potassium channels. Eight-week-old female and male Wistar rats weighing 220-300 g were divided into sham-operated and gonadectomized groups (n=9−12 animals per group). The baseline coronary perfusion pressure (CPP) was determined, and the vasoactive effects of 10 μM E2 were assessed by bolus administration before and after endothelium denudation or by perfusion with NG-nitro-L-arginine methyl ester (L-NAME), indomethacin, clotrimazole, L-NAME plus indomethacin, L-NAME plus clotrimazole or tetraethylammonium (TEA). The CPP differed significantly between the female and sham-operated male animals. Gonadectomy reduced the CPP only in female rats. Differences in E2-induced relaxation were observed between the female and male animals, but male castration did not alter this response. For both sexes, the relaxation response to E2 was, at least partly, endothelium-dependent. The response to E2 was reduced only in the sham-operated female rats treated with L-NAME. However, in the presence of indomethacin, clotrimazole, L-NAME plus indomethacin or L-NAME plus clotrimazole, or TEA, the E2 response was significantly reduced in all groups. These results highlight the importance of prostacyclin, endothelium-derived hyperpolarizing factor, and potassium channels in the relaxation response of coronary arteries to E2 in all groups, whereas nitric oxide may have had an important role only in the sham-operated female group.

Processamento mínimo em goiabas 'Paluma' e 'Pedro Sato': 2. Avaliação química, sensorial e microbiológica

Foram utilizados frutos de goiabeiras das cultivares "Pedro Sato" e "Paluma", provenientes de pomar comercial, no estádio de maturação "de vez", correspondente à coloração verde-mate e considerados "ótimo para o consumo". Os frutos foram inicialmente imersos em solução de hipoclorito de sódio (150mg de cloro.L-1) por 5 minutos, para desinfecção superficial. Pessoas treinadas, utilizando proteção adequada e equipamentos desinfetados descascaram os frutos, cortaram-nos longitudinalmente ao meio e eliminaram a polpa com as sementes, em ambiente a 12°C. Após enxágüe com água clorada (20mg de cloro.L-1) foram embalados em contentores de tereftalato de polietileno (PET) com tampa. Estas unidades foram armazenadas a 3°C por 10 dias. Foram realizadas análises microbiológicas ao longo do período. Determinaram-se quimicamente os conteúdos de lignina, ácido ascórbico, acidez total titulável, sólidos solúveis totais e porcentagem de solubilização das pectinas, bem como as variáveis sensoriais de textura, sabor e preferência. A textura tornou-se mais frágil e os conteúdos de ácido ascórbico se reduziram, ao longo do período de armazenamento, nos produtos de ambas as cultivares. Durante este período houve aumento no conteúdo de lignina e manutenção dos conteúdos de sólidos solúveis totais (SST), acidez total titulável (ATT) e da relação SST/ATT. O produto da cultivar "Pedro Sato" apresentou menor perda de textura que o da 'Paluma', sendo considerado pelos provadores como o mais saboroso e, portanto, o mais preferido. Devido aos cuidados higiênicos tomados durante o processamento o produto apresentou baixa contagem microbiana (< 10³ UFC.g-1), em todas as avaliações efetuadas.

Materiaalisuunnittelun mittaaminen elintarviketeollisuudessa

Työssä tutkitaan materiaalisuunnittelun mittausta elintarvikealan yrityksessä. Tut-kimuksella on kolme tavoitetta, jotka ovat miten materiaalisuunnittelua mitataan yrityksessä, toimivatko mittarit oikein ja mittaavatko mittarit haluttuja asioita. Työ on tapaustutkimus, joka koostuu teoreettisesta kirjallisuusselvityksestä ja empiirisestä tapaustutkimuksesta. Empiirinen aineisto on hankittu sekä kvalitatii-visin että kvantitatiivisin menetelmin. Työn tutkimustavoitteiden saavuttaminen edellytti materiaalisuunnittelijoille ky-selyn tekemistä. Kyselyn ja tavoitteiden avulla määriteltiin mittarit, joita halutaan seurata. Käytössä olevalle mittaristolle etsittiin vaihtoehtoa toiminnanohjausjär-jestelmästä. Vertailemalla mittaristoja päädyttiin käyttämään vanhaan mittaristoa. Materiaalisuunnittelun mittaaminen on tärkeätä, koska hankinnat vaikuttavat yri-tyksen tulokseen ja ilman mittausta materiaalisuunnittelu ei voi kehittyä. Valittu mittaristo täyttää kaikki sen vaatimukset ja siitä löytyvät mittarit, joita halutaan seurata. Mittariston avulla yritys pystyy seuraamaan materiaalisuunnittelun tavoit-teiden saavuttamista eli varastoihin sitoutuneen pääoman kehitystä. Mittariston tulee olla yhteydessä yrityksen hankintastrategiaan ja –tavoitteisiin. Mittariston oikeellisuutta tulee tarkastella säännöllisin väliajoin, jotta varmistutaan siitä, että mitataan seurattavia asioita.

Conteúdo de miricetina, quercetina e kaempferol em chás comercializados no Brasil

Os teores de miricetina, quercetina e kaempferol foram determinados em uma marca de ban-chá, duas de chá verde e quatro de chá preto. Analisaram-se três lotes para cada marca em duplicata por cromatografia líquida de alta eficiência. Quercetina (2,5-3,4 mg/g folha seca) predominou em todas as amostras, seguida por kaempferol (1,0-2,0 mg/g folha seca), com exceção de uma amostra na qual kaempferol e miricetina tiveram teores iguais. Houve variação entre os tipos de chás e mesmo entre marcas do mesmo tipo. Miricetina (traços - 1,9 mg/g folha seca) foi o flavonol, que mais variou e que esteve em menor nível nos chás pretos. Outros chás muito consumidos no Brasil também foram investigados. A miricetina não foi encontrada em chás de frutas (maçã e morango) e de ervas (erva doce, camomila, erva cidreira, hortelã, boldo, mate e erva mate), enquanto que quercetina foi encontrada em quatro chás (camomila, boldo, morango e erva mate) e kaempferol, em dois chás (boldo e erva-mate), em concentrações de 0,4 a 2,5 e 0,4 a 2,6 mg/g de folha seca, respectivamente. Concluiu-se que estes chás são fontes de flavonóis na dieta brasileira, embora com teores menores que em chás verde e preto.

Teores de catequinas e teaflavinas em chás comercializados no Brasil

No presente estudo, foram determinados os teores de catequinas e teaflavinas em três marcas de chá verde e quatro de chá preto comercializadas no Brasil. A metodologia analítica consistiu de extração aquosa bastante simples e cromatografia líquida de alta eficiência. Foi utilizada uma coluna de fase reversa Novapak C18 (3,9 mmx150 mm, 4 µm) com um gradiente de água e metanol ambos em ácido fórmico como fase móvel. Em chás verdes, os conteúdos de catequinas (em mg/g de folha seca) variaram substancialmente: catequina, 0,8 a 2,8; epigalocatequina, 8 a 44; epigalocatequina galato, 11 a 50; epicatequina, 2,3 a 8,5 e epicatequina galato, 3,1 a 7,3. No caso dos chás pretos, as concentrações (mg/g de folha seca) de catequinas estiveram nas faixas de: 10 a 50 de epigalocatequina, 14 a 37 de epigalocatequina galato, 5 a 9 de epicatequina e 10 a 21 de epicatequina galato. As teaflavinas apresentaram variação menor: entre 5 (para teaflavina 3'-galato) e 13 mg/g (para teaflavina 3,3'-digalato) de folha seca. Amostras de chás muito consumidas no Brasil (erva doce, camomila, erva cidreira, hortelã, boldo, mate, erva mate, maçã e morango) também foram investigadas, não sendo encontrada nenhuma catequina ou teaflavina.

Commercial spices and industrial ingredients: evaluation of antioxidant capacity and flavonoids content for functional foods development

The aim of this work was to evaluate spices and industrial ingredients for the development of functional foods with high phenolic contents and antioxidant capacity. Basil, bay, chives, onion, oregano, parsley, rosemary, turmeric and powdered industrial ingredients (β-carotene, green tea extract, lutein, lycopene and olive extract) had their in vitro antioxidant capacity evaluated by means of the Folin-Ciocalteu reducing capacity and DPPH scavenging ability. Flavonoids identification and quantification were performed by High Performance Liquid Chromatography (HPLC). The results showed that spices presented a large variation in flavonoids content and in vitro antioxidant capacity, according to kind, brand and batches. Oregano had the highest antioxidant capacity and parsley had the highest flavonoid content. The industrial ingredient with the highest antioxidant capacity was green tea extract, which presented a high content of epigalocatechin gallate. Olive extract also showed a high antioxidant activity and it was a good source of chlorogenic acid. This study suggests that oregano, parsley, olive and green tea extract have an excellent potential for the development of functional foods rich in flavonoids as antioxidant, as long as the variability between batches/brands is controlled.

Antioxidant activities of kombucha prepared from three different substrates and changes in content of probiotics during storage

Kombucha is a health-promoting fermented beverage worldwide. The present study compared the free-radical scavenging abilities and total reducing power (TRP) of kombucha prepared from low-cost green tea (LGTK), black tea (BTK), and tea powder (TPK). LGTK had the highest scavenging abilities against 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion and hydroxyl radicals, while BTK showed the highest TRP. Changes in content of probiotics in LGTK were investigated during storage as well. The number of acetic acid bacteria decreased moderately up to 10 days of storage. The number of lactic acid bacteria (LAB) decreased significantly, and their survival rate was only 0.98% at the 8th day of storage.

Peroxidase activity and sensory quality of ready to cook mixed vegetables for soup: combined effect of biopreservatives and refrigerated storage

Enzymatic senescence processes and browning of fresh cut vegetables negatively affect their sensory properties and nutritional value and finally result in the rejection of affected products by consumers. In order to prevent quality decay, the combined effects of natural antioxidants and storage temperature on peroxidase activity and sensory attributes (overall visual quality, browning and odor) of individual and mixed vegetables for soup (butternut squash, leek and celery) were evaluated. Fresh cut vegetables were treated with antioxidant solutions as tea tree essential oil (15 μl/mL), propolis extract (15 μl/mL) and gallic acid (2 mg/mL) and stored at optimal (5 °C) and abusive (15 °C) temperature for a maximum of 14 days. The application of natural preservatives, plus optimal storage conditions, exerted significant inhibitory effects in peroxidase activity of squash, celery and mixed vegetables throughout the storage. Furthermore, propolis treatment applied on mixed vegetables retarded browning appearance and preserved the visual quality for a longer period when compared to untreated product.

Utilizing analytic modelling process in industrial retail context

Nykypäivän monimutkaisessa ja epävakaassa liiketoimintaympäristössä yritykset, jotka kykenevät muuttamaan tuottamansa operatiivisen datan tietovarastoiksi, voivat saavuttaa merkittävää kilpailuetua. Ennustavan analytiikan hyödyntäminen tulevien trendien ennakointiin mahdollistaa yritysten tunnistavan avaintekijöitä, joiden avulla he pystyvät erottumaan kilpailijoistaan. Ennustavan analytiikan hyödyntäminen osana päätöksentekoprosessia mahdollistaa ketterämmän, reaaliaikaisen päätöksenteon. Tämän diplomityön tarkoituksena on koota teoreettinen viitekehys analytiikan mallintamisesta liike-elämän loppukäyttäjän näkökulmasta ja hyödyntää tätä mallinnusprosessia diplomityön tapaustutkimuksen yritykseen. Teoreettista mallia hyödynnettiin asiakkuuksien mallintamisessa sekä tunnistamalla ennakoivia tekijöitä myynnin ennustamiseen. Työ suoritettiin suomalaiseen teollisten suodattimien tukkukauppaan, jolla on liiketoimintaa Suomessa, Venäjällä ja Balteissa. Tämä tutkimus on määrällinen tapaustutkimus, jossa tärkeimpänä tiedonkeruumenetelmänä käytettiin tapausyrityksen transaktiodataa. Data työhön saatiin yrityksen toiminnanohjausjärjestelmästä.

Omistajuuden vaikutus yrityksen toimintaan ja tulokseen

Tämän tutkimuksen tavoitteena oli selvittää miten omistajuus vaikuttaa yrityksen toimintaan ja tulokseen ja syventää näkökulmaa omistajan vaikutuksesta yrityksen menestymiseen. Tutkimus tehtiin kvalitatiivisen ja kvantitatiivisen tutkimusmenetelmän yhdistelmällä, joka mahdollistaa laajemman näkökulman tutkittavaan asiaan ja tuo tutkimukseen luotettavuutta. Omistajan kompetenssilla toimia aktiivisena omistajana sekä hallita erilaisia liiketoimintariske-jä on suuri merkitys. Kestävä menestys, uudistuminen ja yrityksen arvon luominen vaativat niin asiantuntemusta, näkemyksiä, sitoutunutta riskirahoitusta kuin vuorovaikutustaitoja. Omis-tajalla on vaikutusvalta osaavan johdon ja henkilöstön motivaation ylläpysymiseen ja tahtoti-laan toimia yrityksen etujen mukaisesti ja yritykseen sitoutuneena. Ymmärtämällä omistamansa yrityksen kilpailuedut ja hallitsemalla organisatoriset riskit, omis-taja voi vaikuttaa valitsemansa roolin kautta yrityksen menestykseen ja tuottoarvon kasvuun. Tutkimuksen tulokset viittaavat myös siihen, että henkilöstön kokemat positiiviset psykologi-sen ja sosiaalisen omistajuuden tunteet selittävät onnistumisia.