Gary Griggs, Our Ocean Backyard: Fukushima radiation: What's going on now?
Nearly five years ago, on March 11, 2011, a massive 9.0 magnitude earthquake off Japan generated a large tsunami, which struck the coastline of Japan in less than an hour. Waves reached elevations of up to 128 feet above sea level and moved inland as far as 6 miles, flooding more than 200 square miles of low-lying coastal land. The earthquake and tsunami led to the deaths of nearly 21,000 people, mostly from drowning.
At the Fukushima Daiichi nuclear plant, tsunami waves overtopped a 33-foot high seawall protecting the backup cooling facility, flooding and disabling the system. This lead to the loss of cooling water, explosions, meltdown of three of the plant's six reactors and the uncontrolled release of radiation into the adjacent ocean.
A major effort was undertaken over the next several years to halt and collect the flow of groundwater beneath the damaged plant that was carrying contaminated water to the sea. In October 2015, contractors completed a one-half mile long shielding wall to reduce the amount of contaminated water leaking into the ocean. Sampling within one-half a mile of the coastline at the Fukushima plant in October revealed radiation levels that are thousands of times lower than immediately after the accident, but indicated that there was still continued release of contaminated water from the plant.
It is projected that complete clean up and decommissioning of the plant will cost tens of billions of dollars and take 10 to 30 years. What happened at Fukushima also had a chilling effect on existing and proposed nuclear power plants around the world.
Concerns about potential radiation exposure along the west coast of the U.S. arose soon after the release of radiation from the plant was reported.
While high levels of radiation were documented in the ocean off Japan almost immediately, movement of that radiation 5,000 miles across the North Pacific to the west coast turns out to be a relatively slow journey.
A chemical oceanographer from Woods Hole Oceanographic Institution has been analyzing water samples intermittently since the March 11 disaster. Just a year ago, they announced the detection of trace amounts of radioactivity that could be used to fingerprint Fukushima because of the presence of cesium-134.
Surface water samples were collected along a 2,000-mile-long transect extending from the Aleutian Islands to Eureka and along the length of the California coast. Cesium-134 was detected in 10 of the water samples analyzed along this transect, all but one of these were from the waters of the current heading towards the Gulf of Alaska. One sample off southern Oregon in November 2014 did reveal the presence of cesium-134.
Continued monitoring in the past year indicates an increased number of sites off the U.S. West Coast showing signs of Fukushima radiation. This includes the highest detected level to date from a sample collected 1,600 miles west of San Francisco.
While this sample had cesium-134 levels 50 percent higher than other West Coast samples, the concentration of this isotope is still more than 500 times lower than U.S. government safety limits for drinking water, and well under the limits of concern for direct exposure while swimming, boating or other ocean activities. The 2015 values are consistent with those of Canadian scientists, who also recently reported that they have found no cesium-134 in fish collected off British Columbia.
The transit across the North Pacific has been quite slow, averaging just several miles a day. This has given the cesium from the damaged nuclear plant, which has a half-life of two years, additional time to decay and dissipate.
This column is the No. 200 I've written over the past 7.5 years, and it seems like a good time to ask any interested reader to send an email with ideas or requests for future columns. Thank you for your input.
Gary Griggs is director of the Institute of Marine Sciences and Long Marine Laboratory at UC Santa Cruz. He can be reached at griggs@ucsc.edu. For past Ocean Backyard columns, visit http://seymourcenter.ucsc.edu/about-us/news/our-ocean-backyard-archive/.


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8. Ontario Bicycling Across The USA
Day 55: Rondeau Park to St. Thomas, ON
1 Comment on Day 55: Rondeau Park to St. Thomas, ON
Colonel Thomas Talbot got a grant of 7,000 acres along the north shore of Lake Erie after retiring from the British military in 1803. He also cut a deal that would give him even more land for every settler he persuaded to prove a homestead there. Talbot made the most of the opportunity, building a 300-mile corduroy road along rich farmland between the Niagara River and the St. Clair River. The area was settled between 1820 and 1840, and Talbot was known as "The Benevolent Dictator of the London District." The region he settled is still one of Canada's most productive agricultural areas. The road also lead to the development of Buffalo and Detroit. But Talbot never married, so the government took his land back when he died.
We saw echoes of the original Talbot Trail in tidy mid-19th century brick farmhouses along the route, as well as homesteads that didn't make it. We slept late on Sunday the 5th because our rides in Ontario should be fairly short, and also because it's too cold now to ride first thing in the morning. The late start gave us a bit of time to walk down the shore of Lake Erie at Rondeau Provincial Park, which was a revelation. I saw fine sand and miles or unbroken woods stretching southward in a peninsula that juts into the lake. Rondeau is one of Canada's oldest parks, so the woods here are near maturity and the land has been managed well for biodiversity. It is a beautiful place, and it was nearly empty of people on a sunny Sunday in October. It would be a great place for a long hike.
After an unsuccessful attempt to find an internet connection in a nearby town, we ate a late breakfast at a diner in Morpeth and started east on the Talbot Trail. The truck was leaking brake fluid, and we learned from Sara that the leak got a lot worse as she drove ahead of us to our destination, Port Stanley. We needed to get it to a mechanic in the morning. So we rode the 45 miles as fast as we could, passing natural reserve areas and the Lake Erie shore in a few places but mostly staying in agricultural land. I could see enough to understand that this would be a great place for birdwatching, hiking, and lying on the beach. But we had a problem to work out.
Port Stanley is an upscale town people describe as "quaint," which meant that it had no affordable lodging or campgrounds. We re-filled the truck with brake fluid and headed north to St. Thomas, a larger town about 15 miles north. We located a car dealer, a Comfort Inn, and a restaurant, which gave our credit cards a small workout. We fell into bed by 10pm, awaiting the morning's verdict on the truck.
Tags Bicycle Touring, Jumbo the elephant, ontario, rondeau parc, St. Thomas
By bradedmondson
Writing about social change and how it happens.
← Days 56 & 57: St. Thomas to Long Point, ON → Through Ontario, Oct. 4 to 10
One reply on "Day 55: Rondeau Park to St. Thomas, ON"
Catherinesays:
Whoop it up in Rochester! Hope you see lots of friends! Wish I could be there.
Hope truck brake repairs are simple and inexpensive!

redpajama_set_name: RedPajamaCommonCrawl


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Gesellschaft bezeichnet in der Soziologie eine durch unterschiedliche Merkmale zusammengefasste und abgegrenzte Anzahl von Personen, die als sozial Handelnde (Akteure) miteinander verknüpft leben und direkt oder indirekt sozial interagieren. Gesellschaft bezieht sich sowohl auf die Menschheit als ganze (gegenüber Tieren und Pflanzen) als auch auf bestimmte Gruppen von Menschen, beispielsweise auf ein Volk oder eine Nation. Gesellschaft kann sich aber auch auf einen räumlich abgegrenzten und strukturierten Zusammenhang zwischen Menschen beziehen, auf Kollektive oder auch auf ein Knäuel (cluster) im sozialen Netzwerk der Menschheit, das sich durch die Netzwerkdichte und Multiplexität der Interaktionen abgrenzen lässt. In den aktuellen Sozialwissenschaften wird die Bezeichnung oft mit ungenauer Bedeutung verwendet.

Der Gesellschaftsbegriff in der Ethnologie bzw. Anthropologie sowie im Staatsrecht wird jeweils anders definiert und verwendet als in der Soziologie. Theoretisch befasst sich die Gesellschaftstheorie mit dem soziologischen Begriff der Gesellschaft.

Begriffsgeschichte
"Gesellschaft bedeutet wörtlich den Inbegriff räumlich vereint lebender oder vorübergehend auf einem Raum vereinter Personen. So ergibt es sich aus der etymologischen Herleitung des Wortes von ahd.  = Raum, ahd.  = Wohnung; heute noch erhalten in: nhd. 'Saal', skand.  = Stockwerk, russ.  = Hof, Landsitz (vielleicht zusammenhängend mit  = Grund und Boden). Geselle, ahd. , ist also der 'Saalgenoss'."

Der Begriff ist unmittelbar wahrscheinlich von Geselle bzw. von Gesellenschaft abgeleitet. Gesellenschaft war der Zusammenschluss von Gesellen zur Durchsetzung von Forderungen zur Änderung der von den Zunftmeistern (der "Meisterschaft") bestimmten Arbeitsbedingungen. Im heutigen Sprachgebrauch deuten Wendungen, wie "sich gesellen" (vgl. "Gleich und gleich gesellt sich gern.") oder "gesellig" (Geselligkeit) auf einen räumlich und zeitlich offenen Zusammenhang von Personen. Der Konflikt zwischen Gesellenschaft und Meisterschaft der mittelalterlichen Zunft ist beiden Begriffen im heutigen Sprachgebrauch fremd. In der neuzeitlichen Rechtsentwicklung hat sich die Rechtssprache als wertungsfreier Gruppenbegriff durchgesetzt. In Schleiermachers Unterscheidung von Gesellschaft und Gemeinschaft klingt dieses Rechtsdenken an.

Die Ideen der Aufklärung von einer vernunftgemäßen Regierung wurden durch die Französische Revolution der breiten Öffentlichkeit bekannt gemacht. Im 19. Jahrhundert wurde dann "Gesellschaft" zur Übersetzung von engl.  und frz.  eingesetzt. Der Begriff der  oder "bürgerlichen Gesellschaft" diente dem aufkommenden Bürgertum als Gegenbegriff zum absolutistischen Fürstenstaat. Fortan wurde der Dualismus der Begriffe Staat und Gesellschaft grundlegend für die politische Philosophie des Liberalismus.

Biosoziologisch gesehen ist der Mensch von Natur aus in Gesellschaft. Mit (bereits) Aristoteles' Worten ist er ein  (), ein auf "Staaten-(Gemeinden-, Poleis-)Bildung angelegtes Wesen".

Soziologische Schulen
Die Bezeichnung "Gesellschaft" ist als zentraler Grundbegriff der Soziologie umstritten. 2011 schrieb Thomas Schwinn in einer Übersicht von "starken und schwachen Gesellschaftsbegriffen". Diese bezeichnete er als "Verfallsstufen eines traditionsreichen Konzepts". Er beschreibt die Theoriegeschichte des Begriffs und unterscheidet u. a. systemtheoretische Ansätze (u. a. Talcott Parsons, Niklas Luhmann), handlungstheoretische Fundierungsversuche (Anthony Giddens, Hartmut Esser) oder Kombinationen aus System- und Handlungstheorie (Jürgen Habermas, Uwe Schimank).

Auch in der heutigen Soziologie ist die Verwendung des Begriffes Gesellschaft umstritten. So fordert z. B. der britische Soziologe John Urry für eine Soziologie des 21. Jahrhunderts die Abkehr von der Analyse von Gesellschaften (Sociology Beyond Societies, London 2000).

Marxismus (Marx)

Nach Karl Marx ist Gesellschaft die Gesamtheit der Verhältnisse zwischen den Menschen, also die Summe der Beziehungen und der Verhältnisse unter den Individuen und nicht die Individuen als solche. Die Gesellschaft wird hier nach dem geschichtlichen Entwicklungsstand der ökonomischen Verhältnisse analysiert, wobei Marx zunächst drei grundlegende Gesellschaftsformationen beschreibt:
 die ursprüngliche primäre oder archaische Formation (Urgesellschaft) auf dem Boden des Gemeineigentums und sozialer Homogenität, die sich auf ihrer letzten und höchsten Entwicklungsstufe mit der Teilung der Arbeit, mit individuell genutztem Gemeineigentum allmählich sozial differenziert und in die sekundäre Formation hinüberzuwachsen beginnt.
 die sekundäre Formation der auf großem Privateigentum basierenden sozialheterogenen Gesellschaften (asiatische Produktionsweise, Sklavenhaltergesellschaft, Feudalismus und bürgerlich-kapitalistische Gesellschaft)
 die kommunistische Gesellschaftsformation mit dem Sozialismus als Vorstufe bzw. Übergangsphase zur klassenlosen Gesellschaft

Soziologische Klassiker

Gemeinschaft und Gesellschaft (Tönnies)
Nach Ferdinand Tönnies ist Gesellschaft eine genau definierte Gruppierung von Personen. Er versteht "Gesellschaft" als Gegensatz zur "Gemeinschaft". Analytisch eingeführt wurde der Fachbegriff Gesellschaft in der sich etablierenden Soziologie durch Tönnies 1887 in seinem Werk Gemeinschaft und Gesellschaft. Danach zeichnet sich Gemeinschaft durch gegenseitiges Vertrauen, emotionale Anbindung und Homogenität aus. Tönnies stellt dem Begriff der Gemeinschaft den Begriff Gesellschaft gegenüber. Der Gemeinschaft bedienen sich nach Tönnies die Akteure mit jeweils individuellen Zielen. Dies führt zu einer nur losen Verknüpfung der Individuen in der Gesellschaft. Beide, Gemeinschaft und Gesellschaft, sind für ihn gemeinsamer Gegenstand der Soziologie. Sein Gesellschaftsbegriff im engeren Sinne ist also axiomatisch abgestützt und streng deduktiv gewonnen. In diesem Sinne wird die Bezeichnung nur noch selten in der Soziologie verwendet. Bei Tönnies ist Gesellschaft eine besondere Form gegenseitiger gewollter Bejahung von Menschen, die sich dieser Form als eines Mittels zur Erreichung ihrer individuellen Ziele bedienen (siehe oben).

Vergesellschaftung (Weber)
Max Weber knüpft mit seinem Begriff Vergesellschaftung noch stark an Tönnies' Merkmale an.

Differenzierung (Simmel)
1890 führte Georg Simmel den Begriff Differenzierung (auch soziale Differenzierung oder gesellschaftliche Differenzierung) in die Soziologie ein. Differenzierung bezeichnet langfristige Veränderungen einer Gesellschaft. Diese Veränderungen können verbunden sein mit der Neuentstehung oder Aufgliederung von Sozialen Positionen, Lebenslagen und/oder Lebensstilen. Mit Differenzierung wird auch das Ergebnis solcher Prozesse bezeichnet, nämlich "soziale Differenziertheit".

Strukturfunktionalismus (Parsons)
Laut Strukturfunktionalismus bildet sich aus Akteuren eine Gesellschaft dann, wenn sie in der Lage ist, mittels bestimmter sozialer Funktionen die menschlichen Bedürfnisse zu befriedigen (vergleiche Talcott Parsons, sowie den Funktionalismus). Funktional darauf ausgerichtet, bilden sich Institutionen, und ohne die Herausbildung von entsprechenden Strukturen ist eine dauerhafte Bedürfnisbefriedigung nicht möglich. Auch ein Robinson Crusoe überlebt nur, weil er die Methoden zur Bewältigung der Welt (Normen, Werte, Fähigkeiten) verinnerlicht hat, weil er die Gesellschaft in sich trägt – beispielsweise wenn er auf seiner einsamen Insel fromm wird. Akteur (oder strittig: Individuum) und Gesellschaft stehen in einem wechselseitigen Abhängigkeitsverhältnis. Langfristig stabilisieren sich Gesellschaften nur, wenn sie sich über Sozialisation, Strukturen und Wertvorstellungen reproduzieren. Ursprüngliche Instanz ist hier durch biologische Determination die Kernfamilie, aber sogar dies ist umstritten.

Systemtheorie (Luhmann)
Niklas Luhmann spricht von einer "Gesellschaft", wenn konformes und abweichendes Verhalten in Bezug auf Normen und Werte festgelegt ist und eine entsprechende Differenzierung von Erwartungen und Reaktionen vorhanden ist.

In der kommunikationstheoretischen Konzeption Luhmanns wird die Gesellschaft als "alle füreinander kommunikativ zugänglichen Ereignisse" beschrieben. Gesellschaft in systemtheoretischen Begriffen ist demnach das umfassendste soziale System – jene Einheit, die keine soziale Umwelt mehr hat und alle anderen sozialen Systeme inklusive Organisationen sowie Verhältnisse und Tatbestände umfasst. Anders ausgedrückt ist Gesellschaft alles, was durch Kommunikation füreinander erreichbar ist.

Praxis und Theorie der Praxis (Bourdieu)
Für Pierre Bourdieu ist Gesellschaft nicht völlig erklärbar. Es gebe aber zwei zu unterscheidende Ebenen: die Ebene der sozialen Praxis, in der sich das Leben in "Gesellschaft stiftender Arbeit" nach Regelmäßigkeiten abspiele, deren Ablauf die Akteure zum großen Teil unbewusst inkorporiert haben, und die Ebene der Theorie der Praxis, wo untersucht werden müsse, die unbewussten, in ihrer Gesamtheit kaum wahrgenommenen Machtverhältnisse der sozialen Praxis aufzudecken, und zwar dort, wo sie  weitestgehend mit den Gewohnheiten des Handelns, des Wahrnehmens und Beurteilens bricht. Bourdieus sehr einflussreiches Werk enthält damit eine gesellschaftskritische Komponente.

Weltgesellschaft

Tönnies' und Luhmanns Ansätze erlauben oder implizieren – wie die vieler anderer soziologischer Makrotheoretiker – auch die Idee einer Weltgesellschaft:
 Karl Marx – globaler Ausbeutungsmechanismus im Kapitalismus;
 Ludwig Gumplovicz – Krieg zwischen Gruppen;
 Ferdinand Tönnies – Welthandel;
 Niklas Luhmann – globale Verbreitung von Medienkommunikation sowie ein (welt)gesellschaftliches Funktionssystem
Diese Ansätze werden interdisziplinär prominent von der Weltsystemanalyse um Immanuel Wallerstein vertreten.

Siehe auch

 Sozialstruktur – Gemeinwesen
 nivellierte Mittelstandsgesellschaft
 Gesellschaftsordnung
 Gesellschaftsmodelle:
 Agrargesellschaft · Arbeitsgesellschaft · Post-Arbeitsgesellschaft · Gute Gesellschaft · Industriegesellschaft · Leistungsgesellschaft · Postindustrielle Gesellschaft · Informationsgesellschaft · Wissensgesellschaft · Digitale Wissensgesellschaft · Netzwerkgesellschaft · Konsumgesellschaft · Freizeitgesellschaft · Erlebnisgesellschaft · Risikogesellschaft · Ego-Gesellschaft · Ständegesellschaft · bürgerliche Gesellschaft · Bürgergesellschaft · Dienstleistungsgesellschaft · Überflussgesellschaft · Wegwerfgesellschaft · Zivilgesellschaft

Literatur
 Norbert Elias: Was ist Soziologie? Juventa, München 1970.
 Johannes Heinrichs: Logik des Sozialen. Woraus Gesellschaft entsteht. Steno, München 2005.
 Karl-Heinz Hillmann: Gesellschaft. In: ders.: Wörterbuch der Soziologie (= Kröners Taschenausgabe. Band 410). 4., überarbeitete und ergänzte Auflage. Kröner, Stuttgart 1994, ISBN 3-520-41004-4.
 Peter Koslowski: Evolution und Gesellschaft. Eine Auseinandersetzung mit der Soziobiologie. 2. Auflage. Mohr Siebeck, Tübingen 1984.
 Jörn Lamla, Henning Laux, Hartmut Rosa, David Strecker (Hrsg.): Handbuch der Soziologie. UVK, Konstanz 2014, ISBN 978-3-8252-8601-9.
 Niklas Luhmann: Die Gesellschaft der Gesellschaft. 1997, ISBN 3-518-28960-8.
 Hartmut Rosa, Jörg Oberthür u. a.: Gesellschaftstheorie. UVK, München 2020, ISBN 978-3-8252-5244-1.
 Peter Ruben: Gemeinschaft und Gesellschaft – erneut betrachtet. In Dittmar Schorkovitz (Hrsg.): Ethnohistorische Wege und Lehrjahre eines Philosophen. Frankfurt am Main 1995.
 Gunter Runkel: Allgemeine Soziologie. Gesellschaftstheorie, Sozialstruktur und Semantik. Oldenbourg, München/ Wien 2005, ISBN 3-486-57708-5 (v. a. Kap. 2: Soziologische Klassiker und ihre Theorien).
 Ferdinand Tönnies: Gemeinschaft und Gesellschaft. 1887; Wissenschaftliche Buchgesellschaft, Darmstadt 2005.
 Bernard Willms: System und Subjekt oder die politische Antinomie der Gesellschaftstheorie. In: Franz Maciejewski (Hrsg.): Theorie der Gesellschaft oder Sozialtechnologie. Suhrkamp, Frankfurt am Main 1973, ISBN 3-518-06101-1, S. 43–77.

Weblinks

 Soziologie schafft Gesellschaft, Armin Nassehi über den soziologischen Diskurs der Moderne. Deutschlandfunk.
 Weltgesellschaft Tagungsintro des Instituts für Weltgesellschaft der Fakultät für Soziologie der Universität Bielefeld und der Zeitschrift für Soziologie.
 Bundeszentrale für politische Bildung Das Unbehagen an der Gesellschaft von Martin Hartmann, Institut für Sozialforschung der Johann Wolfgang Goethe-Universität in Frankfurt am Main.

Einzelnachweise

Soziologie
Philosophische Anthropologie
Rechtsstaat

redpajama_set_name: RedPajamaWikipedia


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Trypanosoma cruzi mitochondrial swelling and membrane potential collapse as primary evidence of the mode of action of naphthoquinone analogues
Kelly Salomão1,
Natalia A De Santana1,
Maria Teresa Molina2,
Solange L De Castro1 &
Rubem F S Menna-Barreto1
BMC Microbiology volume 13, Article number: 196 (2013) Cite this article
Naphthoquinones (NQs) are privileged structures in medicinal chemistry due to the biological effects associated with the induction of oxidative stress. The present study evaluated the activities of sixteen NQs derivatives on Trypanosoma cruzi.
Fourteen NQs displayed higher activity against bloodstream trypomastigotes of T. cruzi than benznidazole. Further assays with NQ1, NQ8, NQ9 and NQ12 showed inhibition of the proliferation of axenic epimastigotes and intracelulluar amastigotes interiorized in macrophages and in heart muscle cells. NQ8 was the most active NQ against both proliferative forms of T. cruzi. In epimastigotes the four NQs induced mitochondrial swelling, vacuolization, and flagellar blebbing. The treatment with NQs also induced the appearance of large endoplasmic reticulum profiles surrounding different cellular structures and of myelin-like membranous contours, morphological characteristics of an autophagic process. At IC50 concentration, NQ8 totally disrupted the ΔΨm of about 20% of the parasites, suggesting the induction of a sub-population with metabolically inactive mitochondria. On the other hand, NQ1, NQ9 or NQ12 led only to a discrete decrease of TMRE + labeling at IC50 values. NQ8 led also to an increase in the percentage of parasites labeled with DHE, indicative of ROS production, possibly the cause of the observed mitochondrial swelling. The other three NQs behaved similarly to untreated controls.
NQ1, NQ8, NQ9 and NQ12 induce an autophagic phenotype in T. cruzi epimastigoted, as already observed with others NQs. The absence of oxidative stress in NQ1-, NQ9- and NQ12-treated parasites could be due to the existence of more than one mechanism of action involved in their trypanocidal activity, leaving ROS generation suppressed by the detoxification system of the parasite. The strong redox effect of NQ8 could be associated to the presence of the acetyl group in its structure facilitating quinone reduction, as previously demonstrated by electrochemical analysis. Further experiments using biochemical and molecular approaches are needed to better characterize ROS participation in the mechanism of action of these NQs.
Chagas' disease (CD), caused by Trypanosoma cruzi, affects approximately eight million individuals in Latin America [1, 2] and is emerging in non-endemic areas due to the globalization of immigration and non-vector transmission routes [3]. The available therapy for CD is based on two nitroheterocycles, benznidazole (Bz) and nifurtimox, and was developed more than four decades ago. Both nitroheterocycles are far from ideal due to substantial secondary side effects, limited efficacy against different parasite isolates, the need for long-term therapy and their well-known poor activity in the late chronic phase. These drawbacks justify the urgent need to identify better drugs to treat chagasic patients [4].
Naphthoquinones account for the largest number of natural naphthalenes, holding a number of different substituents with a variety of structural motifs. They act as vital links in the electron transport chains in metabolic pathways and participate in multiple biological oxidative processes [5]. Quinone-containing plants have been used in diverse cultures as dyes, cosmetics, and food and, especially among Indian populations, for the treatment of different diseases [6, 7]. Naphthoquinones are considered privileged structures in medicinal chemistry due to their structural properties and biological activities [8], especially against tumor cells and pathogenic protozoa [9, 10]. Two major mechanisms of quinone cytotoxicity have been proposed: stimulation of oxidative stress and alkylation of cellular nucleophiles, which are the mechanisms of action common to a large range of biomolecules [11].
Among the simple hydroxylated naphthoquinones, juglone (5-hydroxy-1,4-naphthoquinone), isolated from walnut trees (Juglandaceae), has shown a variety of biological effects, including microbicidal [12], anti-inflammatory [13] and antitumoral [14, 15] effects that are associated with the induction of oxidative stress. As part of our continuing program of screening natural and synthetic quinones for trypanocidal activity, in the present work we investigated the activity and mode of action of naphthoquinones and specific juglone derivatives.
Activity on bloodstream trypomastigotes
In the present work, we initially evaluated the efficacy of sixteen 1,4-naphthoquinones (1,4-NQs) against the infective bloodstream trypomastigote forms of T. cruzi at 37°C in Dulbecco's modified Eagle's medium (Sigma-Aldrich) plus 10% fetal calf serum (DMES) (Table 1). The prototype, 1,4-naphthoquinone (NQ1), was compared with other derivatives that were substituted at C-2 with methyl (NQ2), hydroxyl (NQ3), acetoxy (NQ4), and bromo (NQ5) groups or that were disubstituted at C-2 and C-3, such as 2,3-dichoro-1,4-naphthoquinone (NQ6). In addition, juglone (NQ7) and its derivatives, including those brominated at C-2 (NQ10 to NQ12) or C-3 (NQ13 to NQ15) and 2-methyl-5-hydroxy-1,4-naphthoquinone (NQ16), were also examined. Fourteen compounds displayed an IC50 in the range of 0.16 to 6.51 μM, demonstrating higher activity than Bz (26.0 μM), and the other two tested compounds were less active: NQ3 (563.18 μM) and NQ4 (63.02 μM) (Table 1).
Table 1 Activity of the naphthoquinones on bloodstream trypomastigotes of T. cruzi at 37°C
Among the most active compounds on trypomastigotes at 37°C, four were selected for further studies: the prototype naphthoquinone (NQ1) and three juglone derivatives (NQ8, NQ9 and NQ12) (Figure 1). Interestingly, their activity against trypomastigotes was not decreased when the experiments were performed at 4°C in culture medium, but at this lower temperature in the presence of whole blood, IC50 values higher than 500 μM were obtained (data not shown).
Chemical structures of the studied naphthoquinones.
Activity on the proliferative forms of T. cruziand toxicity to mammalian cells
The selected compounds (NQ1, NQ8, NQ9 and NQ12) were also assayed using the proliferative forms of T. cruzi: axenic epimastigotes and intracellular amastigotes. A dose-dependent effect on epimastigotes was observed, leading to the IC50 values for proliferation inhibition for 1 to 4 days of treatment displayed in Table 2. Comparing the four NQs, the prototype unsubstituted quinone NQ1 was the most active against epimastigotes.
Table 2 IC 50 values (μM) of the naphthoquinones on the proliferation of T. cruzi epimastigotes
The NQs were added to T. cruzi-infected primary cultures of both peritoneal macrophages and HMCs after the removal of non-interiorized parasites. For both host cells the inhibitory effect on the percent infection was in the range of 0.5 to 5.0 μM. Surprisingly, NQ8 and NQ9 caused about a 2.5-fold decrease of infection. For both host cells, the IC50 values after 48 h of treatment used to calculate the endocytic index are displayed in Table 3. NQ8 was the most active compound. Non-infected macrophages and HMCs treated with the compounds for 2 days were tested with the MTT assay to evaluate their toxicity to mammalian cells. For HMCs, the LC50 values were 8 μM for NQ1 and NQ12 and 10 μM for NQ8; NQ9 was the least toxic quinone with values higher than 10 μM. The LC50 was higher than 10 μM in macrophages for all four compounds.
Table 3 IC 50 values (μM) of the naphthoquinones on intracellular amastigotes of T. cruzi
Ultrastructural analysis
Transmission electron microscopy showed that treatment with the NQs induced important alterations in the mitochondrion of the epimastigotes, leading to swelling and the appearance of membranous structures in the organelle matrix (Figures 2, 3, 4 and 5). Autophagic features, such as atypical cytosolic membranous structures (Figures 3, 4, 5) and the appearance of endoplasmic reticulum surrounding reservosomes (Figures 2 and 5), were detected in treated parasites. The naphthoquinones also led to intense cytosolic vacuolization (Figures 4 and 5), the formation of blebs in the flagellar region (Figures 2, 3 and 5) and the induction of loss of the electron-density of the cytosol (washed out aspect) (Figures 3 and 5). The scanning electron microscopy technique demonstrated no important morphological alterations in treated epimastigotes (data not shown).
Transmission electron microscopy analysis of T. cruzi epimastigotes treated with NQ1. (A) Untreated epimastigote showing normal ultrastructural aspect and presenting typical morphologies of the mitochondrion (M), kinetoplast (K), flagellum (F), nucleus (N), Golgi (G), reservosome (R) and cytostome (Cy). (B-E) The concentration of 0.3 μM NQ1 led to swelling in the mitochondrion (*), the formation of abnormal cytosolic membranous structures (white arrowheads) and the appearance of endoplasmic reticulum surrounding reservosomes (white arrows). Blebs (thick black arrows) was formed in the flagellar membrane of treated parasites. Bars = 500 nm (A, B, E) and 200 nm (C, D).
Transmission electron microscopy analysis of T. cruzi epimastigotes treated with NQ8. (A-D) Treatment with 0.8 μM NQ8 led to swelling in the mitochondrion (*) and the appearance of membranous structures inside the organelle (black arrows). The formation of atypical cytosolic membranous structures was also observed (white arrowheads) near to the washed out aspect of cytosol (black star). Blebs containing electron-dense material (thick black arrows) were found close to the flagellar pocket. Bars = 500 nm (A-C) and 200 nm (D).
Transmission electron microscopy analysis of T. cruzi epimastigotes treated with NQ9. (A-C) This naphthoquinone (2.6 μM) induced morphological alterations in the mitochondrion, including swelling (*) and the formation of membranous structures (black arrows) inside the organelle. Parasites treated with NQ9 also presented atypical cytosolic membranous structures (white arrowheads) and intense cytosolic vacuolization (V). Bars = 500 nm.
Transmission electron microscopy analysis of T. cruzi epimastigotes treated with NQ12. (A-E) Parasites treated with 0.5 μM showed a strong mitochondrial swelling (*) with membranous structures in the organelle matrix (black arrows), the formation of flagellar blebs (thick black arrows) and the appearance of endoplasmic reticulum in close contact with the reservosome membranes (white arrows). An intense vacuolization (V) and washed out aspect of the cytosol (black star) were also detected after treatment with NQ12. Bars = 500 nm (A, C-E) and 200 nm (B).
Flow cytometry analysis
This technique was employed to evaluate the mitochondrial membrane potential (ΔΨm) dissipation by labeling epimastigotes with the specific marker TMRE in the presence of 10 μM FCCP. The four NQs, at IC50 levels, induced a significant decrease in the TMRE fluorescence, denoted in Table 4 by the reduction of the IV values (see Methods) from −0.22 to −0.53. NQ8 at the concentration of 8 μM presented the most remarkable reduction in the fluorescence intensity of the marker and totally disrupted the ΔΨm of about 20% of the parasites (Table 4). On the other hand, treatment with NQ1, NQ9 or NQ12 induced no alteration in the percentage of TMRE + epimastigotes, a finding that was quite similar to that observed in control parasites. ROS production was assessed by DHE labeling and incubation with AA, a potent ROS inducer. Only treatment at the IC50 of NQ8 led to a discrete increase in the percentage of DHE + parasites (Table 4). The other three NQs yielded the same labeling pattern as the untreated cells at every dose tested.
Table 4 Flow cytometry analysis of ΔΨm and ROS production in T. cruzi epimastigotes
Initially, the sixteen derivatives were assayed against bloodstream forms of T. cruzi at 37°C (Table 1). The activity of NQ1 was surprising because this compound is the nonsubstituted 1,4-naphthoquinone. The introduction of a hydroxyl at C5 (NQ7, juglone) is detrimental to the trypanocidal activity, which is decreased 8× in comparison with the parent quinone. Among the three simple juglone derivatives, the substitution of a hydroxyl by an acetoxy or methoxy group leads to higher biological activity. The O-methylated (NQ9) and the O-acetylated (NQ8) juglone derivatives were 6.4× and 40×, respectively, more active than juglone (NQ7) itself. Among the 2- and 3-bromojuglone derivatives (NQ10 to NQ15), regardless of the substituent, roughly the same efficacy was observed (IC50 between 1.2 and 2.5 μM), with the exception of NQ14, which displayed trypanocidal activity similar to that of nonsubstituted NQ1. Moreover, NQ12 and NQ15 are very similar and, in both cases, are slightly less effective than the parent methyl ether NQ9. This trend is also valid among the 5-hydroxy derivatives. Thus, NQ10 and NQ13 had similar activity but showed 3-fold higher activity than juglone itself (NQ7).
The effect of the juglone derivatives was previously investigated on Aedes aegypti, the vector of dengue, and on adult Biomphalaria glabrata snails [16]. Concerning the larvicidal activity, NQ10, NQ11 and NQ13 were the most active, with IC50 values of about 4 μM. With respect to their molluscidal effects, NQ11, NQ12, NQ14 and NQ15 had ranges of activity between 1.8 and 3.2 μM. Cytotoxic assays using four human cancer cell lines revealed that NQ9 was the most active, with IC50/72 h values ranging from 1.7 to 4.7 μM, whereas for juglone (NQ7), this range was from 7.6 to over 28.7 μM [14]. The mechanism underlying the cytotoxicity of NQ9 to HL-60 cells involved the activation of caspases leading to an induction of apoptosis independent of mitochondria depolarization [14].
Leaving aside the juglone derivatives, and with the exceptions of NQ3, previously shown by us as inactive against T. cruzi in other experimental conditions [17], and of NQ4, all the compounds displayed IC50 values in the range of 1.37 (NQ5) to 6.04 (NQ2) μM, corresponding to a higher activity in comparison with the standard drug benznidazole, which has an IC50 value of 26.0 ± 4.0 μM. In a study with Bolivian medicinal plants, Fournet and colleagues [18, 19] reported the potent effect of NQ16 (plumbagin), isolated from Pera benensis, against T. cruzi and different species of Leishmania.
The four compounds that were most active on bloodstream forms at 37°C were assayed also at 4°C: in the absence of blood, the lytic effect on trypomastigotes was not decreased, while in the presence of whole blood, IC50 values higher than 500 μM were obtained. These results are consistent with previous reports on the literature regarding the inactivation of the trypanocidal activity of quinones in the presence of blood components [17, 20]. Comparing the susceptibility of the different developmental forms of T. cruzi to the compounds, it was observed that bloodstream trypomastigotes were more susceptible to NQ8, whereas epimastigotes were more susceptible to NQ1. Intracellular amastigotes from heart muscle cells or peritoneal macrophages were at least 2-fold more resistant to treatment with NQ1, NQ8 and NQ12.
For the subsequent investigation of the mode of action of the four selected NQs, electron microscopy and flow cytometry assays with epimastigotes were employed, never exceeding the respective IC50 values. Treatment with these compounds led to remarkable ultrastructural alterations, especially in the mitochondrion. The appearance of different morphological features suggestive of autophagic activity and the interference in flagellar membrane fluidity with bleb formation were also recurrent alterations.
Mitochondrial susceptibility to treatment with naphthoquinones and its derivatives has been extensively reported [21–28]. Mitochondria of trypanosomatids parasites exhibit unique structural and functional features that are remarkably distinct from mammalian counterparts. The absence of efficient mechanisms for ROS detoxification in these parasites make the mitochondrion a good target for drug intervention [29], and functional evaluation of the organelle by ΔΨm measurement represents an important step for the examination of the mechanism of action of novel drugs [22–24, 28]. Here, we assessed ΔΨm by TMRE labeling in epimastigotes treated with NQs. We added FCCP as a control. This ionophore works as an uncoupling agent that impairs ATP synthesis by dissipating the hydrogen ion gradient and consequently stopping oxidative phosphorylation [30]. Flow cytometry revealed a decrease in the mitochondrial potential after incubation with the four NQs at their IC50 values, and in the case of NQ8, even at a concentration 4-fold lower (Table 4). Another parameter analyzed was the percentage of TMRE + parasites. We standardized the negative populations by the addition of 10 μM FCCP, which totally dissipated the ΔΨm in epimastigotes (± 4% TMRE + cells). Interestingly, a reduction of about 20% in the TMRE + population was also observed in NQ8-treated parasites at the IC50. Such a decrease indicates that this naphthoquinone induces the appearance of a sub-population of parasites with metabolically inactive mitochondria.
Previous reports on the effects of several natural quinones, such as lapachol and β-lapachone, against T. cruzi have reported the involvement of free radicals in the trypanocidal activity [9, 31], making the reduction of NQs by parasitic flavoenzymes a promising strategy for the development of trypanocidal drugs [32]. Mechanistically, it was reasonable to postulate that the collapse of the ΔΨm was mediated by ROS generation in the treated parasites. In this context, the fluorescent probe DHE was used for intracellular ROS detection, and AA was added as a positive control because it inhibits the electron flow through the electron transport chain, leading to the accumulation of superoxide [33]. Among the four NQs tested, only NQ8 led to a discrete increase in the percentage of DHE + epimastigotes, giving addition evidence for the strong effect of this quinone on the parasite ΔΨm. Indeed, the pool of anti-oxidant defenses in epimastigotes that includes trypanothione, tryparedoxin peroxidase and other redox enzymes leads to a protective effect in this parasite stage, as previously described [34]. Thus, one plausible hypothesis to explain the absence of oxidative stress triggered by NQ1, NQ9 and NQ12 could be the existence of more than one mechanism of action involved in the trypanocidal activity of these compounds, leaving ROS generation suppressed by the detoxification system of the parasite. Possibly, the strong redox effect of NQ8 could be associated to the presence of the acetyl group in its structure facilitating quinone reduction, as previously demonstrated by electrochemical analysis [35]. Further experiments using different biochemical and molecular approaches must be performed to better characterize ROS participation in the mechanism of action of these compounds.
Electron microscopy evidence of induction of the autophagic pathway by naphthoquinones and their derivatives has also been previously reported [24–26, 28]. The presence of large profiles of endoplasmic reticulum surrounding different cellular structures, such as lipid droplets and organelles, and the appearance of bizarre membranous structures with a myelin-like aspect are the most common characteristics. The autophagic process represents a fundamental constitutive pathway in eukaryotic cells that is responsible for remodeling cellular structures and maintaining homeostasis. In trypanosomatids, other roles for autophagy have been proposed, including in the parasite's differentiation [36]. In a great variety of cell models, the loss of the balance between anabolic and catabolic processes leads to non-apoptotic death [37]. In the last decade, it has been demonstrated that the induction of autophagy in T. cruzi trypanosomatids is triggered by several classes of drugs, in particular naphthoquinones and their derivatives [25, 26, 38]. Our transmission electron microscopy analysis suggested the involvement of endoplasmic reticulum and cytosolic membranous structures in pre-autophagosomal formation, as previously postulated by Yotimitsu & Klionsky [39].
Another ultrastructural alteration observed after the treatment with NQs was the development of blebs in the flagellar membrane, which was also previously reported after treatment with naphthoquinone derivatives [23, 25, 26] and is commonly associated with apoptosis-like death in different cells [40].
Although the autophagic phenotype was the most frequently observed ultrastructural alteration in treated epimastigotes and bleb formation was the unique characteristic of an apoptosis-like process, a hypothesis that there is interplay between the distinct death pathways through a cross-talk signaling mechanism could not be discarded. Similar mechanisms have been demonstrated for other eukaryotic cells in the literature [41]. Especially in T. cruzi, the processes of death regulation are poorly understood and deserve further studies aimed at the development of new therapeutic agents.
The naphthoquinone NQ1 (1,4-naphthoquinone) was purchased from Fluka (Sigma-Aldrich Chemical Co., St. Louis, USA), NQ2 (menadione) and NQ5 were purchased from Sigma-Aldrich, and NQ3 (lawsone) and NQ6 (dichlone) were purchased from Acros Organic (Geel, Belgium). Compound NQ4 was prepared by standard acetylation of NQ3 [14]. All the juglone derivatives (NQ7 to NQ15) were prepared according to methods described in the literature [14]. Juglone (NQ7) is a commercial material and, when needed on a large scale, was prepared according to the method by Tietze et al. [42] and purified by flash chromatography [14, 43, 44]. Acetylation of juglone under standard conditions yielded juglone acetate (5-acetoxy-1,4-naphthoquinone, NQ8) [45]. The methoxy derivative NQ9 (5-methoxy-1,4-naphthoquinone) was prepared by the methylation of NQ7 using methyl iodide and silver (I) oxide [42]. For the 2-bromojuglone derivatives, NQ10 was prepared according to Grunwell et al. [46] by oxidative bromination of 1,5-diacetoxynaphthalene. Starting with NQ10, we obtained NQ11 by standard acetylation and NQ12 by methoxylation [47]. The 3-bromojuglone derivatives were prepared by selective bromination of NQ7 according to Brimble & Brenstrum [48], which yielded NQ13 as the major isomer. From this derivative, either by standard acetylation or methylation, NQ14 [47] and NQ15 [49], respectively, were obtained. NQ16, which combines the structural features of NQ2 and NQ7, was purchased from Sigma-Aldrich (Figure 1).
Stock solutions of the compounds were prepared in dimethylsulfoxide (DMSO), with the final concentration of the latter in the experiments never exceeding 0.1%. Preliminary experiments showed that at concentrations of up to 0.5%, DMSO has no deleterious effect on the parasites [50].
Albino Swiss mice were employed for the trypomastigotes and host cells obtention. This study is in accordance to the guidelines of the Colégio Brasileiro de Experimentação Animal (COBEA) and was performed in biosafety conditions. All the procedures in animal experimentation were approved by the Comissão de Ética em Experimentação Animal (CEUA/Fiocruz), license LW 16/13.
All experiments were performed with the Y strain of T. cruzi. Epimastigote forms were maintained axenically at 28°C with weekly transfers in LIT medium and harvested during the exponential phase of growth. Bloodstream trypomastigotes were obtained from infected mice at the peak of parasitemia by differential centrifugation.
Effect on bloodstream trypomastigotes
The parasites were resuspended to a concentration of 10×106 cells/mL in DMES medium. This suspension (100 μL) was added to the same volume of each of the sixteen naphthoquinones (NQs), which had been previously prepared at twice the desired final concentrations. The incubation was performed in 96-well microplates (Nunc Inc., Rochester, USA) at 4°C or 37°C for 24 h at concentrations in the range of 0.06 to 1000 μM. Benznidazole (Laboratório Farmacêutico do Estado de Pernambuco, Brazil) the standard drug for treatment of chagasic patients was used as control. For experiments performed in the presence of 100% blood, the parasites were resuspended in mouse blood to a concentration of 5×106 cells/mL, and 196 μL of the suspension was added to each well together with 4 μL of the NQs (0.06 to 1000 μM), which had been selected on the basis of the results of previous experiment and had been prepared at a concentration 50 times higher than the final concentration desired. Cell counts were performed in a Neubauer chamber, and the activity of the compounds corresponding to the concentration that led to 50% lysis of the parasites was expressed as the IC50/1 day.
Effect on epimastigotes
The parasites were resuspended in LIT medium to a parasite concentration of 10 × 106 cells/mL. This suspension was added to the same volume of the NQs (NQ1, NQ8, NQ9 and NQ12) at concentrations in the range of 0.06 to 10 μM and then incubated at 28°C in 24-well plates (Nunc Inc.). Cell counts were performed daily (from 1 to 4 days) in a Neubauer chamber, and the activity of the compounds was expressed as IC50, which corresponds to the concentration that leads to 50% proliferation inhibition.
Effect on intracellular amastigotes
Peritoneal macrophages were obtained from mice and plated in 24-well plates (3 × 105 cells/well) (Nunc Inc., IL, USA) for 24 h. Then, the cultures were infected with trypomastigotes (10:1 parasite:host cell) in DMES medium. After 3 h of incubation, the cultures were washed to remove non-internalized parasites, and the selected NQs were added at final concentrations ranging from 0.5 to 20 μM. Alternatively, primary cultures of mouse embryo heart muscle cells (HMCs) [51] were used. Briefly, the hearts of 18-day-old mouse embryos were fragmented and dissociated with trypsin and collagenase in phosphate buffered saline (PBS), pH 7.2. Thereafter, the cells were resuspended in Dulbecco's modified Eagle's medium supplemented with horse and fetal calf sera, chicken embryo extract, CaCl2 and L-glutamine; then, the cells were plated onto gelatin-coated glass coverslips (105 cells/well) and maintained at 37°C in 5% CO2 atmosphere. The HMCs were plated onto gelatine-coated glass coverslips in 24-well plates (105 cells/well) and infected at a 10:1 parasite:host cell ratio after 24 h. Afterwards the cultures were washed, and the NQs (0.5 to 20 μM) were added. At specified intervals, the cultures were fixed in Bouin's solution, stained with Giemsa and counted to assess the following parameters: percentage of cells infected, number of parasites/infected cell and the endocytic index (EI), which refers to the number of parasites/100 cells [52]. The IC50 values for the different days of treatment, corresponding to the concentration that led to 50% inhibition of each parameter, were calculated. To determine the possible toxic effects of the compounds on the host cells, uninfected macrophages and HMCs were incubated at 37°C with the NQs. After 2 days, the viability of the cells was measured using the MTT colorimetric assay [53]. The absorbance was measured at 490 nm with a spectrophotometer (VERSAmax Tunable, Molecular Devices, USA), allowing for the determination of an LC50 value, which is the concentration that reduces cellular viability by 50%.
Transmission and scanning electron microscopy analysis
Epimastigotes (5×106 cells/mL) were treated for 24 h with the selected NQs at their respective IC50/24 h values in LIT medium at 28°C. Afterward, they were fixed with 2.5% glutaraldehyde in 0.1 M Na-cacodylate buffer (pH 7.2) for 40 min at 25°C and post-fixed with 1% OsO4, 0.8% potassium ferricyanide and 2.5 mM CaCl2 in the same buffer for 20 min at 25°C. The cells were dehydrated in an ascending acetone series and embedded in PolyBed 812 resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined in a Jeol JEM1011 transmission electron microscope (Tokyo, Japan). Alternatively, dehydrated samples were dried by the critical point method with CO2, mounted on aluminum stubs, coated with a 20 nm thick gold layer and examined on a Jeol JSM6390LV scanning electron microscope (Tokyo, Japan). Both electron microscopes are located in Plataforma de Microscopia Eletrônica at Instituto Oswaldo Cruz (Fiocruz).
Epimastigotes were treated for 24 h with the NQs at concentrations up to their IC50 values. We then determined the mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) production. For ΔΨm analysis, the parasites were incubated with 50 nM tetramethylrhodamine (TMRE) (Molecular Probes, Carlsbad, USA) for 15 min at 28°C, using 10 μM carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) (Sigma-Aldrich Chemical Co.) as a control for ΔΨm dissipation. Alterations in TMRE fluorescence were quantified using an index of variation (IV), which was calculated using the equation (MT - MC)/MC, where MT is the median of fluorescence for treated parasites and MC is the median of fluorescence of the control parasites. Negative IV values correspond to depolarization of the mitochondrial membrane [54]. To evaluate ROS generation, labeling with 10 μM dihydroethidium (DHE) (Molecular Probes) for 30 min at 28°C was performed, using 22 μM antimycin A (AA) (Sigma-Aldrich) as the positive control. The samples were analyzed in a FACSCalibur flow cytometer (Becton Dickinson, CA, USA) equipped with the Cell Quest software (Joseph Trotter, Scripps Research Institute, La Jolla, USA). A total of 10,000 events were acquired in the region previously established as that of the parasites.
The comparison between control and treated groups was performed using the Mann–Whitney test. Differences with p ≤ 0.05 were considered statistically significant.
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Yorimitsu T, Klionsky DJ: Eating the endoplasmic reticulum: quality control by autophagy. Trends Cell Biol. 2007, 17: 279-285. 10.1016/j.tcb.2007.04.005.
Walker NI, Harmon BV, Gobé GC, Kerr JF: Patterns of cell death methods. Arch Exp Pathol. 1988, 13: 18-54.
Guimarães CA, Linden R: Programmed cell death: apoptosis and alternative death styles. Eur J Biochem. 2004, 271: 1638-1650. 10.1111/j.1432-1033.2004.04084.x.
Tietze LF, Güntner C, Gericke KM: A Diels-Alder reaction for the total synthesis of the novel antibiotic antitumor agent mensacarcin. Eur J Org Chem. 2005, 12: 2459-2467.
Molina MT, Navarro C, Moreno A, Csákÿ AG: Arylation of benzo-fused 1,4-quinones by the addition of boronic acids under dicationic Pd(II)-catalysis. Org Lett. 2009, 11: 4938-4941. 10.1021/ol902084g.
Ortega A, Rincón Á, Jiménez-Aliaga KL, Bermejo-Bescós P, Martín-Aragón S, Molina MT, Csákÿ AG: Synthesis and evaluation of arylquinones as BACE1 inhibitors, β-amyloid peptide aggregation inhibitors, and destabilizers of preformed β-amyloid fibrils. Bioorg Med Chem Lett. 2011, 21: 2183-2187. 10.1016/j.bmcl.2011.03.023.
Fieser LF, Dunn JT: Addition of dienes to halogenated and hydroxylated naphthoquinones. J Am Chem Soc. 1937, 59: 1016-1021. 10.1021/ja01285a017.
Grunwell JR, Karipides A, Wigal CT, Heinzman SW, Parlow J, Surso JA, Clayton L, Fleitz FJ, Daffner M, Stevens JE: The formal oxidative addition of electon-rich transoid dienes to bromonaphthoquinones. J Org Chem. 1991, 56: 91-95. 10.1021/jo00001a020.
Parker KA, Sworin ME: Assignment of regiochemistry to substituted naphthoquinones by bromo juglone derivatives chemical and spectroscopic methods amino-, hydroxy-, and bromojuglone derivatives. J Org Chem. 1981, 46: 3218-3223. 10.1021/jo00329a014.
Brimble MA, Brenstrum TJ: C-Glycosylation of tri-O-benzyl-2-deoxy-D-glucose: synthesis of naphthyl substituted 3,6-dioxabicyclo [322] nonanes. J Chem Soc Perkin. 2001, 1: 1612-1623.
Tietze LF, Gericke KM, Schuberth I: Synthesis of highly functionalized anthraquinones and evaluation of their antitumor activity. Eur J Org Chem. 2007, 27: 4563-4577.
De Castro SL, Pinto MCFR, Pinto AV: Screening of natural and synthetic drugs against Trypanosoma cruzi: I-Establishing a structure/activity relationship. Microbios. 1994, 78: 83-90.
Meirelles MNL, Araujo-Jorge TC, Miranda CF, De Souza W, Barbosa HS: Interaction of Trypanosoma cruzi with heart muscle cells: ultrastructural and cytochemical analysis of endocytic vacuole formation and effect upon myogenesis in vitro. Eur J Cell Biol. 1986, 41: 198-206.
Salomão K, De Souza EM, Carvalho AS, Silva EF, Fraga CAM, Barbosa HS, De Castro SL: In vitro and in vivo activity of 1,3,4-thiadiazole-2-arylhydrazone derivatives of megazol on Trypanosoma cruzi. Antimicrob Agents Chemother. 2010, 54: 2023-2031. 10.1128/AAC.01241-09.
Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth. 1983, 65: 55-63. 10.1016/0022-1759(83)90303-4.
Santa-Rita RM, Henriques-Pons A, Barbosa HS, De Castro SL: Effect of the lysophospholipid analogues edelfosine, ilmofosine and miltefosine against Leishmania amazonensis. J Antimicrob Chemother. 2004, 54: 704-710. 10.1093/jac/dkh380.
Funding was provided by Fundação de Amparo à Pesquisa do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação Oswaldo Cruz (FIOCRUZ) and Spanish MICINN (Project SAF 2009–10399, to MTM).
Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Manguinhos, Rio de Janeiro, RJ, 21040-900, Brazil
Kelly Salomão
, Natalia A De Santana
, Solange L De Castro
& Rubem F S Menna-Barreto
Instituto de Química Médica, CSIC, Juan de la Cierva 3, Madrid, 28006, Spain
Maria Teresa Molina
Search for Kelly Salomão in:
Search for Natalia A De Santana in:
Search for Maria Teresa Molina in:
Search for Solange L De Castro in:
Search for Rubem F S Menna-Barreto in:
Correspondence to Solange L De Castro.
KS and NAS performed the trypanocidal activity assays. MTM synthesized the naphthoquinone derivatives. RFSMB and NAS designed and performed the electron microscopy and flow cytometry assays. SLC contributed to the design and supervision of the experiments. SLC and RFSMB wrote the manuscript. All authors have read and approved the final manuscript.
Salomão, K., De Santana, N.A., Molina, M.T. et al. Trypanosoma cruzi mitochondrial swelling and membrane potential collapse as primary evidence of the mode of action of naphthoquinone analogues. BMC Microbiol 13, 196 (2013) doi:10.1186/1471-2180-13-196
DOI: https://doi.org/10.1186/1471-2180-13-196
Rypanosoma cruzi
Juglone
Experimental chemotherapy
Microbial biochemistry, physiology and metabolism


text:
Add and Remove Sports Teams from the Sports Application on Microsoft Lumia 535
Edited by Thor, Melissa Rae, Jen M
Video: Add and Remove Sports Teams from the Sports Application on Microsoft Lumia 535
Hello, and welcome to VisiHow. In this video, we're going to show you how to add and remove a sports team from the "Sports" application on a Microsoft Lumia 535.
To begin, turn the phone on by pressing the "power" button located on the right-hand side of the phone
Swipe the lock screen up to enter the home screen
Locate and tap the "Sports" application tile
If it's not located on the home screen, search for it in the "App" menu.
Swipe across the screen to scroll through the "Sports" application
It displays the scoreboard, news and "my teams" sections. To add a team, press the white "plus" sign on the bottom of the screen.
Type in the team name
Choose the name from the list that appears.
Press the team's icon under "my teams" to access a database that displays a summary, scores, top stories, a table, team leaders and other aspects
Press the white arrow button on the bottom of the phone to go back
Swipe across to the "scoreboard" page
The team will now be displayed under "favorites". It will offer information about their next game, who they're playing and previous results.
Swipe across the screen to return to the "my teams" section
To remove a team from this section, press the team's icon.
Press the "..." button on the right at the bottom of the screen
Press the "remove" icon that appears as a white circle with a white star and a line in it
To add the team back, press the "star" icon again
It will now have a "plus" sign in it.
This concludes the tutorial on how to add and remove a sports team from the "Sports" application when using a Microsoft Lumia 535
If you have any questions, suggestions or comments about this video, then please leave them in the comment section below, and we shall answer them. You've been watching VisiHow. Thank you for watching. Goodbye.
Categories : Nokia
Recent edits by: Melissa Rae, Thor
Turn Messenger Notifications On or Off from Skype on Microsoft Lumia 535
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Edit or Change My Sports in the Sports Application on Microsoft Lumia 535
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text:
Het Zeitbrücke-Museum is een museum in Gars am Kamp in Neder-Oostenrijk. Het is een streekmuseum dat daarnaast permanente exposities toont over de componist Franz von Suppé, vierhonderd jaar handel en 5000 jaar nederzettingen in de omgeving. Er zijn ook wisselende exposities te zien.

Geschiedenis

In 1898 was er al een klein museum gevestigd in het raadhuis dat olieverfschilderijen, voorwerpen uit het beroepsgilde en documenten toonde. De componist Franz von Suppé (1819-1895) had hier zijn zomerverblijf gehad en zijn weduwe wilde hier na zijn dood een museum over hem oprichten. Door de omstandigheden liet hij echter maar enkele stukken achter in Gars. De expositie in het raadhuis werd in de loop van de jaren telkens weer onderbroken, met onder meer een herstart in 1949.

In 1971 werd opnieuw een herstart gemaakt en op initiatief van hoogleraar Hans Heppenheimer werden meubelstukken aangekocht die van de componist zijn geweest. In 1972 werd de bestaande collectie en de nieuwe stukken ondergebracht in de voormalige woning van Von Suppé aan de Kremserstraße. De expositie van het heemkundige museum vond plaats in het Tauchnerhaus aan de Hauptplatz 4, terwijl gebruik werd gemaakt van het depot in het raadhuis. In 1974 werd het heemkundig museum opgericht met daarin de collectie die aan Von Suppé is gewijd.

In 1979 werd het museum verplaatst naar de eerste verdieping in de Kollergasse, waar het een jaar later werd geopend. Eveneens in 1979 werd in een leegstaande school er tegenover de expositie 5000 jaar nederzetting in Thunau gehouden, met stukken die sinds 1965 in de omgeving waren opgegraven. In 1985 werd een tentoonstelling Handel im Wandel gehouden, waarmee werd teruggeblikt naar het 400-jarige handelsjubileum en het 250-jarig bestaan van de firma Kiennast. Net als de Thunau-expositie kreeg ook deze handelsexpositie een permanent karakter, namelijk vanaf 1989 in het raadhuis.

In 1990 werd de Thunau-expositie opgeheven en werd er voor de collectie van Von Suppé een nieuwe plaats gezocht. Hier werd in 1995 plaats voor gemaakt in het raadhuis. Terwijl dat jaar aan zijn honderdste overlijdensjaar werd herinnerd, bleven veel stukken in het depot staan door plaatsgebrek.

In 2002 verhuisde de collectie van Von Suppé naar het Zeitbrücke-Museum in de Kollergasse. Hier werd in 2006 ook het handelsmuseum naartoe gebracht. Ook is daar de documentatie te zien van de opgravingen in de omgeving.

Zie ook
 Lijst van muziekmusea
 Lijst van musea in Oostenrijk

Museum in Oostenrijk
Bouwwerk in Neder-Oostenrijk


On behalf of Law Offices of Gary A. Costales, P.A. posted in employment contracts on Thursday, February 7, 2019.
Whenever you sign an employment contract, you need to make sure that it contains all the proper elements to define your job. Remember, while your contract can offer you protection from things like a firing without warning, giving you security. It also tells you your obligations and shows what the company must do for you in turn. Never take this lightly or assume you know what a contract offers without reading it.


text:
Die Original Celtics waren eine amerikanische Basketball-Mannschaft, die den Profi-Basketball zwischen den Weltkriegen an der Ostküste prägte und die in die Naismith Memorial Basketball Hall of Fame aufgenommen wurde.

Geschichte
Der Club wurden im Jahr 1914 von Frank McCormack gegründet. Aufgrund der Ereignisse des Ersten Weltkrieges musste sich die Mannschaft wieder auflösen. Im Jahr 1918 baute der Promoter James Furey eine neue Mannschaft auf, die sich an den New York Celtics orientieren sollte. Die Original Celtics bestanden aus den Spielern der New York Celtics und einigen neuen Spielern (darunter Joe Trippe) von der West Side in Manhattan. Die Mannschaft bestand aus vielen Talenten wie dem Forward Dutch Dehnert oder dem Point Guard Nat Holman, der im Jahr 1922 dazukam. In ihrer ersten Saison haben die Celtics 65 Spiele von 69 gewonnen und somit einen neuen Rekord erreicht.

Die Celtics entwickelten einen Vertrag, der den Spielern verbot, die Mannschaft zu wechseln. Am 16. April 1921 spielten die Celtics gegen die New York Whirlwinds vor 11.000 Zuschauern in New York City. Dies war die bis dahin größte Zuschauerzahl bei einem Basketball-Match in den USA.

Im Jahr 1922 konnte der Coach drei weitere Spieler für seine Mannschaft verpflichten: Joe Lapchick, John Beckman und Davey Banks. Joe Lapchick zählte in seiner Basketball-Ära zu den besten Centern der Welt. Der Shooter John Beckman wurde, wie auch Joe Lapchick, Nat Holman und Dutch Dehnert, in die Naismith Memorial Basketball Hall of Fame aufgenommen.

Die Erfolgswelle der Celtics ließ in den nächsten Jahren nicht nach. In der Saison 1922/23 gewannen die Celtics 204 von 215 Spielen und verbesserten ihren alten Rekord. Die Celtics gewannen durchschnittlich 90 Prozent ihrer Spiele und entwickelten sich zu einer der dominantesten Mannschaften des Jahrzehntes. In der Saison 1925/26 gewannen die Celtics 136 von 140 Spielen.

Als bis dahin erste Mannschaften wurden die Original Celtics und das First Team im Jahr 1959 mit der Gründungsklasse in die Naismith Memorial Basketball Hall of Fame aufgenommen. Bisher haben es lediglich zehn weitere Teams in die Hall of Fame geschafft: Die Buffalo Germans, die New York Rens, die Harlem Globetrotters, die Texas Western 1966-Mannschaft, das 1960 United States Olympic Team, das 1992 United States Olympic Team, die All American Red Heads, das 1972-73-74er Immaculata College-Team der Mighty Macs, die Wayland Baptist Flying Queens und die Tennessee A & I State Tigers von 1957 bis 1959.

Die Original Celtics in der ABL
Nach fünf Spieltagen der ABL-Saison 1926/27 ersetzten die Original Celtics die Mannschaft der Brooklyn Arcadians und traten so der American Basketball League (ABL) bei. Dafür nahmen sie den Teamnamen Brooklyn Celtics an. Die Celtics gewannen 32 der verbliebenen 37 Spiele. Im Jahr darauf traten sie unter ihrem alten Namen an und gewannen die Meisterschaft. Im Anschluss an die Saison stimmten die Besitzer der anderen Mannschaften dafür, die Celtics aus der ABL auszuschließen.

Siehe auch
 Liste der Mitglieder der Naismith Memorial Basketball Hall of Fame

Weblinks

 Die Original Celtics in der Naismith Memorial Basketball Hall of Fame Auf: Hoophall-Website; Springfield, MA, 2018. Abgerufen am 19. Januar 2018 (in Englisch).

Literatur
 "Illustrated History of Basketball" (1973), von Larry Fox
 100 Jahre Basketball (2004), Delius Klasing Verlag

Mitglied der Naismith Memorial Basketball Hall of Fame
New York
Sportverein (New York City)
Gegründet 1914
Historische Organisation (New York City)


text:
Kirby Snead (born Kirby Chuck; October 7, 1994) is an American professional baseball pitcher for the Oakland Athletics of Major League Baseball (MLB). He made his MLB debut in 2021 for the Toronto Blue Jays.

Amateur career
Snead attended Santa Fe High School in his hometown of Alachua, Florida. Undrafted out of high school, he then attended the University of Florida, and played three seasons for the Florida Gators. In his freshman season, Snead appeared in 32 games and pitched to a 3–0 win–loss record, 2.40 earned run average (ERA), and 22 strikeouts in 41 innings. As a sophomore, Snead went 1–0 in 28 games played, and posted a 3.15 ERA and 33 strikeouts in 34 innings pitched. In his final season with the Gators, Snead made a NCAA-leading 41 relief appearances, and in 35 innings went 3–1 with a 2.78 ERA and 33 strikeouts.

Professional baseball

Minor leagues
The Toronto Blue Jays selected Snead in the tenth round of the 2016 Major League Baseball draft. He received a $125,000 signing bonus and was assigned to the Advanced-A Dunedin Blue Jays. After a single appearance for Dunedin he was assigned to the Class-A Lansing Lugnuts, where he finished the 2016 season. In 14 total relief appearances, Snead went 0–1 with a 3.62 ERA and 18 strikeouts in 27 innings. He began the 2017 season with Lansing, and was promoted to Dunedin in mid-June. Snead ended 2017 with a 7–2 record, 1.79 ERA, and 56 strikeouts in 42 appearances, and did not yield a home run. In 2018, Snead played with Dunedin and the Double-A New Hampshire Fisher Cats, pitching 51 total innings and posting a 4–5 record, 3.88 ERA, and 51 strikeouts.

Snead continued to progress through the minor league ranks in 2019, appearing for New Hampshire briefly to begin the season before being promoted to the Triple-A Buffalo Bisons. He went 7–2 with a 3.45 ERA and 68 strikeouts in 62 innings between the two teams. The 2020 minor league season was cancelled due the COVID-19 pandemic, and Snead did not play professional baseball at any level that year. He began the 2021 minor league season with the Bisons, and had posted a 2.01 ERA through 31 innings before being called-up.

Toronto Blue Jays
Snead made his Major League debut on July 28, 2021, against the Boston Red Sox retiring the only two hitters he faced. He finished the season with a 2.35 ERA and 7 strikeouts in 7.2 innings.

Oakland Athletics
On March 16, 2022, the Blue Jays traded Snead, Gunnar Hoglund, Kevin Smith, and Zach Logue to the Oakland Athletics for Matt Chapman.

References

External links

1994 births
Living people
Baseball players from Florida
Buffalo Bisons (minor league) players
Dunedin Blue Jays players
Florida Gators baseball players
Lansing Lugnuts players
Major League Baseball pitchers
New Hampshire Fisher Cats players
Oakland Athletics players
People from Alachua, Florida
Toronto Blue Jays players


text:
require 'spec_helper'

describe Answer do
  before do
    @user = Factory(:user)
    @user_1 = Factory(:green)

    @question = Question.new(:user_id=>@user.id, :title=>'question?', :tag_list=>['tag'])
    @question.save

    @answer_1 = Answer.new(:user_id=>@user_1.id, :question_id=>@question.id, :content=>'answer1234')
    @answer_1.save
  end

  describe 'validation' do
    it 'should be created by question' do
      @answer = Answer.new(:user_id=>@user.id, :content=>'answer1234')
      @answer.should_not be_valid

      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should be_valid
    end

    it 'should be created by user' do
      @answer = Answer.new(:question_id=>@question.id, :content=>'answer1234')
      @answer.should_not be_valid

      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should be_valid
    end

    it 'should be created by content' do
      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id)
      @answer.should_not be_valid

      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should be_valid
    end

    it 'content should not be less than 10 words' do
      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer123')
      @answer.should_not be_valid

      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should be_valid
    end

    it 'should only be one answer to the same user with a question' do
      @answer = Answer.new(:user_id=>@answer_1.user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should_not be_valid
    end

    it 'should be answered multiple users with a question' do
      @answer = Answer.new(:user_id=>@user.id, :question_id=>@question.id, :content=>'answer1234')
      @answer.should be_valid
    end
  end

  it 'could be commented' do
    @comment = Comment.new(:user_id=>@user.id, :content=>'comment', :commentable_type=>@answer_1.class.to_s, :commentable_id=>@answer_1.id)
    @comment.should be_valid
  end

  it 'could be voted' do
    @vote = Vote.new(:user_id=>@user.id, :voteable_type=>@answer_1.class.to_s, :voteable_id=>@answer_1.id)
    @vote.should be_valid
  end
end

redpajama_set_name: RedPajamaGithub


A new featurette of Marvel's Captain America: Civil War reveals the real reason behind the epic war between the Avengers. As of now fans know that the impending war between Chris Evan's character and the Iron Man is going to divide the superheroes within the Marvel Cinematic Universe (MCU) but what ignites the war is the centre of the movie's plot.
The movie is set to explore the back story of Steve Rogers, the first Avenger of MCU, and his deep emotional connection with Bucky Barnes aka the Winter Soldier. Evans and executive producer Nate Moore have divulged more details about Bucky's rogue nature and Steve's devotional friendship that will ultimately result in a great disaster.
"I don't think Steve was the kind of guy who grew up as Mr popular. Bucky was the guy who has always been there, who has always had his back. Winter Soldier is his best friend, his only friend in the world. There is an undercurrent plot going on with the Winter Soldier, which sets the maturity of the avengers on its tail," Evans explains in the featurette.
Moore hints that Steve's blind love for his old friend will be the biggest cause of rift among the Avengers. "Caps' allegiance more than anything is to Bucky Barnes, his best friend. And that's the thing that, when we put in the middle of the Avengers, will cause Caps to think twice about keeping the team together."
Captain America: Civil War will premiere in theatres on 6 May 2016, in the US.


These may affect the child's ability to respond to the therapist and activities and carryover skills to new environments.
The amount of time spent on home programming between sessions is also a big factor. If you think about it, a child may see his/her therapist twice a week for 30 minutes at a time. This is a very small percentage of a child's waking hours! Working on home program exercises or assignments daily will increase your child's exposure to the concepts he/she is learning in speech and help your child make progress quicker!
Since the effect of all of these factors can't be predicted through one initial evaluation, offering a fixed amount of time for therapy to work isn't possible. However I tell most parents that we will monitor the child's progress over a period of 3 to 6 months and keep an open dialogue about the need for continued therapy.


Binary Option Trading Explained. After tracking the price movement of EUR/USD for the past hour, a binary option trader believes that the price will rise over the next 5 minutes and decides to invest $100 to purchase a binary call option on EUR/USD expiring in the next 5 minutes.
Free Binary Options Charts The red vertical line indicates when the option expires. Over this timeframe we can see that the overall price trajectory is down, as each move higher is lower than the last, and each move lower reaches a lower price. EUR/USD 15 Minute Chart.
Pro Signal Robot generate binary option trading signals with an up to 94% winning rate & help you to maximize your profit with all binary options platforms. Skip links. Skip to primary navigation; EUR/USD 2. EUR/JPY 3. EUR/GBP 4. USD/CHF 5. USD/JPY 6. GBP/USD 7. NZD/USD 8. AUD/USD.
10/7/2018 · TRADE ANALYSIS. From the video, you can see that the decision to enter the trade was made on EUR/GBP forex (Binary Trade-OTC) chart-weakness. I identified the weakness correctly, and I positioned the trade entry with a capital of USD 3000.


text:
Movie/TV ReviewRed River Theater
·November 10, 2018
Based on the real life of Forrest Tucker, a lifelong criminal who escaped from prison numerous time. The title of the movie comes from a New Yorker profile written by David Grann. The profile and the movie detail Tucker's bank robbery spree at the age of 79 during a time of freedom from incarceration. Robert Redford is charming as Tucker and seems more relaxed in this role than he has in perhaps any previous role. Sissy Spacek is wonderful as his current love interest. There are some great supporting characters as well. The script, however, is a bit strange in that it isn't always clear why the writers chose the particular incidents they did to be part of the movie. There's a particularly odd scene of an interview with Tucker's daughter, played by Elisabeth Moss (who is wonderful as always). There's another odd scene in which Tucker goes to the bank to pay off Spacek's character's mortgage but because the bank won't allow him to do it without her knowledge, he decides not to do it at all. I liked the movie fine because of all the charming characterizations by some wonderful actors. But overall, the movie didn't really stick with me in any way.
At Red River with Pat and Al.
June Movies
May Movies


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Lawson Isles is a subdivision within the city of Davie, Florida. There are currently 3 Homes for Sale within Lawson Isles, with asking prices ranging from $850,000 to $1,300,000. The average list price per square foot of the available inventory in Lawson Isles is $244, which is above the Davie average of $233. The Average Sales price of all sold real estate in this subdivision for the past year is $605,000. Recent comparable homes have been selling for 98.43% of their asking price. Waterfront homes represent 100.00% of the available Lawson Isles inventory. The population of Davie, according to the 2010 Census, is 91,992. The total number of households in the zip code of Lawson Isles is 4,470, with an average household size for of 3.15 people. The total number of renter households within the zip code of 33330 is 310, which means that 93.06% of households in this zipcode are owner occupant households.
4020 S Hiatus Rd, Davie FL 33330 is a Home, with 4 bedrooms which is for sale, it has 36,744 sqft, 235x181 sized lot, and Detached parking. A comparable Home, has 4 bedrooms and 3 baths, it was built in and is located at 4020 S Hiatus Rd Davie FL 33330 and for sale by its owner at $ 850,000. This home is located in the city of Davie , in zip code 33330, this Broward County County Home, it is in the Martha Bright Farms 15-16 Subdivision, and DAVIE WHISPERING PINES are nearby neighborhoods.


Citywest Resort G.C. Results: Sat./Sun. Results 30th/31st. Jan. 2016.
Citywest Resort G.C. in The Daily Mail All-Ireland Foursomes.
Christy Dent/Gary Gregg (8) 41 pts.
Stephen McNevin/Christy Dent (9) 37 pts.
Seamus Grehan/Oliver Lanney (9) 36 pts.
Finian Fox (9) 34 pts.
Class 1. John Keegan (10) 31 pts.
Class 2. Keith Costello (16) 33 pts.
Class 3. John Wade (16) 31 pts.
Stephen Macken on Hole 8. Derek Kelly (2) on Hole 14.


Doubtless you remember and, perhaps, were active in, the grassroots campaign to save the LBA Woods. A last remaining sizeable stand of woodland in southeast Olympia, it is habitat for native plants and myriad birds, both natives and migrants. On this trip, in partnership with Stream Team, expect to hear luxurious spring birdsong and catch glimpses of migrating warblers, wrens, Anna's Hummingbirds, as well as all of our native birds. Our leaders will help you identify and name plants and the birds that are singing, as well as explain the restoration work that volunteers have conducted to date, so that you can get a rich sense of LBA's ecosystem and why it was important to protect it.
Meet at the small LBA lower parking lot at the end of Amherst Street, off Morse-Merriman, at 9:00. Dress for the weather. The paths can be muddy; wear appropriate footwear. Bring binoculars.
Meet at the lower smaller parking lot at the end of Amherst, off Morse-Merriman.


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Cool Boarders 2001
Cool Boarders 2001 Review
For those who think SSX is too simplified and that ESPN X-Games Snowboarding is too difficult, Cool Boarders 2001 is a perfect compromise.
By Shane Satterfield on May 17, 2006 at 3:09PM PDT
Cool Boarders 2001 was released late last year for the PlayStation and was the worst installment of the series yet. Its multitude of gameplay modes was depleted of all fun due to its inexcusably flawed controls that made it impossible to navigate the terrain. Thankfully, Cool Boarders 2001 for the PlayStation 2 is a vast improvement over its PSOne sibling and offers a nice alternative to the competition.
Todd Richards, Michele Taggart, Kevin Jones, and Tara Dakides are just a sample of the 10 professional riders included in Cool Boarders 2001. Idol Minds has also added plenty of its own fantasy riders who would be right at home in SSX. This wealth of riders to unlock somewhat remedies the lack of a create-a-rider option that has been a part of the series for some time. Each rider has two licensed boards to choose from and feature the same graphics, dimensions, and characteristics as the real thing. Billboards from the likes of DC Shoe Co., Burton, Billabong, Vans, and Forum litter the courses and provide an air of authenticity to the game.
Once out on the mountain, it becomes immediately apparent that the jerky, unpredictable controls of the PSOne game have been completely revamped. Gently carving to line up for a big hit is a breeze, and while different from most snowboarding games, the trick system is pretty solid. Performing grabs, flips, and spins are accomplished with a single button press, and the different permutations can produce some interesting results. Strategically placed pipes, downed trees, ski lift cables, and other objects are ripe for grinding. Each rider has several special moves that may be performed by inputting shoulder button sequences while holding the jump button. The controls tend to lag a bit when attempting combos or sequences of tricks, but once the timing is mastered and the moves are memorized, flying down the mountain is a blast.
If you're looking for a variety of events in your snowboarding games, you can't go wrong with the Cool Boarders franchise. Various developers have been piling on the modes for years now--so much so that you begin to wonder just what else could be added. There are nine events included in the career mode, which is broken up into five different categories. The CBX events are reminiscent of SSX in that you must fend off other riders while finding the quickest way down courses with chutes, loops, and death-defying cliff jumps. The downhill events represent the biggest chunk of gameplay. They include a checkpoint mode that asks you to accumulate as many points as possible while racing against a clock and a typical slalom course (against three other riders) called gate run. Boardpark lets you tackle the obstacle-laden stunt courses in search of a high score or hidden sequences. Hidden sequences take a variety of forms, including linked tricks over multiple obstacles and enormous gap jumps. The trickmaster events pit you against one of the game's pros and ask you to duplicate complicated trick combos. The trick events include both the half-pipe and big-air modes.
In each of the five categories, there are frustratingly difficult challenge competitions to complete. Similar to the trickmaster mode, they ask you to mimic one of the real pros as he or she goes through a series of trick combinations on multiple obstacles. The list of combinations can be lengthy, and you're allowed just three failed attempts before you're forced to start again from the beginning. Some of the sequences require precise timing and exacting controls, which correlate into multiple attempts at each challenge. The bot riders often cheat as well--they can miss multiple gates without being disqualified. As you successfully complete each event, your rider receives attribute points that are automatically distributed to four different abilities. After you clear all the courses on the normal difficulty setting, the Vans Triple Crown of Snowboarding opens up, and the serious competition begins. The multiplayer mode for up to two players features all the riders, events, and courses in the game, once they are unlocked in the career mode.
The graphics offered in Cool Boarders 2001 are probably the least impressive of the three snowboarding games released for the system but are by no means ugly. Each character model is built with a healthy number of polygons but not enough to make each character immediately recognizable. The 20 sprawling courses are located on one enormous mountain that is rendered using acclusion culling, so parts of the mountain that are not seen are not hogging up processing power. This lets the game practically eliminate load times. Each course is carefully designed so that none of them look alike. Some feature environmental effects like a setting sun that turns the crests of the drifts a golden color, and others set themselves apart through their unique design--one of the later courses asks you to traverse a bottomless pit using only small, snowy platforms and rails. While some are done better than others, Cool Boarders 2001 includes a wealth of graphical details. Varying depths of snow will cover your rider and board accordingly; real-time shadows skirt around each rider relative to the sun's position; and snow will fall in varying intensities depending upon the course's location on the mountain. Cool Boarders 2001's minor visual discrepancies include clipping that occurs during especially awkward wrecks, snow effects that leave a lot to be desired, and trick animations that could use a few more frames. Thankfully, the game plays with a fluid frame rate even in the split-screen multiplayer mode, and its sense of speed remains intact.
It's no coincidence that the music level is turned down by default. Composed of tracks from a handful of Chemical Brothers, Beastie Boys, and Soundgarden rip-offs, the soundtrack can be grating on the nerves. You can always finish the job and turn it all the way off in the sound options. During the challenge mode, there are voice samples recorded by real pros and riders who give you tips, and competing riders will taunt you when they pass. The sounds of the board edges change as you ride across different snow conditions, and the replication of the low thump that can be heard when someone sticks a trick out on the hill is just right.
Fans of the Cool Boarders series will find a lot to like in this first PlayStation 2 outing. The trick system and controls are deep yet manageable; there is a multitude of events, riders, and courses to unlock; the multiplayer mode is extensive; and the graphics are solid. For those who think SSX is too simplified and that ESPN X-Games Snowboarding is too difficult, Cool Boarders 2001 is a perfect compromise. After a few years spent aloof, the Cool Boarders franchise is finally back on solid footing.
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