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March 14-21, 2020 // Yoga Farm, Costa Rica
March 14-21, 2020 // Yoga Farm, Costa Rica
Join Erica Hartnick, Alex Lanau, and the Yoga Farm Family for 7 days and nights of Buena Vibra: nutrient dense living to nourish the soul and awaken the senses! Celebrate Flavors, Form, and Friends as we explore these realms as the foundations of wellness. The Yoga Farm is a rustic off-the-grid yoga center and sustainable living project located on Costa Rica's southernmost Pacific coast. Set amidst beautiful tropical rainforest, overlooking open ocean, and one of the most biologically diverse places in the world, it is an ideal space for those looking to reconnect with mind, body, and nature. Passionate humans (including the Yoga Trade Founders) and incredible flora and fauna inhabit this magical land. Gather for a week of delicious good vibes! Let's share our wisdom and grow together!
This event is for you if…
6:30-8am Yoga/Movement/Meditation
8am BREAKFAST
9-10:30am Workshop/Activity
12pm LUNCH
2:30-4pm Yoga/Workshop/Activity
6:30pm DINNER
7:30-9pm Restorative/Music/Relax
-7 days/nights shared accommodation at the Yoga Farm
-3 tasty and nutritious vegetarian farm fresh meals a day
-Daily yoga classes, workshops, and scheduled activities
-Beautiful remote beaches and rainforest property to explore
-Network with an amazing community and create new friendships
-Functional knowledge, new recipes, and community building skills
-Continuing Education hours with Yoga Alliance
Check out more details and travel info HERE
$750 Shared Accommodation
$900 Private Accommodation
***Discounts Available to Residentes Centroamericanos
Constantly curious and always exploring, Alex has been the head chef at Pavones Yoga Center, spreading the gospel of nutrient dense foods all day, everyday. Creating simple and inventive plant based meals as a reminder that cooking is not something to be intimidated by. With his practical approach and attention to detail, his edible art has converted countless carnivores to the dark side of the leafy greens. When he is not cooking for hungry yogis in various parts of the world, he is painting murals, eye gazing the sun and sea, and hanging out with his four legged furry son.
Workshops & Wisdom:
Erica spends most her days in California and Costa Rica and enjoys all things wild and free. She is grateful for having the opportunity to explore both the inner and outer worlds. She has worked at a Heli Ski Operation in Alaska, taught yoga on a remote island in the Indian Ocean, and solo backpacked the Tahoe Rim Trail. She is the co founder of Yoga Trade, a RYT 500 and a Continuing Education Provider. Her passion lies in sharing natural beauty with others and inspiring people to choose health, happiness, and peace.
Workshops & Wisdom:


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Trouble with the Curve (2012)
Genres : Drama
Plot Summary : Gus is a baseball scout also regard problems. He can't imagine the baseball reconsider marital plate. His female offspring Mickey wishes to aid plus as a result vegetation her business to press on a taking trip, which may perhaps troth his final if Gus can't style the exact call.

subset: commoncrawl


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Start chatting to our centre now by clicking the button above
Welcome to Williams Stockport BMW. How can we help?COVID-19: SHOWROOMS NOW OPEN
Our Genius is here talk to you about anything BMW, without any sales talk. Whether you already own a BMW, or think you might like to, our Genius is on-hand to ensure your vehicle suits your needs and lifestyle.
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Search our range of new BMWs in stock. Select your preferred model and then confirm your budget to browse the vehicles that match.


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Kuwait Markaz to launch US$171 million energy fund
Markaz Energy Fund to focus on Kuwait, but will also serve wider GCC region and beyond.
Kuwait Financial Centre (Markaz) is to launch the Markaz Energy Fund, a KD 50 million (US$171 million) private equity fund focused on the energy sector of the GCC and neighbouring countries.
The fund will focus mainly on Kuwait because of the unique and promising investment opportunities stemming from huge growth potential in that market. Markaz, which is currently sponsoring and participating in the third Major New Projects Opportunities conference in Qatar, has submitted the fund for approval to the Ministry of Commerce and Industry and the Central Bank of Kuwait, and plans to commence the placement process once all regulatory approvals are obtained.
Manaf Al Hajeri, general manager of Markaz, said: "Markaz Energy Fund has been established to capitalise on the structural changes in the oil and gas industry of the Arab region". He added, "The region will be required to invest approximately US$800 billion over the next 25 years in the upstream and downstream segments of the energy sector to sustain and grow its production capacity. Multinationals are expected to play a larger role in contributing the necessary technical and financial resources to the energy sector; which is expected to lead to strong demand for local/regional contractors, subcontractors, and service companies".
Global energy demand is expected to grow by 1.7% per annum over the next 25 years, driven by the rapidly growing economies of Asia. Arab states' share of world production of crude oil is expected to increase from 26% to 43% by 2030; and their share of world production of natural gas is expected to rise from 9% to 13% by 2030.
Kuwait, the main target market of the fund, is likely to emerge as a main beneficiary of the structural change in the regional energy sector. Markaz believes that Kuwait's political stability, its strategic geographic location between the major oil production centres of Saudi Arabia, Iran and Iraq, and its existing contracting and manufacturing capabilities will enable it to gain a leading role in the development of the sector. This is evidenced by KPC's initiative to double its production capacity over the next 15 years, and expand its refining and downstream processing capabilities through the participation of the private sector.
The Markaz Energy Fund will invest in contracting, existing operating companies and service companies that have the vision, but need the capital to acquire equipment, technology and expertise to benefit from such change. The fund will also enter into strategic alliances with companies to enhance the fund's access to transactions and create synergy with the fund's portfolio of companies.
The Markaz Energy Fund will play an active role in becoming a primary conduit for financial investor to have access to one of the most vibrant and growing sectors in the Region; a sector in which companies and investment opportunities were limited in the past, according to a Markaz press statement.
Markaz has appointed experts from the regional oil companies as technical advisors who will be instrumental generating and evaluating potential investment opportunities for the Fund.
Markaz, one of the largest investment companies in Kuwait, was established in 1974 and is listed on the Kuwait Stock Exchange. It currently manages over US$ 2.5 billion through its various funds investing in money market instruments, private equities, local and international listed equities and real estate. It has one of the leading investment banking operations in Kuwait, and has recently lead several bond and equity issues in 2004 including Kuwait Gulf Link Port International, HumanSoft, Kuwait Privatisation Projects Companies, and Amar among others.


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Broken glass on Galaxy S8+
Hi, my phone still works fine, but the only issue is that my screen is black. I can only find tutorials that require I get a screen with a digitizer, but is there any way to only replace the screen.
Ist dies eine gute Frage?


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Solo Pianist.
Evocative, original Piano music.
New Single Out Now!
"An inspiring album, Melany Thompson's "Memories of Home" brings together eleven original compositions of impressive skill and expansive sound."
-Tom, Broken Records
About Melany
Melany started piano lessons at the age of four, and spent many of her early childhood years performing in music eisteddfods and recitals.
At the age of 11, Melany performed at the Sydney Opera House in a concert for young performers, but after completing her AMEB 6th grade exam at the age of 12, Melany put formal piano lessons on hold to pursue other sporting and creative activities.
Preferring to play for leisure, Melany began composing her own music a few years later. Although she studied music at school and at university for a short time, Melany did not pursue a formal career in music, however it remained a passionate hobby.
A corporate career in marketing and communications, marriage and two kids later, Melany has returned to her passion and fulfilled her dream to record an album of her original music.
Memories of Home is a collection of peaceful and evocative piano pieces, recorded at Underwood Studio in the picturesque Blue Mountains, where Melany grew up. She dedicates the album to her biggest fans – her loving family who have always supported her and provided much inspiration for her songs.
Melany hopes to now inspire her own two children through music. "I want my children to grow up knowing they don't have to wait for permission or for someone to tell them they are good enough before they follow their dreams."
Timeless – The Wedding Song


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Reynolds Gallery is pleased to announce the opening of an exhibition of recent paintings by Ray Kass entitled Oil and Water. The exhibition opens with a reception for the artist on Friday, November 6 and continues through December 23, 2015.
For over thirty years, Ray Kass has derived his muse from nature—the landscape and the garden—and his newest works invoke the same elemental urge to explore the essence of the natural world. Kass's paintings in Oil and Water convey the spiritual and conceptual tendency of nature, inspired by the artist's travels and surroundings. Layering washes of water media, oil emulsion, dry pigment, and smoke, Kass forms bold yet abstracted floral and arboreal imagery. The rag paper surface, covered with shaved beeswax, lends to a soft aesthetic that reflects the natural elements found within the paintings' composition. In a review of Kass's 2012 joint exhibition with Ron Johnson at Reynolds Gallery, critic Dinah Ryan states, "Kass's paintings function more like dissipating clouds, lapping water, or shifting leaves through which things are barely perceptible...Fields of muted color cascade and overlap, modulating from one hue to the next, as patterns and fluid marks slip between them with the elasticity, purpose, and enigma of smoke signals."
Kass has been developing this series of paintings for multiple years, perfecting and expanding upon his process. His previous work began as abstracted, scribble-like paintings en plein air, a direct response to nature. However, it was once Kass returned to his studio that he noticed the continuity of systematic shapes and a sense of pattern in these drawings. Progressing this process, he transformed the geometric, symbolic patterns to compose schematic abstractions rendering landscapes and his environment.
Through the years, Kass has developed inventive methods for producing his paintings that are based in postmodern ideals: chance operations and indeterminacy. The ground of his paintings is created in one of two manners: smoking or staining. Kass developed the technique for smoking paper in the late 1980's while working with the highly influential composer and artist John Cage. In this method, Kass lays wet paper over a fire- the marks and lines left by the smoke cannot be controlled, and thus are random and determined by nature. To stain the paper, Kass pours puddles of medium and pigment over paper, some of which pools and leaves gradients of color and designs once the medium dries. Kass explains "Water has played an essential role in my developing artwork for many years, both strategically and as a source of content or subject. I would like to achieve the dynamic mirror-like transparency and reflectivity on the surface of my paintings that we experience when we look across water. The light that emanates from a mirror reveals a changing, iridescent imagery; it is a revelation of process replete with illuminating layers."
In addition to being recognized nationally as a painter, Kass is also a highly regarded writer and educator. He lives and works in Christiansburg, Virginia and is a Professor Emeritus of Art at Virginia Tech. Kass founded and directs the Mountain Lake Workshop, a collaborative art project drawing on the customs and environmental resources of the New River Valley and Appalachian region. He received his MFA from the University of North Carolina at Chapel Hill in Painting.
Kass is represented not only by Reynolds Gallery, but also by Garvey/Simon: ART ACCESS in NYC. His paintings have been widely exhibited at Allan Stone Gallery, A.V.C Contemporary Arts Gallery, Zone: Chelsea Center for the Arts, Baumgartner Gallery, ir77 Contemporary Art, New York, NY; Japan Art Forum, Kyoto; Awa No Sato Cultural Pavillion, Shikoku, Japan; Art Museum of Western Virginia, Roanoke, VA; Staniar Gallery at Washington and Lee University, Lexington, VA; Boston College, Brookline, MA; Philadelphia Museum of Art, Philadelphia, PA; Anderson Gallery of VCU, Virginia Museum of Fine Arts, both, Richmond, Virginia. He is the recipient of a Virginia Museum of Fine Arts Fellowship and an Individual Artist's Grant from the National Endowment for the Arts. Kass's work is in the collections of the Norton Museum of Art, West Palm Beach, FL; Taubman Museum of Art, Roanoke, VA; University of North Carolina, Chapel Hill, NC; University of Massachusetts, Amherst, MA; Boston Public Library, Boston, MA; Phizer Corporation, the Estate of Willem de Kooning, the Estate of Merce Cunningham and John Cage, all, New York, NY; and Medical College of Virginia, Altria Group, SunTrust, Ethyl Corporation, Markel Corporation, all, Richmond, Virginia.


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How to use:
Enter the four-digit passwords (default 0000), the green lights on, then rotate the knob clockwise to open the door. If you enter the wrong passwords, the red light will be illuminated.
How to Change Password:
Enter the default password 0000, open the box and hold * key input for the password you want to change by # end (in 15s). If you forget your password, you can take off the battery, after 10min you can restore the factory default password 0000.
1. When switch to voice prompt, please enter the next password number after the voice prompt, otherwise the saving box cannot recognize the entering number.
2. If the machine cannot roll the money, open the door, upend the machine, fine the small white gear, then use a screw driver to stir it for several rounds.
We are proud to offer international shipping services that currently operate in over 200 countries and islands world wide. Nothing means more to us than bringing our customers great value and service. We will continue to grow to meet the needs of all our customers, delivering a service beyond all expectation anywhere in the world.
Packages from our warehouse in China will be shipped by ePacket or EMS depending on the weight and size of the product. Packages shipped from our US warehouse are shipped through USPS.
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Wolfcraft Tools
Established by toolmaker and technical draftsman Robert Wolff, the Wolfcraft brand has been supplying innovative products and accessories to tool users since the range was first launched in 1961. Based in Germany, Wolfcraft has a presence in over 16 countries around Europe and supplies around 2000 different products, including machine tables, workbenches and clamping tables, transport systems, clamping tools, wood joints, power tool accessories and hand tools as well as a complete range of project specific solutions.
About The Products
Wolfcraft supply a large range of accessories for use with electric drills; these include nylon and wire brushes, rotary rasps, hole saws, flexible drive shafts, mixer paddles, drill pumps and the Tec Mobil drill guide. Woodworkers are well catered for with Wolfcraft's dowel pins, depth stops and centre points, which are also available as complete sets.
As well as a selection of angle grinder wire brushes and folding lift trolleys, we stock two of the most popular Wolfcraft clamps: the One hand Easy clamp and 40mm Spring clamp.


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NCAA Women's Basketball Tournament 2018: Download a blank, printable bracket
Is this the Louisville women's basketball team's season to win it all?
See if the Cards make it to your Final Four in Columbus this year when you fill out this blank, printable NCAA Women's Basketball Tournament bracket.
Louisville (32-2) received its first NCAA Tournament No. 1 seed and will host No. 16 seed Boise State (23-9) at noon Friday at the KFC Yum Center.
No. 8 Marquette faces No. 9 Dayton at 2:30 p.m. Friday, and the winners advance to play Sunday at a time yet to be determined.
Both of Friday's games will be on ESPN2.
Other No. 1 seeds are UConn (32-0), Notre Dame (29-3) and Mississippi State (32-1).
Click here to download a printable bracket and join Courier Journal's Danielle Lerner at 1 p.m. to follow along as she makes her picks here and facebook.com/courierjournal.


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On Monday, Scoot launched a plane, specially designed for SG50.
Named Maju-lah, the plane is painted with big red balloons featuring names of local companies, the SG50 logo along and bursts of confetti. It also has the iconic Merlion and caricatures of Scoot cabin crew members wishing the world "Happy 50th Birthday Singapore".
In total, twenty local company names will be featured on the balloons. These are the likes of BreadTalk, CapitaLand, Gleneagles Singapore, Metro, UOB, Mount Faber, Frasers Centrepoint, Raffles Education Group and Zalora.
"Scoot is so very proud to call Singapore home. This young nation has achieved remarkable success in a short time through vision and hard work, and is a great inspiration to a young airline like Scoot. We are proud to join the SG50 celebrations and show that pride on an aircraft that will be seen around the world for months to come," Scoot CEO, Campbell Wilson said.
However, a quick check on the Scoot's Facebook page showed that consumers were not very forgiving towards the design. While a few have said the new design works, most of the remarks seem negative.
According to Katie Ewer, strategy director of JKR, special commemorative liveries such as this one play the same role as limited edition packs in the world of FMCG – they need to strike the right balance between the fresh and the familiar.
"Coke does it exceptionally well, with each limited edition they release bringing something new to the market whilst still feeling on-brand. The biggest problem with the Scoot plane, apart from the fact that it's aesthetically offensive, is that it doesn't feel like Scoot. It feels like a plane a bad outbreak of sponsorship measles with a heavy dose of cultural clichés thrown in for good measure," Ewer said.
(Read also: Have we gone overboard with SG50 branding?)
Aside from showcasing Singapore's jubilee, Maju-lah will also host Scoot's "National Day In-Flight Party", being held in the run up to National Day. Featuring Kumar and others, members of the public can join this celebration of Singapore at 40,000 feet by winning themselves a seat through our series of Garang for Singapore challenges.


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Daily Archives: 1st September 2018
Fuel Prices hit Fresh Record High
Fuel prices continued to rise on Saturday, with petrol and diesel prices touching a new record high in several metropolitan cities, including Delhi and Mumbai.
Odisha IAS Officers' Association Condemn mob Attack
Odisha IAS Officers Association on Friday condemned the mob attack on two officers in Maharashtra's Nandurbar in August. The officers also requested Maharashtra government to order an enquiry.
Drug Smugglers Arrested in Jammu, 1.8 kg Heroin Seized
The Directorate of Revenue Intelligence (DRI), Ludhiana Zonal Unit, has busted a gang of narcotic drugs' smugglers and seized a consignment of heroin being sneaked into Kashmir.
Jain Monk Tarun Sagar Dies at 51
Jain monk Tarun Sagar passed away on Saturday at Delhi's Radhapuri temple after a prolonged illness.
EU to Recommend Abolishing Daylight Saving Time
The European Commission President Jean-Claude Juncker on Friday said that following a large-scale public survey, the European Union (EU) is planning to propose ending twice-yearly clock changes.
Jinson Johnson, Arpinder Singh Back From Asian Games With Gold
Indian athlete Jinson Johnson, who clinched a gold medal in Men's 1500m athletics event at the 18th edition of the Asian Games, arrived in India on Friday.
Palestine to Send 'Ghost Hunting' to Oscars
Palestine has officially chosen documentary film 'Istiyad ashbah' (Ghost Hunting) for submission at the 91st Academy Awards.
Trump Criticises Media After Off-the-record Comment Leaked
The United States president Donald Trump on Friday lashed out at media for "dishonest reporting" after off-the-record comments concerning North American Free Trade Agreement (NAFTA) were leaked.
Nadal Defeats Khachanov, Enters Round of 16 of US Open
After a four hours and 22 minutes long epic clash, World No 1 Rafael Nadal on Friday defeated 22-year-old Karen Khachanov (Russia) to qualify for the round of 16 of the US Open at the Arthur Ashe Stadium here.
Rainfall, Landslides Form 100 Meter Long Lake in Uttarakhand
As a consequence of the continuous rainfall and landslides in various parts of Uttarakhand, a 100 meter long and 50 meter deep lake has been formed near Tehri Garhwal-Dehradun border, forcing the people living in nearby houses to vacate the area, on the instructions of the authorities.
Hollywood Actress Vanessa Marquez Killed by Cops
Hollywood actor Vanessa Marquez was shot and killed by police in Pasadena city.
Anushka Unveils new Poster of 'Sui Dhaaga'
Bollywood actor Anushka Sharma unveiled a new poster of her upcoming film 'Sui Dhaaga'.
Balochistan's Resources Belong to Baloch People, Says BNC
"Balochistan's resources belong to Baloch people and Pakistan and China have no right to loot, plunder and exploit the Balochistan's resources against the will and consent of Baloch people", said Dr. Wahid Baloch, the president of Balochistan National Congress (BNC), in a statement.
Indian Woman Dies in Saudi, Family Suspects Foul Play
A 41-year-old woman from Hyderabad, who was trafficked to Saudi Arabia's Riyadh, died under mysterious circumstances.
One More Aasra Shelter Home Inmate Dies in Hospital in Patna
One more girl from the Aasra Shelter Home passed away during treatment at Patna Medical College and Hospital (PMCH) on Friday.
US Decides to end Funding to UN for Palestinian Refugees
The United States (US) has decided to end all the funding for the United Nations (UN) for the Palestinian refugees and called for a reduction in the number of Palestinians considered to be refugees.
Class 6 Student Molested at Private School in Mumbai
Parents protested outside a private school in Mumbai's Charkop area after a class 6 student was allegedly molested by another student in the premises on Thursday.
Here is how Kerala Floods Broke the Religious Barriers
Amidst the devastation due to floods in Kerala, instances of unity have come forward wherein people together are giving their best to help bring life back to normal.
NIA Seizes Rs 48 Lakh in Terror Funding Case in Manipur
The National Investigation Agency (NIA) has seized Rs 48 lakh in cash from a woman during an investigation into the activities of proscribed terrorist organisation Kangleipak Communist Party (KCP).
Brihanmumbai Municipal Corporation Seizes 17,084 kg Plastic
The Brihanmumbai Municipal Corporation (BMC) has seized 17,084 kg plastic and collected Rs 97,75,000 as fine since the plastic ban was implemented in Maharashtra.
Antigua's Attorney General to Discuss Choksi's Extradition With Director of Public Prosecutions
Antigua and Barbuda's Attorney General Steadroy Benjamin has said that he is planning to discuss the possible extradition of fugitive diamantaire and Punjab National Bank (PNB) accused Mehul Choksi with the Director of Public Prosecutions (DPP) Anthony Armstrong.
Abductions In South Kashmir, Chaos And Confusion Among People
all abducted persons were released on Friday after making a video of each of them, who not only warn their parents, relatives but also requested to DGP S P Vaid that do not harass the families of the militants. in the video, militants also warn that this time we are releasing you and if harassment did not stop, our approach would be different.


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After Ashley announces their banns in church, Laurel shocks him by declaring that she wants to marry him that day. After a bit of persuasion Ashley agrees and says that he wants the bishop to do the service so Laurel goes off to find him. The bishop is dubious about the sudden wedding but on hearing Laurel say that Seth's death has taught her to live for the moment, he agrees. Nicola allows Laurel to drive the bishop to Emmerdale in the two-seater, leaving her desperately trying to get a taxi. Nicola eventually flags down a mini-bus and finds herself surrounded by residents from a local old folks' home, telling them that if they give her a lift they can come to the wedding. Back at the village, Laurel is touched when Betty digs out the dress she wore when she married Wally, saying that she'd love Laurel to wear it. Laurel says that she would have loved Seth to have given her away and Betty offers instead. As the service is about to begin there is surprise when Nicola turns up with a gaggle of pensioners. Meanwhile, one of the elderly men, Sandy, notes Ashley's name on the church and asks Victoria where Reverend Thomas lives.
Regular cast
- Sandy Thomas - Freddie Jones
- Laurel Potts - Charlotte Bellamy
- Ashley Thomas - John Middleton
- Nicola Blackstock - Nicola Wheeler
- Emily Kirk - Kate McGregor
- Len Reynolds - Peter Martin
- Lisa Dingle - Jane Cox
- Ivan Jones - Daniel Brocklebank
- Betty Eagleton - Paula Tilbrook
- Belle Dingle - Eden Taylor-Draper
- Louise Appleton - Emily Symons
- Paul Lambert - Mathew Bose
- Rodney Blackstock - Patrick Mower
- Zak Dingle - Steve Halliwell
- Victoria Sugden - Hannah Midgley
- Simon Meredith - Dale Meeks
- Gabrielle Thomas - Annelise Manojlovic (uncredited)
Guest cast
- First appearance of Sandy Thomas.
- Viewing Figures: UK broadcast - 8,210,000 viewers (16th place).


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One of the brain sites more directly related with learning and memory processes is the hippocampus. We recorded, in conscious mice, the activity-dependent changes taking place at the hippocampal CA3–CA1 synapse during the acquisition, extinction, recall, and reconditioning of an associative task. Mice were classically conditioned to evoke eyelid responses using a trace [conditioned stimuli (CS), tone; unconditioned stimuli (US), shock] paradigm. A single electrical pulse presented to the Schaffer collateral–commissural pathway during the CS–US interval evoked a monosynaptic field EPSP (fEPSP) at ipsilateral CA1 pyramidal cells. The slope of evoked fEPSPs increased across conditioning sessions and decreased during extinction, being linearly related to learning evolution. In contrast, fEPSPs were not modified when evoked in control mice in the absence of a conditioning protocol. Long-term potentiation (LTP) evoked by high-frequency stimulation of Schaffer collaterals prevented acquisition, extinction, recall, or reconditioning, depending on the moment when it was triggered. Learning and memory impairments evoked by LTP induction resulted probably from the functional saturation of the CA3–CA1 synapse, although an additional disturbance of the subsequent information transfer toward postsynaptic circuits cannot be discarded. CGP 39551 [(E)-(±)-2-amino-4-methyl-5-phosphono-3-pentenoic acid ethyl ester] (an NMDA antagonist) prevented LTP induction in behaving mice, as well as the acquisition of an eyelid learned response, and the synaptic changes taking place at the CA3–CA1 synapse across conditioning. In conclusion, the responsivity of the CA3–CA1 synapse seems to be modulated during associative learning, and both processes are prevented by experimental LTP or NMDA-receptor inactivation. Our results provide evidence of a relationship between activity-dependent synaptic plasticity and associative learning in behaving mice.
Acquired learning abilities are assumed to be stored in the form of functional and/or structural changes in synaptic efficiency (Hebb, 1949). Although there are many excellent studies in vitro of the electrophysiological processes and molecular events supporting activity-dependent synaptic changes (Bliss and Collingridge, 1993; Engert and Bonhoeffer, 1999; Malenka and Nicoll, 1999; Lynch, 2004), not much information is available on synaptic plasticity during actual learning in behaving animals. Functional changes evoked by learning should be susceptible to being detected at synapses relevant to the acquisition process. For example, it has been reported that inferior olive synaptic contacts on posterior interpositus neurons are potentiated during the acquisition of delay eyeblink conditioning (Gruart et al., 1997).
Another basic tenet is that long-term potentiation (LTP) is the mechanism underlying certain forms of learning (Bliss and Lømo, 1973; McNaughton et al., 1978). LTP is evoked by high-frequency stimulation (HFS) of selected afferent pathways resulting in a long-lasting enhancement of synaptic efficacy. The experimental induction of LTP will surely disturb the synaptic changes taking place during associative learning in selected neural sites. Indeed, it is known that LTP saturation of hippocampal circuits disrupts spatial learning (Barnes et al., 1994), and that place representation in hippocampal networks can be modified experimentally by LTP (Dragoi et al., 2003).
The hippocampus has been implicated in a variety of learning paradigms, including classical conditioning of eyelid responses (Berger et al., 1983; Sanchez-Andres and Alkon, 1991; McEchron and Disterhoft, 1997; Múnera et al., 2001). Bilateral hippocampal lesions impair the acquisition of trace eyeblink conditioning, whereas they do not alter delay conditioning (Thompson, 1988; Moyer et al., 1990). Apparently, whereas trace conditioning requires a conscious knowledge (Clark and Squire, 1998) and/or declarative or explicit memory (Eichenbaum, 1999) of relevant relationships between conditioned (CS) and unconditioned (US) stimuli, delay conditioning does not. Moreover, mice are capable of acquiring classically conditioned eyelid responses using trace paradigms (Takatsuki et al., 2003; Domínguez-del-Toro et al., 2004).
As already shown for dentate granule cells (Weisz et al., 1984), we studied here whether the acquisition of associative learning modifies the synaptic strength of the hippocampal CA3–CA1 synapse. We used classical conditioning of eyelid responses, with a trace paradigm, presenting a tone as CS and an electrical shock as US. We recorded, in vivo, the field EPSP (fEPSP) evoked at hippocampal CA1 pyramidal cells by the electrical stimulation of the Schaffer collateral–commissural pathway. The fEPSPs were evoked at CS–US intervals during habituation, acquisition, extinction, recall, and reconditioning of eyelid responses. LTP was experimentally evoked before selected conditioning sessions to determine its effects both on the acquisition of the associative learning and on the profile of evoked fEPSPs. Finally, (E)-(±)-2-amino-4-methyl-5-phosphono-3-pentenoic acid ethyl ester (CGP 39551) (an antagonist of the NMDA receptor) was administered in vivo to determine the involvement of this glutamate receptor on the three phenomena (i.e., LTP, synaptic enhancement, and associative learning) considered in this study (Bliss and Collingridge, 1993; Malenka and Nicoll, 1999). The present results provide substantial evidence of the intrinsic relationships between LTP, activity-dependent synaptic plasticity, and associative learning in mammals.
Materials and Methods
Experiments were performed on 120 C57BL/6 male adult mice (3–5 months old; 28–35 g) obtained from an official supplier (University of Granada Animal House, Granada, Spain). Before surgery, animals were housed in separate cages (n = 10 per cage). The mice were kept on a 12 h light/dark cycle with constant ambient temperature (21 ± 1°C) and humidity (50 ± 7%). Food and water were available ad libitum. Electrophysiological and behavioral studies were performed in accordance with the guidelines of the European Union Council (86/609/EU) and Spanish regulations (BOE 67/8509–12, 1988) for the use of laboratory animals in chronic experiments. Experiments were also approved by the Institution Committee for animal care and handling.
Animals were anesthetized with 0.8–3% halothane (AstraZeneca, Madrid, Spain) delivered via a home-made mask. Halothane was administered from a calibrated Fluotec 5 (Fluotec-Ohmeda, Tewksbury, MA) vaporizer at a flow rate of 1–4 L/min oxygen. Once anesthetized, animals were implanted with bipolar stimulating electrodes on the left supraorbitary branch of the trigeminal nerve and with bipolar recording electrodes in the ipsilateral orbicularis oculi muscle (see Fig. 1A). Electrodes were made of 50 μm, Teflon-coated, annealed stainless steel wire (A-M Systems, Carlsborg, WA), and with their tips bared of the isolating cover for ≈0.5 mm. The electrode tips were bent as a hook to facilitate a stable insertion in the upper eyelid. Stereotaxic coordinates (Paxinos and Franklin, 2001) were followed to implant animals with bipolar stimulating electrodes aimed at the right (contralateral) Schaffer collateral–commissural pathway of the dorsal hippocampus (2 mm lateral and 1.5 mm posterior to Bregma; depth from brain surface, 1.0–1.5 mm) and with two recording electrodes aimed at the ipsilateral stratum radiatum underneath the CA1 area (1.2 mm lateral and 2.2 mm posterior to Bregma; depth from brain surface, 1.0–1.5 mm). These electrodes were made of 50 μm, Teflon-coated tungsten wires (Advent Research Materials, Eynsham, England). The final position of hippocampal stimulating and recording electrodes was determined under recording procedures explained below. A 0.1 mm bare silver wire was affixed to the skull as a ground. The eight wires were connected to two four-pin sockets (RS Amidata, Madrid, Spain). The ground wire was also connected to the recording system with a single wire. Sockets were fixed to the skull with the help of two small screws and dental cement (Domínguez-del-Toro et al., 2004).
Classical conditioning procedures.
For recordings, two animals at a time were placed in separate small (5 × 5 × 10 cm) plastic chambers located inside a Faraday box (30 × 30 × 20 cm). Classical conditioning was achieved using a trace paradigm (see Fig. 1B). For this, a tone (20 ms; 2.4 kHz; 85 dB) was presented as a CS. The US consisted of a 500 μs, 3× threshold, square, cathodal pulse. The US started 500 ms after the end of the CS. Unless otherwise indicated, a total of 4 habituation, 10 conditioning, and 5 extinction sessions were performed for each animal (see below). A conditioning session consisted of 60 CS–US presentations, and lasted ≈30 min. In 10% of the cases, the CS was presented alone. CS–US presentations were separated at random by 30 ± 5 s. For habituation and extinction sessions, only the CS was presented, also for 60 times per session at intervals of 30 ± 5 s. For recall (i.e., retrieval), animals underwent two additional conditioning sessions 7 d after the 10th one. For pseudoconditioning, unpaired CS and US presentations were performed for 10 sessions (60 times/session). Pseudoconditioned animals also received four habituation and five extinction sessions, as indicated above (Domínguez-del-Toro et al., 2004).
Recording and stimulation procedures.
Recordings were performed using Grass P511 differential amplifiers with a bandwidth of 0.1 Hz-10 kHz (Grass-Telefactor, West Warwick, RI). Hippocampal recordings were performed with a high impedance probe (2 × 1012 Ω; 10 pF). Reflex blinks were collected before conditioning sessions and evoked by single 50 μs, 2× threshold, square, cathodal pulses presented at a rate of 1/30 s (see Fig. 1A). As criteria, we considered a "conditioned response" the presence, during the CS–US period, of electromyographic (EMG) activity lasting >10 ms and initiated >50 ms after CS onset. In addition, the integrated EMG activity recorded during the CS–US interval had to be at least 2.5 times greater than the averaged activity recorded immediately before CS presentation (Porras-García et al., 2005).
Electrodes were surgically implanted in the CA1 area using, as a guide, the field potential depth profile evoked by paired (20–50 ms of interval) pulses presented at the ipsilateral Schaffer collateral pathway. The recording electrode was fixed at the site where a reliable monosynaptic field EPSP was recorded (see Fig. 1A). Synaptic field potentials in the CA1 area were evoked during habituation, conditioning, and extinction sessions by a single 100 μs, square, biphasic (negative–positive) pulse applied to Schaffer collaterals 300 ms after CS presentation. Stimulus intensities ranged from 50 to 350 μA. For each animal, the stimulus intensity was set well below the threshold for evoking a population spike, usually 30–40% of the intensity necessary for evoking a maximum fEPSP response (Gureviciene et al., 2004). An additional criterion for selecting stimulus intensity was that a second stimulus, presented 20–50 ms after a conditioning pulse, evoked a larger (>20%) synaptic field potential (Bliss and Gardner-Medwin, 1973). For evoking LTP, each animal was presented with five 200 Hz, 100 ms trains of pulses at a rate of 1 per second. This protocol was presented six times in total, at intervals of 1 min. The 100 μs, square, biphasic pulses used to evoke LTP were applied at the same intensity used for the single pulse presented after CS presentation. On occasions, some animals (n = 9) presented abnormal EEG responses after HFS; those animals were rejected from the study.
The chemicals used were (E)-(±)-2-amino-4-methyl-5-phosphono-3-pentenoic acid ethyl ester (CGP 39551) and 2, 3-dioxo-6-nitro-1, 2, 3, 4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium salt (NBQX disodium salt) from Tocris Bioscience (Avonmouth, UK). The two drugs were dissolved in saline and administered intraperitoneally at doses of 6.5 and 15 mg/kg, respectively. Control animals received the same volume of saline.
Experimental groups.
As illustrated in Figure 1B, animals were divided into eight groups. Unless otherwise indicated, and for the sake of homogeneity, all experimental groups were composed of 10 successful animals. (1) Controls were habituated and conditioned as indicated. After conditioning, half of them were submitted to extinction sessions and reconditioned (Control-1), whereas the other half underwent two recall sessions. (2) The HFS-1 group received two HFS sessions 10 min before the beginning of the first two conditioning sessions. (3) The HFS-2 group received two HFS sessions 10 min before the fifth and sixth conditioning sessions. (4) The HFS-3 group received two HFS sessions 10 min before the ninth and 10th conditioning sessions. These animals had no extinction sessions, but (like the Control-2 group) underwent two recall sessions. (5) Animals of the HFS-4 group received habituation and conditioning, but not extinction, sessions. Seven days later, they received two HFS sessions 10 min before the two recall sessions. (6) The HFS-5 group received a training similar to that of the Control-1 group, but including two HFS sessions 10 min before the first two reconditioning sessions. (7) A group of animals was injected with CGP 39551 (6.5 mg/kg, i.p.) 30 min before each conditioning session. (8) The pseudoconditioned group received 4 habituation, 10 pseudoconditioning, and 5 extinction sessions. (9) A group of animals was presented with 60 stimuli/d at Schaffer collaterals, across 19 d, but without CS or US stimuli. Stimuli were presented during 30 min, at 30 ± 5 s intervals. Finally, animals (n = 17) initially included in groups HFS-1 to HFS-5 that did not present a reliable potentiation (<20% increase in fEPSP slope, lasting <30 min) after the two HFS sessions were also conditioned. Data for this group are shown in Figure 7C and D. Another 20 animals were used for preliminary trials, to calibrate the recording set-up, determine selected drug doses, and fix the recording procedures.
At the end of the experiments, mice were deeply reanesthetized (sodium pentobarbital; 50 mg/kg) and perfused transcardially with saline and 4% phosphate-buffered paraformaldehyde. Selected sections (50 μm) including the dorsal hippocampus were mounted on gelatinized glass slides and stained using the Nissl technique with 0.1% toluidine blue to determine the location of stimulating and recording electrodes (see Fig. 1C, D).
Data analysis.
EMG and hippocampal activity, and one-volt rectangular pulses corresponding to CS and US presentations, were stored digitally on a computer through an analog/digital converter (CED 1401 Plus; Cambridge Electronic Design, Cambridge, UK), at a sampling frequency of 11–22 kHz and an amplitude resolution of 12 bits. Commercial computer programs (Spike 2 and SIGAVG, from Cambridge Electronic Design) were modified to represent EMG and extracellular synaptic field potential recordings. Data were analyzed off-line for quantification of conditioned responses and the fEPSP slope with the help of homemade representation programs (Gruart et al., 1995, 2000; Domínguez-del-Toro et al., 2004; Porras-García et al., 2005). The slope of evoked fEPSPs was collected as the first derivative (i.e., volts per second) of fEPSP recordings (volts). For this, five successive evoked field synaptic potentials were averaged, and the mean value of the slope was determined for the rise time period (i.e., the period of the slope between the initial 10% and the final 10% of the evoked field potential). Computed results were processed for statistical analysis using the SPSS (Chicago, IL) for Windows package. Unless otherwise indicated, data are represented by the mean ± SD. Collected data were analyzed using a two-way ANOVA test, with time or session as repeated measure, coupled with contrast analysis when appropriate. One-way ANOVA allowed checking the statistical differences between different groups. Regression analysis was used to study the relationship between the fEPSP slope and the percentage of conditioned responses.
In vivo recordings of eyelid responses and of CA3–CA1 synaptic field potentials
In a first series of experiments, we checked the functionality of the proposed experimental model. The electrical stimulation (2× threshold) of the supraorbitary branch of the trigeminal nerve (n = 10 animals) evoked an early EMG activation of the orbicularis oculi muscle at a latency of 5.3 ± 1.2 ms (mean ± SD), followed by a second EMG activation with a latency from the stimulus of 18.6 ± 3.2 ms (Fig. 1A, bottom recordings). These successive muscle activations correspond to the R1 and R2 components already described in humans (Kugelberg, 1952) and other species of mammals, including mice (Gruart et al., 1995, 2000; Domínguez-del-Toro et al., 2004). Electrode implantation did not seem to disturb the normal kinematics of spontaneous and reflexively evoked eyelid responses.
The electrical stimulation of Schaffer collaterals sometimes evoked an afferent volley into the CA1 area, usually appearing as a small triphasic (positive-negative-positive) potential, with a latency of 1.45 ± 0.5 ms (n = 15). The afferent volley was followed, 1.5–2 ms later, by a large negative wave when recorded at the stratum radiatum (i.e., on the apical dendrites of the CA1 pyramidal cells). The location of stimulating and recording electrodes in the hippocampus was checked at the end of the experiments and is illustrated in Figure 1C and D. In approximately one-fourth of cases, the wave recorded in the CA1 area presented a positive shape, corresponding to a more-dorsal location of the recording electrode (i.e., near the pyramidal cell layer) (Schwartzkroin, 1986).
In a preliminary study, we checked whether, in the absence of classical conditioning and/or HFS, the slope of fEPSPs evoked by the stimulation of Schaffer collaterals was stable across the maximum period of time used in this study (i.e., 29 d). For this, a group of animals (n = 10) was stimulated daily (60 single pulses for a 30 min period, with a time interval of 30 ± 5 s), during 29 d. Quantitative analysis showed that the percentage of variation in fEPSP slopes across the last 25 d, as compared with the mean value (100%) computed from fEPSP slopes collected during the first four recording days, was ≤12.5%, with no statistically significant tendency toward a decrease or increase (p = 0.71).
Evolution of CA3–CA1 synaptic field potentials during classical conditioning of eyelid responses
A total of 20 animals (i.e., Control-1 and Control-2 groups) (Fig. 1B) were classically conditioned using a trace (CS, tone; US, shock) paradigm (Fig. 2A). The time interval between the end of the CS and the beginning of the US was 500 ms. The experimental design included the presentation of a single electrical pulse to Schaffer collaterals 300 ms after CS presentation. Figure 2A shows single recordings of the EMG activity of the orbicularis oculi muscle and of the fEPSP evoked in a well trained animal during the seventh conditioning session. Although the stimuli presented to Schaffer collaterals disrupted the regular theta rhythm recorded in the local EEG for 100–200 ms, the rhythm reappeared in phase. As illustrated in Figure 2B, the learning curve presented by these conditioned animals was normal when compared with previous descriptions in mice, using similar trace conditioning procedures (Takatsuki et al., 2003; Domínguez-del-Toro et al., 2004). Animals presented a mean of 37 ± 6% responses during the first conditioning session, and reached asymptotic values from the seventh session onward (70–75% of conditioned responses). The mean percentage of conditioned responses collected for control animals was significantly larger [F(18, 162) = 192.7; p < 0.001 for the 10 conditioning sessions compared with values reached by the pseudoconditioned group (n = 10)] (Fig. 2B).
Field EPSPs evoked in controls by the electrical stimulation of Schaffer collaterals increased progressively in slope (taking the slope of fEPSPs collected during the four habituation sessions as 100%) across conditioning, to a maximum of 120% during the eighth and ninth sessions. During extinction, the fEPSP slope decreased to a minimum of 82.5% by the fifth session. In contrast, fEPSP values collected from the pseudoconditioned group did not present any tendency for a change in slope during either conditioning or extinction sessions. Values of fEPSP slopes for the control group were significantly different from those for pseudoconditioned animals for the 5th to the 10th conditioning and 3rd to 5th extinction sessions (F(18, 162) = 48.2; p < 0.001).
As illustrated in Figure 3, the slope of fEPSPs evoked by Schaffer collateral stimulation at the CA3–CA1 synapse was linearly related (r ≥ 0.66; p < 0.001) to the percentage of conditioned responses across sessions of conditioning (slope, 0.55) and extinction (slope, 0.65), but not habituation. The fact that the slopes of the two regression lines were similar suggests that the activity-dependent plasticity at the CA3–CA1 synapse functioned as a continuum for both acquisition and extinction processes. This interesting finding prompted us to hypothesize that evoking LTP experimentally at the CA3–CA1 synapse could interfere with the acquisition of associative learning using classically conditioned procedures.
Evoking LTP in behaving mice
In a pilot study, we evoked LTP in behaving mice (n = 5). To obtain a baseline, animals were stimulated every 5 s for 15 min at Schaffer collaterals. The stimulus consisted of a 100 μs, square, biphasic pulse. Pulse intensity (50–350 μA) was set at 30–40% of the amount necessary to evoke a maximum fEPSP response. For evoking LTP, each animal was presented with five trains (200 Hz; 100 ms) of pulses at a rate of 1 per second. This protocol was presented six times in total, at intervals of 1 min. To avoid evoking a population spike and/or unwanted EEG seizures, the stimulus intensity was set at the same amount used for generating the baseline recording. After HFS, the same single stimulus was presented at the initial rate (12 per minute) for another 15 min. As illustrated in Figure 4B, with this experimental approach we were able to evoke LTPs lasting 4 d. In accordance, and to obtain LTPs lasting as long as the conditioning sessions (i.e., ≈10 d), in an additional pilot study (n = 5 animals), we used the same procedure for evoking LTP, but this time the HFS protocol was presented for two successive days. As illustrated in Figure 4A and B, this procedure evoked LTP (with above-baseline values) that lasted up to 10 d.
Evolution of CA3–CA1 synaptic field potentials and acquisition of conditioned eyelid responses after LTP induction
As illustrated in Figure 1B, we prepared five experimental groups (HFS-1 to HFS-5) to be conditioned as indicated for controls, but which received two HFS sessions 30 min before the first and second (HFS-1), fifth and sixth (HFS-2), or ninth and 10th (HFS-3) conditioning days, during two recall sessions (HFS-4), or during the first and second reconditioning sessions (HFS-5).
As shown in Figure 5A, HFS applied during the first two conditioning sessions (HFS-1 group; n = 10) produced a noticeable LTP that remained above baseline values for up to 9 d (F(18, 162) = 32.8; p < 0.001). Animals of this group were unable to present a normal learning curve. In fact, the second HFS produced a noticeable decrease in the percentage of conditioned responses (Fig. 5B). Along conditioning, the percentage of learned responses remained F(18, 162) = 15; p < 0.001). HFS-1 animals presented a definite extinction of conditioned responses in simultaneity with a decrease (as in controls) of the fEPSP slope during the extinction sessions. In accordance, fEPSP slopes and the percentage of conditioned responses were not linearly related for HFS-1 animals during the ten conditioning sessions (Fig. 3B); however, the same parameters collected during the five extinction sessions showed a statistically significant linear relationship (r = 0.79; p < 0.001).
Animals (n = 10) of the HFS-2 group received the two HFS sessions 30 min before the fifth and sixth conditioning sessions (Fig. 5C, D). In this case, the slope of the fEPSP remained above baseline values for the 6th to the 10th conditioning sessions and during the five extinction sessions (F(14, 126) = 7.58; p < 0.001). These animals presented a gradually increasing learning curve up to the day before HFS presentation (52.5% of conditioned responses by the fourth conditioning session; F(14, 126) = 43.2; p < 0.001) (Fig. 5D). However, the two HFS sessions decreased the number of conditioned responses to values <40% from the 5th to the 10th conditioned sessions. Interestingly, these animals were unable to extinguish the acquired level of conditioning, probably because extinction sessions took place during the presence of a significant LTP. In accordance, no linear relationship between fEPSP slopes and the percentage of conditioned responses across conditioning and extinction sessions was observed for the HFS-2 group (Fig. 3C).
Animals (n = 10) of the HFS-3 group received the two HFS sessions during the last 2 d of conditioning (Fig. 6A, B). The HFS provoked an LTP that was still evident during the two recall sessions (i.e., it lasted at least 11 d; F(7, 63) = 16.8; p < 0.001). LTP induction evoked a significant decrease in the learning curve achieved by these animals (Fig. 6B). The percentage of conditioned responses during recall was increased with respect to the last two conditioning sessions, during which the HFS protocol was presented (F(7, 63) = 162.4; p < 0.001). No linear relationship between fEPSP slope and the percentage of conditioned responses across conditioning and recall sessions was observed for the HFS-3 group (Fig. 3D).
In contrast to results collected from the HFS-3 group, those from animals (n = 10) of the HFS-4 group (Fig. 6C, D), which received the two HFS sessions during recall, showed a significantly (F(5, 45) = 16.9; p < 0.001) lower percentage of conditioned responses than those obtained during the last two conditioning sessions. In this situation, it was of interest to determine the retention index [i.e., the ratio between the mean percentage of condition responses (CRs) during the last two conditioning sessions and the two recall sessions] for controls (Control-2 group) and for the HFS-3 and HFS-4 groups (Fig. 6E). The retention index for the HFS-4 group (0.69 ± 0.11) was significantly lower (p ≤ 0.01) than that collected from controls (1.09 ± 0.2) and from the HFS-3 group (1.32 ± 0.2). These results suggest that the recall of an earlier experience is a rather unstable situation, susceptible to being blocked by LTP evoked experimentally at the CA3–CA1 synapse. Nevertheless, and according to the present data, the effects of evoking LTP during classical conditioning were greater when evoked in the presence of a large percentage of conditioned responses (i.e., +10% for the HFS-1 group, −20% for the HFS-2 group, −29% for the HFS-3 group, −28.5% for the HFS-4 group, and +14.5% for the HFS-5 group).
Evolution of CA3–CA1 synaptic field potentials and acquisition of conditioned eyelid responses during reconditioning for controls and after LTP induction
Figure 7 represents data collected from the Control-1 group (n = 10; i.e., from control animals that received 10 reconditioning sessions), and from the HFS-5 group (n = 10; i.e., from animals that received the two HFS sessions during the first 2 d of reconditioning). During the 10 d of reconditioning, HFS-5 animals were unable to exceed values reached during the last extinction session, indicating that the evoked LTP completely blocked the possibility of reacquiring eyelid conditioned responses, in contrast to the case of controls. Thus, the percentage of conditioned responses presented by animals from the HFS-5 group for the 10 conditioning sessions was significantly lower than that achieved by controls (F(13, 117) = 207.4; p < 0.001). The slope of fEPSPs evoked in the CA1 area by Schaffer collateral stimulation increased steadily in controls across reconditioning sessions (Fig. 7A), reaching a maximum (128.3%) above baseline values (100%) during the 10th conditioning session. In contrast, the slope of fEPSPs evoked in HFS-5 animals was clearly dominated by the effect of the induced LTP, with no sign of modulation by (or during) the acquisition process (Figure 7A) (F(13, 117) = 6.09; p < 0.001). In this regard, no linear relationship between fEPSP slopes and the percentage of conditioned responses across reconditioning sessions was observed for the HFS-5 group (Fig. 3E).
It is important to point out that the learning deficits described here were the effect of LTP induction and not of the HFS by itself. Thus, some mice (n = 17 initially included in groups HFS-1 to HFS-5) received the two HFS sessions, but no significant potentiation (<20% increase in fEPSP slope, lasting <30 min) was observed in them. Data collected from these animals were not included in the analysis for groups HFS-1 to HFS-5, but were used to determine whether the deficits in associative learning were caused by the HFS or the evoked LTP. Indeed, all of these animals presented acquisition curves (F(18, 162) = 175.6; p = 0.23) and fEPSP slopes (F(18, 162) = 68; p = 0.41) nonsignificantly different from those shown by controls (Fig. 7C, D). In accordance, the learning deficits described in this and the preceding sections cannot be ascribed to any damage of hippocampal circuits produced by the HFS, but to the proper induction of LTP.
Involvement of NMDA channels in the acquisition of trace conditioning
Although the involvement of AMPA and NMDA receptors in LTP phenomena evoked at the CA3–CA1 synapse is well known (Collingridge et al., 1983a, b; Bliss and Collingridge, 1993), an attempt was made at a precise determination of the relationships between LTP, NMDA receptors, and the acquisition of associative learning using a trace classical conditioning paradigm. As illustrated in Figure 8A, the administration of NBQX (a selective and competitive antagonist of the AMPA receptor; 15 mg/kg, i.p.; n = 5) significantly decreased (by some 40%; p < 0.01) the slope of fEPSPs evoked in the CA1 area by single pulses applied to Schaffer collaterals, whereas the addition of CGP 39551 (a selective and competitive NMDA receptor antagonist; 6.5 mg/kg, i.p.; n = 5) had no additional effect, indicating that the contribution of the NMDA receptor to the fEPSP was almost negligible in this situation (Harris et al., 1984). However, and as illustrated in Figure 8B, HFS of Schaffer collaterals was unable to evoke an LTP in the CA3–CA1 synapse in the presence of CGP 39551 (6.5 mg/kg, i.p.; n = 5) as compared with controls (n = 5).
In an additional experiment, animals (CGP 39551 group; n = 10) were conditioned after the daily injection of CGP 39551 (6.5 mg/kg, i.p., 30 min before each conditioning session). Field EPSPs evoked in the CA1 area by the electrical stimulation of Schaffer collaterals were not modified in slope across conditioning sessions in the CGP 39551 group (Fig. 8C, second set of recordings) as compared with controls (Fig. 8C, first set of recordings). Significantly, animals from the CGP 39551 group did not present any evidence of eyelid conditioned responses, as illustrated by the two recordings from the eighth session collected from a control and from a CGP 39551-injected animal (Fig. 8D). Finally, Figure 8E illustrates the evolution of conditioned responses and of the slope of fEPSPs collected across conditioning from the CGP 39551 group. The percentage of conditioned responses in the control group was significantly larger than that in the CGP 39551 group from the 2nd to the 10th conditioning sessions (F(13, 117) = 235.6; p < 0.001). Moreover, fEPSP slopes were significantly larger in controls than in the CGP 39551 group for the last four conditioning sessions (F(13, 117) = 45.9; p < 0.001).
General remarks
We have shown here that the hippocampal CA3–CA1 synapse is involved in the acquisition, extinction, recall, and reconditioning of eyelid CRs using a trace paradigm. These synaptic elements underwent a slow potentiation, or decrease, in synaptic strength (Hebb, 1949) in parallel with the acquisition and/or extinction of eyeblink conditioning. We have also shown that LTP, evoked by HFS of Schaffer collaterals, interferes with both the acquisition of CRs and the linear relationships between learning scores and fEPSP slopes. The saturation of the CA3–CA1 synapse by LTP, at different stages across conditioning, prevented additional synaptic changes in plasticity (Barnes et al., 1994; Otnæss et al., 1999), causing both anterograde and retrograde amnesia. This experimental disturbance of the hippocampal pattern of synaptic strengths is apparently enough to interfere with the functional states necessary for the proper acquisition, storage, and retrieval of eyeblink conditioning, as already demonstrated for spatial learning (Barnes et al., 1994; Brun et al., 2001). Finally, we showed that the administration of an NMDA-receptor antagonist is not only able to prevent LTP induction in vivo, but also to hinder both the formation of eyelid CRs and the functional changes in strength at the CA3–CA1 synapse. Thus, the functional transformations taking place at CA1 pyramidal cells seem to be necessary for the proper acquisition, extinction, recall, and reconditioning of eyelid CRs.
Modulation in the synaptic strength of the CA3–CA1 synapse during trace conditioning
In a seminal study, Weisz et al. (1984) demonstrated a change in the efficacy of synaptic activation of dentate granule cells by perforant pathway axons during the acquisition of nictitating membrane CRs. This modulation in synaptic strength during the learning process has been confirmed here for the CA3–CA1 synapse, and it has been extended to the extinction process, suggesting that the two phenomena are equally active (Inda et al., 2005) and require an activity-dependent modification of synaptic responsivity for learning to occur (Weisz et al., 1984). Moreover, it has also been shown that the evoked changes in synaptic efficacy present a linear relationship with the amount of acquired, or extinguished, learning. The increase in fEPSP slope during the acquisition process explains the increased firing in hippocampal CA1 areas across conditioning, described for both rabbits and cats (McEchron et al., 2003; Múnera et al., 2001).
The involvement of hippocampal unitary activity in classical conditioning of the nictitating membrane/eyelid response is very well known (Berger et al., 1983; Moyer et al., 1990). For example, it has been shown, using unitary in vivo recordings (McEchron and Disterhoft, 1997), that hippocampal pyramidal cell firing to CS presentation increases several sessions in advance of behavioral conditioning. Although the discharge rate of hippocampal CA1 pyramidal neurons does not seem to encode the kinetic peculiarities of conditioned eyelid responses, it has been shown that CA1 firing is linearly related with the progressive acquisition of the eyelid learned response, with a gain of ≈0.035 spikes per second per trial, as measured in behaving cats during trace (tone-shock) conditioning (Múnera et al., 2001). Interestingly, this slow building up of hippocampal neuronal responses across conditioning is similar to the small increase in the slope of the fEPSP evoked at the apical dendrite of CA1 pyramidal cells by single pulses applied to Schaffer collaterals (i.e., a 0.03% increase in fEPSP slope/trial) reported here. It appears that fEPSP slopes reached an asymptotic value during the last conditioning sessions, in parallel with the evolution in the percentage of CRs. Because only five extinction sessions were performed here, similar observation has not been made during the extinction process. Nevertheless, the two phenomena presented linear regression lines of the same slope when plotted versus the amount of CRs, suggesting that the modulation in synaptic strength is similar for both processes, moving in the increasing (or decreasing) direction depending on the learning situation.
The modulation in CA3–CA1 synaptic strength reported here during acquisition, extinction, recall, and reconditioning seems to be a slow process originated by changes in the probability of releasing synaptic vesicles by CA3 terminals and/or by subtle modifications in the number of presynaptic active zones and/or postsynaptic receptor sites (Bolshakov et al., 1997) (i.e., by synaptic plastic changes also related to LTP) (Nicoll, 2003). From a functional point of view, the increased (or decreased) responsivity of CA1 pyramidal cells to Schaffer collateral stimulation suggests that, during the CS–US interval, they are in a facilitated (or disfacilitated) state, evoked by changes in coincident inputs of different (entorhinal, septal, etc.) origins (Weisz et al., 1984; Fontán-Lozano et al., 2005). Thus, it is possible that several mechanisms are acting in parallel to enhance CA3–CA1 synaptic transmission across learning. In any case, the plastic changes in synaptic strength evoked during the learning process are noticeably much smaller (in volts per second of the fEPSP) and slower (in time) than those evoked by the experimentally induced LTP (i.e., the two phenomena have a quite different time scale).
Anterograde and retrograde amnesia evoked by experimentally induced LTP
Humans with hippocampal lesions present both anterograde and (immediate) retrograde amnesia (Squire and Alvarez, 1995). Indeed, it has been shown that spatial learning is prevented when saturating LTP is evoked in the perforant pathway or when evoked by the repeated stimulation of a large number of hippocampal synaptic contacts (McNaughton et al., 1986; Casto et al., 1989; Moser et al., 1998; Dragoi et al., 2003). The effects of LTP evoked at the hippocampus are more evident for the acquisition of new learning skills and on recently acquired knowledge, but not for remote memory retrieval (Morris et al., 2003). Present results essentially confirm those findings, but using trace conditioning (i.e., a task which requires the active participation of hippocampal circuits) (Thompson, 1988).
It has been reported that, for spatial learning, pretraining prevents the deficits evoked by LTP (Otnæss et al., 1999), a fact not confirmed here for trace conditioning. In fact, the retrograde deficits were more evident with additional training, because, in all cases (Figs. 5⇑–7), the amount of evoked CRs decreased to the same level (≈40%) after LTP. Perhaps relational learning is more dependent on the functional state of hippocampal circuits than is spatial learning. It is still possible that this 40% of residual learning is hippocampally independent (i.e., acquired by subcortical and/or cerebellar circuits).
The present results show that when LTP values returned to baseline physiological values (i.e., 100–120% from baseline fEPSP) (Fig. 7A, B) learning did not recover. This may indicate that the induction of LTP through high frequency stimulation resulted in a permanent physiological disruption which produced both anterograde and retrograde memory deficits.
According to the present results, the retention index during recall plus HFS (HFS-4 group) was lower than in controls, or even in the group (HFS-3) that received HFS 7 d before the two recall sessions. In this regard, it has been reported that electroconvulsive shock or systemic drug administration given after memory reactivation might produce a severe amnesia for the original learning (Nader et al., 2000; Sara, 2000; Nader, 2003; Inda et al., 2005). These results indicate that recent memories become somewhat labile when they are recalled; thus, they require a proper reconsolidation, which is blocked in the HFS-4 group by the simultaneous presentation of the two HFS sessions. Thus, hippocampal firing seems to be necessary during conditioning paradigms in which conscious knowledge is required, namely, for trace paradigms (Clark and Squire, 1998). These findings suggest that the hippocampus is perhaps not necessary after memory consolidation (Squire and Alvarez, 1995; Morris et al., 2003).
Relationships between LTP, NMDA channels, and functional modifications of synaptic strength
The generation of CRs is a slow process requiring a large number (<300) of paired CS–US presentations in mice, rabbits, and cats (Thompson, 1988). Thus, the neural processes underlying its generation cannot be directly related to LTP mechanisms, by which an almost immediate acquisition of the learned response is expected (Ito, 1989; Bliss and Collingridge, 1993). The motor output expected from associative learning is not a sharp, sustained increase in neuronal firing of a pool of related neural centers, but a distributed and limited increase in the number of neurons recruited to respond to an initially irrelevant sensory stimulus (i.e., the CS) (Woody, 1986). But LTP has the property of associability, indicating that a weak input can still be potentiated if activated simultaneously to (or within a given time window of) a strong stimulus evoked through a separate but convergent input (McNaughton et al., 1978; Levy and Steward, 1983; Bliss and Collingridge, 1993).
However, it is known that hippocampal NMDA receptors are involved in the acquisition of eyelid CRs (Kishimoto et al., 2001; Sanders and Fanselow, 2003; Mokin and Keifer, 2005). As confirmed here, the administration of CGP 39551, a competitive antagonist of the NMDA receptor (Maren et al., 1992; Servatius and Shors, 1996; D'Hooge et al., 1999), prevents the acquisition of an eyeblink conditioning and also blocked the fEPSP potentiation observed in controls across learning. CGP 39551 also prevented the induction of LTP in the hippocampal CA1 area after HFS of Schaffer collaterals (Maren et al., 1992), but had no noticeable effects on the monosynaptic fEPSP evoked in the CA1 area by single pulses presented to the ipsilateral Schaffer collaterals. In contrast, the fEPSP evoked in CA1 was attenuated by NBQX, a selective and competitive AMPA-receptor antagonist (Parada et al., 1992; Namba et al., 1994), indicating that CA3–CA1 synapses are normally activated by the opening of AMPA channels.
This work was supported by Spanish Ministerio de Ciencia y Tecnología Grant BFI2002-00936 and Junta de Andalucía Grant CVI-122. We thank María Sutil for her technical assistance and Roger Churchill for his editorial help.
- Correspondence should be addressed to Dr. Agnès Gruart, División de Neurociencias, Universidad Pablo de Olavide, Ctra. de Utrera, Km. 1, 41013 Sevilla, Spain. Email: agrumas{at}upo.es


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When Mt. St. Helen's snow is on, it's ON!!! - and on this day, it was on! I estimated about 60" of fresh untouched, snow was on the ground. No rocks or tree's were poking through. This is the first significant snowfall we've seen here on the south side of MSH in a few years. There was enough snow to get into some of the classic lines I used to love to ride.
Kris, Luke, and myself left my house at 6am and rallied the Suby, with fresh studded tires up to Marble Mountain. The road was plowed to the parking lot, something I haven't seen ever up here. Whoever the plow crew is deserves a pat on the back - no chunder, just smooth, perfect, ice packed roads to travel on. Nice work fella's. We were the 2nd group of people to the lot. Conditions were 2 day old, cold pow, and sun! The skin line was already in place from a few people breaking track the few days prior (Smith, probably being one of them), so it made for easy travel to the 4,800ft mark.
We cut up and across putting our own skin-line in to about the 6,800ft mark, until we reached a nice bench to take lunch, bull shit, rejoice in the surroundings and have some beers. We were high on the ridge just above the SE treeline that was sheltered from the blast in 1980.
We put our splitboards together and rode some of the best snow, and coverage I've ever ridden on Mt. St. Helen's. When conditions are this good here, there's now place I'd rather ride. Deep natural half-pipes in the worm flows, nice little rock drops, some mellow run-outs, and even some pretty steep descents towards the bottom of our run-out. I dipped in first, so once again - there are no riding pictures of me. Photog needed. lol
Being out in these mountains with good people is truly a blessing! I can't wait to get back out!!!


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How to Sell Vintage Print Ads on eBay
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A retired baker, looking to expand his eBay business, engages in a new niche--selling vintage magazine ads on eBay. He shares his initial steps of progress as well as positive results.
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Social responsibility in promoting vintage and contemporary print ads on eBay is just as important as social responsibility in the drinking of alcohol beverages. This hubber/eBayer considers it a privilege to convey this public service message.
The Premise
Are you retired and looking for a way to supplement your fixed income?
Are you in college, hoping to chip away at your mounting student debt?
Are you a young mom wanting to establish an online small business venture?
Are you an ex-felon, your skin toughened by one too many doors slammed in your face?
Are you a teenage entrepreneur, the next aspiring Donald Trump (without the iconic hairdo)?
Are you an established brick and mortar businessman looking for another viable stream of income?
Are you a homeless person, sick and tired of being sick and tired with the looks people give you as you sit on the corner outside the Safeway store, your pride and self-respect even more tattered and foul-smelling than the rags on your back?
Are you a HubPages writer in search of a niche business and the golden possibility of linking both passions to each other?
If you answered YES to one or more of the above, this article may be of some help to you. I don't promise anything magical. What I will provide you is a reason to leave your couch of self-pity, anxiety, boredom, or despair and give you impetus on your road to success.
Again, just so we understand each other, I want to make one thing very clear: This is NOT a get rich quick scheme! I'll provide you with the map of the route I took, but you, my friend, will have to do the walking! If you follow my advice, tweaking the process, of course, to fit your unique personality, style, and circumstances, you'll find this to be a profitable endeavor. If you slack on commitment and work ethic, you'll absolutely fail. I guarantee either side of this scenario. The proverbial adage holds true: You reap what you sow. So let's sow good seed on fertile ground, my friend.
And what am I getting out of this, you ask?
Alright, it's a fair enough question. And here's my fair enough answer.
This is the part where I talk to you about my mission statement in life. In the weeks and months ahead, you'll find me repeating, in the future articles yet to be conceived and created, this core element of what makes me tick.
My mission statement in life can be capsulized in a short and simple prayer--
May I be a positive difference in someone else's life today.
From that root intention stems all kinds of possibilities and opportunities on a daily basis, and my responsibility is to be mindful of when those openings occur.
That's what I get out of this.
And now...a moment of clarification...
Before we go any further, the purists among you will be fidgeting because you'll have this unspoken statement on the tips of your perfectionistic tongues: Yeah, but how in the world does he expect the homeless person to access a computer?
Let me tell you about homeless people, my friends. The emphasis is on the second word, not the first. They're people, and I respect them very, very much. But for the grace of God, I am one step removed from being homeless at any given second, and I never forget that for one moment. Should that misfortune ever happen to me, I will still be a human being with dignity and self-respect.
People who happen to be homeless are the most resourceful people I have ever been blessed to know. Given the misplaced priorities of our municipal, county, state, and national governments, they HAVE to be. I have no doubt in my mind that where there's a will, there's a way.
Okay, 'nuff said! Let's proceed!
The Modus Operandi
- Find Vintage Magazines There are a variety of places where you can find old magazines. Yard sales, estate sales, church rummage sales, thrift stores, consignment stores, and homes of friends and relatives are great places to look for old magazines. You can do a search on eBay, Amazon, Etsy, Craigslist, and other online sites. While ads can be gleaned from all kinds of magazines, I've had a good amount of sales with ads from Life, National Geographic, Saturday Evening Post, Collier's, Esquire, Fortune, Boy's Life, Photography, and Scientific American.
- Prep Work Carefully remove the three or four staples from the spine of the magazine with a staple remover. You may also need a pair of pliers to remove the old and sometimes rusty staples. Stack the pages with ads that you want to post, and toss out the other pages.
NOTE: At this point, my waste not, want not eBay chum, Heather (aka mommymay on HubPages) would want me to tell you to toss those superfluous pages into a cardboard box to be used as filler packing material when shipping fragile objects.
- Invest in a Good Rotary Trimmer Sure, you could trim the raggedy edges of your ads with a pair of scissors, but why torture yourself? Besides, with a good rotary trimmer, you'll save yourself hours of labor. Trust me--I learned this the hard way! The process is very simple. I place an unassembled cardboard box on a bed and slice all of my pages in five to ten minutes. You don't have to precisely measure each edge and mark it with pen or pencil for guidelines. Simply eyeball the entire edge for the amount you want to trim. As you do this, you can simultaneously do your quality control work, throwing away pages that are excessively oxidized, overly wrinkled, or have large holes. If you come across a tiny chip or two, you don't necessarily have to throw the ad away, but be sure to make mention of it in your description when you list your ad. Because a lot of customers matte and frame these ads, any blemishes on the outer edges are moot issues.
- What about ads that are less than a full page size? I make snap decisions about the smaller ads. If my gut tells me that something looks unusual, quirky, campy, politically incorrect by today's standards, aesthetically appealing, iconic, or unique in any other way, I cut them out, too. Some sellers charge less for ads that are smaller. I don't do that. Taking into account that the smaller ads ironically take more work to prepare (including cutting them out by hand), I charge the same amount straight across the board. Respecting oneself is key, in my opinion, to respecting the buyer, the transaction, and the entire gamut of the eBay experience.
- What about half-page ads? With half-page ads, I sometimes cut them to size. My recent research, however, has determined that many sellers leave the half-page ad intact on the full page sheet. If and when I follow suit, I'll mention in my listing the specific size of the ad as well as the dimensions of the full page. I also state that by leaving the ad intact, the buyer has the discretion of determining how wide he wants the specific uncut margin to be prior to matting and framing. Presenting options as win-win situations contributes to the overall goodwill and success of an eBay transaction.
Please Rate This Hub! Aloha and mahalo!
The Bottom Line
I have been selling vintage print ads gleaned from old magazines for approximately eighteen months. In that period of time, I've sold over 360 ads. That breaks down to an average of twenty ads per month. The prices have ranged from $2.95 to when I was first starting out to my current $12.48 per ad. I pay the shipping, so after expenses and fees, I'm netting roughly $8.19 per ad. Currently, I have 884 vintage ads and 188 contemporary ads listed. In addition, I have scores of articles and prints, also removed from the same magazines, that I am selling at $12 or more a pop.
There are sellers of these vintage ads that have been doing this for decades. Their inventory numbers are in the tens of thousands. On a daily basis, a lot of their items are unsold, but it doesn't matter because they're selling double-digit items every single day! It will take me years to match their inventory. I'm not sure that I want to because it takes too much time away from what I truly want to do--my writing. However, a point can be made that amassing vintage print ads over time will enable one to make a comfortable one-niche career on eBay.
I share all of this with you to underscore what I said earlier--If you follow my advice...you'll find this to be a profitable endeavor. By no means is it a get rich quick scheme. As promised, it is a gift of impetus to get you motivated and moving!
E Komo Mai! Welcome to My Other Hubs!
This is an interesting idea. I had no idea.
When you get the time would you email me buddy...email@example.com....I have some questions about all of this...Lizzy is trying to talk me into doing this stuff.
Thanks my friend!
Joe that was a GREAT answer! Thank you.
Hawaiianodysseus, your post is written in an absolutely marvelous manner! Awesome! You did teach me an eBay trick or two after all! Thank you! :)
You really know what you are doing. It is really creative and you can really get money out of it. Thanks for sharing yoru wonderful ideas.
Have a good day! :-)
This may not be a get rich quick scheme but it certainly sounds like a useful second income plan. It is great that you are prepared to share such a lucrative niche market with those who ned to supplment their income. Voted up and useful.
Wow, Joe! This hub needs to get there at the top with the hub of the day! I respect your noble intentions of teaching those of us who need guidance on venturing out into the Ebay business and what a great set of ideas you have laid out for us.
Thanks and I'll be following you on this mission to educate the likes of me who'd like to do something different and succeed. Thanks for showing the way.
Voted up, useful and awesome. Sharing this as well.
Aloha, my friend.
This sounds like an excellent idea. Thanks so much for the idea. I have a couple of magazines here from the 40's. Now I know what I have been saving them for.
This hub was very interesting to me, Joe, because I'm a former vintage ad and other ephemera collector. (I had to stop because the collection was taking over my house! I've since either sold or given away most of those items, keeping only my favorites.)
One of my favorite ads is a fashion illustration, complete with matching Borzoi hounds, from the 1920 issue of Circe, a French publication. I had it carefully framed with acid-free paper and glare-proof glass, and I enjoy looking at it in passing every day.
Your tips are great for niche selling on eBay (or etsy). Thanks for sharing.
Voted Up++
Hi Joe. How creative. You are one clever guy. Who knew that there was money to be made from old print ads? You for one, and good for you. This hub really had me fixated to your every word. Really, you have a way of keeping your readers interest right up to the last word. Just excellent. Voted up, shared, pinned, etc....
I meant continued success on ebay, though sometimes good luck helps. But your success is what counts. know what I mean?
Very interesting hub. I have to look into this. Thanks for the info.
You have done another incredible hub and step by step how to do it right.
And to think when I moved from Wisconsin to Alabama, I left behind a lot of National Geographics. I really have to search my packrat house to see if I have any such things.
Thanks for the tremendous write-up and best of continued luck in your ebay venture.
Where were you when I threw out my old magazine ;) I have to agree with Bill, you made this so interesting to read and added your own warmth and humor. Now if I can only get moving!!
I do have one quick question though, how do you determine shipping costs for your eBay items?
Voted up, useful, and interesting.
Hi Joe,
Good informative hub. There must also be a big buyer's market on eBay.
Great advice and a great hub. Thanks for the tips my friend. I have never tried selling vintage ads on eBay........yet. Your hub answers my questions and I just might give it a try....thanks.
Hi my friend, another great informative hub and loaded with all valuable information . Well done !
Vote up and more !!! Sharing !
Oh my gosh...that's amazing!!! I never knew vintage ads could be sold on Ebay. I had no idea people were even interested in vintage ads. CRAZY!!! Thanks for giving me more fuel for my fire about selling on Ebay. And I love the way you started this hub, describing people that may be interested in it. Quite entertaining reading my friend! Voted up, useful and interesting!
You're not gonna believe this--well, you probably will--but my dad gave me a huge stack of old Life magazines a couple of years ago to do just this and I gave them back last summer. It seemed like an impossible task to go through the magazines without my little one "getting involved", making things too aggravating. Now, I think I could do this without too much trouble, so I'm going to get some of those magazines back and see how I do. Thanks for always sharing such good eBay advice.
One thing I do want to ask: how do you ship them? Do you use a cardboard mailer, or do you sandwich the ad between pieces of cardboard and inside a bubble mailer? Or something else altogether?
You are so wise, my friend. I love your perspectives on life.
Great hub with lots of very specific info! You are a man of compassion, ingenuity and talent for writing. I have found no vintage magazines as of yet, but this will push me forward to look for some. Is there a reasonably priced trimmer you would suggest?
Thanks for sharing this info, Joe!
Very interesting article Joe!
You know... I have somewhere a newspaper from 1968 with the first flight to the moon..... from Belgium! Maybe I should look what it's worth :-) The newspaper is not in super shape as it moved transatlantic... and then 3 other moves.
Have a great day sorting and clipping those adds :-)
Aloha, Joe!
How very clever my friend. I had no idea one could make that kind of money doing this. Bravo!
Above and beyond, you added just enough humor to keep this from being another "how to" hub, and you tossed in a personal mission statement that immediately won me over.
In other words, this hub is excellent and reflects very kindly on the man and writer that you are.
Have a great day my friend.
Blessings coming from sunny Olympia


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President Abdulla Yameen Abdul Gayoom has visited the "Hithaanee Ufaa" camp organized by Sadhaqaath Foundation being held in Rehendi School, Hulhumalé. The camp is organized to perform circumcisions of boys during the school holidays.
The President was accompanied by First Lady Fathimath Ibrahim, Minister of Economic Development Mohamed Saeed, some members of the parliament and senior officials of the government.
During the visit, the President and First Lady met with the boys and their families. In addition the President and First lady participated in a cake cutting with the children, and distributed gifts.
Full details are available at the link below:


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