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The Blackhawks Have Playoff Potential
Questions abound in the net and on defense. But the Hawks, who open tonight in Ottawa, can score.
It would be an understatement to say there are question marks surrounding the Blackhawks's 2018-2019 season, which opens tonight in Ottawa. The Hawks did, after all, miss the postseason last year for the first time in a decade. And the two years prior, they were knocked out in the first round.
Chief among the Hawks's uncertainties is whether veterans like Patrick Kane can help the organization's young players adapt to the pace and level of talent in the NHL. There are obstacles beyond youth and inexperience with this team, though, that bear examining.
In the net
Two-time Stanley Cup champion Corey Crawford hasn't played in a game since December 23 in New Jersey. Crawford has been dealing with lingering concussion symptoms, but progressed nicely in on-ice workouts during training camp and began practicing with the team on Monday for the first time since February.
"I'm still not 100 percent clear, but it's almost there," Crawford said after the practice. "We're pretty close."
It's likely the goalie could miss the first handful of regular season games, but he'll be traveling with the team for practicing purposes to start the season. The longer their starter is out of the net, the worse off the team will be.
Goalies Cam Ward and Anton Forsberg have yet to prove they can rack up wins on their own, so expect Crawford back sooner rather than later.
Will the blueline be better?
The Hawks defense is as uncertain as their goaltending, if not more. Duncan Keith and Brent Seabrook appear to have had offseason success recovering and training. The alternate captains will do everything in their power to show they've regained the steps they lost last season to today's younger, speedier NHL.
Rookie Henri Jokiharju, who Chicago picked 29th overall in the 2017 NHL Draft, will need all the help he can get. The 19-year-old Finnish defenseman has an arsenal of weapons — scoring prowess, skating ability, puck-handling — but for a player his age, growing pains are inevitable.
Jokiharju suffered a few costly losing battles in the preseason, but finished camp strong. If he continues to pair up with Keith once the games start counting, he'll learn quickly. There's tremendous upside to the young blueliner, it may just be a bit till we see it.
"I think he's good, I think he's smooth," Seabrook said of Jokiharju following Saturday's 4-1 preseason victory over the Blue Jackets. "His skating, it's a lot different than mine. I've got to play a little more positionally. He can get himself in trouble a little bit and make up for it. So it's impressive to see a kid come in at an age like that and play as well as he has in training camp."
Can they score?
Jonathan Toews has looked lighter on his skates in training camp, and if Alex DeBrincat stays on his line, the center will greatly benefit from his play-making.
Patrick Kane also put serious work in this offseason. Along the way, the dynamic winger worked to develop a dangerous chemistry with forward Nick Schmaltz that opponents ought to fear.
Meanwhile, the Hawks need Brandon Saad to rebound offensively — and if he's placed on one of the Hawks' bottom two lines, like he was at times during the preseason, that could prove difficult. In the Hawks' final preseason game Saturday, Saad played on the second line alongside center Nick Schmaltz and opposite Patrick Kane, and he was there again at practice on Monday. If he sticks with Kane and Schmaltz, rebound he will.
Barring a fluke, it'll be the Blackhawks' offense that makes them a playoff contender this season. But it could come down to the wire.
"I think we want to play fast," Seabrook said of this year's group. "We want to play quick and get guys in front of the net. We want to be a hard working team that doesn't give up much defensively and plays both sides of the ice."
Toews is equally optimistic.
"We've had a lot of turnover these last number of years. But I think we got a lot of young guys that have shown a lot in training camp, and guys that are excited to take part [in] a team that's looking to bounce back."
Kane says the team has a lot to prove, which could be easy given their still somewhat foreign underdog status.
"I like it," he said. "It's a young group, but you look around the room [and] a lot of the younger guys have a lot to prove, [and so do] the older guys. You kind of embrace the underdog role."
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text: A chemical spill at the mail distribution center in Coppell has caused a major disruption in the mail headed to East Texas. A mercury spill caused a Hazmat situation there. The incident is mostly affecting letters and magazines and some area residents have not received mail for two days. There is no official word as to when normal mail service will be reestablished.

subset: commoncrawl


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On June 2, NewOak joined with hundreds of local volunteers for this annual day of volunteerism to contribute to a variety of service projects throughout Greater Danbury and New Milford.
This year, projects focused on supporting ALICE® (Asset Limited, Income Constrained, Employed) households in an effort to help close the widening achievement gap in the state between children from low-income households and their more affluent peers. With studies showing that nearly 80% of the achievement gap may be attributable to summer reading loss, volunteers took steps in addressing the issue by supplying summer reading materials to create "Camp Wannaread" bags – i.e., a reading summer camp in a bag – for area second graders.
With education-focused initiatives being top-of-mind with NewOak, the firm was excited to donate popular children books to United Way's summer reading cause. NewOak also completed a second project at Ability Beyond Home for the disabled where the activities included landscaping and grounds maintenance.
NewOak is committed to enriching the communities where we operate. In 2015, NewOak made this commitment official by forming our philanthropic arm NewOak Cares. Through NewOak Cares, we invest time, energy and financial resources to develop area youth and contribute to community improvement projects in Western Connecticut. Locally, we have donated supplies, sponsorships and time to support Hillside Food Outreach, United Way of Danbury, Wolcott Crossroads, Parkinson Foundation, American Heart Association and Toys for Tots.


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Hi everyone, welcome to the new look Keeping Healthy Getting Stylish, I hope you like the new design! I decided that since I was going to be out of normal action for a while due to the wisdom teeth coming out, I would make the most of things and spend some time updating the blog. I felt that the old design was a bit out of date and I wanted it to feel a bit more 'me'. I also wanted to streamline the side bar and links and arrange my sponsors logos better to give more space for bigger images within my blog posts. I would love your feedback on the new design! I have also updated my 'About Me' page, please take a look and let me know what you think :-)
On Friday evening I felt really poorly but yesterday I started to pick up and even decided to make a trip to the gym. I actually felt so much better for some light cardio. I just did an hour on the bike, cross trainer and step machine. I would have loved a run but anything high impact is well and truly off the agenda for now. This morning I decided to suck it up so I could make my new kettle bell class. This is a new class that my gym is offering.
It was 45 minutes made up of a warm up followed by an exercise done repeatedly for 1 minute before moving on to another, then some of the toughest abs moves I've ever done and a cool down. I really enjoyed it! You use the kettle bell in swinging motions or passing it around your body or similar to a dumbbell for tricep kickbacks and rows etc. Its definitely going to become a regular part of my training plan!
I have to admit I'm feeling a bit down in the dumps with my recovery, its a lot worse than I expected it to be. I had worried that working out would make things worse, but I've worked out twice now and felt better for it each time so I'm going to try and keep to my usual training plan but just dial it down a bit and ditch the high impact running for a while. I also can't see me being able to eat solid foods for at least a few more days yet :-( The pain seems to come and go but its the swelling that's really getting to me. Oh well, I might as well make the most of it and keep on enjoying making delicious smoothies – as they say 'when life gives you lemons, make lemonade'! I hope I'm not boring you with these smoothie combos, but here's a few I've had over the weekend:
Spinach, frozen banana, frozen mango, midori greens, plain hemp powder, manuka honey, bee pollen, water topped with more bee pollen and honey
Based on the 'Bunny Spice' recipe from Raw Food Real World: home made almond milk, carrot and ginger juice, cinnamon, maple syrup – so delicious!
Greens, half a large pineapple, Thai young coconut, bee pollen, honey, water, ice – very tropical!
Based on the 'Black Sesame Jewel' recipe in Ani's Raw Food Kitchen: frozen banana, 2 tbsp of tahini, 2 tbsp of black sesame seeds, 1 tbsp of manuka honey, water topped with more sesame seeds, bee pollen and honey. This was utterly delicious, so creamy and full of good stuff!
Cabbage, courgettes, 2 scoops of vanilla sun warrior, maca powder, midori greens, ground flax, water, ice, maple sugar – this was way better than it looked, very creamy!
Kale, frozen cherries, cacao nibs, cacao powder – topped with coconut – so sweet with no need for additional sweeteners
Greens, apple, pineapple, nectarine, dates, coconut water, water, ice – tasted lovely and cleansing
I've also added in some different things including some protein pudding:
Just 1 scoop of chocolate sun warrior protein powder mixed with water, a teaspoon of cashew butter and some agave syrup.
This morning for breakfast I defrosted some frozen raspberries and mashed them with some Viva Pure cacao chai chia pudding and agave syrup:
This was perfect, just the right consistency, I'll be eating more mashed raspberries me thinks!
I've also been having juices, just my fave green juice blend and beet, carrot, cucumber, ginger blend. I've started to try and eat some of the smoothies with a spoon out of a bowl as I read that sucking through a straw may not be best when your gums are healing. I have to say its a bit strange, on one hand I feel awesome from all the goodness I'm getting from the juice and smoothies, then I feel crap when I feel how swollen and sore I am!
Today I've made a big pan of my Lentil and Spinach Soup and blended it down so I'm going to have that for dinner plus I can't wait to make a shake with my remaining Thai young coconut and raw cacao!
I also wanted to mention that I have a new Goodness Direct discount code for you – so even if you've used my old code, you can use this one. It gives you £10 off and free delivery on all orders over £35 – just use code 322345GH at the check out!
So what do you think of the new blog design and my 'About Me' page? I really hope you like it and would love any feedback! Hope your all having a brilliant weekend!


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Plane - Multi blue
Fly high up into the skies and rush down through the tree tops. bObles Plane is full of flying speed and fun.Lie on the stomach and fly, rock from side to side or stand on the tilting wings – can you stay in the air? There is hours of play in bObles Plane for young and old, who can balance, jump, walk and lie, while challenging their motor skills.
bObles tumbling products are made of firm EVA foam without any dangerous phthalates. The material is soft, easy to clean and does not leave any marks in the floor. Tested for children.
Size: h 25, l 47, b 24 cm
All our products are finished by hand and none of them are exactly alike. The size may vary a bit.


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We have a wide range of interviews recommending books on the history, politics and culture of Russia.
Andrei Maylunas chooses his best books on pre-Revolutionary Russia. Roland Chambers chooses his best books on the Russian Revolution and Thomas Keneally chooses his best books on Revolutionary Russia. Both recommend A People's Tragedy: The Russian Revolution by Orlando Figes Francis Spufford chooses his best books on the country in the 20th -century and Robert Conquest chooses his best books on Communism. Stephen Lucas chooses his best books on Soviet Law and Lyubov Vinogradova chooses her best books from the KGB Archives. Robert Service chooses his best books on totalitarian Russia. Anna Reid chooses her best books on the Siege of Leningrad.
Turning to the more recent history and politics of the country, Simon Pirani chooses his best books on Putin's Russia and Martin Sixsmith looks at why Russia isn't a democracy. Thomas de Waal looks at conflict in the Caucasus. A number of other interviews also deal with Russia's relationship with its periphery. Vanora Bennett chooses her best books on Chechnya and the poet, Nigan Hasan-Zadeh, chooses her best books on Azerbaijan.
On more cultural and literary themes, Michael Nicholson chooses his best books on Solzhenitsyn and Maxim D Shrayer his best books on Vladimir Nabokov and on Vasily Grossman. Rosamund Bartlett chooses the best Russian short stories.
The best books on The Russian Revolution, recommended by Roland Chambers
The Russian revolution was the beginning of the modern age, says award-winning author Roland Chambers. He tells us what Solzhenitsyn imagined Lenin was like, and about the children's author who led a double life as a spy in Bolshevik Russia.
Russia at War
by Alexander Werth
A Writer At War: Vasily Grossman with the Red Army 1941-1945
by Vasily Grossman, translated by Antony Beevor and Luba Vinogradova
Reflections on the Russian Soul
by Dmitry Likhachov
Less Than One
by Joseph Brodsky
Conversations with Stalin
by Milovan Djilas
The best books on The Siege of Leningrad, recommended by Anna Reid
The Best Books About Aleksandr Solzhenitsyn, recommended by Michael Nicholson
Russian literature specialist Michael Nicholson, Emeritus Fellow at University College, Oxford, talks us through the best books to learn more about the great Soviet dissident and winner of the 1970 Nobel Prize in Literature, Aleksandr Solzhenitsyn.
The Best Tales of Soviet Russia, recommended by Robert Chandler
Robert Chandler, one of the best known translators of Russian literature, recommends some of his favourite tales of Soviet Russia. There's the one about a dog in space and the one about the Soviet café which stocked nothing but champagne and Mars bars…
The best books on 20th Century Russia, recommended by Francis Spufford
Reading about Russia's 20th century is like finding another vision of how the world might have been. Francis Spufford, author of Red Plenty, recommends books that tell the story of Russia in the last century — from Soviet science fiction set in capitalist wastelands to Khrushchev as raconteur.
The best books on Totalitarian Russia, recommended by Robert Service
Robert Service, Professor of Russian Studies at Oxford, when forced to choose between Lenin, Trotsky, and Stalin, says Stalin was definitely the worst of the lot. He takes a look at the dynamics of totalitarian Russia, gleaning insights from Thucydides to Orwell.
The best books on Revolutionary Russia, recommended by Thomas Keneally
Best-selling author Thomas Keneally explains that the Cold War biographies couldn't afford to say that Stalin was attractive, or that Lenin was magnetic, but they were, because otherwise people wouldn't have followed them. He picks some great introductions to Revolutionary Russia.


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Circus Party
Take our word for it: there are few things in this world quite as fun as a circus themed party. This is a colourful, lively theme for boys, girls, babies and grown-ups, meaning that it works equally as well at a 1st birthday as it does at a wedding. Circus themed parties even come ready made with their own options for catering and activities that are easy and timelessly entertaining for guests.
We've got harlequin paper plates, paper hot dog trays, retro popcorn tubs, striped paper straws, party hats and more. Mix and match from our large range of circus themed party supplies to create all the fun with none of the hassle!
Items 1-50 of 86
- Yellow Paper Straws Mix£3.49
- 12 Paper Popcorn Bags£1.99
- 12 Retro French Fry Bags£1.99
- 10 Red Dot Paper Straws£0.99
- Red Diamond Paper Straws£2.49
- Red Plastic Tablecloth£1.99
- 5 Large Yellow Latex Balloons 17"Now Only £1.99 Regular Price £2.99
- Giant Red Balloon 2 ft£2.49
- Red Honeycomb Ball 20 cm£1.99
- Giant Blue Balloon 2 ft£2.49
- Circus Party Invitations£4.50
- Pink Carnival Stripes Paper BuntingNow Only £1.99 Regular Price £3.99
Items 1-50 of 86


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mlw50 replied to mlw50's topic in Evernote IntegrationsThanks for the reply. I have seen a few Evernote to *** but hardly anything that will take *** to Evernote. It may turn out that I'll just have to run the two side by side. I'll keep looking.
Importing into Evernote
mlw50 posted a topic in Evernote IntegrationsI use a product called Web Research Professional (http://www.macropool.com/en/products/webresearch/professional/) which allows me to cpature web pages, much like Evernote does. I've been using it for quite some time and I am looking for a way to import this data into Evernote. Web Research allows me to export as an MHT, in the original HTML or as a CHM file. The problem is that when I try to import an HTML, Evernote will only import a URL. Does anyone know of another way to import? Thanks...


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Leptospira is the etiologic agent of leptospirosis, a bacterial zoonosis distributed worldwide. Leptospiral lipopolysaccharide is a protective immunogen, but the extensive serological diversity of leptospires has inspired a search for conserved outer membrane proteins (OMPs) that may stimulate heterologous immunity. Previously, a global analysis of leptospiral OMPs (P. A. Cullen, S. J. Cordwell, D. M. Bulach, D. A. Haake, and B. Adler, Infect. Immun. 70:2311-2318, 2002) identified pL21, a novel 21-kDa protein that is the second most abundant constituent of the Leptospira interrogans serovar Lai outer membrane proteome. In this study, we identified the gene encoding pL21 and found it to encode a putative lipoprotein; accordingly, the protein was renamed LipL21. Southern hybridization analysis revealed the presence of lipL21 in all of the pathogenic species but in none of the saprophytic species examined. Alignment of the LipL21 sequence from six strains of Leptospira revealed 96 to 100% identity. When specific polyclonal antisera to recombinant LipL21 were used, LipL21 was isolated together with other known leptospiral OMPs by both Triton X-114 extraction and sucrose density gradient membrane fractionation. All nine strains of pathogenic leptospires investigated by Western blotting, whether culture attenuated or virulent, were found to express LipL21. In contrast, the expression of LipL21 or an antigenically related protein could not be detected in nonpathogenic L. biflexa. Infected hamster sera and two of eight human leptospirosis sera tested were found to react with recombinant LipL21. Native LipL21 was found to incorporate tritiated palmitic acid, consistent with the prediction of a lipoprotein signal peptidase cleavage site. Biotinylation of the leptospiral surface resulted in selective labeling of LipL21 and the previously known OMPs LipL32 and LipL41. These findings show that LipL21 is a surface-exposed, abundant outer membrane lipoprotein that is expressed during infection and conserved among pathogenic Leptospira species.
Leptospira is a genus of spirochetal bacteria and the causative agent of leptospirosis, a zoonotic disease with a global distribution. An international survey of human leptospirosis deduced that approximately 100,000 severe cases (requiring hospitalization) occur annually (42). This figure is likely to be a significant underestimate due to the lack of diagnostic and epidemiologic resources in many areas of endemicity.
The distribution of the approximately 230 recognized serovars of pathogenic leptospires may be restricted geographically (13). The extensive serovar diversity has been attributed to differences in the structure and composition of lipopolysaccharide (LPS) (6). Much work has focused on the role of leptospiral LPS in immunity (14, 29), and the genetics of LPS biosynthesis have been partly elucidated (6, 7, 10, 11). Preparations of leptospiral LPS can elicit protective immunity, but this immunity is generally serovar specific (13). Consequently, the focus of research on protective antigens has shifted toward conserved outer membrane proteins (OMPs), which may be able to stimulate heterologous immunity.
Three classes of leptospiral OMPs have been identified. The most abundant class comprises the outer membrane lipoproteins and includes the major OMP and immunodominant protein antigen LipL32 (15, 18), the in vivo-down-regulated protein LipL36 (4, 19), LipL48 (20), and the surface-exposed protein LipL41 (39). Consistent with the theory of paucity of transmembrane proteins postulated for Treponema pallidum (34) and despite much investigation, only one rare transmembrane OMP has been identified. This protein has been designated OmpL1 and is thought to function as a heat-modifiable porin (17, 38). The third class, comprising the peripheral membrane proteins, contains one member, P31LipL45, which exploits the lipoprotein secretion pathway to target itself to both the inner and the outer membranes (32).
Most of the genes encoding leptospiral OMPs have been identified in independent studies by probing of gene libraries with degenerate oligonucleotide probes or hyperimmunized rabbit sera. Recently, a more systematic approach was used to identify novel leptospiral OMPs (8). In that study, outer membrane preparations from leptospires grown under a variety of environmental conditions were separated by two-dimensional gel electrophoresis and the resolved proteins were analyzed by mass spectrometry. However, the usefulness of this approach is limited by the availability of a genome sequence against which to compare the mass spectrometry data. Currently, there are at least four leptospiral genome sequencing projects in the final stages of sequence annotation or publication. Analysis of the ensuing genome data will provide a subset of putative membrane proteins from which a comprehensive set of OMPs may be determined experimentally.
Immunoprotection studies with recombinant antigens in animal models of leptospirosis have met with only partial success (5, 21). Thus, the identification of additional OMPs, particularly those that are surface exposed, is of prime importance for the development of recombinant subunit vaccines that will elicit heterologous immunity.
In this study, the gene encoding the second most abundant protein in the Leptospira interrogans serovar Lai outer membrane proteome was identified and designated lipL21. The LipL21 sequence is well conserved among pathogenic leptospires but could not be detected in saprophytic strains. LipL21 is surface exposed and immunogenic, being recognized by immune sera from humans and hamsters infected with Leptospira.
Bacterial strains and media.The following strains of Leptospira were grown at 30°C in EMJH (26) or bovine serum albumin-Tween 80 medium (Bovuminar PLM-5 microbiological medium; Intergen): L. biflexa serovar Patoc strain Patoc I, L. borgpetersenii serovar Hardjo strains HB-15B/93U and L550, L. inadai serovar Lyme strain 10, L. interrogans serovar Bratislava strain AS-05, L. interrogans serovar Canicola strain CA29, L. interrogans serovar Copenhageni Li-133, L. interrogans serovar Lai strain Lai, L. interrogans serovar Pomona strains PO-01 and RZ11, L. kirschneri serovar Grippotyphosa strain RM52, L. kirschneri serovar Mozdok strain 5621, L. noguchii serovar Fort Bragg strain Fort Bragg, L. noguchii serovar Proechymis strain LT796, L. santarosai serovar Bakeri strain LT79, L. wolbachii serovar Biflexa strain Codice, and L. weilii serovar Celledoni strain Celledoni. Escherichia coli DH5α was used for cloning and was cultured in Luria-Bertani (LB) broth or on 1.5% LB agar at 37°C overnight. E. coli BL21-SI (Invitrogen) was used for protein expression and was cultured at 37°C in LB medium without NaCl but containing the appropriate antibiotics.
Oligonucleotides.Oligonucleotides used in this study were synthesized by GIBCO BRL and are listed in Table 1.
DNA manipulations.Chromosomal DNA from Leptospira was prepared by using the cetyltrimethylammonium bromide precipitation method (3), while plasmid DNA was isolated by using a QIAprep spin miniprep kit (Qiagen). The lipL21 sequence was amplified by high-fidelity PCR for double-stranded DNA sequencing with an Expand high-fidelity PCR kit (Roche). DNA sequencing was performed at the Yale/Keck Core DNA Sequencing Facility. Standard methods for molecular biology were performed essentially as described by Sambrook et al. (36).
DNA and protein sequence analyses.Sequence data were analyzed by using Sequencher 3.1 (GeneCodes), while DNA and protein database comparisons were made by using the BLAST programs of Altschul et al. (1, 2). Multiple sequence alignments and hydrophophilicity analysis were performed by using the Lasergene (DNAstar) software package.
Southern hybridization.After electrophoresis, EcoRI-digested genomic DNA was transferred to a positively charged nylon membrane (Roche) by capillary transfer in 20× SSC (1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate) (pH 7.0). Overnight hybridization and subsequent washings were performed under high-stringency conditions at 68°C as recommended in the Roche digoxigenin labeling and detection kit instructions. The lipL21 probe was labeled by PCR amplification in the presence of digoxigenin DNA labeling mix (Roche) as specified by the manufacturer.
Gel electrophoresis and immunoblotting.Samples were solubilized in a final sample buffer consisting of 50 mM Tris-HCl (pH 6.8), 100 mM dithiothreitol, 2% sodium dodecyl sulfate (SDS), and 0.1% bromophenol blue in 20% glycerol. Proteins were separated in 12% PAGEr Gold precast Tris-glycine gels (BioWhittaker Molecular Applications) or SDS-12 or 15% polyacrylamide gels by using a discontinuous buffer system essentially as described previously (30). After electrophoresis, each gel was stained with 0.2% Coomassie brilliant blue R-250 (in 10% acetic acid-45% methanol) or transferred electrophoretically to a polyvinylidene difluoride membrane (Millipore). The membranes were incubated with rabbit (diluted 1/5,000 to 1/20,000), hamster (diluted 1/2,000), or human (diluted 1/1,000) serum in blocking solution (5% skim milk powder and 0.05% Tween 20 in phosphate-buffered saline [PBS]); washed three times with PBS containing 0.1% Tween 20 (PBS-T); for 30 min; incubated with donkey anti-rabbit (Amersham), goat anti-hamster (Jackson ImmunoResearch Laboratories), or sheep anti-human (Silenus) immunoglobulin-horseradish peroxidase conjugates at a dilution of 1/5,000 in blocking solution for 30 min; and again washed three times with PBS-T. The membranes were developed with an enhanced chemiluminescence Western blot detection system (Amersham), and bands were visualized with Hyperfilm (Amersham).
Recombinant protein expression and purification.PCR was used to amplify lipL21 from L. kirschneri serovar Grippotyphosa strain RM52 with primers designed to engineer attB sites into the final product (Table 1). BP clonase (Invitrogen) was used to mediate λ site-specific recombination between the attB-flanked PCR product and the attL-containing plasmid pDONR201 (Invitrogen). lipL21 was then transferred from the entry vector pDONR201 to the protein expression vector pDEST17 by using LR clonase (Invitrogen). Plasmid DNA was isolated from the lipL21 pDEST17 clone and used to transform the expression strain E. coli BL21-SI. Cultures of the expression strain were grown to an absorbance at 600 nm of 0.6 and induced for 3 h with 0.3 M NaCl. The bacteria were lysed by sonication, and inclusion bodies containing the fusion protein were solubilized in 8 M urea-10 mM Tris-HCl-100 mM NaH2PO4 (pH 8.0). An N-terminal hexahistidine-tagged fusion protein was subsequently purified by using an Ni2+-nitrilotriacetic acid affinity column according to the manufacturer's instructions (Qiagen). LipL32 and LipL36 were prepared as described previously (18, 19).
Production of polyclonal antisera.Purified hexahistidine-tagged LipL21 was loaded onto a preparative SDS-12% polyacrylamide gel and allowed to migrate into the separating gel during electrophoresis. A band containing 1.5 mg of fusion protein was excised from the gel, divided into six equal pieces, and desiccated. Dessiccated gel pieces corresponding to 250 μg of recombinant protein were ground to a powder, dissolved in 1 ml of water, mixed with 1 ml of complete Freund's adjuvant (Sigma), and inoculated subcutaneously and intramuscularly into New Zealand White rabbits (Harlan Sprague-Dawley) that were free of leptospiral antibodies. Additional immunizations with similar amounts of fusion protein in powdered acrylamide gel mixed with incomplete Freund's adjuvant (Sigma) were administered 2 and 4 weeks after the primary immunization. Serum was obtained from the rabbits 5 weeks after the primary immunization (23). Antisera to LipL31 (20), LipL36 (19), LipL41 (39), and ImpL63 (20) were prepared as described previously.
Hamster infection.Sera were collected from hamsters infected with culture-derived or host-derived L. kirschneri serovar Grippotyphosa strain RM52 as described previously (4). Briefly, 5-week-old golden Syrian Hamsters (Harlan Sprague-Dawley) were inoculated intraperitoneally with 105 or 106L. kirschneri organisms from a liquid culture (culture derived) or with leptospires recovered from the liver tissue of moribund weanling hamsters (host derived).
MAT.The microscopic agglutination test (MAT) (12) was perfomed by using viable serovars Australis, Canicola, Copenhageni, Grippotyphosa, Hardjo, Pomona, Tarrasovi, and Zanoni as antigens and sera at final dilutions (including antigens) of 1/50 to 1/32,000.
Triton X-114 extraction. L. kirschneri serovar Grippotyphosa strain RM52 outer membrane material was extracted by using a previously described method (18). Briefly, leptospires were washed in PBS-5 mM MgCl2 and then extracted in the presence of 0.5% protein-grade Triton X-114 (Calbiochem)-150 mM NaCl-10 mM Tris (pH 8)-1 mM EDTA at 4°C. The insoluble material was removed by centrifugation at 17,000 × g for 10 min. After centrifugation, 20 mM CaCl2 was added to the supernatant. Phase separation was performed by warming the supernatant to 37°C and subjecting it to centrifugation for 10 min at 1,000 × g. The detergent and aqueous phases were then separated and precipitated with acetone.
Alkaline plasmolysis and membrane fractionation.Membrane vesicles were prepared as described previously (20). Briefly, leptospires were washed, resuspended in 20 mM Tris (pH 9)-1 M NaCl-27% (wt/vol) sucrose-2 mM EDTA buffer, and subjected to high-speed vortexing with a microstir bar. After treatment with DNase and RNase and removal of the insoluble material by centrifugation, the supernatent was loaded onto a 27 to 55% (wt/vol) sucrose gradient. Appropriate fractions were then collected for subsequent analyses.
Radioimmunoprecipitation and fluorography.A total of 200 μCi of [3H]palmitic acid (Amersham) was added to 25 ml of L. kirschneri serovar Grippotyphosa strain RM52 culture at a cell density of 107 cells/ml. When a cell density of 109 cells/ml was obtained, the cells were harvested by centrifugation. A total of 1010 cells were resuspended in boiling buffer (2% SDS, 50 mM Tris-Cl [pH 8.0]), boiled for 10 min, and lysed by sonication. Insoluble material was removed by centrifugation at 12,000 × g. Native LipL21 was immunoprecipitated from this mixture by the addition of 0.2 ml of LipL21 antiserum and 0.2 ml of protein A slurry, incubation at 4°C for 30 min, and centrifugation at 12,000 × g. The pellet was washed three times in radioimmunoprecipitation buffer (1% Nonidet P-40, 0.5% deoxycholic acid, 0.1% SDS, 50 mM Tris-Cl [pH 8.0]) before being resuspended in final sample buffer for SDS-polyacrylamide gel electrophoresis (PAGE). The sample was separated by SDS-PAGE, and the gel was treated with Amplify solution (Amersham). An image was obtained by incubating the dried gel in a film cassette with preflashed Hyperfilm-MP (Amersham) at −70°C for 1 week.
Surface biotinylation. L. kirschneri serovar Grippotyphosa strain RM52 was grown to mid-log phase, washed twice in PBS (pH 8.0), and resuspended in PBS containing 0.5 mg of sulfosuccinimidyl-6-(bitinamido)hexanoate (Sulfo-NHS-LC-Biotin) (Pierce)/ml. The labeling reaction was allowed to proceed for 1 min before the residual Sulfo-NHS-LC-Biotin was quenched by the addition of Tris (pH 8.0) to a final concentration of 50 mM. Inactivated Sulfo-NHS-LC-Biotin was removed by two washes with PBS (pH 8.0). LipL21 was immunoprecipitated from a lysate of 109 surface biotinylated cells as described above. Lysates of 109 surface biotinylated cells or immunoprecipitated LipL21 were resolved by SDS-PAGE and transferred electrophoretically to a polyvinylidene difluoride membrane. The membrane was blocked overnight in 5% (wt/vol) skim milk buffer, washed twice in PBS-T, and incubated with streptavidin-horseradish peroxidase conjugate (Amersham) at a dilution of 1/20,000 in PBS-T for 30 min. The membrane was washed six times with PBS-T and developed with the enhanced chemiluminescence Western blot detection system before visualization with Hyperfilm (Amersham).
Nucleotide sequence accession numbers.The nucleotide sequences of lipL21 from L. interrogans serovar Lai, L. interrogans serovar Pomona, L. interrogans serovar Bratislava, L. kirschneri serovar Grippotyphosa, L. kirschneri serovar Mozdok, and L. borgpetersenii serovar Hardjo have been deposited in GenBank under the accession numbers AY187271 , AY187273 , AY187269 , AY187270 , AY187272 , and AY187274 , respectively.
Identification and distribution of lipL21.In a previous study, Cullen et al. obtained the peptide sequence from pL21, the second most abundant constituent of the L. interrogans serovar Lai outer membrane, by electrospray-ionization tandem mass spectrometry (8). This peptide sequence was used to search the in-progress genome sequence of L. borgpetersenii serovar Hardjo. A match to a translated open reading frame of 564 bp, which encoded a protein of 188 amino acids with a predicted molecular mass of 19.8 kDa, was identified (GenBank accession number AY187274 ). At the time of the initial identification (8), no proteins with significant similarity were identified in the public databases. Subsequently, a homologous sequence was identified in the L. interrogans serovar Lai genome sequence (GenBank accession number NP_710192 ). The pL21 amino acid sequence was found to contain a 17-amino-acid signal peptide and a putative lipoprotein signal peptidase (LSP) cleavage site (F−3A−2A−1C+1) (16, 41). In accordance with standard nomenclature for leptospiral lipoproteins, the protein was renamed LipL21.
A 400-bp lipL21 probe was synthesized and used to detect the presence of similar DNA sequences in a panel of pathogenic and environmental leptospires by high-stringency Southern hybridization (data not shown). A single copy of the lipL21 gene was present in all five pathogenic Leptospira species analyzed (L. interrogans serovar Lai, L. interrogans serovar Pomona, L. kirschneri serovar Grippotyphosa, L. kirschneri serovar Mozdok, L. borgpetersenii serovar Hardjo, L. santarosai serovar Bakeri, and L. noguchii serovar Proechymis). However, no hybridizing bands were detected when genomic DNAs from the environmental leptospires L. biflexa serovar Patoc and L. wolbachii serovar Biflexa were probed. Oligonucleotide primers flanking the sequence were designed to amplify the lipL21 open reading frame from five other strains of pathogenic Leptospira for DNA sequencing. The translated lipL21 sequences from L. interrogans serovar Lai, L. interrogans serovar Pomona, L. interrogans serovar Bratislava, L. kirschneri serovar Grippotyphosa, L. kirschneri serovar Mozdok, and L. borgpetersenii serovar Hardjo were found to share 96 to 100% identity (GenBank accession numbers AY187271 , AY187273 , AY187269 , AY187270 , AY187272 , and AY187274 , respectively).
The sequence encoding the mature protein (without the hydrophobic signal peptide) was cloned into the protein expression vector pDEST17. Introduction of the construct into BL21-SI cells resulted in high-level expression of the N-terminal hexahistidine-tagged fusion protein. Recombinant LipL21 was purified by immobilized metal affinity chromatography and used to raise polyclonal antiserum in New Zealand White rabbits. To assess the level and distribution of LipL21 expression, immunoblot analysis was performed on a panel of Leptospira species with LipL21 antiserum (Fig. 1), which reacted with a single band of 21 kDa. Three virulent low-passage isolates of Leptospira were included in the panel (Fig. 1, lanes 1 to 3) to assess whether there were variations in the levels of expression between virulent and culture-attenuated organisms. LipL21 could be detected in all seven species of pathogenic leptospires examined, although the levels of expression in serovars Bakeri and Fort Bragg were slightly reduced (Fig. 1, lanes 6 and 9, respectively). No significant difference in the levels of LipL21 expression was observed between virulent and culture-attenuated leptospires. No LipL21 was detected in nonpathogenic L. biflexa serovar Patoc.
Cellular localization of LipL21.Two independent methods of cellular fractionation were used to confirm the cellular location of LipL21. Due to the association of peptidoglycan with the cytoplasmic membrane rather than the outer membrane in spirochetes (25, 28), the outer membrane can be solubilized with low concentrations of Triton X-114 (0.1 to 1%) without disruption of the cytoplasmic membrane. Subsequently, the contaminating hydrophilic periplasmic proteins can be removed by temperature-induced phase separation with TX-114. Recently, an alternative method for isolating leptospiral outer membranes was developed (20). This method involves the generation of membrane vesicles by alkaline plasmolysis and their subsequent separation by sucrose density gradient ultracentrifugation. Western blotting of these membrane vesicle and TX-114 fractions with antisera to several proteins with known cellular locations as well as LipL21 was performed. Antisera to the inner membrane proteins LipL31 and ImpL63 were included to show that the outer membrane fractions were free from contamination with inner membrane material. Antisera to two previously characterized leptospiral OMPs that differ in their properties during TX-114 extraction were included for comparison (Fig. 2). Both the TX-114 detergent phase and the outer membrane vesicle fractions contained the OMPs LipL36 and LipL41 and the novel protein LipL21. Notably, the inner membrane markers LipL31 and ImpL63 were absent from the TX-114 detergent phase and the outer membrane vesicle fractions. Unlike that of LipL36, solubilization of LipL21 by TX-114 was incomplete, as some material remained in the protoplasmic cylinder fraction, a behavior also exhibited by the leptospiral OMPs LipL41 (39) and OmpL1 (17).
Immunological reactivity of LipL21 with hamster and human sera.To determine whether LipL21 is expressed during infection and is immunogenic, sera were collected from four hamsters inoculated with leptospires from infected hamster liver, three hamsters challenged with virulent L. kirschneri RM52, and one uninfected hamster. In Western blotting experiments, all seven infected hamster sera recognized recombinant LipL21, while the uninfected control sera showed no reaction. Controls included recombinant forms of immunodominant protein antigen LipL32 and in vivo-down-regulated OMP LipL36 (Fig. 3a). Hamster sera recognized recombinant LipL21 and LipL32 but not LipL36. When the blotting membrane was stripped of antibody and reprobed with pentahistidine antibody, all of the recombinant proteins could be detected. Eight MAT-positive human sera were selected randomly from a bank of sera from Australian leptospirosis patients. In Western blotting experiments with this panel of sera, two of the eight sera recognized recombinant LipL21 (MAT titers: Copenhageni, 1,600; Pomona, 800; and Hardjo, 200) (Fig. 3b). No reactivity with LipL21 was observed when sera from eight nonleptospirosis patients were similarly examined (data not shown).
Lipid modification of LipL21.Analysis of the amino acid sequence of the protein encoded by lipL21 revealed a putative LSP recognition sequence. Leptospires grown in the presence of [3H]palmitic acid and separated by SDS-PAGE were shown to selectively incorporate the radiolabeled fatty acid into proteins of 21, 24, 32, 41, 45, 60, 62, 86, and 96 kDa (Fig. 4). The four most prominent bands appeared at 21, 25, 32, and 41 kDa, excluding the diffuse material below 14 kDa that may represent incorporation of the labeled fatty acid into LPS. When LipL21 was immunoprecipitated from leptospires grown in the presence of [3H]palmitic acid and separated by SDS-PAGE, it was shown to incorporate the labeled fatty acid and to resolve at a mass similar to that of the prominent 21-kDa band from the whole-cell material.
Surface exposure of LipL21.To determine whether LipL21 was exposed on the leptospiral cell surface, intact cells were surface labeled in the absence of organic solvents with the water-soluble reagent Sulfo-NHS-LC-Biotin. After labeling, the residual nonreacting biotinylation reagent was quenched with Tris and removed by several washes before the cells were lysed. When the samples were separated by SDS-PAGE, protein bands of 17, 21, 24, 30, 32, 36, 41, 48, 55, 60, 62, and 70 kDa were selectively biotinylated (Fig. 5). The four most prominently labeled proteins resolved at 21, 24, 32, and 41 kDa; the last two corresponded in mass to the previously characterized OMPs LipL32 and LipL41. When LipL21 was immunoprecipitated with a specific antiserum from a lysate of surface-labeled L. kirschneri and separated by SDS-PAGE, a single biotinylated band of 21 kDa was obtained. This band resolved at a mass similar to that of the prominently labeled 21-kDa band from the whole-cell material.
In this study, we have characterized LipL21, the second most abundant protein in the L. interrogans serovar Lai outer membrane proteome. Previous studies showed no variation in the expression of this protein when leptospires were cultured at 20, 30, or 37°C or at 37°C in the presence of serum or in iron-depleted medium (8). Preliminary data from our laboratories suggested the presence of variations in the relative abundances of leptospiral OMPs between strains (unpublished data). Western blotting with LipL21 polyclonal antisera revealed similar levels of expression of LipL21 among a selection of pathogenic leptospires, with the exception of serovars Bakeri and Fort Bragg, which exhibited slightly lower expression levels. Pathogenic leptospires lose virulence after an undefined number of passages in laboratory culture media and undergo quantitative and qualitative changes in their OMP profiles (22). However, the expression of LipL21 did not appear to vary between the virulent and culture-attenuated strains included in the Western blotting experiments. The uniform expression of LipL21 may indicate that this protein endows the pathogenic members of the genus Leptospira with a selective advantage both in the mammalian host and in laboratory cultures.
Southern hybridization and Western blotting experiments did not indicate the presence of the lipL21 gene or a protein antigenically similar to LipL21 in the nonpathogenic leptospires. An alignment of the LipL21 sequences from six strains (encompassing three species) of pathogenic Leptospira revealed 96 to 100% identity, indicating that LipL21 is a new member of the small but growing family of conserved leptospiral OMPs that are unique to the pathogenic strains and that have no significant similarity with proteins of other organisms (including the other pathogenic spirochetes). Other conserved leptospiral OMPs include the porin OmpL1 and the lipoproteins LipL32 and LipL41. Given that these proteins are unique to the pathogenic leptospires, it is likely that they play a specific role in leptospiral pathogenesis. Although it is interesting to speculate about the functions of these proteins, without the technology to generate isogenic mutants of pathogenic Leptospira species, definitive studies cannot be performed.
A study investigating the expression of leptospiral antigens during infection identified an antigen that had a molecular mass similar to that of LipL21 and that was designated p22 (4). p22 was observed to react with sera from hamsters challenged with culture-derived leptospires and not with sera from hamsters challenged with host-derived leptospires. In a later study investigating the humoral immune response to leptospirosis, p22 was found to react with pooled human sera from Brazilian leptospirosis patients (15). In our study, sera from hamsters challenged with both host- and culture-derived leptospires were found to react with recombinant LipL21. However, p22 was observed to react only with sera from hamsters challenged with culture-derived leptospires, raising the question of whether p22 and LipL21 are the same. The observation that p22 and LipL21 resolve at approximately the same isoelectric point and molecular mass during two-dimensional electrophoresis suggests that they are the same and that the experimental discordance is due to variations in antibody responses between hamsters. When a random selection of serum samples from MAT-confirmed leptospirosis patients was used in Western blotting experiments, serum samples from two of eight patients recognized recombinant LipL21. The variable patterns of LipL21 recognition observed in humans may reflect differences in the time of collection of sera after the initial infection, the severity of the infection, the genetic heterogeneity of the human host, or even variable masking of LipL21 by other leptospiral surface components. The recognition of recombinant LipL21 by serum samples from leptospirosis patients and hamsters challenged with host-derived organisms suggests that LipL21 is expressed during infections of both hamsters and humans.
In eubacteria, lipoproteins are synthesized with an N-terminal hydrophobic signal peptide that is cleaved from the mature polypeptide by LSP prior to covalent linkage of fatty acids (usually palmitate) (24). The putative LSP recognition site of LipL21 (I-4F−3A−2A−1C+1) differs from the spirochetal consensus sequence (16) at positions −4 and −2 and differs from the E. coli consensus sequence (41) at position −3. Thus, the LSP recognition site of LipL21 resembles more closely that of E. coli. However, when whole cells were grown in the presence of labeled palmitic acid, the 21-kDa band was one of the four most abundant labeled proteins, suggesting that the variation in the LSP recognition site sequence did not affect the efficiency of LipL21 lipidation. The abundant 21-kDa band from the whole-cell material was shown to be LipL21 by immunoprecipitation of a labeled protein with the same molecular mass by LipL21 antiserum.
To examine the cellular location of LipL21 in Leptospira, two techniques based on different rationales were used. The first method involves solubilization of the leptospiral outer membrane in situ with TX-114 and the subsequent removal of released periplasmic proteins by phase partitioning. This technique has been used successfully numerous times for a number of spirochetal membrane proteins (9, 22, 40, 43). However, on one occasion the TX-114 method was shown to selectively release a treponemal cytoplasmic membrane lipoprotein (35). Concerns that the TX-114 procedure was releasing cytoplasmic membrane proteins resulted in the development of an alternative procedure for leptospiral membrane fractionation (20). This technique relies on the generation of outer membrane vesicles by alkaline plasmolysis and separation by sucrose density ultracentrifugation. A comparison of the TX-114 detergent fractions and the outer membrane vesicle fractions by two-dimensional electrophoresis revealed similar protein profiles (20), suggesting that both methods result in relatively pure outer membrane fractions. Cellular fractionation with these methods demonstrated the presence of LipL21 in the TX-114 detergent phase and outer membrane vesicle fractions, which were both shown to be free from cytoplasmic membrane markers (Fig. 2). These results unequivocally demonstrate that LipL21 is a component of the leptospiral outer membrane.
Interestingly, during membrane fractionation, virtually no LipL21 could be detected in the cytoplasmic membrane fraction, unlike OMPs LipL36 and LipL41. In the E. coli background, it is thought that the amino acid directly after the cysteine of the LSP recognition site sequence determines to which membrane the lipoprotein will localize, presumably by its interaction with the lipoprotein shuttle LolA (37). The amino acids aspartic acid, proline, and tryptophan at this position are thought to cause retention in the cytoplasmic membrane, while phenylalanine, glycine, and tyrosine serve as ambiguous signals, with the resultant lipoprotein being distributed in both the cytoplasmic and the outer membranes. The two amino acids following the cysteine in the LipL21 sequence are both serines, as is also the case in the major outer membrane lipoprotein of E. coli (33). The +2 amino acids in the LipL36 and LipL41 sequences are lysine and alanine; by default, these would be trafficked to the outer membrane in an E. coli background. However, both the outer membrane and the cytoplasmic membrane fractions contain LipL36 and LipL41, suggesting that lysine and alanine may serve as ambiguous sorting signals in Leptospira. The TX-114 method also reveals some LipL41 associated with the protoplasmic cylinder; however, LipL36 appears to be completely absent from this fraction. This anomaly may be due to variability in the solubility of different proteins in TX-114, where proteins that are poorly solubilized in TX-114 are partially extracted and therefore appear in both the cytoplasmic membrane and the outer membrane fractions. The experimental findings of this study suggest that although the TX-114 cellular fractionation technique may provide an accurate representation of the constituents of the outer membrane, this technique does not provide an accurate account of which OMPs may also be represented in the cytoplasmic membrane. When this variability between the two cellular fraction techniques is taken into account, the novel lipoprotein LipL21 appears to be exclusively located in the outer membrane of Leptospira.
The identification of proteins located on the surface of the cell is of the utmost importance, as they are likely to be involved in the interaction of leptospires with the external environment, including host cells or extracellular matrix components. In addition, leptospiral surface molecules may serve as targets for immune-mediated clearance. We have shown that LipL21 is exposed on the leptospiral surface by virtue of its susceptibility to the biotinylation of intact cells (Fig. 5). This experimental approach has some clear advantages in determining surface exposure of proteins. Because a water-soluble biotinylation reagent was used, the labeling reaction could be carried out in the absence of organic solvents, such as those which were used in other studies (31) and which may permeabilize the outer membrane. Subsequent immunoprecipitation of the labeled proteins, after quenching of the biotinylation reaction, demonstrates specific labeling of the protein under investigation. Only two other leptospiral proteins have definitively been shown to be exposed on the leptospiral cell surface. The rare outer membrane porin OmpL1 was shown to have surface-exposed epitopes by immunogold labeling of whole cells with specific OmpL1 antiserum (17), while surface immunoprecipitation was used to demonstrate the surface exposure of LipL41 (39). Quizzically, when the surface immunoprecipitation method was applied to LipL36, it appeared not to be surface exposed, although it has been definitively localized to the outer membrane. Experiments visualizing freeze-fractured liposomes have suggested that spirochetal lipoproteins take up a conformation external to the membrane bilayer (27), yet specific proteins seem to have a bias for the inner or outer leaflet of the outer membrane. Nothing is known of the topological mechanisms that orchestrate this process in Leptospira; however, a run of six negatively charged residues at the LipL36 N terminus (16) may direct this protein to the inner leaflet of the outer membrane. These residues are notably absent from the surface-exposed proteins LipL21 and LipL41. Future work will investigate whether LipL21 can stimulate protective immunity to leptospirosis.
This work was supported by a grant (to B.A.) from the National Health and Medical Research Council, Canberra, New South Wales, Australia; by VA Medical Research Funds (to D.A.H.); and by Public Health Service grant AI-34431 (to D.A.H.).
We thank James Matsunaga and Yolanda Sanchez for assistance and helpful suggestions. We gratefully acknowledge the excellent technical assistance of Ian McPherson and Vicki Vallance.
- Received 16 December 2002.
- Returned for modification 24 January 2003.
- Accepted 6 February 2003.
- Copyright © 2003 American Society for Microbiology


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The romantic scene for Buffy and friends amps up in unexpected ways. But personal lives are soon put aside to deal with demons, both real and intangible. Spike struggles with vivid dreams of murder, and Buffy journeys into his mind, discovering the terrifying new face of evil…
In this volume, everyone is dealing with their relationship issues. Dawn and Xander are still trying to figure things out, only realizing that the more they try, the worse things seem. Meanwhile Giles is struggling to come to terms with his new situation. As a adolescent teen, he finds it difficult to truly pick up his life where he left it. Willow is busy trying to find herself and she realizes her relationship with Aluwyn isn't going to work out. Meanwhile Buffy and Spike are discovering that despite their best efforts to be friends, the underlying feelings are just too hard to ignore.
This is probably the first time I felt a volume in this series was purely filler. There's something odd going on, as Spike has a vision of killing two people, who actually wind up dead. But for the most part, the majority of this story is all about the relationships within the Scooby Gang. I'm kind of torn, because I like seeing them trying to figure things out, but I also want more and this book was lacking on the action part.
Sadly, I have to wait until the next volume comes out in a few months. But at least this isn't quite the end, just yet.
Pick up your copy on Amazon: Love Dares You by Joss Whedon


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Some say that the health claims of agave are just hype. I disagree. If you're trying to minimize your family's sugar intake, in my opinion organic blue agave is the way to go. If you watch the Dr. Oz show, you know that the doc recommends consuming raw Organic Blue Agave as a sugar substitute. I concur. My alternative health doctor also recommends using it in place of sugar. She's been treating patients for years successfully helping them beat sugar addictions.
I've tried Splenda, Stevia and other sugar substitutes and nothing compares to agave.
What is Agave?
Agave nectar comes from a plant commonly found in Mexico. It's the same plant that is used to make tequila. The nectar from the agave plant is extracted to create a sweetener that's very low on the glycemic index. That's great for diabetics. It's also sweeter so you don't need to use as much as you would sugar.
Agave syrup is available in different colors: light, nectar and amber.
Not All Agave is The Same
Just like with any food, how it's processed and where it comes from is important. You don't want to consume agave that's been processed with chemicals or other pollutants. There's also white agave and blue agave.
The first brand of Agave I tried was from Trader Joe's. It was absolutely delicious. It was a blue agave and costs slightly over $3 for an 8 ounce bottle.
The next brand I tried was Madhava Agave Nectar. I'm not sure if this is white or blue agave but it was good. It was the most expensive of the 5 brands I've tried though.
I also tried Whole Foods brand of organic blue agave and this was by far the worst tasting in my opinion. It was also expensive.
The fourth brand I tried was by a company called Izzy's Organic. This brand was the best tasting. It's right up there with Trader Joe's and it's only $4.99 for a 23 ounce bottle, making it the most cost effective. Hence, this is my brand of choice.
The last brand of organic blue agave I tasted was by a company called Wholesome. This was similar in taste and price to the Whole Foods brand. I would never buy either of these again. An important reason is these two brands were dark in color and therefore processed longer. The longer processing brings out a stronger flavor.
While no brand of agave is exactly the same due to extraction methods and other factors, you should try various brands for yourself. You can find the various brands in supermarkets. This way you'll get a taste for what you prefer. I prefer the lighter nectars for taste.
For more information on agave, here's a site with more detail: http://blog.xagave.com/tag/agave
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It's funny, cause yesterday POLAR won the COTD. He did. For rizzle. But due either to my post Rose Bowl hangover (not likely) or the gerbils in the hamster wheel eating the brown acid (ding ding ding!) that post is lost forever. I loved the title, too. "Commenter of the Day: Salty Ferrous Oxide Edition." Oh well, turns out POLAR gets the award today as well.
In fact, not one, but two Jalopnikers IM'd me to recommend POLAR's sing-songy commentsfor COTD status. And both were from the same post — the DOTS Plymouth Fury III. Of course, I'm an ass-man, so here's why POLAR wins:
Oh, my, god. Murilee, look at her butt. Its so big. *scoff* She looks like, one of those rap guys' gang cars. But, y'know, who understands those rap guys? *scoff* They only drive things like her, Because, she looks like a total bad ass, 'kay? I mean, her butt, is just so big. *scoff* I can't believe its just so long, its like, out there, I mean - gross. Look! She's just so ... BIG!
I like big Fury's
And i can not lie
you other brothers can't deny
That when a Fury drives in
It lacks such taste
and its looks are in your face
you get sprung,
want to pull out your tongue
'cause you notice the butt
was stuffed
Deep in the sheet metal she's wearing
I'm hooked and I can't
stop staring
Oh Fury, I want to get in ya
and take your picture
My Jalops tried to warn me
but with that butt you got
makes me wanna beep horny
Ooh, Rump-o'-smooth-bends
you say you want to get in my hands?
Well, use me, use me,
'Cause you ain't got average groupy
I've seen that suspension dancin'
The hell with romancin'
She's sweat, wet,
Don't need to be going like a turbo 'Vette
I'm tired of magazines
Sayin' small butts are the thing
Take the average Jalop man
ask him that.
She gotta pack much back
So, fellas! (yeah) Fellas!(yeah!)
your wagons and camino's got the butt?
(Hell Yeah!)
Tell 'em to shake it!
(shake it!)
Shake it!
(shake it!)
Shake those handy, fill-able butts!
Fury got back!
Fury got back!
I like 'em square, and big
and pack you up when I'm throwing gig
I just can't help myself, I'm actin' like an animal
Now here's my scandal
I wanna get you home
And ugh, double-up, ugh, ugh
I ain't talkin' bout Civics
'Cause silicone parts
are made for toys
I want 'em real thick and juicy
So find that juicy double
This Jalop's in trouble
Beggin' for a piece of that Fury
'cause I ain't in no hurry
Small trunk econos
Can't touch what she holds
You can have them minivans
I'll keep my cars like Flo Jo
A word to the thick DOTS,
I wanna get with ya
I won't let other cars hit ya
But I gotta be straight when I say I wanna
Cruise Till the break of dawn
Baby got it goin' on
A lot of Jalops may not like this song
'Cause them punks like to
hit it and quit it
And I'd rather stay and play
'Cause the Fury's long, and I'm strong
And I'm down to get the friction on
So, ladies! {Yeah!}
Ladies! {Yeah}
If you wanna role in my Fury {Yeah!}
Then turn around! Stick it out!
'cause this white boy's got to shout
Fury got back!
Fury got back!
Tomorrow we're expecting some Furry action. You heard me.


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FULL OF SPIRIT and COCktails
This spring in Downtown Chandler, more than 2,000 21+ fans of Arizona's burgeoning booze scene will mix, mingle and make new friends and contacts while sampling dozens of diverse alcoholic beverages fit for fans of all types, from the alcohol apprentice to the cocktail connoisseur.
From Sweets to Sour plus a little wine and craft beer. We will even have alcohol kambucha and mead.
April 25, 2020
178 E. Commonwealth Ave
1/4-mile south of Chandler Blvd.
4Pm - 10pm
Looking to get your BRAND in front of thousands of thirsty fans? Sign up below to participate.
Sell Fun Shirts, Have information People could use?. Have great Food?! sign Up Below. Note: WE are sold out for Savory Food Vendors. Email us for waiting list.
TIKI LOUNGE * Speakeasy * MUSIC * BOTTLE PURCHASE
Music Lineup: COMING SOON
2020 AZ Craft Spirits Competition Entry Form
Hosted in conjunction with the Arizona Craft Distillers Guild, the 2020 Arizona Craft Spirits Competition is a true double-blind competition where professional judges award medals based on spirit quality. Open to all domestic and international spirits producers, the competition follows TTB standards and honors the best of the best in multiple categories.
Please Direct all Media Inquiries and General Event Questions to
Tiffany Shultz
The Boss & Event Coordinator


text:
Definitions for "Objects"
Keywords: freestanding, scrolling, chunk, terminology, applet
In HTML programming, building blocks of a page (paragraph, image, sound, ActiveX components...)
The basic components of a database application: forms, tables, queries, macros, reports and modules.
In programming terminology, a freestanding chunk of code that defines the properties of some thing. For example, this Netdictionary applet is an object that contains button objects, scrolling list objects, window objects, URL objects, definition objects, etc. Not all programming languages use this object model; the ones that do are said to be object-oriented.
Netsize Active Gateway proposes a set of objects which can contain all data types. There are three object's types: slots, frames and arrays.!-- if (navigator.appName=="Netscape") { document.write("img src='separation.bmp' x-maintain-ratio='TRUE' height='14' width='687' border='0'");} else { document.write("img src='separation.bmp' x-maintain-ratio='TRUE' style='border-style: none; height: 14px; float: none; width: 687px;' height='14' width='687' border='0'");
an extensive graphical resource about VBA Object Models from AutoCAD to Visio
Graphical entities that can be dynamically created or loaded from model files. Many functions act upon them: • Tasks: each object performs a task per time frame; • Hierarchies: objects can be "linked" together; • Sensors: objects can be connected to sensors; • Modify: geometry, color, texture, scale, etc; • Collision Detection: between objects and polygons.
Keywords: actionscript, iff, predefined, instance, clock
an instance of the union of two classes iff it is an instance of either class
are collections of properties and methods; each object has its own name and is an instance of a particular class. Built-in objects are predefined in the ActionScript language. For example, the built-in Date object provides information from the system clock.
Lines, shapes, text boxes, clips, drawings, and other items you insert in a slide.
An area designator for spells and magical effects. This type of spell does not allow the caster to select individual targets. Rather, it affects some or all eligible objects within a designated area ( burst, cone, or other shape).
Elements that make up the Options Assistant (i.e. Questions, Components, Loops, etc.)
SGML elements, entities, attributes of elements, public identifiers, notations, and standard tagging schemes.
Run-time entities that contain both data and operations.
Keywords: plural
In Access, objects are the tools you need to manage your database
In the Network Installation Management (NIM) environment, an entry in the NIM database that represents a machine, network, or resource.
Keywords: perceptible, something, environment
Keywords: charity, motors, document, industrial, considered
are the purposes of the charity and are set out in the governing document.
physical objects like motors in an industrial plant; also, well-defined actions like using a purchasing document can be considered Objects.
Keywords: ornamental, regularly, lace, flowers, appearing
Ornamental devices (such as flowers) appearing regularly in various parts of a piece of lace.
Keywords: edit


text:
New terminal at Kangaroo Island Airport taking shape | Photos, video
The impressive new terminal building at Kangaroo Island Airport is taking shape.
The state-of-the-art, high-tech building will not only be the first thing that greets inbound passengers, it's also being touted as a facility for all Islanders to use for functions and events.
The Islander was given a tour of the building this week by Kangaroo Island Council CEO Andrew Boardman.
The building is being fitted out with the final touches and it is hoped it will become operational in the week of Monday, May 14 when passengers will be moved over from the old terminal building.
There will be a grand opening with ribbon cutting ceremony and dignitaries sometime in June.
The new terminal component makes up about $8 million of the $18 million airport upgrade project, funded jointly by the State and Federal governments.
The $3.2 million cost over runs we have reported on pertain specifically to the air side or runway component and Mr Boardman said the terminal building project, primarily completed by Mossop Construction + Interiors, had come in under budget and on time.
"It needed to be IATA Class C compliant, which meant it had to be of a certain international standard," he said. "But we also wanted to have something unique to the Island and something with a wow-factor that greeted people flying in for the first time."
Mr Boardman explained the decision was made to keep the existing terminal building and it include it in the new layout, as it was in good shape and would add further cost savings.
It will house Departure Gates 3 and 4, used for the smaller flights to Adelaide that will not require passenger screening.
Departure Gates 1 and 2 in the new terminal building have been designed for the larger, interstate flights, where passenger security screening is required for flights of over 50 passengers and 20 tonnes.
There will be carry-on luggage X-ray screening and passenger metal detection area, ahead of passengers going into the secure area of the departure lounge.
Significant cost savings had been realised by securing some of this security equipment from the Esperance airport in WA that had been downgraded, and this included large checked-bag X-ray machine that cost the project $160,000 instead of $600,000 new.
Similarly, 70 per cent of the luggage carousel system had come from the old international terminal building at the Adelaide airport.
The airport has all the high-tech, state-of-the-art of equipment you would expect at a modern international airport, including digital screens listing flight departures and arrivals, USB charging stations throughout and efficient, energy saving lighting, heating and cooling systems.
The airport also has an open feel with lots of windows to the outside, a fireplace and an open deck.
"I wanted the airport to have an open feel and we have big windows and the deck so that people arriving can easily see the landscape we have on the Island," Mr Boardman said.
There will be tourism promotional displays, including a touch-screen display, and KI branding throughout. Southern Ocean Lodge even has its own closed-off lounge.
The building will be filled with local artwork and while the council's parks and gardens crew will be looking after the landscape featuring all native plants and even a mini sculpture garden.
The airport cafe included a fully, functional commercial kitchen, and Mr Boardman said the idea was that contract to operate the cafe was going through the KI Food and Wine Association, who could then secure local chefs and eateries to make use of the facilities.
The airport building will be able to be used a conference and event centre outside of flight times in the evenings, with the cafe, interior and seating areas an extra attraction for the Island, he said.
The Islander plans on bringing you a series of follow-up articles on the new terminal building in the lead-up to its official opening, looking at the its high-tech features, artwork and tourism promotions contained within and finally its use as a conference and dining facility.
Discuss "New terminal at Kangaroo Island Airport taking shape | Photos, video"
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Regular price
This is a handmade bracelet and a gift from East Asia. When you wear it, you can experience the traditional art and culture.
No Gold Mining
Gold mining can pollute the land, contaminate surface and groundwater, and destroy pristine environments and ecosystems. There is no gold mining during the production process of this item.
Dimensions: adjustable from 10 cm in diameter
Weight: 5 g
Materials: Thread
Vendor: Eco-Kay
Sold in association with Mytheast.com.


text:
Download T2embed.dll and fix "dll not found" error!
What is T2embed.dll?
T2embed.dll - dll file called "t2embed" is a part of Microsoft t2embed program developed by Microsoft Corp..
Some applications or games may need this file to work properly. If t2embed.dll is missing, whenever you start the application/game you may experience various kinds of errors. To fix those errors, please read the Recommended Solution below.
- File version: 0.2.0.81
- File size: 209.91 KB
- MD5 file sum: CE3608B8EC45A0873FA0E52308C0D473
- SHA1 file sum: 319D786ED8DDC710D9711A464B0FFBE063FD1106
T2embed.dll Error Screenshot
Possible error messages regarding this file:
- t2embed.dll is missing
- t2embed.dll error loading
- t2embed.dll crash
- t2embed.dll was not found
- the procedure entry point t2embed.dll
- t2embed.dll could not be located
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Recommended Solution to Fix Dll Error
To fix errors related with .DLL file you need to download t2embed.dll and copy it to the installation folder of the application or game, or copy it into the Windows system folder and it should fix the error. If you don't know how to install this file, please read our tutorial about How to install DLL files.
This page is available in other languages: português | Deutsch | français
You can read more information about t2embed.dll on WikiDll.com


text: Quillows can be made any size but I made mine to have a finished size of 45 x 65? which is long enough to cover a full size adult but not designed for use as a bed comforter. They do however make a nice throw pillow to be used as a bed accessory. Learn how to make them here.


text:
Oster® Heated Microsuede Pet Pad
STPP27-001Oster STPP27-001 https://www.osteranimalcare.ca/on/demandware.static/-/Sites-master-catalog/default/dw62d30372/images/highres/STPP27-001-1.jpg
Provide your dog a softer, warmer and more comfortable sleep surface with the Oster® Heated Microsuede Pet Pad. Designed for all sizes of cats and dogs and features soft cushioned support with a machine washable RoyalMink™ microsuede cover. See More...
Additional Information
Oster® Heated Pet Beds and Pads are ideal not only for the chillier climates but also for offering soothing warmth to your pets muscles and joint year round. They warm to a mild and safe 102.5° F - since the human body temperature is 98.6° F, it will not feel that warm to you, but to your four legged friend, it feels wonderful. The Oster® Heated Microsuede Pet Pad is designed for all sizes of cats and dogs and features a soft sided pad with a machine washable RoyalMinkTM microsuede cover.
- Warms to a Vet recommended 102.5° F
- Thick orthopedic foam offers support to hips and joints
- Microsuade, zippered cover is machine wahable
- Warms up quickly for faster heat-up time
- 36" w x 27" d x 1" h


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By SHERIDAN ROY
The Southington High School wall of honor proudly recognizes graduates of Southington schools who have gone on to accomplish wonderful things in their careers and communities. Just outside of the auditorium, a brick wall is lined with dozens of plaques that current students pass by each day.
Board of education member and former SHS teacher, Bob Brown, works each year to compile and sort through nominations, connect with nominated individuals and/or their families, research nominees who are deceased, coordinate to purchase plaques, and more. This year, he hopes to have a helping hand from current SHS students.
After conversations with incoming SHS principal Frank Pepe, Brown is hopeful that school administration can coordinate the wall of honor into a special project for students, having them assist in research, and design new and improved posters and materials to mount on the wall.
"They walk by this wall every day, but most of them probably aren't even aware of exactly what it is," said Brown. "Having the students involved will help draw their attention to the whole concept."
Brown said the wall of honor can be an inspiration for current students, offering a glimpse of what they, too, can accomplish in their lives beyond high school.
"We have terrific artists in our schools who could make these posters look wonderful," Brown said.
Brown also will focus on fundraising efforts this year to help the project enhance and continue. Looking ahead, he has several big ideas for the wall, such as a mobile wall that can be displayed in public places around the community. He hopes to install a wall-mounted television near the wall, as well, that can display information about honorees and also be used for school announcements.
He also looks to eventually throw a gala at the Aqua Turf Club and invite all of the past honorees and their families at no cost to them.
Several past honorees have been generous in making donations to the program. Donations can be made to the SHS Wall of Honor Foundation, Inc., a tax-deductible nonprofit, by contacting Brown at email@example.com.
"I think the wall is something the community is proud of," said Brown. "Our community is very generous, and I'm hopeful we'll get some support."
Nominations for the upcoming year are being accepted at this time through the end of October. Selection of honorees will be notified in November, and a ceremony will be held in the spring.
Criteria for nominations include: a career or lifetime achievement, outstanding awards received, humanitarian work, or community service. Honorees must qualify for at least one of those categories.
Nomination forms can be found at the high school, or online by visiting southingtonschools.org. Under "schools," click "Southington High School." On the side, click "SHS Celebrations," then "Wall of Honor."
For more information, contact Brown at email@example.com.


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Title: Indebted Heart
Written by: Measha Stone
Release Date : July 28 2016
She works so hard to earn a place in the world, but all he wants is for her to find a place in his heart.
Alyssa Sanders has taken care of herself since she was too young to do so. But she's all grown up and ready to get her life on the right track. She heads to Chicago with her life savings and a few prospects. She expects to get a job, find an apartment and get on her way of settling down. What she doesn't expect is to find her new boss not only severely attractive and Dominant, but the son of an old friend.
Alex Trebelli would do anything for his father, so when he's asked to escort an old friend's daughter to dinner he doesn't hesitate to agree. Except the daughter isn't the small town young woman his father described. She's all grown up, full of beauty and curves, and a submissive nature that draws out his dominant side.
When Alex explains he won't be her friend, he can't just be her friend, he wants more, he wants to be her dominant, her boyfriend, she has to decide if she's going to take the leap. She's never been good at a full time D/s relationship, but Alex makes her feel safe and secure. She finds him easy to be with, even when he's being overprotective and bossy.
Can she manage to put aside her fears of failure, and self-doubt and give them a real chance, or will she chose the safer route and keep clear of his sexy smile, and dominant ways?
The silence made her nervous; she wanted his words, his touches, and the lack of both worried her. Just as she was about to break her silence he stood in front her. His hands dug into her hair and he pulled her head back to look up at him. The pleasure and joy she expected to see alluded her. Instead firm, frank eyes glared down at her.
"Where did you get the cuffs?" he asked.
"The foot locker in the closet," she said, searching his expression. Anger did not lurk behind his words, but displeasure shone through his eyes.
"What were your instructions?" he asked, releasing her hair and moving down to his haunches, keeping their eyes locked.
"To come in here, strip, and kneel with my hands behind my back and my eyes downcast," she answered and before she could continue, he spoke.
"And in those instructions where did I say to go into the closet and dig out a pair of cuffs?"
"Nowhere, but I—"
"You are trying to control the situation." His words were given in a soft tone, but the implication was harsh.
"What? No. I was trying to add to it, to give you more of what you wanted." She wished they could have the discussion clothed.
"What I want, what I desire, is your submission, your obedience. If I want your hands cuffed, I'll give you the instructions or I'll do it myself. You wanted your hands bound. You wanted to feel the cuffs on your wrists, bound, and waiting for me. Those were your desires."
"Don't my desires matter?" she asked with an edge that she quickly regretted at the instant raise of his eyebrow.
"Of course they matter. And I will always take your desires into account. When you submit to me, you trust me to give you want you want, what you need, and push your limits, but you don't control what happens."
A woman given up on love, second guesses her solitary life when meeting a man who won't take no for an answer and opens her eyes to an entire world of pleasure she didn't know existed before him.
A headstrong, fiery woman seeks to fulfill her fantasies in all the wrong places, and it's up to the security expert to tame her, and keep her safe.
Buy Links:
Blushing Books: http://www.blushingbooks.com/index.php?l=product_detail&p=3917
Measha Stone lives in the western suburbs of Chicago with her husband and their children. She is a member of Romance Writers of America, and graduated Summa Cum Laude with a degree in Creative Writing from Southern New Hampshire University. Her vanilla writing has been published in the online magazine efiction and the DuPage Writers Group annual journal Possibilities. When she's not writing, she's reading and spending time with her kids- who are just as creative and crazy as her.
Social Media
Facebook: https://www.facebook.com/measha.stone.5
Twitter: @measha_stone
Instagram: https://www.instagram.com/meashastone/
Blog: https://www.meashawrites.com/blog
Author Questions
What do you like to do when you're not writing? I still have a day job, but in the off hours, if I'm not writing, or thinking about writing, I'm usually curled up with a book. If my kids happen to actually be home instead of off playing with friends, we'll spend time together watching old sitcoms on Netflix, or having a family game night. .
Which body part would you choose to get a tattoo done? I celebrated my first book with Blushing Books by getting a tattoo of a heart on my right wrist. I'm planning to get two more, another heart design on my ankle, and one on the top of my foot the words "Happily Ever After".
The furthest I have traveled was to: Germany. My husband was in the Army when we married and was stationed in Darmstadt, Germany. It was a great time, even though I was far away from family and friends. I was sad to hear the base has been shut down since then.


text:
Introducing The WoolieBow Bag!
A collaboration with Instagram shop @littlelaylaloves and wooliebutton brings you a place to store all your beautiful bows and gives you a photo prop all in one!
You can even choose your bags' design from a selection of sassy bow related quotes!
Flip your bag over to #wooliebutton, snap your pic and tag @wooliebutton on social media to get a discount on your next Woolie order.


text:
What other guests say about AE sailing experiences
An amazing day – fun all the way around and Robert (the skipper) was fantastic – we did everything.
Day Sail – July 2013
We had a relaxing but entertaining time sailing and enjoying the best of the Croatian islands – the water, the food and the warm welcome – a delight for all ages!
Day Sail – Sept 2013
The best way to see the Islands of Croatia!
Day Sail – Sept 2013
Awesome trip! The skipper was super nice, super helpful, and part of our group! He's very passionate about his country as he should be! Its stunning!


text:
If you've ever opened a bottle of wine and only had one glass, you've probably been disappointed a few days later to find the taste is no longer what it was. Opened wine can oxidize, resulting in an off taste within a matter of days. In the past, this could have led to pressure to finish a bottle of wine in one night or risk pouring it (and the money you spent on it) down the drain. Now there is a product on the market that allows you to pour your wine with actually removing the bottle's cork. In other words, your bottle of wine can last you longer without risk of getting stale.
Coravin's Model One Wine System uses a thin needle to gently penetrate the wine bottle's cork to access your wine. Simply position the Coravin System on top of the cork and push the needle through. Then, with the bottle at a 45-degree angle, you can press the trigger to pressurize it and then release it to pour wine into your glass. On average, you can pour a serving of wine in 25 to 29 seconds when using this system. To remove the needle, just pull the Coravin System straight up, and the cork will reseal naturally! Premium Coravin Capsules contain argon gas, which is used in the winemaking process, to protect the wine remaining in the bottle. Each capsule allows you to pour up to 15 five-ounce glasses of wine.
Each Coravin Model One Wine System weighs less than one pound. It comes in a white and blue design. The handle has a soft-touch grip to make it easy to pour your wine. This comes with two Coravin Capsules and a needle cleaning tool. Reviewers love this gadget, whether they personally just drink one glass of wine a night, like to host wine tastings, or like to pair different wines with different courses of their meals. Most found it easy to use. Coravin notes that the resealing only works with natural (not synthetic) cork so keep that in mind. Coravin offers a one-year warranty on this product.


text:
Portbury Burials
Transcribed by Mary Mason.
Dec 3 A poor man named William was buried.
Jan 26 Joane Russell widow late wife of Willm: Russell.
Feb 2 Thomas Hardinge.
Feb 9 William Gayton.
John Hill and William Coxe Church Wardens.
Apr 4 Johane Patch.
Apr 7 Johane Thomas wife of David Thomas.
May 9 Robert Yvans.
Jul 26 Robert Gadon.
Jul 27 Johane Yvans wife to Jenkyn Yvans.
Aug 6 William Godan.
Sep 14 Richard James.
Sep 17 Agnes Harris.
Oct 6 Richard Syms son of Nicholas Syms.
Oct 19 Isabell George wife of Bennett George.
Oct 22 Robert Grayle.
Nov 27 Joane Pickett wife of Robert Pickett..
Dec 16 Alice West widow.
Jan 16 Edward George son of Bennett George.
Jan 28 Johane Syms daughter of Nicholas Syms.
Feb 6 James Teage son of John Teage.
Feb 16 Johane Anderway daughter of John Anderway.
Feb 17 Thomas George son of Bennett George.
Feb 19 Johnae Durban daughter of John Durban.
Mar 6 Johane Baker daughter of John Baker.
Mar 23 Agnes Baker daughter of Thomas Baker.
John West and Jacob Pickett Church Wardens.
Apr 3 Agnes Powell daughter of Johane Willins.
Apr 9 ……ye Grayle wife of John Grayle. (Faded could be Jenye.)
May 23 Jane Dall daughter of Thomas Dall.
The last of May, Joane Hobes wife of William Hobes.
Jun 10 William Blanch son of John Blanch.
Jun 15 John Day son of William Day.
Jun 19 Joane Grayle wife of John Grayle.
Jun 23 John Grayle husbonde .
Jul 3 Robert Dowtinge a lad.
Jul 28 Alice Thomas daughter of John Thomas.
Jul Marteyne Hichins son of Robert Hichins.
Aug 23 William Passer.
Sep 5 William Hardinge husbandman.
Sep 13 Johne Baker son of Richard Baker.
Sep 25 Joane Payne wife of Edmond Payne.
Oct 5 John Smyth son of William Smyth.
Oct 20 Thomas Bryan son to Mathew Bryan vicker. (vicar)
Nov 17 John Perre son of Frances Perre.
Dec 1 Joane Baker daughter to William Baker.
Dec 17 Joane Demock, chrisomer.
The same day Agnes Teage, chrisomer.
Dec 20 William Colyins.
The same day Alice Baker daughter of John Baker.
Jan 8 Margaret Stibons.
Jan 18 Thomas Baker, youngman.
Jan 28 Agnes Richards young child.
Jan 19 John Hill, chrisomer.
Feb 4 Robert Leways.
Feb 15 Joane Spure, chrisomer.
Apr 7 John Stidman christened at home.
May 5 Johan Stybons daughter of John Stybons.
May 3 John Knight, chrysomer.
May 12 Alice Porter wife of Thomas Porter.
Jun 15 Joane Weaver.
Jul 17 Agnes Hardinge, chrisomer.
Aug 20 John Paschal,l chrisomer.
Sep 16 Frances Hawkins was borne, christened and buried.
Oct 22 James Netheway, chrisomer.
Ther same day Robert Grayle son to John Grayle.
Dec 4 Joane Pryan wife to Mathew Bryan, clark vicker here. (vicar)
Dec 20 Elizabeth Lullot daughter to William Lullot.
Jan 1 Joane Lullot wife of William Lullot.
Jan 6 John Picket Bachelor.
Jan 10 Joane Hiscoke wife of John Hiscocke.
The same day, Agnes Gooden daughter of Robert Gooden.
Mar 11 Andrew Lowle.
Mar 16 Isable Stibon.
May 8 Agnes Beney wife of John Beney.
Apr 16 Edith Wylshyer. (Wilshire)
Jun 22 Elizabeth Lullet wife of Richard Lullet.
Jun 27 David Thomas.
Aug 9 John Walis.
Sep 8 Elizabeth Acridge.
Oct 9 David Grayll.
Dec 5 John Stybyn.
Dec 14 Walter Payne.
Jan 1 Phillip Willys.
Jan 6 Isable Blanch.
Mar 3 Richard Grayle.
Mar 5 Margaret Feare wife to Robert Feare.
May 1 Isabell Crocke wife of John Crocke.
May 4 Thomas Sanders.
May 15 Alice Loke wife of Thomas Lok.
Jun 18 John Hanncson.
Jul 28 John Watte.
Aug 1 John Hurtan.
Oct 23 John and Joane Yvans, chrisomars.
Oct 30 Richard Lowle.
Dec 3 John Nethway.
Dec 27 Marye Werne.
Jan 2 Alice Spure.
Jan 24 George Dabney, a stranger.
Feb 4 John Heichclive.
Feb 18 William Baker, chrisome.
Feb 20 Agnes Denys.
The same day Alice Lyrence, chrisome.
Apr 20 Joane Coke, chrisome.
Apt 27 Edith Lullet, chrisome.
Jul 3 Roger Hill.
Jul 16 Mathew Bryan late vicare.
The same day William Handdy.
The same day Mathew Fysher.
Jul17 Edieth Cylly daughter of John Cylly.
Jul 18 Alice Cylly wife of John Cyllye.
The same day Richard Arther.
The same day Richard Lawrence a servant of Thomas Dale.
Jul 20 Joane Smyth wife of William Smyth.
Jul 21 Joane Baker daughter of John Baker.
Jul 23 Joane Hodge wife of Thomas Hodge.
Jul 28 John Cilly son of John Cilly.
Jul 29 John Cilly.
Jul 30 Allice Cottrell.
Aug 6 Thomas Dale.
Aug 11 Joane Stevyns daughter of William Stevyns. (Stephens)
Aug 13 Alice Hodges and Agnes Hodges daughters of Tho: Hodges.
Aug 21 Wlliam Stevyns son of William Stevyns.
Aug 22 William Deygh son in law of William Stevyns.
Sep 3 Thomas Stevyns son of William Stevyns.
Oct 19 John Powell.
The same day Edith Chamber.
Nov 4 Catherine Chamber daughter of John Chamber.
Nov 6 Joane Chamber daughter of John Chamber.
Nov 24 Thomas Heysell. (Hazel)
Dec 13 Margaret Lullett wife of William Lullett.
Dec 16 Joane Lullett daughter of William Lullett.
Dec 19 William Pickereil son of Robert Pickereil.
Jan 1 John Hurne son of Thomas Hurne.
Jan 2 Grace Heydon daughter of M Heydon.
Jan 4 Agnes Lullett daughter of William Lullett.
Jan 8 William Lullett.
Jan 15 Robert Pickarde.
Jan 18 Joane Hurne.
Jan 26 John Tucky.
Jan 28 Robert Pyckerde son of Robert Pickerde.
Feb 5 John Pickerde son of Robert Pickerde.
Mar 21 Lucy Sweat wife of John Sweate.
James Williams son of John Williams.
May 15 Joane Thomas wife of John Thomas.
Aug 12 John Wyte.
Aug 15 Jane a childe of newmans.
Sep 15 Luce Baker.
Oct 26 Cisly Smyns.
The same day Joane Coope.
Dec 5 Francis Pers. (Pearce)
Aug 10 John Averie son of Thomas Averie.
Aug 18 Elizabeth Grayle daughter of Robert Grayle.
Apr 20 Thomas Halyiard son of Paule Halyiard.
Jun 10 Edmond Payne.
Nov 2 John Russell.
Aug 16 John Mulgreie son of William Mulgreie. (Mulgery)
Sep 1 William Weste son of John Weste.
Feb 2 John Cope.
Jan 4 John Cope son of John Cope.
Jan 10 William Cooke son of John Cooke.
Feb 12 John Richards.
Mar 10 John Cooke.
Sep 10 Agnes Humphery.
The last of May, Elizabeth Hitchins.
Jun 4 ?Thomas Gardener.
Jun 28 John Payne son of Edmond Payne.
Aug 2 John Gardener son of Thomas Gardener.
Aug 5 Thomas Whitehande.
Sep 1 William Lullet son of Richard Lullet.
Sep 28 Joane Andrews widow.
The last of Oct, William Russell son of Thomas Russell.
Nov 4 Joane Lowle wife of Richard Loule.
Aug 8 Joane Baker daughter of Thomas Baker.
Sep 10 Joane Davis a kyns woman of John Whelses. (kins woman)
Nov 18 Thomas Taylor.
Mar 16 Elizabeth Thomas daughter of John Thomas.
Mar 28 Marye Lewys wife of Lewys.
The Last of March, Margaret Williams daughter of John Williams.
Apr 11 Joane Baker wife of John Baker.
May 20 John Cotrell.
The last of August Joane Corbet widow.
Sep 1 Joane Smyes.
Sep 10 Agnes Dymock daughter of William Dymock.
Feb 10 John Hicke.
Oct 1 Myles Payne son of John Payne.
Feb 2 Alice Gayner wife of Lewys Gayner.
May 15 Joane Cronie.
May 18 Agnes Paschalle daughter of Thomas Paschall.
Aug 19 John Jones wife of David Jones.
Sep 11 Agnes Blackdon wife of John Blackdon.
Sep 13 John Gardener son of Richard Gardener.
Sep 18 Mathew Baker son of John Baker.
Sep 29 Joane Williams.
Dec 1 John Thomas.
Nov 28 John Payne son of John Payne.
Jan 3 Isable Bynye.
Mar 1 Thomas Atwood.
Mar 5 Joane Russell daughter of Thomas Russell.
May 24 Alice Burnam widow.
Apr 23 William Score the minister.
Mar 18 John Taylor the mason.
May 20 Joane Blysse wife of Harye Blysse.
Dec 20 Joane Russell alias Smyth wife of Thomas Russell.
Dec 26 Joane Blanch.
Jan 26 Alice Woodward.
Apr 13 Richard Kinge.
The last of June Katherin Evans daughter of Thomas Evans.
Jun 10 Agnes Nethway.
Oct 20 Julian Coope daughter of John Coope.
Jan 10 Thomas Mydwynter.
Feb 20 Robert Lokyer.
Feb 25 William Lokyer son of William Lokyer.
Mar 10 Richard Adams.
Apr 3 Maude Thomas wife of Richard Thomas.
Apr 10 William Robins son of John Robins.
Apr 28 Thomas Rothe son of John Rothe.
May 15 Agnes Griffin wife of John Griffin.
Jun 5 John Came a servant of Richard Lowle.
The same day Welthian Gefferys a servant of John Payne.
Jun 12 Ellyn Cronine servant of David Jones.
Sep 7 Agnes Willys wife of Richard Willys.
Sep 1 Margaret Lowle daughter of Richard Lowle.
Oct 2 Joane Basset wife of one Basset.
Oct 5 Margaret Dale widow.
Oct 6 Rose Joyner wife of Lewyes Joyner.
Oct 21 John Hiskox.
The same day Agnes Lullet.
Oct 29 Elizabeth Dymocke wife of one William Dymock.
Dec 3 Joane Paschall daughter of Thomas Paschall.
Jan 13 Richard Paschall.
Mar 15 Richard Browne.
Feb 14 Mary Holbrocke.
Apr 10 Richard Habynton.
Jun 1 Elizabeth Pickerd.
Nov 3 Joane Simber wife of Nicholas Simber.
Nov 10 Joane Durban daughter of William Durban.
Feb 12 Isable Crome.
Feb 21 Mrs Alice Perryvall.
Apr 2 Phillip Gawdyer wife of William Gawdyer.
Apr 7 Alice Bynye daughter of John Benye.
Jun 23 John Philpot.
Sep 22 Alice Hardinge daughter of Richard Hardinge.
Nov 18 Elizabeth Geninge wife of Thomas Geninge.
Jan 26 Thomas Hamons.
Jan 20 Edith Burnam daughter of Richard Burnam.
Jan 30 Juliana More daughter of Thomas More.
Mar 13 Alice Gardener daughter of John Gardener.
Apr 19 Richard Durban cosen to Richard Thomas. (cousin)
May 29 Joane Willis widow.
Jul 20 Katherin Evans daughter of Thomas Evans.
Aug 1 William Standford servant of William Godwyne.
Sep 28 Alice Thomas daughter of Richard Thomas.
Oct 18 Alice Hichinge daughter of Robert Hitchinge.
Oct 21 Alice Andrews daughter of Robert Andrews.
Nov 8 Joane Baker daughter of William Baker.
Feb 18 Robert Smyth.
Feb 28 The wife of John Hobe.
May 8 Agnes West.
May 28 Mary Habinton.
Jun 8 William Morgan.
Jun 10 Jane Jeyne.
Jun 13 Richard Hill.
Jun 15 Frannce Gayne wife of James Gayne.
The same day Thomas Cokoerye and his wife.
Jun 18 Cisly Lullet.
Jun 25 John Neytway.
Jun 29 Joane Champlyn.
Aug 13 William Evanice.
Sep 3 Elizabeth Evance daughter of Evance Bonaventer.
Sep 20 Joane Blane servante to William Dymocke.
Sep 19 Joane Graylle daughter of John Graylle.
The same day Ellyn Hamons.
Sep 24 Lawrence Powell.
Oct 3 Evance Bonaventure his wife and two children.
Nov 18 James Philpot son of John Phillpot.
Nov 26 Robert Pinsyphy.
Dec 7 Joane Grove wife of Edmond Grove.
Dec 13 Agnes Bishope wife of Robert Bishope.
The same day Elizabeth Holbrocke daughter of Thomas Holbrocke.
Jan 3 Phillip and his two children.
The same day Agnes Grove.
Jan 8 John Grove son of Edmond Grove.
Feb 20 John Holbrocke.
Jan 10 Edith Molgray daughter of William Moulgraye
Mar 25 Edith Love wife of Robert Love/Lone.
Apr 10 John Andrewes son of John Andrewes.
The last of August, Joane Katkins wife of John Katkins.
Sep 6 Tobye Katkins son of Thomas Katkins.
Sep 19 Daniel Mason a child.
Dec 24 Alice Nethwaye wife of Richard Nethway.
Dec 10 Agnes Langford daughter of M Langford of Bristol.
The same day Alice George widow.
Feb 1 Wother Dymocke.
Mar 4 Cornelius Gardener.
The last of July, Joane Nicholas widow.
Aug 13 Joane Harding.
Aug 13 John Champion son of William Champion.
Aug 16 Joane Jones daughter of David Jones.
Sep 2 Harry Duckes.
Oct 13 Saray Dymock daughter of William Dymock.
Nov 7 Edith Andrewes.
Nov 30 Lewys Gaynard.
Dec 4 Richard Stevens.
Mar 24 William Come.
Apr 15 Richard Gardener.
Apr 15 Richard Gardener.
Apr 26 Richard Weare.
Jul 1 John Welshe.
Oct 10 John Powell.
Sep 10 Margaret Williams.
Dec 30 Joane Grayle.
Jan 9 Alice Hichins.
Jan 18 Robert Greene.
Feb 18 John Haterthacke.
Feb 28 Thomas Whitting.
Mar 9 Alexander Sheppard.
Apr 21 Margaret Baker wife of William Baker.
Apr 22 William Baker.
Apr 25 Edith Russell.
May 4 John Atkins son of John Atkins.
May 7 John Russell.
The same day Florence Smyth.
May 10 Alice Hicssetor.
May 24 Grace Robbins.
Jul 2 William Sheppard.
Jul 20 Mary Norton.
Aug 5 William Russell.
Aug 15 Joane Messenger.
Aug 20 William Goddine.
Aug 26 John Andrews.
Sep 4 Alice Greene daughter of John Greene.
Oct 13 John Hardinge.
Oct 17 Elizabeth Lawrence.
Nov 17 Julian Russell.
Dec 6 A poore mans child.
Jan 13 Joane Perryne the bastard.
Feb 9 Robert Grayll.
Feb 23 Jane/James Pickear.
Mar 14 Alice Whightinge. (Whiting)
Apr 10 Alice Burnam.
Jul 4 Alice Butler wife of Edward Butler.
Jul 15 William Hawkins.
Jul 18 Richard Coke.
Jul 24 James Williams.
Jul 25 Alice Smyth.
Aug 28 Agnes Willet.
Nov 20 Agenes Lawrence.
Nov 27 John Feare.
Nov 27 Christopher Taylor son of Christopher Taylor.
Mar 24 John Surat.
Mar 27 Thomas Russell.
Apr 4 Joane Mulgraye.
Apr 5 John Dymry son to John Dymry.
Apr 20 Agnes Haule widow.
May 2 Thomas Harding son of Richard Harding.
May 5 Joane Symber.
Jul 21 Margaret Phillipps widow.
Sep 4 Julianis Feare.
Oct 6 Richard Hardinge son of Richard Hardinge.
Jan 2 John Durban.
Jan 3 Edmond Hardwick.
Jan 8 William Richards.
Jan 21 John Tringe.
Mar 28 Joseph Randle.
Feb 14 John Lawrence.
Apr 15 Alice Sweat widow.
Jun 15 Joane Norcot.
Jul 10 Margaret Welsh.
The last of June Alice Willsonne.
Jul 3 John James.
Jul 10 Agnes Randall.
The same day Robert Edsonne.
Aug 20 Edyth Schore widow.
Aug 28 Agnes Merton.
Sep 20 Thomas Hanrze.
Oct 2 Christopher Taylor son of Christopher Taylor.
Oct 3 John Streynbon.
Jul 3 John Coope son of Robert Coope.
Mar 6 David Jones.
Mar 10 Robert Andrews.


text:
Learning Spanish in Quito
Home to more than seventy language schools, Quito is the most popular place in South America to learn Spanish, partly because rates are so cheap – $4–10 per hour for one-to-one classes – and partly because Spanish is spoken much more clearly here than in many other countries, or even in Guayaquil and the coast. Ecuadorian serranos (highlanders) tend to speak slowly and pronounce all the letters in each word (elsewhere, consonants are frequently dropped), making them easy to understand.
Nearly all schools offer one-to-one lessons or classes in small groups, but not all use books or visual aids. It's a good idea to ask if you can rotate your teachers, as this keeps the lessons fresher and allows you to compare techniques and decide what suits you best. Visiting a school and enquiring about facilities or methods, or perhaps even asking to sit in on a lesson, will give you the best idea if it's right for you before enrolling.
Some schools will encourage you to sign up for seven hours a day, but most students find that exhausting – four hours a day is a better bet, whether studying for just a few days or several weeks. The majority of schools offer the option of staying with a family (usually $10–15 per day, with meals included), while others now offer daily classes as part of full tours to the jungle or coast. Many schools also offer activities such as cookery classes, dancing lessons or day and weekend trips – all good ways to meet other students.
- The old townchevron_right
- Colonial religious art and the Quito Schoolchevron_right
- The new townchevron_right
- Learning Spanish in Quitochevron_right
- Fiestas in Quitochevron_right
- Accommodationchevron_right
- Quito addresseschevron_right
- Restaurants and caféschevron_right
- Nightlifechevron_right
- Arts and entertainmentchevron_right
- Shoppingchevron_right
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