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Healthy Weight Loss Program at Sparkling Hill Covers Fitness, Nutrition and Detox
Catherine Dunwoody | 9/18/2012
Sparkling Hill Resort's new weight loss program includes spa treatments at KurSpa
Vernon, BC's dazzling jewel on the hilltop, Sparkling Hill Resort, has recently added a healthy weight-loss program to its menu of wellness packages
Certainly no fat-farm, Sparkling Hill's Swarovski-crystal-adorned, family-owned and designed resort has been only around a couple of years, but true to its initial mandate, has kept health at the forefront of its offerings.
The new program does not foolishly promise you'll drop a waist-size in a week, rather, it is designed to "kick-start healthy and sustainable weight-loss" says Dr. Ray Lendvai, naturopathic doctor on staff at the resort's KurSpa.
The eight-day program concentrates on three key points – fitness, integrative medicine and nutrition – and each guest receives a personal program tailored to their specific needs.
For the exercise component, expect a mix of cardio, strength training in the Keiser-equipped fitness room, pool jogs, trail walking and yoga classes.
Cardio and strength training is done in the fitness room at Sparkling Hill Resort (Image: Sparkling Hill Resort)
Treatments scheduled in the resort's KurSpa, which also happens to be Canada's largest spa, are all part of the detoxifying and integrative medicine aspect.
Aromatherapy steams (my personal favourite was the rose aroma – heavenly) and saunas, exercising in the Kneipp Water therapy pool, and swims in the indoor and outdoor pools are part of the plan, plus spa treatments like the signature cold sauna (-110C/-166F) and the Universal Contour Body Wrap. This wrap is included twice in the program, and uses detoxifying clay to pull impurities and toxins from the body.
The rose aromatherapy steam is heavenly (Image: Sparkling Hill Resort)
Food, as in any weight loss program, is the biggie. "Dr. Ray" as he is called at the spa, provides a custom-designed healthy menu for each guest. Expect meals with lean protein, lots of vegetables, whole-grain carbs like brown rice and plenty of cleansing teas and filtered water to flush your system.
Programs like this don't come cheap anywhere in the world, but considering most involve jetting off to Europe, skipping the airfare certainly will save you a bundle. Plus, Vernon's a driveable destination for most BC residents.
This program includes seven nights' accommodation at the resort, three meals per day, wireless Internet and valet parking. It costs $2,900 for single occupancy and $2,500 for double occupancy before tax and gratuity.
Investing in your health isn't the craziest idea, is it?
Catherine Dunwoody, Contributor
Catherine Dunwoody is a regular contributor to BCLiving and TV Week, including penning the 'TV Dinner' column each week. She writes about style, travel, food, the arts, celebrities, décor, & more – for The Vancouver Sun, Nuvo, WestJet's UP!, Flavours, The Globe & Mail, & Jetsetter. She's held editor posts at LouLou, Real Weddings & FASHION magazine, and appears as a 'lifestyle expert' on CTV Morning Live, The Shopping Bags, and Global TV.
Follower her on Twitter @cdunwoody
weight loss, resort, spa, BC travel, Sparklinf Hill Resort, Vernon
Trending / Health & Fitness


We have just added a new page to the navigation menu on the Store. It's a page dedicated to accessories that we are offering for the Ridge Runner Hammock. This includes our Spreader bar storage bag, the Ridge Creek Under Quilt as well as a new accessory the Spreader Bar Buckle Bumpers. The Bumpers can help to protect your tarp from accidentally being poked by the spreader bars.


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Biden tells Xi US and China must not 'veer into conflict'
US president Joe Biden and his Chinese counterpart, Xi Jinping, have spoken in their first phone call for seven months, amid continuing tensions between the world's two largest economies.
During the 90-minute call, which was initiated by Biden, the two leaders discussed their shared responsibility to ensure competition does not "veer into conflict", according to a readout from the White House.
A White House statement said the two leaders had "a broad, strategic discussion" including on "areas where our interests converge, and areas where our interests, values and perspectives diverge." It said Biden and Xi agreed to engage "openly and straightforwardly".
"This discussion, as President Biden made clear, was part of the United States' ongoing effort to responsibly manage the competition between the United States and the PRC," the statement said.
A Chinese readout said the conversation was candid and in-depth. "Whether China and the US can handle their relationship well bears on the future of the world. It is a question of the century to which the two countries must provide a good answer," said Xi, according to the readout.
Xi allowed his officials to work with their US counterparts to "continue their engagement and dialogue to advance coordination and cooperation" on the climate emergency, Covid-19 response and economic recovery as well as on significant international and regional issues, but added that they should be "on the basis of respecting each other's core concerns".
Bilateral relations between two of the world's most significant powers have plummeted since the Trump administration Photograph: Jewel Samad/AFP/Getty Images
Analysts in Washington said there has been ongoing frustration in the US that efforts to find common ground had so far been fruitless. They said China's increasing aggression in the South China Sea and Taiwan Strait, and the growing presence of the US and its increased support of Taiwan, had also increased concerns over the prospect of conflict.
But the Chinese side accused the US of interfering in its domestic affairs by politicising issues such as Hong Kong and Xinjiang – both China's territories. Some Chinese commentators went as far as to suggest the US was attempting to contain China.
Recent efforts to progress relations have stalled. Last week John Kerry met China's foreign minister, Wang Yi. Kerry told reporters he had urged Wang and the Chinese delegation to do more on the climate crisis, which was more important than politics.
Wang, however, countered that bilateral climate cooperation "cannot be separated from the wider environment of China-US relations," and that Washington should "stop viewing China as a threat and a rival" and "cease containing and suppressing China all over the world".
He added that the US saw the two sides' joint efforts against global heating as an oasis. "But surrounding the oasis is a desert, and the oasis could be desertified very soon," he said.
The two leaders also discussed the issue of the climate emergency in their latest call. The Chinese readout said Beijing had "taken the initiative to actively shoulder international responsibilities befitting China's national conditions".
'Deep concerns': US and China trade criticisms at Alaska meeting – video
Bilateral relations between two of the world's most significant powers have plummeted since the Trump administration. A high-level meeting in March including Wang, China's top diplomat Yang Jiechi, US secretary of state Antony Blinken and US national security adviser Jake Sullivan also descended into hostile public rebukes.
According to the White House, Biden initiated Thursday's call. A senior administration official told the Associated Press the White House had been unsatisfied with early engagements with China.
The official, who was not authorised to comment publicly and spoke on condition of anonymity, said White House officials were hopeful that Xi hearing directly from Biden could prove beneficial.
The White House official said Biden made clear to Xi that he had no intention of moving away from his administration's policy of pressing China on human rights, trade and other areas where it believes China is acting outside international norms.

redpajama_set_name: RedPajamaCommonCrawl


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Hearing on lesbian health care denial
Guadalupe Benitez's Southern California fertility doctor refused to do an artificial insemination because Benitez is a lesbian, and the doctor said that it conflicted with her Christian religious beliefs.
Now, the State Supreme Court is set to decide if the doctor broke the law, or if she's protected under religious freedom rights.
"The question is, is whether you can offer a given service and offer that service, and yet say, 'but you, oh, I'm not offering it to you," says Justice Carol Corrigan of the State Supreme Court.
Benitez's lawyers say the doctor did violate the state's anti-discrimination law governing businesses.
"In this case, we're just asking the court to apply existing rules, and again to reject the idea that civil rights laws can be avoided based on religious belief," says Benitez's attorney Jennifer Pizer.
"It doesn't need to be one side versus the other. We can have a win-win situation here," says the doctor's attorney Kenneth Pedroza.
The doctor's lawyers argued a compromise should be struck.
"When a physician feels in good conscience they can't provide care, that physician should go out and help the patient get the care they need," says Pedroza.
But Benitez's lawyers say that it would be illegal if the care were denied because of the patient's race or ethnicity, and being denied on the basis of sexual orientation is no different.
"California doesn't license doctors to be a Whites only doctor or a heterosexual only doctor or a Jew only doctor. It's a license to be a doctor," says Pizer.
"A rule of accommodation would allow individual physicians in a group to do it and others not to and to only refuse when it's a substantial burden to their religious belief," says Pedroza.
Benitez did eventually conceive through in-vitro fertilization with the help of another doctor. Justice Kathryn Werdeger questioned if ultimately, there really was any harm done to Benitez.
"It does a great deal of damage to a person when being discriminated like that, being told you're not worthy of becoming a mother or having a child or having a family, which ultimately the only thing I wanted to do was to be a family and have children and just be a great mom," says Benitez.
The court has 90 days to decide, but that's far from the end of the story. Either way, the case will then go back to the San Diego area trial court. The Supreme Court's decision will determine if the doctor and her practice can use the religious rights argument as a defense.


We are interested in collaborative art making. Collaborations simply for the enjoyment of experiencing the creative process with another person, to create a more inclusive environment for non-artists and to redirect the value of art from finished masterpieces to the explorative and failure-driven process. We imagine that collaborative art making would harness the energy/movement/attitudes of your partner which could be extended to include remote collaboration. The following proposes an Emotive Watercolor Palette to engage partners in painting in a new, collaborative way.
Imagine this scenario: two painters, each with their own an Emotive Watercolor Palette, paintbrush (with FSR and tilt sensors), and a water cup. Partner A is painting and the intensity at which they paint influences the colors available to Partner B. And, vice versa, resulting in a dance between Partner A and Partner B where the actions of one influences the creative process/outcome of the other. Both partners could either be in the same room, or in remote locations*.
Partner B's watercolor palette will have lids covering each paint color. These lids will flap up and down at various rates depending on the intensity read in from Partner A's brush – a faster rate if Partner A is painting with higher intensity and slower rate if Partner B is painting with lower intensity. The flapping lids are meant to indicate life in a rhythmic fashion, similar to playing the keys of a piano or beating a drum.
When Partner B needs more paint, they will dip their paint brush into the water cup. This activates a sensor (either photocell or FSR) which sends a signal to the water palette to pause the up and down flapping motion of the paint lids for approximately 1 minute. (This is so Partner B can actually get their brush into the paint color).
For a middle intensity, YGB would be open while RO IV would be close. RO[YGB]IV.
This gives the painter a range of colors to work with that all have a similar tone to them.
Additionally, a complimentary color would be shown. For example, if [ROY] is open, [B] would also remain open to give Partner B encouragement to use a color that may be out of their comfort zone (this could be an educational tool for novices).
Meanwhile, this same process is happening for Partner A.
We plan to make the water color palettes on a wooden box such that the colors from the paint will seep into and color the wood over time. This will serve as an artifact of the intensity state of one's partner over time.
All team members intend to continue to participate in all aspects of design and creation. Although, we have identified leaders for each aspect to ensure organized development. Dina will lead research, Jake will lead electronics, and Andrea will lead the coding.
* We intend to focus first on synchronous interactions, but plan to consider asynchronous interactions, time permitting.
** We will test the FSR to see if we can detect the degree of the painter's intensity by pressure on the brush bristles, or the pressure at which someone holds the brush.
In trying the HTC Vive, the thing I liked the most was using the Tilt Brush – drawing and creating in a 3D space. I liked how it forced me to draw something in its entirety, not just the flat 2D perspective. For example, if I were to draw a house and then walk inside of it, I'd have to think about what the house looked like from the inside, the outside, and all 4 walls. As someone creating on a 2D space, I force the viewer to observe the piece in a certain way. In the 3D space I lose a lot of that ability. What I also liked was the ability to play back someone else's drawing. This places so much more emphasis on the process and observing someone else at work and making mistakes. I like that I can literally put myself in someone else's shoes and follow their actions step by step. To expand on the experience, I would want the ability to stop, slow, rewind, etc. in following someone else's process to really use it as an educational tool. I would also want the ability to look at something in the real world and try to draw it. For example, if I go to draw a mountain in the VR realm of Tilt Brush, I am only drawing the mountain from my memory, as opposed to drawing the mountain by looking at it.
The piece of VR that I liked the least was the limited sensory feedback to go with the very immersive visual environment. For example, I wanted to be feeling the textures, heat, and weight of something as I held it. I wanted to feel the swoosh in the water as the whale tale flew by me. When skiing down the mountain, I wanted to feel the pull of gravity as I flew off the cliff. The design team could create a room where you do VR to give you the illusion of some of these senses. For example, a room could have wind and/or rain functionality. A room could have different forces (ex: tipping side to side) to give the illusion of the various forces that the user is experiencing in the game. The user could also have gloves or a bodysuit that changes heat or somehow gives varying senses of texture. As shown in the video and discussed in the reading, physical blocks can be used in the physical world, paired with image manipulation to give the user the illusion that they are playing with multiple blocks and physically touching them, but actually only playing with a single block.
Uses a DC motor to spin a thread spool to put the thread on it. A pot can be used to adjust the speed of the spinning, similar to how someone would use a foot pedal of a sewing machine to adjust the speed of the thread spinning.
Lady using the rail intended for holding onto on public transport train to perform gymnastics.
When people are transporting between places, it takes time, but is not necessarily productive time. This lady is making use of the train structure and turning it into an urban gym. One potential solution here would be to replace the majority of the seats in the train with areas for people to do mini exercises. It would be an interesting experiment to see if such a layout of the train would be used. Sometimes people don't want to exercise in front of the general public, and usually people on the train are wearing street clothes as opposed to workout clothes. Maybe if exercise equipment was available on a train people would start wearing the appropriate clothes and the cultural norms would break down. All of these would have to be considered if introducing exercise equipment to the design of train cars.
Last year, Target had an Open House set up in its downtown San Francisco city store. The Open House was a very early stage prototype of what a smart home might look like. The coffee maker talked to the baby monitor, which talked to the lights, to the curtains, to your cell phone, etc. Everything in your home was smart, connected, and could communicate with each other. So, for example, the baby monitor would hear your baby cry in the morning, which would signal to the coffee maker to start brewing, which would signal to your alarm clock to wake you up. We've seen this type of functionality in movies, but Target was prototyping a physical space for potential users to explore. Instead of us, the users, seeing a video or a movie, we got to explore and experience the environment first hand.
One thing aspect of the smart home I was surprised by was these text bubbles that were projected on the walls with the conversations that were being passed from baby monitor to coffee maker to alarm clock. The texts were peripheral information for the human in the scene to pay attention to if they so choose, but their main purpose was to inform the user of what was going on and that devices were in motion/action. The text bubbles gave the illusion that the human was writing the messages themselves delegating and giving orders to the devices. While I didn't like the text bubbles on the wall because I found them unaesthetic and distracting, it did make me think about what types of information I would want to know about when devices are fully connected such that they can truly act as full personal assistants. Having a personal assistant that tells me everything they are doing is almost more distracting than me just doing all of the actions myself. But, having no visibility into actions that are going on between devices could potentially be dangerous or concerning depending on the device/situation. Of course, these thresholds of how much information would like to know/not know varies for each user. But, I thought this was such a great way to "provoke insights into important functional and emotional issues and inspire thoughts about how to deal with them" (Experience Prototyping, pg. 426). I don't think I would have had such a strong reaction and thoughts on the idea if I hadn't experienced it for myself.
When the cap of a bottle is screwed on, the bottle is not enchanted. But, when the lid of the bottle is unscrewed, the contents inside the bottle light up and change different colors!
Placing an FSR on a charcoal pencil, so that when I draw, the pressure I use in gripping the pencil maps to the color of a brush stroke on a processing visualization. The intention is to create a physical drawing, but also create a completly different digital drawing that reflects my process and maps to the same intensities I was using throughout the drawing. The next iteration will have an FSR on my eraser, also mapping to a specific color on the Processing image. This will show when I erase or backtrack in my work.
The Processing image is a spiral, meant to reflect the iterative and never finished aspects of drawing. Multiple slightly curved marks are used side by side to look like a brush stroke. Slight +/- randomness is put into the mark size to give a more organic effect. There is also a +/- randomness to the angle of each mark, so as the spiral is painted, it gives the effect of a brush moving along as some marks appear to be following other marks.
String portname = "/dev/cu.usbmodem1411"; // or "COM5"
float c = .1; //any constant. higher = radius gets larger faster.
float r = 0.0; //changes the trailing effect with the radians randomly going plus or minus.
Make lights blink faster or slower by turning one of the pots. Make the lights dim by turning one of the pots.
Fishkin's taxonomy does not capture TUIs where the input is remote from the output. For example, the Materiable TUI that we saw in class. In this TUI, the user gives input to the square pegs and then the output is reflected on the square pins, but in a remote location. Fishkin's taxonomy, on the embodiment axis includes 'distant', but the Materiable input/output doesn't fit in this category because the user conducting the input isn't shifting their gaze to the output at some distant location, but rather a second user is receiving the output in an un-seeable, remote location. I would add 'remote' to the embodiment scale in the taxonomy.
Additionally, Fishkin claims that TUIs that are at higher levels of embodiment and metaphor are 'more tangible'. Yet, a TUI can be multiple levels on the embodiment scale at once (ex: 'environmental' and 'full'), indicating that they the levels are not hierarchical. This flaws the logic of thinking about the two axes as scales. I agree that a TUI can be multiple of these levels, so I wouldn't change that in the taxonomy, but I would more simply talk about them as categories and a TUI can fall into multiple categories along one axis.


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Home Features Disadvantaged Canadian alpine teams embark on 'catch up' tour
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Disadvantaged Canadian alpine teams embark on 'catch up' tour
Gordie Bowles
The good news is, they're training — actually on snow, training. The bad? The game of "catch up" has just begun. It's like starting a best-of-seven NHL playoff series down a game or two. The series isn't over but the chips are stacked against you.
Canada's national alpine teams have been on snow in Saas Fe, Switzerland and Stelvio, Italy since late July with plans to move over to Zermatt with the speed team in September, picking up where they left off nearly four months earlier when the season was terminated due to the COVID-19 pandemic and travel restrictions.
I'm back, and I couldn't be happier!!🤩 Tellement heureuse d'être de retour sur mes skis!!
A post shared by Valérie Grenier (@valeriegrenier) on Jul 25, 2020 at 9:57am PDT
When the FIS recently announced it had moved up the World Cup openers in Soelden by a week, it put the Canadian alpine teams at a further disadvantage.
"We will definitely be behind the schedule but not to a place where I would say that we're way off track," said Phil McNichol, Alpine Canada's high performance alpine director.
The former U.S. ski team men's head coach with 42 World Cup victories during his tenure, McNichol is no stranger to adaptation and creative programming to achieve world-class results. And he has chosen the glass-half-full option in looking toward the season.
"We will need to work on training volumes and additional training opportunities but we should be pretty close. As things are playing out we will be right around 30 (days) going into Soelden now that it's been moved up. You're going to see Erik Read and others get into the start gate and they'll be behind many nations who were able to stay on their trajectory of around 60 days (on snow)."
Nothing quite compares to that early morning light on the glacier #CANSkiTeam #WeAreSkiing #MissionWealthAdvisors #WiegelePowder #SaasFee #Sunrise 📸: @twphilp
A post shared by Erik Read (@erik.read) on Aug 10, 2020 at 7:09am PDT
With the premature end to the 2019-20 season and the huge gap of no on-snow training, the Canadian teams may need to look at quality over quantity.
"From the outset we knew it was going to be a big challenge," he said. "Not only were we facing some daunting financial challenges at the end of the season, but everything was shut down. As we were adjusting and looking at what else could be done in Canada it became quite apparent that it wasn't feasible to get on snow in Canada and the window started to rapidly close."
After shifting focus to training options in Europe, McNichol and the coaching staff successfully negotiated a special exemption with the EU, which would allow for early admission, but prior to departure, the EU announced its opening. McNicol aims to keep the document "in our back pocket if things get wonky moving forward."
Revised race calendar the best route to fair play
It's no surprise that the FIS continues to make European-friendly decisions without accounting for the ski nations who are unable to train within its own borders. The last active ski glacier in Canada, the Horstman Glacier in Whistler, B.C., shut down permanently and the T-bar removed in July due to deteriorated and unsafe conditions caused from glacial melt. And with the borders to the U.S. closed, the popular option for provincial and club programs, Mount Hood, is no longer an option.
"I'm working hard with the NorAm committee to delay the NorAms this year," McNichol said. "But there's still some continental debate about early racing, but I feel relatively confident that the majority of the NorAm calendar will come out starting late January to April."
Morning view from the office. 🌄🙏 – – – Vue de notre bureau le matin. 🌄🙏 . . #CANskiteam #MorningView #GoodMorning #HappyToBeHere
A post shared by Alpine Canada Alpin (@alpinecanada) on Aug 13, 2020 at 9:12am PDT
McNichol is in the early stages of developing an integrated national alpine structure in Canada, including the Next Gen program, a roster of 12 up-and-coming skiers who will continue within the provincial ski team – or NCAA – systems.
"All of those athletes need to use the NorAm's as their focal point to matriculate forward in the national team structure and in their quest to make the World Cup. So that gives them some latitude to train in November and December and a good part of January to focus on the 25-30 NorAm starts."
According to McNichol, the Quebec team is training now in France, with Ontario, Alberta and B.C. provincial teams all preparing for a Europe training block soon.
'Level playing field' not a reality
When asked if it would be realistic for FIS to get more involved in regulating the rules of engagement, i.e., the number of days on snow, in an effort to create a level playing field, McNichol provided a thoughtful response to the inner workings of the FIS.
"[The FIS] always have two things at play: the economics and marketing that's driving things and then the athletic side, or what is better for the athlete in terms of development and preparation," he explained. "Athletically it would be far better if the World Cup started later when the whole world is ready."
McNichol believes the worldwide pandemic "shines a light" on some of the structural challenges within the FIS.
"People are thinking, 'Who's controlling this thing?' … haven't they noticed that some teams are training as in any normal year and then there are those on the sidelines? In general, the FIS operates in this amateurism. You look at the NHL, they are governed as professionals. FIS doesn't want to take on that responsibility or liability just as they don't want to be proactive with decisions right now as it's easier for them if each of the race organizers and countries who are hosting World Cup races take on that decision-making.
Missed this feeling #CANSkiTeam #WeAreSkiing #WiegelePowder #MissionWealthAdvisors #Norquay 📷: @bor_mat
A post shared by Erik Read (@erik.read) on Jul 27, 2020 at 11:13am PDT
"It would have been more of an NHL-style picture if the FIS said early on that they're going to push the season back, start in Val d'Isere in mid December and move the end of season to April 15. This would give everyone an extra month-and-a-half of training. We know that Norway is training, Sweden's training up north and we know that the Europeans fired their glaciers back up early. And in the U.S. they can organize some special stuff in Copper Mountain and use Mount Hood."
McNichol also identifies sourcing local on-snow glacier venue as an important path forward for the Canadian alpine structure.
"Our board is shifting some of their thinking and energy towards the exploration of Canadian glaciers. That need became very apparent in a country that has the most glaciers in the world and we're the only ones who can't ski (in the offseason).
"But developing a glacier opportunity is a big endeavor and we have a lot of work to get ready for Beijing and Cortina in the next two Olympic cycles, so we're going to have to do things that immediately impact us – and for me, the low hanging fruit is that we have to improve our possibilities in Whistler-Blackcomb, in Sunshine and Panorama, even in Sima [Yukon]. We need to turn over every stone with resorts big and small in how we can train as long as possible in the spring and as early as possible in the fall at home in Canada."
In the meantime, McNichol's half-full glass and creative leadership should help weather the storm.
"Passion and motivation are not easily taught. This situation has maybe taught us how to appreciate what we have and get the best out of it."
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Longtime editor, publisher, writer, producer & ski racing administrator and volunteer, Gordie Bowles grew up with the ski club in Fernie, B.C., and eventually joined the NCAA circuit with the University of Nevada, Reno, in the mid 1990s. Gordie lives in Vancouver managing a content marketing agency. Favourite places to ski: Fernie, Squaw Valley, Alta, Lake Louise, Whistler, Mount Bachelor.
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Scott Wilson August 14, 2020 At 2:01 pm
Best wishes Phil


The first hint request has arrived!
But, in the course of composing a suitably cryptic reply, I realized there's another technical suggestion that might make your life easier, if you're playing along.
The cookies are used only for the game. We never look at them. If you have your browser set to delete all cookies at the end of a session (as I do), then you'll have to retrace your steps each time you play. (That's what I did when we were beta-testing the game. So I've played it through, from beginning to end, more than once.) But if you remember what you've already done, recovering your work is very easy. Promise.


Brooke. . HD Wallpaper and background images in the Lipstick Jungle club tagged: brooke shields.
This Lipstick Jungle fan art contains diner jurk, diner toga, formele, avondjurk, formeel, and diner-jurk. There might also be japon, jurk, strapless, blote schouders, and balldress.


I also love that a guest book can often double as an activity at the event. If you' re planning a wedding, this is a must- have for your reception!
Just click on the images to be taken to the. We have the largest free bridal shower party games collection. Pdf template of the free printable votive candle wrap that you are interested in.
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A lot of times, we get the chance to talk with our customers about how they plan to use their guest book at their wedding, and lately we' ve been hearing that our books are a great vessel for wedding mad libs that have been filled out by lightful free printable baby shower games to play and entertain your guests while you host a party to celebrate the arrival of a new baby boy or girl.
Your guests will have so much fun playing these free printable baby shower games and they are easy to make! Simply print out the baby shower games and follow the instructions on how to make and play each ee download and free printable Azure Floral Wedding Invitations, introducing Azure floral pack made with watercolour shades of blue. Perfect for: wedding invitations, branding, logos, or.


The Mayfly, a beautiful, fragile-appearing, almost fairy-like insect with transparent wings, lives from six weeks to three years as an underwater stage and then emerges as an adult to live only one or two days. Because of this very brief life (as an adult) the Mayflies as a group are classified in the insect family Ephemeridae, which comes from the Greek word ephemeros (meaning daily or on a day or approximately a day) and the Greek word for the mayfly actually is "ephemeron". These beautiful creatures are not to be confused with the tiny black fly (also called mayfly) which comes out in hordes in the spring in northern temperate areas looking for mammalian blood.
Anything short-lived or with a short-term usefulness can be referred to as ephemeral. Years ago, probably in the early nineteenth century, collectors began referring to printed, paper material, which was never bound in the form of a book or pamphlet and which had a limited, short-term life, as "ephemera". This non-book printed material includes calendars, labels, bookplates, advertising cards, letters (correspondence), deeds & other documents, broadsides, posters, transportation time tables, newspapers, sheet music, stock certificates, matchbooks, business brochures, rewards of merit, tickets, invitations, greeting cards, playing cards, paper toys, etc. Decades ago the concept evolved to include most forms of pamphlets — especially those with short-term usefulness such as magazines, theater program & other event programs, almanacs, trade catalogs, promotional material, investment prospecti, annual reports of towns & businesses, comic books, pamphlet cookbooks, museum exhibition catalogs, etc.
Today, the only printed paper material, which seems to be excluded from the concept of ephemera is hard-bound books. Most collectors would include printing on cloth in the form of menus, tickets, broadsides and other similar uses as ephemera. Some ephemera collectors even include short-lived items printed on metal such as tobacco tins and license plates and on wood or plastic such as yard sticks or meter sticks.
A few collectors believe the term "ephemera" as used to describe printed material was coined in the 1970's, but one has only to look through early 20th century antiquarian book catalogues to see the use of the term to describe "non-books". In fact, we have a copy of a Catalogue issued by Goodspeeds Book Shop in Boston (when the business was located at 7 Ashburton Place) entitled "Ephemera Americana, A collection of manuscripts, printed and engraved documents, and small prints, illustrating phases of American business, social, and political life from 1638 to 1875" which, although the catalogue is not dated, is from the mid 1930's. This catalogue offers incredible ephemera at prices which would be considered a dream today.
Most of us would love to order all four lots and we would be very pleased to pay the prices asked for these interesting pieces of ephemera in the 1930's.
These "scraps" of paper are a historical record of a country or region's culture including every aspect of society such as business, social events, politics, entertainment, literature, art, music, food, drink — all aspects of everyday living!
The study of printing history — ephemera was produced prior to the advent of printing, during the incunable period of printing (the first decades), through the advent of various printing presses, during the use of woodcuts, wood engraving, copperplate engraving, lithography, stereotyping, steelplate engraving, chromolithography to modern offset and laser printing.
The study of design — the art work, illustrations, typefaces, color inks, types of paper and other media. One can study specific periods of style such as early Victorian, Art Nouveau, Art deco and follow the evolution of design in all forms of ephemera including the recent advent of the use of holograms on ephemera.
If you stop for a few minutes to think about the vast quantity and variety of modern ephemera that you have used during the past month, you would easily understand how ephemera is a record of everyday life. You have probably used newspapers, magazines, receipts from all sorts of purchase, tickets to a movie or concert, food packaging, slips to pick up processed photographs or laundry, receipts from banking, advertising brochures, grocery store coupons, fast-food paper place mats, posters or handbills you have read, mail sent and received, birthday cards, invitations to a friend's social event and the list goes on.
One can easily get caught up in the "frenzy" of collecting ephemera — people do collect modern ephemera such as place mats, cereal boxes, matchbooks, tickets, movie posters, and even restaurant doggie bags. Other collectors collect only antiquarian ephemera and often only in a very narrow area such as rewards of merit, advertising or trade cards, postcards, valentines, letterheads, labels, tickets, clipper ship cards, theater programs, etc. Many book collectors look for antiquarian ephemera related to their collecting interest(s) to supplement their collection and often end up building separate collections of ephemera. Collecting ephemera is very challenging and can be a rewarding hobby.
There are a number of ephemera collecting fields that have been pursued by many collectors for many years and have developed as separate, well-established hobbies: postcard collecting, stamp collecting, map collecting, autograph collecting, comic book collecting, baseball and other sportcard collecting are a few examples. These areas have developed self-supporting infrastructures and there are organizations and clubs and sales events or shows or swap-meets and exhibitions and although there is considerable overlap between the collectors in these established fields with ephemera collectors in general, there is also a large degree of separation. At general ephemera shows/exhibitions you will find some comic book dealers and some postcard dealers, but the bulk of activity remains away from the general ephemera events and is well-sustained within the established venues.
Although there are trade or advertising card collector, newspaper collector, and stock certificate collector associations and other similar organizations, the majority of these ephemera collectors are also very involved in general "paper" or ephemera events and organizations. In the remainder of this part we will provide links to some ephemera organizations, ephemera fairs/shows or exhibitions and examples of some dealers as well as web sites specializing in ephemera.


The capacity of bone marrow mesenchymal stromal cells (BMSCs) to be induced into chondrocytes has drawn much attention for cell-based cartilage repair. BMSCs represent a small proportion of cells of the bone marrow stromal compartment and, thus, culture expansion is a necessity for therapeutic use. However, there is no consensus on how BMSCs should be isolated nor expanded to maximize their chondrogenic potential. During embryonic development pluripotent stem cells differentiate into chondrocytes and form cartilage in a hypoxic microenvironment.
Freshly harvested human BMSCs were isolated and expanded from the aspirates of six donors, under either hypoxic conditions (3% O2) or normoxic conditions (21% O2). A colony-forming unit fibroblastic (Cfu-f) assay was used to determine the number of cell colonies developed from each donor. BMSCs at passage 2 (P2) were characterized by flow cytometry for the phenotypic expression of cell surface markers on mesenchymal stem cells. BMSCs at P2 were subsequently cultured in vitro as three-dimensional cell pellets in a defined serum-free chondrogenic medium under normoxic and hypoxic conditions. Chondrogenic differentiation of the BMSCs was characterized by biochemical and histological methods and by quantitative gene-expression analysis.
After 14 days of culture, the number of BMSC colonies developed under hypoxia was generally higher (8% to 38% depending on donor) than under normoxia. BMSCs were positive for the cell surface markers CD13, CD29, CD44, CD73, CD90, CD105 and CD151, and negative for CD34. Regardless of the oxygen tension during pellet culture, hypoxia-expanded BMSC pellets underwent a more robust chondrogenesis than normoxia-expanded BMSC pellets after three weeks of culture, as judged by increased glycosaminoglycan synthesis and Safranin O staining, along with increased mRNA expression of aggrecan, collagen II and Sox9. Hypoxic conditions enhanced the mRNA expression of hypoxia inducible factor-2 alpha (HIF-2α) but suppressed the mRNA expression of collagen X in BMSC pellet cultures regardless of the oxygen tension during BMSC isolation and propagation.
Taken together, our data demonstrate that isolation and expansion of BMSCs under hypoxic conditions augments the chondrogenic potential of BMSCs. This suggests that hypoxia-mediated isolation and expansion of BMSCs may improve clinical applications of BMSCs for cartilage repair.
Articular cartilage covers the end of long bones in articulating joints where it provides near frictionless movement. Unfortunately, articular cartilage has a very limited capacity to repair after injury. If left untreated, cartilage defects progressively lead to more extensive lesions and, ultimately, require joint arthroplasty. Cell-based strategies using culture expanded autologous chondrocytes from non-loading regions of articular cartilage are currently used to treat focal cartilage defects . However, there is some evidence of progressive degenerative changes in the joint using this technique . Furthermore, there is evidence that the matrix-forming capacity of expanded chondrocytes is compromised due to de-differentiation processes [3, 4]. Thus, there is interest in other cell sources for cartilage repair.
Adherent bone marrow stromal cells or bone marrow mesenchymal stromal cells (BMSCs) have received much interest for cartilage repair because of their multipotent capacity to differentiate into different cell types including chondrocytes [5–10]. While there has been much study related to the potential of BMSCs to form cartilaginous tissue, there has been a limited number of reports of the implantation of human BMSCs for cartilage repair . The reason for this is unclear but may be related to hypertrophic differentiation or the lack of a consensus on how human BMSCs are to be cultured for reproducible and optimal chondrogenic differentiation. Human BMSCs have been estimated to account for a mere 0.001% to 0.01% of the total bone marrow mononuclear cells (MNCs) in the stromal compartment of bone marrow [5, 12, 13]. Thus, in vitro culture expansion is a requisite for increasing cell numbers for research and clinical applications. Since the first published report of Friedenstein and co-workers , describing the isolation and expansion of an adherent and spindle-shaped population of cells from whole human bone marrow aspirates, little has changed in the methodology of isolation and expansion of BMSCs. While it is practiced within the art that human BMSCs are isolated after initial cell adherence to tissue culture plastic ware and subsequent cell expansion under normal mammalian conditions of air containing 21% oxygen tension (normoxia), there is increasing evidence that BMSCs are adapted to limiting metabolic conditions . In agreement with this observation, hypoxic (3% O2) conditions have been reported to favor the multi-potentiality of a subpopulation of human bone marrow stromal cells over osteogenic differentiation . Accordingly, human BMSCs showed enhanced proliferative activity under hypoxic (1.5% to 3% O2) conditions relative to normoxia [16–18]. While these studies demonstrated the effect of hypoxia on human BMSC expansion in vitro, the downstream effect of hypoxic conditions during isolation and expansion on human BMSCs' chondrogenic differentiation capacity was unexplored. D'Ippolito et al.  and Grayson et al.  investigated osteogenic differentiation of BMSCs after hypoxia mediated expansion, while the study of Martin-Rendon et al.  investigated chondrogenesis on commercially acquired human BMSCs that lacked initial isolation and cell expansion culture history prior to further cell propagation under normoxia and subsequent chondrogenic differentiation under normoxia and hypoxia for their studies; hypoxia enhanced BMSC chondrogenic differentiation potential.
Ovine BMSCs have been isolated and propagated under hypoxia (5% O2) and shown to increase in proliferation rate relative to cells expanded under normoxia. Moreover, it was demonstrated that ovine BMSCs isolated and expanded under hypoxic conditions and subsequent chondrogenic differentiation under normoxia displayed an enhanced chondrogenic phenotype compared to their counterparts after normoxia mediated isolation and expansion . However, it is not clear whether hypoxia isolated and propagated ovine BMSCs would have similarly displayed improved chondrogenesis had they been differentiated under hypoxic conditions.
While it has been shown that normoxia isolated and expanded non-human and human BMSCs undergo enhanced chondrogenic differentiation under hypoxic conditions [9, 17, 20], it is unknown whether human BMSCs isolated and expanded under hypoxia are predisposed towards improved subsequent chondrogenesis regardless of the oxygen tension. The study of Mueller et al. , showed that hypoxic conditions during expansion culture of human BMSCs resulted in improved chondrogenesis even under normoxic conditions. However, the study used BMSCs that had been initially isolated and expanded under normoxia prior to further culture expansion under hypoxia and subsequent chondrogenic differentiation culture under hypoxia and normoxia. Perhaps the improved chondrogenesis noted by Mueller et al. was not surprising because, during embryonic development, cartilage formation occurs when mesenchymal stem cells condense and differentiate into chondrocytes during an avascular period in a low oxygen tension microenvironment . Thus, oxygen tension is known to play a significant role in the fate of mesenchymal stem cells.
In this study, we investigated the effect of oxygen tension from the onset of human BMSCs isolation and expansion on subsequent in vitro chondrogenesis under normoxic and hypoxic conditions. We hypothesized that persistent hypoxic culture during human BMSC isolation and expansion enhances the chondrogenic differentiation capacity under normoxia and hypoxia by altering the expression of genes that facilitate chondrogenesis.
Typically, in vitro chondrogenesis of BMSCs is performed in 3D cell pellet culture, in order to mimic mesenchymal cell condensation during chondrogenesis in embryonic development, in the presence of serum free chondrogenic factors comprising TGF-β1 or -β3, dexamethasone, ascorbate and ITS+1 under normoxic conditions [6, 9, 10, 19, 22–25]. In this study, we adopted the pellet culture method to investigate in vitro chondrogenesis of both normoxia and hypoxia isolated and expanded human BMSCs. In vitro chondrogenesis was implemented under normoxic conditions only, in order to isolate the downstream effect of cell isolation and expansion under hypoxic conditions on BMSC chondrogenesis; several studies have reported that hypoxic culture conditions enhance chondrogenic differentiation [9, 20, 26–32].
Bone marrow aspirates were obtained from surgically discarded material after approval and a waiver of informed consent of the local ethical committee of the University of Alberta (Edmonton, Canada) during orthopedic procedures from the iliac crest of six donors (Table 1; one woman, 34 years old, and five men, 43 to 62 years old). The number of nucleated cells in the aspirates was determined by crystal violet nuclei staining and cell counting using a hemacytometer. Thereafter, 15 million mono-nucleated cells (MNCs) were seeded per 150 cm2 tissue culture flask. The culture medium was αMEM supplemented with 10% heat inactivated fetal bovine serum, penicillin-streptomycin, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), sodium pyruvate (all from Invitrogen, Mississauga, Ontario, Canada) and 5 ng/ml basic fibroblast growth factor (bFGF or FGF2; from Humanzyme, Medicorp Inc., Montreal, Quebec, Canada) in order to maintain pluri-potency . Nucleated cells were allowed to adhere and grow for seven days before the first media change under normoxia (ambient oxygen tension of 21%) or hypoxia (low oxygen tension of 3% O2) at 37°C in a humidified incubator with 5% CO2. Thereafter, the media were changed twice per week until 70% to 80% cell confluence was obtained. However, during feeds hypoxia cultivated cells experienced short periods (< 5 minutes) of re-oxygenation. These adherent bone marrow stromal cells (BMSCs) were detached using 0.05% trypsin-ethylenediaminetetraacetic acid (EDTA) (Sigma-Aldrich, Oakville, Canada) and expanded under normoxia or hypoxia until passage 2 prior to experimental use. The time taken from plating of nucleated cells (P0) to reach approximately 80% confluence at passage 2, before experimental use, varied three to four weeks and was donor and oxygen tension dependent. Both normoxia- and hypoxia-expanded BMSCs from the same donor were kept in culture for the same duration until experiments. The experimental set up for BMSC isolation, expansion and subsequent chondrogenic differentiation is illustrated in Figure 1.
Experimental set up for isolation, expansion and in vitro chondrogenesis of bone marrow mesenchymal stromal cells (BMSCs).
The proportion of adherent cell population within each bone marrow aspirate specimen was assessed by Cfu-f assay. Cfu-f was performed by plating 2.5 × 105 nucleated cells in 100 mm sterile petri dishes in triplicate (Becton Dickinson, Mississauga, Ontario, Canada). The cells were cultured as described before using αMEM supplemented with 10% heat inactivated fetal bovine serum, penicillin-streptomycin, HEPES, sodium pyruvate and 5 ng/ml FGF-2 for two weeks under normoxia or hypoxia. After the first week, the non-adherent cell population was removed by aspiration and culture media were replenished every three days until two weeks of culture. During feeds, hypoxia cultivated cells experienced a short period (< 5 minutes) of exposure to normal oxygen tension. After two weeks the developed cell colonies were visualized after fixing with 4% phosphate buffered formalin, washing by PBS and staining using 0.25% crystal violet solution (Sigma-Aldrich). The number of cell colonies developed was recorded as well as their respective diameters. Data were compared in a Student t-test and a significant difference between the two culture conditions was considered when P < 0.05.
All primary monoclonal antibodies used herein were directly conjugated antibodies to fluorescein isothiocyanate (mAb-FITC) or to phycoerythrin (mAb-PE). Antibodies were either from BD Pharmingen or Invitrogen-see Table 2. The cells were analyzed on a FACScan flow cytometer (Becton Dickinson) after detachment from the culture flask by 0.05% w/v trypsin-EDTA (Sigma). Staining buffer was prepared with PBS containing 1% w/v BSA (Sigma). Normoxia and hypoxia expanded BMSCs were re-suspended in 4°C cold staining buffer at 5 × 106/ml. The cells were dispensed into sample tubes (12 × 75 mm polystyrene round-bottom tubes, Becton Dickinson) in 20 μl aliquots and incubated for 15 minutes with the antibodies at 4°C. All incubations were implemented in 5 ml dilution tubes at room temperature in the dark and all washing steps were performed by a combination of centrifugation (400 g, 5 minutes) and aspiration of supernatant. Staining buffer (200 μl) was then added to the tubes and the cells were incubated for 10 additional minutes. After removal of the supernatant by centrifugation, cells were washed with PBS and kept cold before analysis by flow cytometry. Non-specific staining was assessed using relevant isotype controls. Single color immunofluorescence analysis for the different surface markers was performed with mAb-FITC and mAb-PE. Data acquisition was performed with Cellquest software (Becton Dickinson). FITC emission was measured at FL1 and PE at FL3. For each sample a region for live cells was defined, according to the Forward Scatter (FSC) and Side Scatter (SSC) signals, which excluded aggregated cells from the analysis. Data analysis was performed with Cyflogic version 1.2.1, Perthu Terho & ©CyFlo Ltd., Finland. For each surface marker analyzed, the percentage of positive cells and the level of marker expression were calculated. The percentage of positive cells was calculated as the percentage of cells having a measured fluorescence greater than that of 99.5% of the cells stained with each associated isotype control. Cells were considered positive for a surface marker when the percentage of positive cells for that surface marker was ≥ 6%. The level of expression of each marker was calculated as the ratio between geometric mean fluorescence intensity (MFI) of samples and that of the isotype control.
am, mouse; r, rat. BMSC, bone marrow mesenchymal stromal cells; CAT.#, catalogue number.
BMSCs at passage 2 (P2) were re-suspended in chondrogenic culture medium consisting of high glucose (D)MEM containing 4.5 mg/ml D-Glucose, 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, 100 mM HEPES buffer, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin, 0.29 mg/ml L-glutamine (all from Invitrogen) supplemented with 0.1 mM ascorbic acid 2-phosphate, 10-5 M dexamethasone, 1x ITS+1 premix (Sigma-Aldrich), and 10 ng/ml TGFβ1 (Humanzyme-Medicorp Inc.). For chondrogenesis in pellet culture, a total of 2.5 × 105 cells were spun in 1.5 ml sterile conical polypropylene microfuge tubes (Enzymax LLC, Kentucky, USA) at 1500 rpm for three minutes to form spherical cell pellets. The final volume of chondrogenic media was 250 μl per pellet. Medium change was performed three times a week. Hypoxia cultivated pellets experienced a short period (< 5 minutes) of exposure to normal oxygen tension during media changes. The pellets were cultured for three weeks under normal or low oxygen tension to allow appreciable matrix accumulation. Thereafter, the pellets were processed biochemically for glycosaminoglycan (GAG) and DNA contents, histologically, immunohistochemically and gene expression analysis via quantitative reverse transcription polymerase chain reaction (qRT-PCR) for cartilage specific matrix gene and proteins expression.
In vitro cultivated pellets were rinsed in PBS (Invitrogen) and were digested in proteinase K (1 mg/ml in 50 mM Tris with 1 mM EDTA, 1 mM iodoacetamide and 10 mg/ml pepstatin A; all from Sigma-Aldrich) for 16 hours at 56°C. The sulfated GAG content was measured by 1,9-dimethymethylene blue binding (Sigma-Aldrich) using chondroitin sulfate (Sigma-Aldrich) as standard. The DNA content was determined using the CyQuant cell proliferation assay kit (Invitrogen) with supplied bacteriophage λ DNA as standard. Statistical differences between test groups were evaluated by one-way analysis of variance (ANOVA) with Tukey's multiple comparison post-tests. Statistical analyses were performed using SPSS (version 18). A significant difference was considered when P < 0.05.
Tissues generated from the pellet cultures were fixed in 4% phosphate buffered formalin, processed into paraffin wax, sectioned at 5 μm and stained with 0.1% safranin O and counterstained with 1% fast green to reveal sulfated proteoglycan (GAG) matrix depositions. Other sections were probed with antibodies raised against collagen types I and II. Sections were treated with trypsin and then incubated with antibodies against collagen I (MAB3391, Millipore, Temecula, California, USA) or collagen II (II-II6B3 from Developmental Studies Hybridoma Bank at University of Iowa, USA). Immuno-localized antigens were visualized with goat anti-mouse IgG biotinylated secondary antibody (Dako Canada Inc, Mississauga, Ontario, Canada) and a streptavidin-horseradish peroxidase labeling kit with 3,3'-diaminobenzidine (Dako). Images were captured using an Omano OM159T biological trinocular microscope (Microscope Store, Virginia, USA) fitted with an Optixcam summit series 5MP digital camera and Optixcam software and assembled in Adobe Photoshop (Adobe Systems Inc. San Jose, USA).
Total RNA was extracted from pellets using Tri-Reagent (Sigma-Aldrich) after grinding with Molecular Grinding Resin (Geno Technology Inc. St Louis, USA) in combination with the use of an Aurum Total RNA Fatty and Fibrous Tissue Kit (Bio-Rad, Mississauga, Ontario, Canada) and after removal of contaminating genomic DNA from the pellets by DNase treatment. In order to mitigate changes in gene expression, the caps of pellet cultures were closed before removal from the low oxygen tension incubator and cell pellets were immediately transferred into Tri-Reagent. Total RNA (100 ng) in a 40 μl reaction was reverse transcribed to cDNA using GoScript reverse transcriptase (Fisher Scientific, Whitby, Ontario, Canada) primed in the presence of oligo dT primers (1 μg). Real-time quantitative polymerase chain reaction (qRT-PCR) was performed with a DNA Engine Opticon II Continuous Fluorescence Detection System (Bio-Rad) using hot start Taq and SYBR Green detection (Eurogentec North America Inc, San Diego, CA, USA). Primer sequences (Table 3) were taken from previously published work or were custom designed using the Primer Express software (Applied Biosystems, Foster City, California, USA) [23, 34, 35]. All primers were obtained from Invitrogen. Gene (mRNA) expression levels for each primer set were normalized to the expression level of human β-actin [9, 34, 36] by the 2-Δct method . Statistical differences between test groups were evaluated by one-way ANOVA with Tukey's multiple comparison post-tests. Statistical analyses were performed using SPSS (version 18). A significant difference was considered when P < 0.05.
All studies were implemented with stem cells propagated from bone marrow aspirates taken from surgical discards of patients undergoing routine orthopedic procedures after approval and a waiver of informed consent of the local ethical committee of the University of Alberta (Edmonton, Canada).
To investigate the colony forming characteristics of adherent BMSCs cultured under normoxia and hypoxia, the number and diameter of the cell colonies developed after 14 days of culture of bone marrow MNCs was evaluated (Figure 2). The number of adherent cell colonies developed was consistently higher under hypoxic conditions by 8% to 37% albeit with dependence on the donor (BM68 (13%), BM69 (9%), BM73 (14%), BM74 (37%), BM79 (8%) and BM84 (23%)) (Figure 2b). There was no significant difference between the measured diameters of the cell colonies under normoxia and hypoxia (Figure 2c).
(A) Representative photograph of colony forming unit fibroblastic (cfu-f) assay of passage 0 (P0) BMSC after 14 days of culture under normoxia or hypoxia. Bone marrow mono-nucleated cells (MNCs) were seeded into 100 mm petri dish in triplicate (n = 3) at 250,000 MNCs per dish. Culture media was α-MEM comprising 10% FBS and 5 ng/ml FGF-2. Media were not changed within the first seven days. Thereafter, the media were changed twice a week. Developed cell colonies were visualized by crystal violet staining. (B) The number of cell colonies developed in the Cfu-f assay as described in A. The data represent mean ± standard error of colony counts derived from six donors (n = 3). (C) The mean diameter of cell colonies developed in the Cfu-f assay as described in A. The data represents mean ± standard error of colony counts derived from six donor specimens. Not significant (n.s.) = P > 0.05 or * P < 0.05 in normoxia versus hypoxia, Student's t test. BMSCs, bone marrow mesenchymal stromal cells; FBS, fetal bovine serum; FGF-2, fibroblast growth factor-2; MEM, modified Eagle's medium.
To investigate the immuno-phenotype of the adherent population of BMSCs isolated and expanded under normoxia and hypoxia, we analyzed the cell surface molecule expression from three different donors: BM68, BM73 and BM84. A panel of cell surface markers that are typically used to characterize mesenchymal stem cells was selected [25, 38]. Both normoxia and hypoxia expanded BMSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105 and CD151. Both normoxia and hypoxia expanded BMSCs were negative for CD34. The positive cell surface markers expressed on the adherent BMSCs displayed a single fluorescence peak binding profile by flow cytometry (Figure 3a). However, quantitative expression levels (MFI) of cell surface markers were dependent on donors and on the oxygen tension during cell expansion. For example, the expression level of CD90 was consistently and significantly lowered in hypoxia-expanded BMSCs compared to normoxia-expanded BMSCs (Figures 3b, c, d and 3e).
Flow cytometry analysis of cell surface markers on passaged 2 (P2) BMSCs after culture in α-MEM comprising 10% FBS and 5 ng/ml FGF-2. (A) Representative fluorescence intensity histograms are presented showing expression of cell surface markers of normoxia- and hypoxia-expanded BMSCs at P2 of a 43-year-old man (BM74). (B-D) The mean fluorescence intensity (MFI) of cell surface markers on BMSCs from: (B) a 34-year old woman (BM68), (C) a 43-year-old man (BM74), (D) a 62-year-old man (BM84). (E) Represents the mean fluorescence intensity ± standard error of three donors; BM68, BM74 and BM84. *P < 0.05 in normoxia versus hypoxia, Student's t test. BMSCs, bone marrow mesenchymal stromal cells; FBS, fetal bovine serum; FGF-2, fibroblast growth factor-2; MEM, modified Eagle's medium.
Safranin-O staining for sulfated GAG matrix deposition of pellets derived from four donors (BM68, BM73, BM74 and BM84) showed that the pellets derived from hypoxia-expanded BMSCs stained more strongly and more uniformly throughout the pellet than their counterparts from normoxia-expanded BMSCs, albeit with dependence on the donor (Figure 4). Furthermore, most pellets derived from normoxia-expanded BMSCs increased in safranin O staining under hypoxia, while pellets derived from hypoxia-expanded BMSCs remained intensely stained with safranin O. Quantitative GAG matrix normalized to DNA content of the pellets was determined (Figures 4e, j, o and 4t). The pellets formed from hypoxia-expanded BMSCs had a 1.4 to 3.5 fold increased GAG per DNA content than their counterparts from normoxia-mediated expansion, depending on the donor, when pellets were cultivated under normoxia while the pellets derived from hypoxia-expanded BMSCs had a 1.2 to 7.0 fold enhancement in GAG per DNA content than pellets from normoxia-expanded BMSCs when cultured under hypoxic conditions. These results correlated with the more uniform and intense safranin O staining observations in the pellets derived from hypoxia cultivated BMSCs. In the pooled data set, the mean GAG per DNA content of the pellets from hypoxia-expanded BMSCs was 1.7-fold (P < 0.05) higher than their normoxia counterparts under normoxic condition while under hypoxic condition. There was a linear-inverse relationship between the GAG per DNA contents of pellets and age of BMSC donors under a specific culture condition (data not shown).
Paraffin wax 5 μm-section pellets were stained with safranin-O/fast green stain. All photomicrographs represent low (4x objectives) images. (A-D) 34-year-old woman (BM68), (F-I) 51-year-old man (BM73), (K-N) 43-year-old man (BM74) and (P-S) 62-year-old man (BM84). Scale bar is 100 μm. Biochemical assay of pellets for sulfated glycosaminoglycan (GAG) per DNA content. Pellets were cultured for three weeks in a defined serum-free chondrogenic medium under low (3%) or normal (21% O2) oxygen tension. Data are mean ± standard error of triplicates. (E) 34-year old woman (BM68), (J) 51-year-old man (BM73), (O) 43-year-old man (BM74) and (T) 62-year-old man (BM84). Data represent mean ± standard error of triplicate pellets; one-way ANOVA with Tukey's post hoc test: n.s. (not significant), * = P < 0.05, ** = P < 0.001 and *** = P < 0.0001. ANOVA, analysis of variance.
Quantitative RT-PCR was performed for gene expression analysis on pellets derived from four donors (BM68, BM73, BM74 and BM84) after three weeks of chondrogenic culture. The mean relative gene expression of aggrecan (AGG), collagen I (Col1a2), collagen II (Col2a1), collagen X (Col10a1), cartilage oligomeric matrix protein (COMP), HIF-1α and HIF-2α, Sox9, TGFβ1-receptor I (TGFβ1-RI) and TGFβ1-receptor II (TGFβ1-RII) in the pooled pellets is presented in Figures 5 and 6. The mRNA expression levels of Col2a1 (2.44-fold) and Sox9 (2-fold) were significantly higher in pellets formulated from hypoxia-expanded BMSCs than in pellets derived from normoxia-expanded BMSCs after culture under normoxic conditions (Figure 5). The mean mRNA expression of AGG was not significantly higher in the pellets derived from hypoxia isolated and expanded BMSCs under normoxic conditions when compared to those derived from normoxia-expanded BMSCs (Figure 5). Under hypoxic culture condition, the mean mRNA expression level of AGG and Col2a1 were 2-fold higher while the mRNA expression of Sox9 was 3-fold higher in pellets derived from hypoxia-expanded BMSCs compared to their counterparts from normoxia-expanded BMSCs (Figure 5). The mean mRNA expression level of Col1a2 was significantly higher in pellets formulated from normoxia expanded BMSCs after culture under normoxic conditions while the expression of Col10a1 and COMP were not significantly different between the pellets from normoxia- and hypoxia-expanded BMSCs (Figure 5). In contrast, under hypoxic conditions of pellet culture the expression of Col1a2, regardless of the oxygen tension during BMSC expansion, was not significantly different between pellets (Figure 5). The expression of COMP increased under hypoxic pellet culture conditions relative to expression in pellets cultivated undr normoxia. However, the increment was not stastically significant. Col10a1 decreased significantly under hypoxic conditions of pellet culture. Col10a1 expression declined by 2225-fold in pellets formed from normoxia-expanded BMSCs and by 43-fold in pellets derived from hypoxia-expanded BMSCs when compared to their respective expression under normoxic conditions (Figure 5).
(A) Real-time PCR analysis of cDNA derived from pellet cultures of normoxia- and hypoxia-expanded BMSCs. Pellet cultures were performed under normoxic or hypoxic conditions for three weeks in the presence of defined serum-free chondrogenic factors as described Materials and Methods text. Real-time PCR analysis was via SYBR Green detection. Presented data represent the mean ± standard error of pellets pooled from four donors (that is, BM68, BM73, BM74 and BM84) in triplicate (N = 4, n = 12 per experimental group). Data represent mean ± standard error of triplicate pellets per donor; one-way ANOVA with Tukey's post hoc test: n.s. (not significant), * = P < 0.05, ** = P < 0.001 and *** = P < 0.0001. Gene expression is presented as relative mRNA level normalized to mRNA expression of human β-actin; y-axis. ANOVA, analysis of variance; BMSCs, bone marrow mesenchymal stromal cells; PCR, polymerase chain reaction.
(A-H) Immuno-labeling of 5 μm-sections of pellets with anti-collagen types I and II. All photomicrographs represent low (4x objectives) images. Pellets were cultured for three weeks in chondrogenic medium under low (3%) or normal (21% O2) oxygen tension. Images were taken from pellets derived from BM73. Scale bar is 100 μm. (I) Real-time PCR analysis of cDNA derived from pellet cultures of normoxia- and hypoxia-expanded BMSCs. Pellet cultures were performed under normoxic or hypoxic conditions for three weeks in the presence of defined serum-free chondrogenic factors as described in the Materials and Methods text. Real-time PCR analysis was via SYBR Green detection. Presented data represent the mean ± standard error of pellets pooled from four donors (that is, BM68, BM73, BM74 and BM84) in triplicate (N = 4, n = 12 per experimental group). Data represent mean ± standard error of triplicate pellets; one-way ANOVA with Tukey's post hoc test: n.s. (not significant), * = P < 0.05 and *** = P < 0.0001. Gene expression is presented as relative mRNA level normalized to mRNA expression of human β-actin; y-axis. ANOVA, analysis of variance; BMSCs, bone marrow mesenchymal stromal cells; PCR, polymerase chain reaction.
Immunohistochemical staining (brown coloration) of pellets from the four donors confirmed that the pellets formulated from normoxia- and hypoxia-expanded BMSCs expressed types I and II collagens. Representative photomicrographs of pellets (from donor BM74) labeled with antibodies to collagen I and II are shown in Figure 6a-6h. The pellets derived from hypoxia-expanded BMSCs were strongly and uniformly labelled with anti-collagen type II (Figures 6f (under normoxia) and 6h (under hypoxia)). In contrast, the pellets formulated from normoxia-expanded BMSCs labeled only strongly with anti-collagen type II at the pellet periphery and weakly in the central core (Figures 6e (under normoxia) and 6 g (under hypoxia)). Collagen type I was uniformly distributed in all pellets regardless of the oxygen tension during pellet culture or whether the pellets were formed from normoxia or hypoxia-expanded BMSCs (Figure 6a-6d). Primary antibody and non-specific IgG controls showed no false positive staining (data not included).
TGFβ-RI and TGFβ-RII have been implicated in the reduced chondrogenic potential of adipose derived mesenchymal stem cells under standard chondrogenic culture conditions using TGF-β1 or TGF-β3 . We therefore investigated the expression of these receptor proteins in our pellets. The mRNA expression of TGFβ-RI and TGFβ-RII were statistically similar between the pellets derived from normoxia- and hypoxia-expanded BMSCs under normoxic conditions. However, under hypoxic conditions the expression of TGFβ-RI in pellets derived from hypoxia-expanded BMSCs were significantly up-regulated by 3.5-fold relative to its expression in pellets formed from normoxia-expanded BMSCs. Similarly, the mRNA expression of TGFβ-RII was increased under hypoxic condition in pellets derived from hypoxia-expanded BMSCs; however, the increment was not statistically significant relative to its expression in pellets formed from normoxia-expanded BMSCs (Figure 6i).
The cellular response to low oxygen tension in many mammalian cells is regulated by the transcriptional activity of HIF-1, a heterodimer of HIF-1α and HIF-1β . HIF-2, a heterodimer of HIF-2α and HIF-1β, has also been identified as a regulator of the response of mammalian cells to low oxygen tension [41, 42]. In addition, both HIF-1 and HIF-2 have been implicated in hypoxia-mediated enhancement of chondrogenesis of mesenchymal cells [29, 30, 41–43]. Thus, we investigated the gene expression of HIF-1α and HIF-2α in our study pellets after cultivation under normoxic and hypoxic conditions. The mRNA expression level of HIF-1α was similar in pellets regardless of whether the pellets were formed from normoxia- or hypoxia-expanded BMSC or cultivation of pellets under normoxic or hypoxic conditions (Figure 6i). The expression of HIF-2α was the same in pellets derived from normoxia- and hypoxia-expanded BMSCs under normal oxygen tension. However, under low oxygen tension the expression of HIF-2α increased regardless of whether the pellets were derived from normoxia- or hypoxia-expanded BMSCs. The expression of HIF-2α increased significantly by 3.2-fold in pellets derived from hypoxia-expanded BMSCs when compared to its expression in pellets under normoxic conditions. In contrast, the increment determined (1.6-fold) in pellets derived from normoxia-expanded BMSCs under hypoxic conditions relative to its expression in the same pellets under normoxic conditions was not statistically significant (Figure 6i).
In this study we have compared the chondrogenic potential of human BMSCs obtained under low (3%) and normal (21%) oxygen tension. The BMSCs were isolated via plastic adherence and cell culture mediated propagation. We have used flow cytometry to characterize the cell surface protein expression of the BMSCs and used an in vitro pellet model of chondrogenesis with TGF-β1 to investigate their capacity to undergo chondrogenic differentiation.
Our data showed that the BMSCs obtained after plating of bone marrow nucleated cells and subsequent cell culture propagation under low oxygen tension expressed a panel of conventionally used mesenchymal stem cell surface markers but with a consistently reduced concentration (that is, reduced mean fluorescence intensity) of CD90. The BMSCs obtained via propagation under low oxygen tension underwent a more robust chondrogenesis than their counterparts under normal oxygen tension albeit with dependence on the donor. The reduced CD90 expression in the BMSCs harvested under hypoxia may be associated with improved chondrogenesis. A link between CD90 expression and chondrogenic potential of BMSCs has not been reported in the literature. However, the lack of CD90 expression in in vitro expanded human articular chondrocytes has been observed with chondrocytes with higher chondrogenic potential . Thus, there is reason to speculate on a potential link between its expression and the chondrogenic capacity of BMSCs. Furthermore, in vitro expanded human articular chondrocytes have also been reported to display plasticity features that are similar to those of mesenchymal stem cells . Regardless of the oxygen tension during pellet culture, the BMSCs isolated and propagated under hypoxic conditions displayed a superior chondrogenic capacity than their counterparts from normal oxygen tension. The superior chondrogenic capacity was characterized by higher GAG per DNA content, enhanced transcript expression of a panel of chondrogenic genes (aggrecan, Col2a1 and Sox9) as well as the intense and uniform distribution of collagen II and safranin O staining for sulfated proteoglycans. Our data mirrors the findings of Tew et al. , that demonstrated that pellets derived from de-differentiated and Sox9 transduced articular chondrocytes displayed a phenotype that was consistent with a more robust chondrogenic response (increased safranin O staining and Col2a1 expression) compared to control pellets formed from non-transduced and de-differentiated articular chondrocytes. Furthermore, our data of increased mRNA expression of Sox9 and aggrecan supports previously reported evidence that transcriptional activity of Sox9 enhances the gene promoter activities of aggrecan in chondrocytes [46–49]. Transcriptional factors Sox9, L-Sox5 and Sox6 have been reported to be essential for regulating the expression of Col2a1 as well as other genes involved in chondrogenesis [48, 50, 51].
The chondrogenic capacity of all pellets in our study improved further under low oxygen tension. The improvement was characterized by a marginal (that is, non-statistically significant) increase in transcript expression of aggrecan, Col2a1 and COMP. Furthermore, and more significantly, the improvement in chondrogenic potential under hypoxia was accompanied by a concomitant suppression in Col10a1 expression, a marker of hypertrophic chondrogenesis and terminally differentiated chondrocytes. This finding is in accordance with the observation of Hirao et al. that hypoxia (that is, 5% O2) supports commitment of C3HT10T1/2 (a pluripotent mesenchymal cell line) to chondrogenic differentiation rather than osteogenesis through mechanisms involving a concomitant down-regulation of Col10a1 and Runx2 activity via Smad suppression and histone deactylase 4 activation (HDAC4) .
Our results indicate that the response of BMSCs to hypoxic conditions involves up-regulation of the transcriptional expression of HIF-2α rather than HIF-1α, which remained unperturbed. This finding was surprising since hypoxia has been reported to induce chondrocyte-specific gene expression (aggrecan, Col2a and Sox9) in mesenchymal cells (mouse ST2 stromal cells and C3HT10T1/2) through transcriptional activity of HIF-1α . Furthermore, HIF-1α, has been implicated in hypoxia-mediated inhibition of senescence and maintenance of human mesenchymal stem cell properties . Our finding, therefore suggests that the response of the BMSCs to hypoxia was mediated by the transcriptional activity of HIF-2α. The involvement of HIF-2α here is in accordance with a report that hypoxia (that is, 5% O2) enhances the expression of several chondrogenic genes including Col2a1, aggrecan and Sox9 during chondrogenesis of adipose derived mesenchymal stem cells . However, in contrast to the hypoxia-mediated down-regulation of Col10a1 noted in our present study, hypoxia (5% O2) enhanced the expression of Col10a1 during the chondrogenic differentiation of adipose derived mesenchymal stem cells . A plausible explanation for the differential outcome may depend on the difference in oxygen tension or cell type. Nonetheless, our data opens the perspective of a possible mechanistic link between the transcriptional activity of HIF-2α and collagen X expression.
TGFβ-RI and TGFβ-RII have been implicated in the reduced chondrogenic potential of adipose derived mesenchymal stem cells relative to the chondrogenic potential of BMSCs. Thus, we investigated the expression of these receptor proteins in our BMSCs pellets. Our data showed that the expression of these receptors was generally higher in pellets derived from hypoxia-expanded BMSCs after culture in hypoxic conditions. However, it was surprising that the expression of TGFβ-RI was statistically higher in pellets formulated from hypoxia-expanded BMSCs during pellet culture under hypoxic conditions.
We have shown that the isolation and expansion of BMSCs under hypoxic conditions of 3% oxygen tension increases the propensity of the BMSCs to undergo a more robust chondrogenesis under normoxic and hypoxic culture conditions relative to BMSCs isolated and propagated under normoxic conditions. Our results also show that the response of these cells to low oxygen tension is mediated by HIF-2α. Taken together, our finding highlights the need to isolate and propagate BMSCs under hypoxic conditions for improved in vitro chondrogenesis and for in vivo cartilage repair and/or regeneration. The outcome of this study suggests that oxygen tension is an important factor in the determination of the chondrogenic differentiation fate of adherent populations of BMSCs.
We would like to thank Dr Thomas Churchill and Jacek Studzinski (Department of Surgery, University of Alberta, Canada) for histological assistance. Financial support was provided in part by: Edmonton Orthopaedic Research Committee, University of Alberta Hospital Foundation to AA, Edmonton Civic Employee's Charitable Fund to AA and new investigator startup fund by Department of Surgery, University of Alberta to AA.
transforming growth factor β receptor II.
ABA conceived and designed the study, performed experiments, data acquisition and analysis, manuscript writing and supervision of the entire study. AMS performed experiments and data acquisition. NMJ was responsible for bone marrow procurement, data analysis and manuscript writing. All authors read and approved the final manuscript.


Since God is loving and in control of all things, why does he allow bad things to happen?
Why doesn't God get rid of sin and only create good people?
By nature, we are all anti-God. In fact, even people whom we respect and adore the most on earth have the capacity to do the greatest type of evil. So, why does God allow this world to continue in its present sinful condition?
If God created humans with the capacity for doing evil, was it worth the risk?
God's creation of the world was "risky" in that it involved him becoming human flesh and dying a horrible death on the cross. He loved the world so much that he was willing to take upon himself all the pain, rejection and the consequences sin produces so that we may receive an eternal relationship with him. Jesus died on the cross so humans could live eternally in the peace and joy of God (heaven).
I just got home from Wal-Mart where my wife and I bought a new 18-speed bicycle for my daughter.
Have you ever experienced fear in sharing the message of Christ? Do you battle with guilt from missed opportunities to share our faith? A neighbor makes a flippant comment about the Christian faith and we are tongue-tied. A conversation about Jesus Christ around the water-cooler makes us nervous and awkward. Knowing eyes turn toward us for a response and we only mumble a feeble reply. Mormon missionaries knock on our door at just the wrong time. We're tired, we're busy and we make excuses, hoping they will go away. Doesn't this sound familiar? We get nervous, fearful, and struggle with feelings of inadequacy and the guilt of being unprepared. How do we overcome this?


text:
MLA apologises for Facebook comment which appeared to compare immigrants to dogs
Jenny Palmer MLA
Published: 09:55 Tuesday 06 September 2016
An MLA has apologised for an online comment in which she appeared to liken refugees to dogs.
UUP Lagan Valley MLA Jenny Palmer had denied her remark had been racist, although she said that it "could easily have been misinterpreted", and pledged to give more thought to the language which she uses on the internet in the future.
The controversy reportedly began on Facebook, when one user of the social media service had questioned why "refugees" are not housed in the home of an MLA.
Mrs Palmer had then responded by saying that "I have two dogs, don't need any more thank you".
Her reply ended with the letters "lol" – slang for "laugh out loud".
Shortly afterwards, in response to a web user who criticised her, Mrs Palmer said there had been a misunderstanding about what she meant.
The offending remark was reportedly made at the weekend.
Speaking on the BBC's Nolan Show on Tuesday, commentator Malachi O'Docherty said that the existence of the refugee crisis itself is an "obscenity greater than the obscenity of her [Jenny Palmer's] joke".
However, he added: "For a politician to show so little sympathy and so little understanding of one of the major crises of our time that she can be that glib about it, I think pretty much disqualifies her from being taking seriously as a politician at all."
In a statement, she said: "I would like first of all to apologise for any hurt that my careless words may have caused.
"These comments and actions do not reflect my own views.
"My words, made in reply to a Facebook comment, were intended as a joking remark regarding the busy-ness of my own home.
"The intention was to imply that between myself, my husband, our dogs and my daughter, we would struggle to accommodate anyone else.
"This was meant as a simple joke. However, as a later comment pointed out, my reply could easily have been misinterpreted as a racist or xenophobic comment.
"Particularly if taken without context or knowledge of my home.
"As soon as I was made aware of the offensive nature of the comment, I rectified my actions.
"I would again stress that those words do not reflect my viewpoint whatsoever. My actions and comment were genuine mistakes.
" While these actions were mistakes, I take responsibility for them.
"As an elected official, I must hold myself to higher standards.
"I apologise unambiguously and will endeavour to ensure that my social media activity is more carefully considered in future."
In September last year, then-Prime Minister David Cameron agreed to help resettle 20,000 Syrian refugees in Northern Ireland following calls from charitable groups and others.
The UN refugee agency has reported that, at time of writing, more than 289,000 people had arrived in Europe by sea so far this year – with by far the biggest group (30%) from Syria.
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text:
St. Paul Fire Department welcomes 15 new firefighters
Brett Hoffland
Updated: January 14, 2022 07:39 PM
Created: January 14, 2022 05:30 PM
More than a dozen new firefighters have officially joined the St. Paul's Fire Department.
The virtual ceremony took place Friday afternoon at the Wellstone Center.
It's a day that some of the graduates have waited for since they were little.
"I remember being 4 years old and wanting to pursue being a firefighter," said Austin Johnson, a fire academy graduate.
KSTP
Together 15 men and women make up the newest St. Paul firefighters, including Sergio Sahlstrom, a former a bodybuilder and personal trainer.
"In personal training, it's all about helping somebody achieve their goals, and throughout my career I always felt there was something bigger I could do to help others," Sahlstrom said.
Even though it might not sound like it, Sahlstrom thinks the two careers have similarities.
"You have to be able to talk to people and I think that's definitely going to attribute to my work as a firefighter," Sahlstrom said.
Johnson is the son of a St. Paul firefighter.
"We definitely will see each other at work," Johnson said. "I just remember as a kid spending Thanksgivings at the stations when he has to work or Christmas Eves even and just falling in love with it."
Cidney Ellis was one of the department's first fire medic cadets.
"It's actually something I've always wanted to do as a kid, it just took me a long time to get here," Ellis said.
During Friday's virtual ceremony, Chief Butch Inks talked about the hiring challenges brought on by the pandemic and civil unrest. But he commended these men and women for staying committed and answering the call after 14 weeks of vigorous training.
"In the midst of everything going on in the world, this group said yes to this job," Inks said.
Now the real work begins, and this group is ready to join the family that makes up the St. Paul Fire Department.
"I'm excited to grow our family and give back to the people of St. Paul," Ellis said.
Copyright 2022 - KSTP-TV, LLC A Hubbard Broadcasting Company
As one Lake Street business closes, others ready to ride out 'rebuilding phase'


Understanding the life of Jesus is a necessary component to comprehending the message he proclaimed.
"Understanding the land of Jesus is a necessary component to comprehending the message he proclaimed. From the beginning of the four Gospels until their end, the Evangelists assume that we possess an intimate knowledge of the historical and geographical stage onto which Jesus stepped.
For most Christian readers this is unfortunately not true. Many have not had the opportunity to visit the Holy Land. Even for those who have, it can prove to be a confusing experience. Much about life in this land has changed over the course of two millennia.
Happily, however, the contours of the land, which influenced where people settled and the routes they travelled, have remained the same. With some basic knowledge of the physical settings we can once again trace Jesus' steps.
The Growth of Herod's Kingdom - 10 to 4 B.C.
The Church in the First Century A.D.
About the Authors: Michael Avi-Yonah (1904-74) was Professor of Archaeology and History of Art at The Hebrew University of Jerusalem. He wrote numerous books and papers, among them In the Time of Rome and Byzantium, A Histoical Geography of the Holy Land from the Return to Zion until the Arab Conquest, and Gazetteer of Roman Palestine. He planned and contructed a model of Second Temple Jerusalem at 1:50 scale, now on display at the Israel Museum, Jerusalem.
R. Steven Notley is Distinguished Professor of New Testament and Christian Origins on the New York camput of Nyack College and director of its graduate programs in Ancient Judaism and Christian Origins. He is at the cutting edge of modern New Testament research, combining his philological training with an intimate firsthand knowledge of biblical geography.


text:
Merrick Garland's nomination to be attorney general advances to full Senate
Published Mon, Mar 1 20211:23 PM EST Updated Mon, Mar 1 20213:15 PM EST
Tucker Higgins@in/tucker-higgins-5b162295/@tuckerhiggins
The Senate Judiciary Committee advanced federal Judge Merrick Garland's nomination to be attorney general to the full Senate on Monday, paving the way for his confirmation to head the Department of Justice.
Garland's hearings before the committee last week focused on the sprawling investigation into the Jan. 6 riot at the U.S. Capitol.
The largely cordial question-and-answer session indicated that the nomination of the centrist former Supreme Court nominee was likely to be approved on a bipartisan basis. It was, 15-7.
Attorney General nominee Merrick Garland testifies during his confirmation hearing before the Senate Judiciary Committee, Washington, DC, February 22, 2021.
Drew Angerer | Pool | Reuters
The Senate Judiciary Committee on Monday advanced federal appeals court Judge Merrick Garland's nomination to be attorney general to the full Senate, paving the way for his confirmation to head the Department of Justice.
The bipartisan vote was 15-7. A full Senate vote has not been scheduled but could come early this week.
Garland's hearings before the committee last week focused on the sprawling investigation into the Jan. 6 riot at the U.S. Capitol. The largely cordial question-and-answer session indicated that the nomination of the centrist former Supreme Court nominee was likely to be approved on a bipartisan basis.
The nominee is expected to be crucial to accomplishing President Joe Biden's agenda on a number of fronts, particularly with regard to civil rights and criminal justice reform, but also on antitrust enforcement, climate change progress, financial regulation and other areas.
The attorney general is seventh in line of presidential succession, after the Defense secretary.
Garland is known to the public largely as the face of Senate Republicans' successful bid to stonewall former President Barack Obama's effort to replace Justice Antonin Scalia on the nation's highest court.
In 2016, Obama nominated Garland, a longtime judge on the D.C. Circuit Court of Appeals, to fill the vacancy, but Garland never received a hearing for the role in the GOP-dominated chamber of Congress. Republicans kept the seat open until President Donald Trump nominated Neil Gorsuch in 2017.
At his hearings last week, Garland pledged to remain independent from Biden when it comes to the Justice Department's investigations and prosecutions. Garland said he had not spoken to the president about an investigation into Biden's son, Hunter Biden, related to the younger man's tax affairs.
Garland also spoke about his role overseeing the prosecutions that stemmed from the 1995 Oklahoma City far-right terrorist bombing, saying he would draw from the experience as he oversaw the investigations into the riot at the Capitol. Garland said the probe into the attack would be his No. 1 priority.
"We begin with the people on the ground, and we work our way up to those who were involved and further involved," Garland told senators.
The hearings foreshadowed what could be more difficult processes for other Justice Department nominees.
Sen. Mike Lee, R-Utah, targeted Kristen Clarke, who has been nominated to lead the Justice Department's civil rights division, and Vanita Gupta, whom Biden picked to hold the No. 3 spot in the department as associate attorney general.
Clarke and Gupta have both been the subject of attack by conservative groups.
Clarke is under fire for a letter she co-wrote that was published in The Harvard Crimson in 1994 in which she said that Black people were in some ways physically superior to white people. She has since said the comments were designed to show the absurdity of racist views against Black people. Clarke is the former president of the Lawyers' Committee for Civil Rights Under Law.
Gupta has been targeted by the conservative Judicial Crisis Network, which has accused Gupta, without evidence, of wanting to defund the police. Those arguments were largely defanged, however, after major policing groups, including the Fraternal Order of Police, came out in support of the civil rights leader last month.
"Vanita Gupta has always worked with us to find common ground even when that seemed impossible," the national FOP, which twice endorsed former President Donald Trump, said in a statement.
Garland defended both women at his own hearing, telling Lee, "I have complete faith in them."
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We are bunch of Bioinformatics enthusiasts [Sharib, Rajeev, Kamal, Dhwani and Animesh] from JNU Bioinformatics program [Advanced post graduate diploma in Bioinformatics], 2001 Batch. We have created this group to disperse information amongst us and the community at large. Any research group who is interested in having a working knowledge of tools and algorithms used presently in the Bioinformatics research community [Molecular and Medical] can contact us. Generating money is not the agenda, just gaining and sharing knowledge, is the motto :) [Though the requester has to bear the minimum expenses like travel, stay and food]. Our email addresses are our-names suffixed with@fuzzylife.org .


In 3 1/2 or 4-quart slow cooker, combine chicken, beans, onion, garlic, oregano, salt, soup and water; mix well.
Cover; cook on low setting for 8 to 10 hours or until beans are tender and chicken is no longer pink.
Just before serving, stir in cumin, hot pepper sauce and chiles. Serve with additional hot sauce if desired. Garnish with sage.
Tip: This recipe was tested in slow cookers with heating elements in the side and bottom of the cooker, not in cookers that stand only on a heated base. For slow cookers with just a heated base, follow the manufacturer's directions for layering ingredients and choosing a temperature.