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Home Latest News Heather Graham, 50, Recreates Her 'Roller Girl' Days From 'Boogie Nights' In...
Heather Graham, 50, Recreates Her 'Roller Girl' Days From 'Boogie Nights' In Cute New Video — Watch
Heather Graham was feeling totally 'inspired' and decided to put on her skates and hit the pavement in a new post she made on her TikTok account! Check out the 'OG Rollergirl' tearing it up to music by The Emotions!
Heather Graham gave her fans a major blast from the past with her September 8 TikTok video! In the clip, seen below, the stunning actress, 50, donned her vintage roller skates and took a spin on the road while wearing a cute, multi-colored mini-dress. Heather looked absolutely flawless grooving to "Best Of My Love" by The Emotions, and she showed that she could still rollerblade just as well as she did in the 1997 film Boogie Nights where she played Rollergirl!
@imheathergraham
I'm inspired by the cool Rollergirls & their good vibes so I put on my skates. Remembering being the OGRollergirl in Boogie Nights. Duet me! #goskate
♬ Best Of My Love – The Emotions
"I'm inspired by the cool Rollergirls & their good vibes so I put on my skates. Remembering being the OGRollergirl in Boogie Nights. Duet me," she captioned her TikTok video. Heather also added the hashtag "go skate," encouraging her followers to get out and get active in the last few days of summer. TikTok is a brand new platform for the celeb, with the video above being her very first post on the social media platform!
But what isn't new for the veteran screen star is her ability to rollerblade like a pro. Heather showed off that same skillset in Boogie Nights, where her character wore roller skates the entire film! After a number of credits in TV shows like Twin Peaks, Heather's star completely skyrocketed when she played the young adult film star in the critically lauded film directed by Paul Thomas Anderson.
At the age of 27, Heather starred alongside Mark Wahlberg, Julianne Moore, Burt Reynolds, John C. Reilly, Philip Seymour Hoffman and more in the ensemble film. The story chronicled the lives of adult film stars in the late '70s and early '80s in California. Boogie Nights went on to earn three Oscar nominations at the 1998 Academy Awards ceremony, including Best Actor in a Supporting Role (Burt Reynolds), Best Actress in a Supporting Role (Julianne Moore), and Best Writing, Screenplay Written Directly for the Screen (Paul Thomas Anderson).
Since her time donning her iconic rollerblades, Heather has gone on to have a flourishing career. She's starred in Austin Powers: The Spy Who Shagged Me, The Hangover, and her 2018 directorial debut Half Magic! Next, Heather appears in the romantic comedy Love, Guranteed opposite Rachael Leigh Cook and Damon Wayans Jr. We cannot wait to see what Heather cooks up next on social media and in her career!
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Pittman Motors is part of the Ametek Precision Motion Control division. Pittman brush and brushless DC servo motors are used in a wide variety of high tech motion applications including lab automation, medical devices, communications equipment, semiconductor processing equipment, aerospace systems, and many other applications where precision motion is critical.
The Pittman motor line spans a wide variety of dc motor sizes and technologies ranging from tiny brushless slotless dc motors used in high speed medical applications to large NEMA frame dc servo motors used in sophisticated automation equipment. If there's a need for precision motion, more than likely there's a Pittman motor to meet your requirements.


Making the decision to build your own home in Scottsdale means dedicating at least a few months to the process. You'll need to choose a builder depending on your timeline, budget, and level of customization. Custom builders and production builders might seem similar, but they offer different pros and cons. Consider each and compare them to your wishlist before making your final decision.
Custom builders create your Scottsdale home, taking you all the way from a blank sheet of paper to a turnkey property. You'll get exactly what you want, but it can also mean a less predictable process. Here are some of the major pros and cons of going custom.
Production and custom builders in Scottsdale each have their pros and cons, so it's a personal choice. Would you rather have the home of your dreams, even if it's a little more expensive? Or is price your true bottom line? By deciding which factors are most important to you, you can choose the type of builder that makes the most sense for you, your family and your build.


text:
est un daimyo de l'époque d'Edo à la tête du domaine de Tokushima.

Clan Hachisuka
Daimyo
Naissance en janvier 1737
Décès en août 1754
Décès à 17 ans

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When I started playing the guitar in 1979, spiral guitar cables were common. Many players favoured these spiral – or coil(ed) – cables because it is very handy not to have meters of cables lying on the floor between your feet and your effects. The spiral cable shortens automatically to a few feet, while it allows distances up to several meters when stretched out, and they don't wrap themselves after turning around a few times such as normal cable does. Last not least, I always found they look cool.
I am not really sure why they got out of fashion a few years later, so much that for decades it was almost impossible to get a long high-quality spiral cable. I was surprised to see them back a few years ago, when Vox presented their Vintage Coil Cord. Fender also has one again. In fact the Vox cable looks pretty much exactly as the cable that Mark Knopfler played live when he started Dire Straits, so I tried to get hold of them. Unfortunately it turned out that they were not available in Germany, and shipping them from the US meant high additional costs. I finally got some from the German distributor that were left over after a music fair. I use them live and for recording since then.
What I like about this cable is the look, the feel, and the sound. It is really true that different cables sound different. This is not vodoo thinking but based on a few technical facts about pick-up design. With these cables the sound becomes smoother with some nice bite in the upper mids. I am not sure if this has to do with Vox's multi-core design forthis cable – different cores for different frequencies – or if it is just the result of the electrical specs. Anyway, it sounds good.
The sound of the cable is especially important when using two of these, before and after a passive effect, like Mark Knopfler apparently did in the late 7oies. With an active effect device, only the cable before the device matters, but with e.g. the Morley volume pedal, both will affect the sound, to be concrete, the capacity of both cables adds and transforms the pick-up's resonance peak, adding some high mids.
This and the next picture prove that one spiral cable was before and one after the Morley volume pedal.
The cable is very reliable. I guess spiral cables were phased out because they can be damaged when you step on them very hard, thus bending the coiled cable to some radius smaller then allowed, which makes the cable break or the shielding loose. The Vox cable, however, is very strong so that it will not be harmed when stepping on it (something I nevertheless try to avoid). Even after using it for a few years now, none of mine makes noises when moving the cable, or shows any other problems.
I recently got some of these great cables which I can offer at a great price in this site's online shop. Check them out!
Anything better than a red Schecter Strat? – Yes, two of them.
Nice article. I have experienced with many coiled cables (vox, fenders, vintage old etc..) I stayed with one now that is magical to me compare to the new ones. I agree as you say Ingo that the vox is very good quality cable with nice tone especially from the new market. More acoustic tone to me compare to others.
Excellent cable that Vox. I have it. I'm wondering if the bullet coil cable sounds compareto the vox. The plug on the bullet seems like Mark's too. Who knows if mark uses bullet back then..
I'm not a guitar player but I think viral cable is a convenient cable to use. I would prefer using this if I was a guitar player.
I still have this spiral cord with me and it still works but I don't use it that much anymore.
Wow, these kind of cords are very rare now. I had a chance to use this kind of cable for an acoustic guitar while on practice at a local studio here and it's very convenient to use. I noticed less tangling of the wires and convenient to walk through the studio while carrying guitar. I also haven't seen one on stores like this one.
Awesome article. I'd be very curious to know how these Vox curly guitar cables sound on bass. In fact, I think I'll test this out and report back!


When you pick up your plans at the Permit and Information Center, your permits will be printed for you and attached to your approved set of plans. The permits will list all required inspections for your project listed in the typical order of construction.
Homeowners and contractors can schedule inspections for their construction projects online. Online scheduling shows all outstanding inspections for a project and allows multiple inspections to be scheduled at one time. If an inspection cannot be scheduled the system explains the reason why.
Schedule before 7 a.m. to request an inspection for that day.
You will receive a confirmation email when your inspection has been scheduled.
Make sure to leave a phone number where we can reach you in case we need to contact you about the inspection.
The IVR system is no longer available for scheduling inspections. If you do not have access to a computer or smart phone, inspections can be scheduled by contacting inspection support staff at 541-682-5283.
Inspections are assigned based on geographic area and availability of inspectors. We do not make appointments for inspections. If you need to meet the inspector on site, it is possible to leave a voice mail message for the inspector to have them call you 20 minutes before the inspection. The inspectors are available by cell phone from 7:30 a.m. to 4 p.m. daily. Their phone numbers can be found on our Staff Contacts page.


If you are looking for a one-stop shop for your electrical system needs you have to go the best experts. MAP Electrical Contractors offer a complete range of services for industrial electrical work while guaranteeing safety.
in all kinds of wiring jobs.
Electrical wiring is an invaluable part of distributing power to various power signals and destinations. MAP Electrical Contractors have specialised in large distribution of power that is necessary for industrial settings.
Our experts are well-versed in single phase wiring that is used in most homes. The phase provides ample power for medium-sized customers, including residential homes and small, nonindustrial businesses as it runs motors up to 5 HP. We also install three-phase wiring, which is ideal for industrial and manufacturing settings as it allows lower voltages and is less expensive to run.
We also install, replace and repair various types and kinds of cable containment like cable and conduit systems. Cable tray wiring systems are simple, flexible, affordable and quickly adapt to complex configurations. MAP Electrical Contractors also install conduit systems that reduce electromagnetic interference.
MAP Electrical Contractors also provide portable appliance testing to ensure the safety of all electrical appliances and equipment for use. Most electrical safety faults are evident through visual examination, but our experts conduct visual checks and tests to ascertain the credibility of the appliances.
Additionally, our specialists help you keep records that aid in reviewing the effectiveness of the company's maintenance policy.
With age and continued use, electrical installations are prone to wear and tear. MAP Electrical Contractors conduct periodic inspections and testing to verify whether the systems are in satisfactory conditions.
After the inspection, we issue a report that highlights all overloaded circuits, potential risks of electric shocks, defective electrical wiring and the adequacy of earthing and bonding. The contractors will repair or replace any electrical systems found defective. They also provide certification, depending on the scope of inspection and testing performed.
Most industrial buildings require emergency lighting in the event of power cuts or fires. MAP Electrical Contractors install emergency lighting according to the current building regulations and any local authority rules.
Our experts are knowledgeable about the intricacies that come with installing fire alarm systems in industries. As such, they provide a complete range of services that includes designing the systems to meet the company's unique needs, programming and conducting final connections and tests to ensure functionality.


Changes are coming to Sharp Hall. The renovations will allow the Woodbrook Players to offer their theater productions three times a year and provide performance space for the Senior Center, Preschool, and other renters, while maintaining the room as a meeting and activity space for all. The project includes construction of a stage at the south end of the room, removing the architectural arch, installing blackout curtains over the window wall that can be opened and closed, installing a stage curtain, converting current storage into a backstage area, converting the recessed area and folding wall on the right side of Sharp Hall to new storage for tables and chairs, providing permanent access from Sharp Hall to the Senior Center activity room, installing stage lighting, and repainting the hall.
What process has the Committee followed?
The Woodbrook Players Committee first met on March 18, 2018. The Committee worked to move the Woodbrook Players to Govans, purchase and install a storage shed, research alternative performance spaces, define requirements for a stage and additional modifications to Sharp Hall, create sketches of the desired future state, send a Request for Proposal (RFP) to three architecture firms, review those proposals, and select one firm (Carl Schwartz) to move forward.
Financial support of the Woodbrook Players was specified in the merger Plan of Union. Session has approved draft sketches of the stage along with a $150k fund, which was allocated from the proceeds of the Brown Memorial Woodbrook estate.
The Committee is composed of Session members, Church members, and key members of the Woodbrook Players company. The following people have participated in meetings and contributed in various ways to this process: Stephen Gaede (Chair), Beth Bryant, Peter Conrad, Mike Field, Tom Harris, Ron Oaks, Sherry Peck, Ashley Short, Jocelyn Taylor, Wayne White, Bob Wong, Elsie Wong, and Maria Wong.
How often does the Woodbrook Players hold performances?
The Woodbrook Players hold performances three times a year – typically a play in the fall, a fundraiser in the winter (with no set), and a musical in the spring.
Have the Woodbrook Players looked at alternative places to hold performances?
N. Baltimore Mennonite Church on Roland Park Ave – they have a nice stage but the rental fee was $27/hr.
What changes is the Committee proposing to make to Sharp Hall?
Building a stage 28 in. high behind a proscenium arch and 6 ft. thrust in front of arch at same height.
The stage will be 24 feet deep, measured from the windows into Sharp Hall.
Removal/renovation of storage closets to right of stage to create a wing and off-stage storage.
Remove false ceiling above stage for light and sound equipment.
Adjust or remove built-in areas behind the proscenium arch and at the arch for the stage hangings etc.
Ability for actors to enter the stage from the audience on the left or right side of the stage via steps, 4 steps 7 inches high.
Storage under the stage for folded and stacked tables, to be slid underneath using carts or other system. This space may alternatively be used for prop storage.
Black curtain across the back window wall that can open to reveal light when the stage is not in use.
Sky drop 5 ft. from back window wall and width of stage that can be drawn open or closed (if allowed per fire code).
Two rows of black teasers across the top of the stage to hide light and sound equipment.
Two sets of black tormentors on either side of the stage to block entrances.
Curtain at the proscenium arch, 17' 4" from the back window wall.
A pole or other support hanging from the ceiling 20 feet from stage for lights.
Wiring for all sound and lighting equipment in theatre area and hall for running shows.
Creation of a closet in Sharp Hall for chair/table storage in the enclosure between the Hall and the Senior Center.
Creating an entrance between the Senior Center and Sharp Hall.
What will happen to the window wall and natural light?
As seen in the proposed changes above, the window wall will be preserved, but the bottom 28" will be hidden by the stage. During productions the windows will be covered by black curtains. When the stage is not in use the curtains can be opened to allow natural light from the window wall. In addition, sets can be designed to open up/fold and reveal the window wall.
What will happen to the decorative arch?
The decorative arch will be removed, transformed into a proscenium arch which will frame the stage and hide curtains.
Will the stage limit table space for gatherings? Will the stage impact the ability to accommodate rentals such as weddings?
The floor space will be large enough to accommodate 16 tables with 96 people. If more space is needed, we can place tables and chairs on the stage. Unfortunately, Tom has mentioned that large wedding rentals are becoming less common as people are opting to get married outside of churches.
The Woodbrook Players would like to perform the musical 1776 the second week of May. If permits, bidding and construction go according to schedule we will have a stage in place by April 15. If there are delays then we will address them along the way, prioritizing construction accordingly.
Are we confident that the hall can support the weight of a stage and lights?
We will have a structural engineer review the requirements, looking at the floor and ceiling to ensure that the space can accommodate the stage and lighting system.
What about electricity load and lighting?
We will have an electrician review existing infrastructure as well to ensure we can accommodate lights.
Are we taking into account acoustics?
Although we are not working with an acoustical engineer, we will ensure the space works well acoustically.
Do we have enough money in the budget - is $150k a strict budget?
This budget was characterized as "comfortable" by two of the architecture firms who submitted proposals, including Carl and the builder he is recommending. We aim to fit within that budget but understand there may be unknowns and overruns as we get further into construction. We won't really know how tight the budget is until we receive bids back from contractors.
When the stage is complete, will the stage be truly functional so that the Woodbrook Players will be happy with the results now and in the future?
Woodbrook Players Director Ron Oaks has been involved with the process from the beginning and he is optimistic that the final outcome will be a practical/workable solution.
What about use by the Senior Center and Pre-School?
The stage will be available to be used by more than the Woodbrook Players, including the Pre-School, Senior Center, and church committees/events (such as Summer Vacation Bible School). Having a stage will be a real asset for the church which will engage current members and potentially attract new ones to our congregation.
Everyone is welcome here. And we mean everyone as in all colors, cultures, orientations, identities, abilities, levels of perfection, or lack thereof. And we mean welcome as in, please become a full, participating member of our community and share your gifts at any level of leadership. Everyone is welcome here.
NOTE: Occasionally this schedule changes when we come together for a combined 10:30 a.m. worship service. Check the homepage for updates.


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EBOLA CURE: Kalbar vet's vital role in finding a treatment
Home » EBOLA CURE: Kalbar vet's vital role in finding a treatment
Daisy Bernstein
A KALBAR facility has played a vital role in finding the cure for Ebola.
More than 11,000 victims died and 30,000 were infected by Ebola in just two years during the virus outbreak in 2014.
The largest recorded outbreak of Ebola virus occurred primarily in West Africa from 2014 to 2016, with exported cases in Europe and North America.
Like many people local vet Shane Belford watched in horror as news story after news story showed the effects the outbreak was having on thousands.
He got the chance to help the dire situation when he received a call in October of 2014 which was the catalyst for his journey into vital Ebola research.
Shane works for Plasvacc, a facility in Kalbar that manufactures and distributes high quality blood plasma products for use in veterinary medicine.
He was part of the international team led by Queensland researchers who developed the effective and economical treatment for Ebola.
The research was led by University of Queensland School of Chemistry and Molecular Bioscience's Professor Alexander Khromykh and QIMR Berghofer Medical Research Institute's Professor Andreas Suhrbier.
It involved injecting horses with a specially-developed vaccine.
The horses then produced antibodies which were used to create the Ebola treatment.
"It was October 2014 when I was first approached to be part of the team," Mr Belford said.
"Our role was small but vital. We were actually making the active ingredient for the serum.
"QMI developed the vaccine which allows the horses to produce antibodies against a specific protein in Ebola. I then injected the vaccines into the horses and they did the rest. It provoked an immune response and they produces adequate amounts of antibodies. Those antibodies were successful in treating the Ebola virus."
The research paper was officially published in February this year.
"It was exciting. It was great to be involved in good research and was fantastic to be a part of the solution," Mr Belford said.
"We have a track record in producing equine-based antibodies as an active pharmaceutical ingredients for humans already. We help produce snake anti-venom plasma so if you go to hospital after a snake bite it's a 50% chance the antibodies came from us.
"Horse antibodies have been used in many products. Who knows what we can achieve if we keep going down this path of treating disease with immune therapy."
Mr Belford said the potential for using a similar method to treat future diseases was exciting.
"It was great to know we have the ability if we were suddenly exposed to some unusual virus we have a track record on being able to combat it," the vet said.
"Hopefully it leads to more work in that field for our business. The CSIRO contacted me and asked to do more work for them in the future so we will try and work together on future projects.
"It proves you don't have to live in a big city to make a difference in the scientific world."
Professor Khromykh said the post-exposure treatment made with antibodies from horses could be used in the next Ebola outbreak.
"This is a cost-effective treatment that can be used in low-income countries in Africa where equine production facilities are already in operation for producing snake-bite antivenin," he said.
"It's the first time that equine antibodies have been shown to work effectively against Ebola infection.
"The research resulted from a strong collaboration between Australian, French and Russian scientists and a local Queensland company (Plasvacc)."
Professor Khromykh said this led to the development of monoclonal antibodies that were used in the UK to treat infected health workers returning from Africa.
"The down side is that monoclonal antibodies require considerable investment for scale-up and manufacture, and are expensive," he said.
"Equine antibodies are a considerably cheaper alternative, with manufacturing capacity already in place in Africa.
"Antibodies from vaccinated horses provide a low-cost alternative, and are already in use for rabies, botulism and diphtheria."
As well as Plasvacc, the study included researchers from UQ, the State Centre for Virology and Biotechnology "Vector" in Russia, CSIRO, INSERM and Claude Bernard University in France and the United Nations Medical Service in New York.
UQ School of Chemistry and Molecular Biosciences Head Professor Paul Young, who was part of the research team, said the finding offered great hope as a rapid treatment option for Ebola patients
"It's a significant advance on the way we think about responding to urgent disease threats, and could be applied to the treatment of other infectious diseases," he said.
"It is also a far more appropriate option for resource-poor settings."
Professor Khromykh's team has previously developed an experimental Ebola vaccine made using an Australian virus called Kunjin that might also help in the fight against the deadly Ebola virus.
The Kunjin virus-derived vaccine vector first constructed by Professor Khromykh in 1997 has been used to develop several vaccine candidates, including one for Ebola.
The research, published in the Nature series journal Scientific Reports, was funded by the Australian Infectious Diseases Research Centre.
This material was created specially for WorldHealthNews project by Daisy Bernstein.
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Katarzyna Devraj was interviewed by business TV portal "Capital TV". The topic was "Feng shui in business". You can listen to the entire interview by clicking on the link below. The interview is in Polish language only.


There are benefits to both private and group lessons. In private lessons, students get personal attention for the entire class, and each lesson is tailored specifically to the individual student's needs. The classes are dynamic, entertaining, and with the right amount of challenge for each student.
This method is very effectively taught in groups where students get the added benefit of learning through interacting with each other. This is often inspiring, motivating, and fun.
Group lessons range from 30 minutes for two students to 40 minutes for three students. Students are matched according to their ages and skill levels.


This page shows business listings of all kinds in Ayr beginning with the letter B. You can also view listings by type of business or by area of Ayr.
Do It Yourself Shops (Shops & other Retail Outlets) in Sanquhar Farm Road, Ayr.
Architects (Business at Home) in Bellevue Crescent, Ayr.
Joiners & Carpenters (Business at Home) in Northfield Avenue, Ayr.
Night Clubs (Sports, Leisure, Entertainment) in Main Street, Ayr.
Plasterers, Screeders & Dry Lining Contractors (Business at Home) in Low Road, Ayr.
Restaurants - English (Shops & other Retail Outlets) in Dunure Road, Ayr.
Health Authorities (Sports, Leisure, Entertainment) in Highfield Drive, Ayr.
Garage Services (Workshop & Repair Centres) in Maybole Road, Ayr.
Electricians & Electrical Contractors (Head Office) in Green Street Lane, Ayr.
Charities & Voluntary Organisations (Office & Administration) in Viewfield Road, Ayr.
Shoe Shops (Shops & other Retail Outlets) in High Street, Ayr.
General Stores (Shops & other Retail Outlets) in Dalmellington Road, Ayr.
Bathroom Equipment & Fittings (Shops & other Retail Outlets) in Alloway Street, Ayr.
Hotels (Shops & other Retail Outlets) in Queens Terrace, Ayr.
Cafes, Snack Bars & Tea Rooms (Shops & other Retail Outlets) in Seafield Road, Ayr.
General Stores (Shops & other Retail Outlets) in Beech Grove, Ayr.
Guest Houses (Shops & other Retail Outlets) in Prestwick Road, Ayr.
Carpets & Rugs - Retail (Shops & other Retail Outlets) in Arran Mall, Ayr.
Carpets & Rugs - Retail (Shops & other Retail Outlets) in Old Bridge Road, Ayr.
Estate Agents (Office & Administration) in Citadel Place, Ayr.
Nail Technicians (Shops & other Retail Outlets) in Sandgate, Ayr.
Hotels (Shops & other Retail Outlets) in Belleisle, Ayr.
Golf Courses & Clubs (Sports, Leisure, Entertainment) in Belleisle, Ayr.
Nursing Homes (Hospitals & Medical Establishments) in Racecourse Road, Ayr.
Leisure & Recreation Centres (Sports, Leisure, Entertainment) in Bellsbank Road, Dalmellington.
Fish & Chip Shops (Shops & other Retail Outlets) in Merrick Drive, Dalmellington.
Libraries (Police, Fire, Ambulance) in Craiglea Crescent, Dalmellington.
Hotels (Shops & other Retail Outlets) in Bellsbank Road, Dalmellington.
Education Agencies & Authorities (Office & Administration) in Craiglea Cresent, Dalmellington.
Post Office Services (Shops & other Retail Outlets) in Merrick Drive, Dalmellington.
Schools - Local Authority (School & Educational Establishments) in Craiglea Crescent, Dalmellington.
General Stores (Shops & other Retail Outlets) in Merrick Drive, Dalmellington.
Convenience Stores (Shops & other Retail Outlets) in Merrick Drive, Dalmellington.
Adult Education Centres (School & Educational Establishments) in Craiglea Crescent, Dalmellington.
Schools - Local Authority (School & Educational Establishments) in Belmont Road, Ayr.
Guest Houses (Business at Home) in Park Circus, Ayr.
Newsagents (Shops & other Retail Outlets) in Belmont Road, Ayr.
Post Office Services (Shops & other Retail Outlets) in Dalmellington Road, Dalmellington Road.
Car Body Repairs (Workshop & Repair Centres) in Green Street, Ayr.
Beds, Bedding & Blankets (Shops & other Retail Outlets) in Liberator Drive, Heathfield Industrial Estate.
Chemists - Dispensing (Other) in Beresford Terrace, Ayr.
Chiropodists (Podiatrists) (Hospitals & Medical Establishments) in Dalblair Road, Ayr.


This 2015 Harley-davidson Fxsb Break sustained All Over damage. This unit is confirm to run and drive. This unit is being sold with a salvage title. You are more than welcome to contact us to schedule an appointment for vehicle inspection. Mon-Fri 8am-4pm.


text:
You asked: How did Jesus call Simon?
How was Simon Peter called?
What did Jesus say about Simon?
Why did Jesus call Simon Peter the rock?
What does the name Cephas mean?
What is the 12 disciples name?
What did Jesus say to Peter after he denied him?
What did Jesus say to Peter about denying him?
John the Baptist (1:35) and had heard John indicate that Jesus was the Lamb of God—left John and introduced Peter to "the Messiah," who at that time gave him the name (or title) Cephas (i.e., Peter, or Rock).
Again Jesus said, "Simon son of John, do you truly love me?" He answered, "Yes, Lord, you know that I love you." Jesus said, "Take care of my sheep."
"I think that Jesus was telling Peter that he (Peter) was the rock because his name means 'rock,'" says Hillary, 12. The rock upon which Jesus would build his church could refer to Peter, since Jesus changed Peter's name to "petros" meaning "rock." This would make Peter the foundation of the church.
The name Cephas is a boy's name of Aramaic origin meaning "rock". The apostle Simon was called Cephas by Jesus because he was to be the rock upon which the Christian church was to be built. In most versions of the New Testament, Cephas is translated into Petros in Greek, Peter in English.
IMPORTANT: How do I pray for my family?
When morning came, he called his disciples to him and chose twelve of them, whom he also designated apostles: Simon (whom he named Peter), his brother Andrew, James, John, Philip, Bartholomew, Matthew, Thomas, James son of Alphaeus, Simon who was called the Zealot, Judas son of James, and Judas Iscariot, who became a …
Jesus restored Peter to fellowship after Peter had previously denied him and told Peter to feed Jesus' sheep.
Jesus answered, "I tell you, Peter, before the rooster crows today, you will deny three times that you know me." John 13:31–38. When he was gone, Jesus said, "Now is the Son of Man glorified and God is glorified in him.


Masao Hayashi, M.D.; Sumedha G. Penheiter, Ph.D.; Tetsuzo Nakayama, M.D.; Alan R. Penheiter, Ph.D.; David O. Warner, M.D.; et al Keith A. Jones, M.D.
* Research Fellow, † Research Technologist, ‡ Assistant Professor, § Professor.
There are conflicting reports concerning the efficacy of β2-receptor agonists when administered in the presence of volatile anesthetics. In several studies using a variety of experimental models, halothane impaired β2receptor–mediated cell function,15,16 e.g. , halothane markedly attenuated isoproterenol-induced relaxation of vascular smooth muscle.16 This effect was considered to be due to inhibition of the β2receptor and/or its cognate stimulatory heterotrimeric guanosine-5′-triphosphate (GTP)–binding protein (G protein), because smooth muscle relaxation induced by activation of the signaling pathway distal to the G protein was not inhibited by halothane.16 On the other hand, β2-receptor agonists relieve bronchospasm during halothane anesthesia in animal models of asthma.17 In addition, we found that halothane did not affect the ability of isoproterenol to relax isolated canine tracheal smooth muscle contracted with acetylcholine.18 However, because halothane alters numerous other signaling systems in intact cells, this latter study could not be unambiguously interpreted in the context of direct anesthetic effects on β2receptor–Gαscoupling. Because halothane inhibits the coupling between the muscarinic receptor and the heterotrimeric G protein Gq/11,19 similar effects on the β2receptor–Gαscomplex could occur.
The purpose of the current study was to investigate whether halothane affects the functional coupling between the β2receptor and its cognate heterotrimeric G protein, Gs. We tested the hypothesis that halothane does not affect isoproterenol-promoted guanosine nucleotide exchange at the α subunit of Gs(Gαs) and hence would not affect isoproterenol-induced decreases in Ca2+sensitivity in permeabilized airway smooth muscle. Experiments were also conducted to assess the effect of halothane on the biochemical coupling between the β1receptor and Gαsas a positive control, because halothane is known to inhibit isoproterenol binding to this receptor20 and hence should inhibit β1–Gαscoupling.
After obtaining approval from the Mayo Foundation Institutional Animal Care and Use Committee (Mayo Foundation, Rochester, Minnesota), porcine tracheas were procured by euthanasia of research animals. The animals were first anesthetized by intramuscular injection of Telazol (10 ml/kg) (Fort Dodge Animal Health, Fort Dodge, IA) and xylazine (6 mg/kg) and intravenous injection of Nembutal (400–600 mg) (Ovation Pharmaceuticals Inc., Deerfield, IL), and then killed by exsanguination via bilateral transection of the carotid arteries. Then, the extrathoracic tracheas were excised and immersed in chilled physiologic salt solution of the following composition: 110.5 mm NaCl, 25.7 mm NaHCO3, 5.6 mm dextrose, 3.4 mm KCl, 2.4 mm CaCl2, 1.2 mm KH2PO4, and 0.8 mm Mg2SO4. After removal of fat, connective tissue, and epithelium, the tracheal smooth muscle was cut into strips (0.8–1.2 cm long × 0.25–0.5 mm wide) for isometric force measurements.
COS-7 cells (American Type Culture Collection, Manassas, VA) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (10% DMEM) and penicillin and streptomycin (50 U/ml each). The day before transfection, confluent cells were trypsinized and seeded in 10-cm tissue culture dishes so as to reach 90% confluence in 24 h (approximately 4.5 × 106cells per 10-cm dish). The cells were then transiently transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) as per the manufacturer's recommendation. Briefly, for each 10-cm plate transfection, the complementary DNA (cDNA) constructs were each mixed (5 min, room temperature) with 1.5 ml Opt-MEM I (Invitrogen) in a 50-ml falcon tube. For transfection of Gαsonly, 5 μg cDNA plus 5 μg of the vector plasmic cDNA 3.1 were used for each 10-cm plate transfection; for cotransfection experiments, 5 μg β1or β2cDNA and 5 μg GαscDNA were used. Lipofectamine 2000 was mixed in another falcon tube with 1.5 ml Opti-MEM (2.5 μg/μl cDNA to be transfected). The two solutions were then mixed and allowed to stand for 20 min at room temperature to promote DNA–Lipofectamine complex formation. Three milliliters of the transfection mixture was added to each 10-cm plate with 5 ml 10% DMEM without penicillin/streptomycin. The transfection mixture was replaced with 7 ml fresh 10% DMEM plus penicillin/streptomycin after 12 h. Twenty-four hours after transfection, the cells were washed twice with phosphate-buffered saline, scraped in ice-cold phosphate-buffered saline, transferred to 1.5-ml microfuge tubes, and pelleted by centrifugation at 500g (2 min at 4°C). The cells were flash-frozen in liquid nitrogen and stored at −80°C until they were used to prepare crude membranes.
Crude membrane was prepared as previously described for tissue with several modifications.19,28 Frozen cells from three 10-cm plates were suspended for 15 min in ice-cold lysis buffer (500 μl per plate) composed of 20 mm HEPES (pH 8.0), 1 mm EDTA, 0.1 mm phenylmethysulfonyl fluoride, 10 μg/ml leupeptin, and 2 μg/ml aprotinin and then gently homogenized on ice by repeated passage through a 27-gauge needle (approximately 10–12 times). The lysate was then subjected to low-speed centrifugation (400g , 10 min, 4°C) to remove intact cells and nuclei and then to ultracentrifugation (87,000g , 30 min, 4°C) to pellet the crude membrane. The membrane pellet was washed with lysis buffer and resuspended in assay buffer (100 μl per plate) composed of 50 mm Tris-HCl (pH 7.4), 100 mm NaCl, 4.8 mm MgCl2, and 1 μm GDP, and the membrane protein concentration was determined.29 The crude membrane suspension was then diluted with assay buffer to a protein concentration of 2 mg/ml, frozen in liquid nitrogen and stored at −70°C until used for the assay.
Gαsnucleotide exchange was assayed using methods originally described by Barr et al. 30 and previously used by our laboratory.28 Reaction mixtures containing 10 μg membrane protein, 100 mm NaCl, 4.8 mm MgCl2, and 1 μm GDP, with or without halothane and with or without isoproterenol, in a total volume of 62 μl were preincubated for 5 min at 30°C. The assay was initiated by the addition 5 μl of the radioactive, nonhydrolyzable form of GTP, [35S]GTPγS, to the reaction mixture (1,250 Ci/mmol; 10 nm final concentration in assay). Reactions were terminated at times according to experimental design, and the supernatant was subjected to the immunoprecipitation step of the assay. Radioactivity was quantified by scintillation counting. Background radioactivity measurements were determined by performing tandem experiments with the same amount of protein except that the assay was immediately terminated with 600 μl ice-cold immunoprecipitation buffer. The amount of background radioactivity was less than 30% of the radioactivity of the basal Gαsnucleotide exchange measurements. Halothane had no effect on this nonspecific background radioactivity. Data were normalized to the amount of protein and the specific activity of the [35S]GTPγS in the assay, and each experimental condition was assayed in triplicate.
β1and β2were quantified in the crude membranes prepared from COS-7 cells by saturation binding with the tritium-labeled β1/β2receptor antagonist [5,7-3H](−)CGP-12177.31,32 Specific binding was determined in triplicate assays with a single saturating concentration of [5,7-3H](−)CGP-12177 of 6 nm. Nonspecific binding was determined in the presence of 5 μm propranolol. Aliquots of membranes containing 20 μg protein from COS-7 cells cotransfected with β1and Gαs, β2and Gαs, or untransfected COS-7 cells were incubated (90 min, 25°C) in a total volume of 0.5 ml containing 6 nm [5,7-3H](−)CGP-12177, with or without 5 μm propranolol, 50 mm Tris, and 10 mm MgCl2(pH 7.4). After the 90-min incubation, reactions were applied to prewetted Whatman GF/B filters using a Brandel cell harvester (Gaithersburg, MD) and washed three times with 10 ml Tris, 50 mm, pH 7.4, and 10 mm MgCl2. Bound radioactivity on the filters was quantified by scintillation counting.
For physiologic studies of permeabilized airway smooth muscle strips in the superfusion apparatus, halothane was added to the aqueous buffers via a calibrated halothane vaporizer by vigorous aeration of the solutions as previously described.11,12,33 For biochemical studies of Gαsnucleotide exchange, saturated aqueous stocks of halothane were prepared and added directly to the reaction tubes in volumes that produced the desired final anesthetic concentration.34–37 To account for the unavoidable rapid loss of halothane upon mixing in the assay tubes, tandem experiments were conducted under the same assay conditions where anesthetic concentrations were measured after hexane extraction by gas chromatography using an electron capture detector (Hewlett-Packard, Waltham, Massachusetts, model 5880A) according to the method of Van Dyke and Wood38 and previously used by our laboratory24,28,35,39,40 In preliminary studies, we found that after an initial loss of approximately 30% due to transfer and mixing, the amount of halothane in the reaction tube was relatively stable, with less than 5% additional loss during the longest assay time in this report (5 min).
Effect of Halothane on Isoproterenol-induced Inhibition of Ca2+Sensitivity.
An example of the experimental protocol is shown in figure 1. After permeabilization of a pair of strips obtained from the same animal, one strip was superfused with relaxing solution containing 0.73 ± 0.17 mm halothane (2.7 ± 0.7 MAC for pigs),37 which was maintained for the duration of the protocol. The second strip was not exposed to halothane and served as a control for the effect of isoproterenol on isometric force. Then, both strips were contracted by superfusion with solution containing 180 nm free Ca2+plus 1 μm GTP for 15 min, which, in preliminary studies, induced contractions (referred to hereafter as "initial force") of 40–50% of maximal force that were stable for up to 2 h. Concentrations of GTP less than 5 μm do not induce changes in isometric force induced by free Ca2+alone (preliminary data not shown). Finally, concentration–response curves were generated for isoproterenol (0.1–100 μm) with both strips during constant activation with 180 nm free Ca2+plus 1 μm GTP by increasing the isoproterenol concentration in the superfusate every 15 min. The effect of isoproterenol on isometric force is expressed as a percentage of the initial forces induced by 180 nm free Ca2+.
Fig. 1. Representative tracings of the experimental protocol designed to determine the effect of halothane on isoproterenol-induced inhibition of isometric force in porcine tracheal smooth muscle strips permeabilized with Staphylococcus aureus α-toxin. See text for description of the experimental protocol .
Effect of Isoproterenol on Gαs[35S]GTPγS–GDP Exchange.
These studies were conducted to determine the time course for [35S]GTPγS incorporation into Gαsand the concentrations of isoproterenol that produce half-maximal or maximal promotion of Gαs[35S]GTPγS–GDP exchange. These data were then used to guide the design of subsequent protocols to examine the effect of halothane on Gαs[35S]GTPγS–GDP exchange. To determine the time course for Gαs[35S]GTPγS–GDP exchange, crude membrane prepared from β1–Gαsor β2–Gαscotransfected COS-7 cells were incubated for 5 min without (for basal [35S]GTPγS–GDP exchange measurements) or with (for isoproterenol-promoted [35S]GTPγS–GDP exchange) 10 nm isoproterenol, and then the reactions were initiated with [35S]GTPγS. The reactions were terminated after 1, 2, 5, or 10 min, and then the samples were subjected to the immunoprecipitation step of the assay.
To determine the isoproterenol concentration that produced half-maximal and maximal promotion of Gαs[35S]GTPγS–GDP exchange, crude membrane prepared from β1–Gαsor β1–Gαscotransfected COS-7 cells were incubated without or with various concentrations of isoproterenol (0.01–1,000 nm) for 5 min. The reactions were then terminated 10 min after initiation with [35S]GTPγS, and the samples were subjected to the immunoprecipitation step of the assay. The isoproterenol-promoted increase in Gαs[35S]GTPγS–GDP exchange was expressed as a percentage of the difference between the basal exchange values (measured in the absence of isoproterenol) and those measured in the presence of the isoproterenol concentration that produced the maximal effect.
Characterization of GαsNucleotide Exchange Assay.
To determine the dependence of isoproterenol-promoted Gαs[35S]GTPγS–GDP exchange on expression of the human β1or β2adrenergic receptor (referred hereafter as β1or β2, respectively), measurements were made using crude membrane prepared from COS-7 cells transfected with cDNA for human Gαsonly, or cotransfected with the cDNAs for β1and Gαs, or β2and Gαs. The crude membrane was incubated with or without 10 nm isoproterenol for 5 min, and then the reactions were initiated with [35S]GTPγS. The reactions were terminated after 5 min, and the samples were subjected to the immunoprecipitation step of the assay.
Effect of Halothane on [35S]GTPγS–GDP Exchange.
Crude membrane prepared from β1–Gαsor β2–Gαscotransfected COS-7 cells were incubated without or with 10 nm isoproterenol for 5 min. Then, the samples were incubated with 0.75 or 1.5 mm halothane (approximately 2.8 and 5.6 MAC for pigs, respectively)37 for an additional 5 min, and the reactions were initiated with [35S]GTPγS. Finally, the reactions were terminated after 5 min, and the samples were subjected to the immunoprecipitation step of the assay.
The cDNAs for the human β1and β2receptors and human Gαsin the expression vector plasmic cDNA 3.1 were obtained from the University of Missouri-Rolla cDNA Resource Center.1Adenosine-5′-triphosphate disodium salt was purchased from Research Organics, Inc. (Cleveland, OH). Halothane was purchased from Ayerst Laboratories, Inc. (New York, NY). Staphylococcus aureus α-toxin and rabbit nonimmune serums were purchased from Calbiochem (EMD Biosciences, Inc. Affiliate, San Diego, CA). The Gαsrabbit polyclonal antiserum was produced by Covance Research Products (Denver, PA) using a synthetic decapeptide that corresponds to the carboxy-terminal sequence for human Gαs(RMHLRQYELL). This antiserum is highly specific for Gαsand does not cross-react with recombinant, purified Gαior Gαqprotein. Protein A-agarose beads were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Lowry protein assay kits were purchased from Bio-Rad Life Science Research Produces (Hercules, CA). All other chemicals were purchased from Sigma Chemical Company (St. Louis, MO). A23187 was dissolved in dimethyl sulfoxide (0.05% final concentration). All other drugs and chemicals were prepared in distilled, filtered water.
Data are reported as mean ± SD; n represents the number of animals studied or independent times an assay was performed. For concentration–response curves, EC50and maximal agonist concentrations were determined by nonlinear regression analysis as described by Meddings et al. 44 In this method, a dependent variable (y ), such as isometric force or Gαs[35S]GTPγS–GDP exchange, for any concentration of drug (c ) is given by the equation y =vc /(EC50+c ), where v represents the maximal response and EC50represents the concentration that produces a half-maximal response for that drug. Nonlinear regression analysis was used to fit values of v and EC50to data for y and c for each condition studied. For the time course curves, the data for Gαs[35S]GTPγS–GDP exchange were fit with the equation y =a (1 −e −kt ) using nonlinear least squares fitting. The independent variable is time (t ), the dependent variable is the amount of [35S]GTPγS-bound Gαsimmunoprecipitated from solution (y ), the parameter k is the rate of Gαs[35S]GTPγS–GDP exchange, and the parameter a vertically scales the curve and is the maximal value. Repeated-measures analysis of variance with post hoc testing performed using the Student-Newman-Keuls test was used to compare values of k and a and to determine the effect of halothane Gαs[35S]GTPγS–GDP exchange. For all statistical comparisons, a value of P < 0.05 was considered significant.
Increasing the free Ca2+concentration in the superfusate from 1 to 180 nm in the presence of 1 μm GTP caused a sustained increase in isometric force to 47.7 ± 9.3% of maximal force induced by 10 μm free Ca2+(fig. 1). The presence 0.73 ± 0.17 mm (2.7 ± 0.7 MAC) halothane in the superfusate had no significant effect on this contraction (41.6 ± 9.6% of maximal isometric force; P = 0.40). The subsequent addition of isoproterenol to the superfusate caused a concentration-dependent inhibition of isometric force, which was not significantly attenuated by halothane (figs. 1 and 2). The EC50values for inhibition of Ca2+sensitivity by isoproterenol were 0.38 ± 0.15 and 0.25 ± 0.05 nm for curves generated in the absence and presence of halothane, respectively (P = 0.13). Likewise, there was no significant effect of halothane on maximal inhibition of Ca2+sensitivity by 100 μm isoproterenol (17.1 ± 4.3 and 13.2 ± 6.6% of initial force for curves generated in the absence or presence of halothane, respectively; P = 0.36).
Fig. 2. Effect of halothane (0.73 ± 0.08 mm; 2.7 ± 0.9 minimum alveolar concentration) on isoproterenol-induced inhibition of isometric force in porcine tracheal smooth muscle permeabilized with Staphylococcus aureus α-toxin. See text for description of the experimental protocol, which is depicted in figure 1. Effect of isoproterenol on isometric force is expressed as a percentage of the initial force induced by 0.18 μm free Ca2+plus 1 μm GTP. Data are mean ± SD; n = 5 .
In the absence of isoproterenol, membranes prepared from COS-7 cells cotransfected with β1–Gαs(fig. 3A) or β2–Gαs(fig. 3B) incorporated [35S]GTPγS into Gαsin a time-dependent manner with apparent rate constants of k appof 0.045 ± 0.054 and 0.053 ± 0.061 min−1, respectively. There was no significant difference in k appbetween the two preparations (P = 0.8), whereas the maximal exchange value, a , measured at 10 min was significantly greater with membrane prepared from the β2–Gαscotransfected cells (fig. 3).
Fig. 3. Time-dependent change in exchange of the radioactive, nonhydrolyzable form of guanosine-5′-triphosphate (GTP), [35S]GTPγS, for guanosine-5′-diphosphate (GDP) ([35S]GTPγS–GDP exchange) at the α subunit of the Gsheterotrimeric G protein. Gαs[35S]GTPγS–GDP exchange was measured in the absence and presence of 10 nm isoproterenol using crude membranes prepared from COS-7 cells cotransfected with the complementary DNAs encoding for β1and Gαs (A ) or β2and Gαs (B ). The reactions were terminated 1, 2, 5, and 10 min after initiation of the assays with [35S]GTPγS. Data are mean ± SD; n = 4 .
The presence of 10 nm isoproterenol in the assay buffer promoted Gαs[35S]GTPγS–GDP exchange, increasing k appin both preparations by approximately fourfold to fivefold to 0.294 ± 0.024 and 0.351 ± 0.066 fmol/min for membranes prepared from β1–Gαsand β2–Gαscotransfected cells, respectively; these values were not significantly different (P = 0.25). The value of a was significantly greater in membrane prepared from the β2–Gαscotransfected cells, whereas the relative magnitude of the maximal increase in Gαs[35S]GTPγS–GDP exchange above basal values was similar between the two preparations, approximately threefold. For both preparations, the isoproterenol-promoted increase in Gαs[35S]GTPγS–GDP exchange above basal exchange was concentration dependent, with EC50values of 2.8 ± 0.4 and 2.4 ± 0.4 nm for membranes prepared from β1–Gαs(fig. 4A) and β2–Gαs(fig. 4B) cotransfected cells, respectively; these values were not significantly different (P = 0.39).
Fig. 4. Concentration-dependent effect of isoproterenol (ISO) on the exchange of the radioactive, nonhydrolyzable form of guanosine-5′-triphosphate (GTP), [35S]GTPγS, for guanosine-5′-diphosphate (GDP) ([35S]GTPγS–GDP exchange) at the α subunit of the Gsheterotrimeric G protein. Gαs[35S]GTPγS–GDP exchange was measured in crude membranes prepared from COS-7 cells cotransfected with the complementary DNAs encoding for β1and Gαs( A or β2and Gαs (B )). Assays were performed in the absence and presence of isoproterenol (0.01–1,000 nm), and the reactions were terminated 5 min after initiation of the assays with [35S]GTPγS. The isoproterenol-promoted increase in Gαs[35S]GTPγS–GDP exchange was expressed as the percentage of the difference between the values measured in the absence of isoproterenol and that measured in the presence of the isoproterenol concentration that produced the maximal effect. Data are mean ± SD; n = 4.
The endogenous levels of β1/β2receptor in the crude membranes prepared from untransfected COS-7 cells were insignificant, expressed at levels barely above the nonspecific background of the assay (< 30 fmol/mg protein). The amount of β1or β2receptor in the crude membranes prepared from COS-7 cells cotransfected with β1–Gαsor β2–Gαswere similar (approximately 15 pmol/mg protein each). In membranes prepared from COS-7 cells transfected with the cDNA for Gαsonly, a significant increase in Gαs[35S]GTPγS–GDP exchange was measured above that of the nonspecific background measurements (data not shown). This increase in Gαs[35S]GTPγS–GDP exchange was not promoted by the inclusion of 10 nm isoproterenol in the assay buffer (fig. 5). This was also true in studies using membranes prepared from COS-7 cells transfected with the β1or β2receptors only (i.e. , no Gαstransfection; preliminary data not shown). In membranes prepared from COS-7 cells cotransfected with β1and Gαsor β2and Gαs, Gαs[35S]GTPγS–GDP exchange in the absence of isoproterenol was significantly greater than that measured in membranes prepared from Gαs-only transfected cells. The inclusion of 10 nm isoproterenol in the assay buffer caused an additional approximately threefold increase in the magnitude of this exchange with both membrane preparations (fig. 5).
Fig. 5. Effect of isoproterenol on the exchange of the radioactive, nonhydrolyzable form of guanosine-5′-triphosphate (GTP), [35S]GTPγS, for guanosine-5′-diphosphate (GDP) ([35S]GTPγS–GDP exchange) at the α subunit of the Gsheterotrimeric G protein. Gαs[35S]GTPγS–GDP exchange was measured in crude membranes prepared from COS-7 cells transfected with the complementary DNAs encoding for Gαsonly, or β1and Gαs, or β2and Gαs. Gαs[35S]GTPγS–GDP exchange was measured in the absence or presence of 10 nm isoproterenol. The reactions were terminated 5 min after initiation of the assays with [35S]GTPγS. Data are mean ± SD; n = 3. * Significant difference from Gαs[35S]GTPγS–GDP exchange measured using crude membranes prepared from COS-7 cells transfected with the complementary DNAs encoding for the Gαsonly. † Significant difference from Gαs[35S]GTPγS–GDP exchange measured in the absence of isoproterenol (basal exchange) .
The presence of 2.8 (fig. 6A) or 5.6 (fig. 6B) MAC halothane in the assay had no significant effect on Gαs[35S]GTPγS–GDP exchange when the assays were performed in the absence of isoproterenol with either the β1–Gαsor β2–Gαsmembrane preparation. In crude membrane prepared from β2–Gαscotransfected cells, the presence of 5.6 MAC halothane in the assay significantly inhibited the increase in [35S]GTPγS–GDP exchange promoted by 10 nm isoproterenol, although only by approximately 15% of control (fig. 7). The presence of 2.8 MAC halothane in the assay had no significant effect on isoproterenol-promoted Gαs[35S]GTPγS–GDP exchange. By contrast, halothane was significantly more effective in inhibiting the isoproterenol-stimulated incorporation of [35S]GTPγS into Gαswhen the assays were performed with the β1–Gαsmembrane preparation. In these assays, halothane caused a concentration-dependent inhibition of isoproterenol-promoted Gαs[35S]GTPγS–GDP exchange (fig. 7). These effects were significantly greater at both 2.8 and 5.6 MAC halothane compared with measurements performed with membrane prepared from β2–Gαscotransfected cells.
Fig. 6. Effect of 0.75 mm (approximately 2.8 minimum alveolar concentration; A ) or 1.5 mm (approximately 5.6 minimum alveolar concentration; B ) halothane on the exchange of the radioactive, nonhydrolyzable form of guanosine-5′-triphosphate (GTP), [35S]GTPγS, for guanosine-5′-diphosphate (GDP) ([35S]GTPγS–GDP exchange) at the α subunit of the Gsheterotrimeric G protein. Gαs[35S]GTPγS–GDP exchange was measured in the absence of isoproterenol using crude membranes prepared from COS-7 cells cotransfected with the complementary DNAs encoding for β1and Gαs (A ) or β2and Gαs (B ). The reactions were terminated 5 min after initiation of the assays with [35S]GTPγS. Data are mean ± SD; n = 9 .
Fig. 7. Effect of 0.75 mm (approximately 2.8 minimum alveolar concentration) or 1.5 mm (approximately 5.6 minimum alveolar concentration) halothane on the exchange of the radioactive, nonhydrolyzable form of guanosine-5′-triphosphate (GTP), [35S]GTPγS, for guanosine-5|′-diphosphate (GDP) ([35S]GTPγS–GDP exchange) at the α subunit of the Gsheterotrimeric G protein. Gαs[35S]GTPγS–GDP exchange was measured in the presence of 10 nm isoproterenol (ISO) using crude membranes prepared from COS-7 cells cotransfected with the complementary DNAs encoding for β1and Gαs, or β2and Gαs. The isoproterenol-promoted increase in Gαs[35S]GTPγS–GDP exchange was expressed as the percentage of the difference between the values measured in the absence of isoproterenol or halothane, and that measured in the presence of the 10 nm isoproterenol. The reactions were terminated 5 min after initiation of the assays with [35S]GTPγS. Data are mean ± SD; n = 9. * Significant difference from Gαs[35S]GTPγS–GDP exchange measured in the absence of halothane. † Significant difference from Gαs[35S]GTPγS–GDP exchange measured at the same halothane concentration in membranes prepared from COS-7 cells cotransfected with the complementary DNAs encoding for β1and Gαs .
The major finding of this study is that halothane does not inhibit the biochemical coupling of the β2-receptor to its cognate heterotrimeric G protein, Gαs. This finding is consistent with the observation that halothane also had no effect on isoproterenol-induced inhibition of Ca2+sensitivity in airway smooth muscle.
To more directly assess whether halothane inhibits β2receptor coupling to Gαs, we used both a physiologic and a biochemical model (the permeabilized airway smooth muscle model and the Gαsnucleotide exchange assay, respectively). In contrast to studies of living animals or intact, isolated tissue, the experimental protocol using the permeabilized airway smooth muscle model minimized the possible confounding influence of halothane effects on other systems, because halothane has no effect on isometric force in this preparation when induced by free Ca2+alone19,22,24,26,39,41; this was confirmed in the current study (fig. 1). Using these experimental models, approximately 2.8 MAC halothane had no significant effect on the decrease in Ca2+sensitivity induced by isoproterenol. In addition, there was no effect of 2.8 MAC halothane on Gαs[35S]GTPγS–GDP exchange when measured either in the absence or presence of isoproterenol using membranes prepared from β2–Gαstransfected COS-7 cells. These results are consistent with a ligand binding study of human lymphocytes (which express predominantly β2receptors) showing that the β2receptor and a measure of its immediate interactions with the Gsheterotrimer (i.e. , high-affinity ligand binding) were largely unaffected by halothane.48 There was a small but statistically significant effect of 5.6 MAC halothane on isoproterenol-promoted Gαs[35S]GTPγS–GDP exchange, but this concentration is beyond that achieved during the management of severe bronchospasm. It is plausible that the conflicting observations on the effects of halothane seen in physiologic studies15,16 may be due to differences in indirect effects on the β2receptor as proposed by Saito et al. ,49 including halothane-induced β2-receptor phosphorylation and desensitization via activation of intracellular receptor kinases.
The results of our nucleotide exchange assays showing differences in the sensitivity to halothane of isoproterenol-promoted coupling of β1compared with β2coupling to Gαsmay have mechanistic implications for the effects of volatile anesthetics on heptahelical receptor–heterotrimeric G protein complexes. We observed an approximately threefold greater inhibition of β1–Gαscoupling at both halothane concentrations. An early radioligand binding study by Bohm et al. 50 demonstrated that 1–2 MAC halothane inhibited high-affinity binding (i.e. , the active receptor conformation) of isoproterenol to the β1receptor in human myocardial membranes but had a minimal effect on low-affinity ligand binding determined in the presence of 100 μm of the GTP analog GPP(NH)P. These results were interpreted as an inhibitory effect on β1receptor–Gsheterotrimer interaction without a direct effect on the receptor itself. In a more detailed competitive radioligand binding study on membranes prepared from cardiac myocytes, 1–2.8 MAC halothane, in a concentration-dependent manner, also significantly reduced the percentage of receptors in the high-affinity conformation.20 However, contrary to the study of Bohm et al. , this study found that halothane also inhibited isoproterenol affinity in the presence of GPP(NH)P (i.e. , also inhibited low-affinity ligand binding). The fact that an effect was seen in the presence of GPP(NH)P would seem to indicate a direct effect of halothane on the β1receptor conformation independent of the Gsheterotrimer. However, care must be taken when interpreting allosteric effects on competitive binding assays, because an unnoticed effect of halothane on the radiolabeled antagonist binding can lead to incorrect conclusions about the contribution of the G-protein heterotrimer to the effect.
Our results clearly demonstrate an inhibitory effect of halothane (approximately 2.8 MAC) on isoproterenol-promoted Gαs[35S]GTPγS–GDP exchange in measurements made using membranes prepared from the β1–Gαscotransfected cells, as would be predicted from the previous radiolabeled ligand binding studies that showed halothane effects on high-affinity isoproterenol binding to the β1receptor.20,50 This inhibitory effect may be due to a direct effect on the β1receptor, as previously demonstrated for the rhodopsin receptor,51 or its local membrane environment. This type of receptor-dependent effect would be consistent with the apparent greater effect of halothane on β1versus β2receptor, which are both coupled Gαs, although a differential sensitivity of the β2receptor to a halothane effect on the Gsheterotrimer could also explain our results. Alternatively, it is possible that a unique anesthetic binding region created at the interface between the β-receptor isoform and Gαsconfers sensitivity to halothane.
The clinical significance of the findings of this study is readily evident. The relative balance between the neurologic and hormonal mediators that induce airway smooth muscle constriction and relaxation determines airway smooth muscle tone. Numerous types of receptors mediate airway smooth muscle constriction, whereas the β-adrenergic receptor is the best-characterized receptor that mediates airway smooth muscle relaxation. Combined with the previous findings that halothane inhibits muscarinic receptor–heterotrimeric G protein coupling19,28 and hence the increase in Ca2+sensitivity and contraction of isolated airway smooth muscle induced by muscarinic receptor agonists,12–14,24,26,33,39–41 the current findings that β2receptor coupling to Gαsand hence the downstream signaling pathways that decreases Ca2+sensitivity is spared make volatile anesthetics an ideal therapeutic agent for the treatment of perioperative bronchospasm. Signaling systems that induce bronchoconstriction are attenuated, whereas β2receptor–mediated signaling induced by bronchodilators are unaffected by halothane.
In summary, the current study provides evidence that concentrations of halothane typically achieved during treatment of acute perioperative bronchospasm or exacerbation of asthma do not prevent the biochemical coupling between the β2receptor and its associated heterotrimeric G protein, Gαs. This is supported by the physiologic studies showing that halothane had no effect on the inhibition of Ca2+sensitivity induced by isoproterenol and indicate that halothane would not prevent the efficacy of β2agonists when used to treat perioperative bronchospasm.
Masao Hayashi, Sumedha G. Penheiter, Tetsuzo Nakayama, Alan R. Penheiter, David O. Warner, Keith A. Jones; Halothane Does Not Inhibit the Functional Coupling between the β2-Adrenergic Receptor and the GαsHeterotrimeric G Protein. Anesthesiology 2006;104(4):754-762.


text:
Yakima River, WA
Sometimes You Can Go Home Again
By Michael Hamilton
Once in a while, revisiting something old to find out something new is worthwhile. In the case of the Yakima River, Washington's only true-blue catch-and-release fishery, the combined elements of salmon, high water, habitat, and hatches merit a closer look. The Yakima River—or "Yak," as locals call it—displays multiple personalities (some might add "disorders" to complete the phrase). The Yak can be very kind to anglers at times and relentlessly cruel at others. It reminds me of my favorite fly-fishing shirt. Ignore the frayed cuffs. Overlook the permanent pocket stain. Forget the missing button. Regardless of how it looks or what others think, I keep returning to it time and time again.
Located in an algae-corroded irrigation culvert at the south end of Lake Keechelus, on the east slope of Snoqualmie Pass in the Cascade Range, the Yakima River's unromantic and humble origin is a lot like a Charles Dickens character who's forced to begin life in a miserable workhouse. Fortunately, these beginnings are short lived. Quickly gaining tempo and swelling with personality as it goes, the Yakima starts a 214-mile dodge through pine-scented forests, towering cottonwoods, slender white-barked aspens, flatlands planted with potatoes and Timothy grass, craggy basalt cliffs, humpback hills of sage and canyon scrub, and endless miles of fertile valley before widening into the Yakima River Delta and reaching its destination, the mighty Columbia River.
The Yak's history is rich with oral tradition and noteworthy facts. It was named for the indigenous Yakama people (note the spelling difference, which was adopted by the tribe in 1993), and the first western explorers to see it, 215 years ago, were Lewis and Clark, in October of 1805. The expedition camped at the confluence of the Yakima and Columbia but never went upriver. If it had, it would have encountered the Yakama people catching wild salmon with nets and wooden fish traps. Today, it is the Yakama Nation's past that informs their present efforts to restore depleted stocks of spring and summer chinook, coho, and sockeye to the Yakima River and its tributaries. My relationship with the Yak began in 1993 under the watchful eye of Bob Aid, one of only a handful of fly-fishing guides whose names may sound familiar to baby boomers and totally foreign to millennials. With the late Tim Irish in the lead, others, such as Aid, Larry Graham, Jack Mitchell, John Farrar, and Chuck Cooper, led the early cry of collective fly-fishing voices in the 1990s attempting to convince the Washington Department of Fish & Wildlife (WDFW) to designate approximately 75 miles of the upper reaches as catch-and-release. Every river has its legends, pioneers really, who did the heavy lifting so the rest of us could follow in their footsteps.
Floating from high country to lower canyon, anglers seldom encountered another drift boat. But times change. Today, more than two decades since the river went "wild," five guide shops along with a handful of independents work the river year-round from Easton to Roza Dam.
The upper Yakima's 75 miles designated as a catch-and-release fishery boasts optimal habitat for trout: clear, cold water; rocky bottoms in riffle and run areas; as many pools as there are riffles; areas of slow, deep water; abundant and stable banks with a lot of plant growth. Furthermore, Yakima trout use undercut banks, overhanging vegetation, rocks, deep pools, water turbulence, and woody debris as cover to protect them from predators.
A Game of Give and Take
Currently, according to Joe Rotter of Red's Fly Shop and Ted Truglio of Troutwater Fly Shop, stonefly populations on the Yakima remain as strong as these guides have seen in their careers. Both cite the Skwala stoneflies as being—in the words of Truglio—an exciting "big-meal, season-starter" insect that draws more eager anglers to the river in late winter than ever. Even while the Skwala stoneflies are still hatching, the Yakima's early-season hatches of little Baetis (Blue-Winged Olive)
mayflies begin.
However, hatches of Blue-Winged Olives in the lower canyon where they used to thrive have been noticeably absent lately. The reason is anybody's guess. Did the high water events of recent years take a toll? "It's a puzzle with many missing pieces," notes longtime Yakima guide Bruce Skotland, owner of Yakima River Angler. Luckily for anglers, the fish adapted to the shifting menu and by all accounts, are doing better than ever.
Skotland—who spends much of his time fishing in the lower canyon—adds that hatches of Pale Morning Duns and March Browns, traditionally strong in the lower canyon, are currently sporadic at best and nowhere near historic levels (although it's worth considering that the codified history of Yakima River hatches doesn't span many years anyway). However, upriver through the farmlands to Cle Elum, other guides tell a different story.
"Fishable dry-fly hatches on the upper river from Easton to Ellensburg are consistent and as diverse as ever," says Jim Gallagher, co-owner of Troutwater Fly Shops in Cle Elum and Ellensburg. Gallagher maintains that he and his guides encounter both sparse and dense seasonal hatches of Baetis, PMDs, March Browns, Green Drakes, and more
Steve Worley, owner of the Worley Bugger Fly Co. in Ellensburg, spends most of his guiding days in the farmlands and upper river. He says that he's personally witnessed, on a regular basis, intense March Brown hatches lasting four hours. "This is a hatch that in past years you could count on being anywhere from 20 minutes to an hour at the most—if you were lucky."
Worley adds that in recent years, he has seen many days when March Browns, Blue-Winged Olives, and Skwala stoneflies were all on the water at the same time for hours, especially through the Thorp and Cle Elum sections of the Yakima River.
Worley and Gallagher both mention that craneflies are a food source that's overlooked by most anglers. Dry and larvae patterns can be effective in both spring and fall. Cranefly dries draw the most attention from trout on softer water. Moreover, while Skwalas seem to be the darlings of the Yakima stoneflies in recent years, anglers who fish the river from May through summer should come armed with flies to match Salmonflies, Golden Stoneflies, and Yellow Sallies.
Other guides on the Yakima also cite impressive hatches, although as Steve Joyce, owner and general manager of Red's Fly Shop, points out, "Talking about hatches is relative to whether the fish are actually keying in on them and not necessarily the number of bugs you see."
Further, he says, "I really think the hatches have shifted a bit from the high-water events that we've been hit with the past several consecutive years."
Eat or Be Eaten
Setting high water aside for the moment as a cause for changing bug populations, another possible factor to consider is the increase in predation. For example, many mayfly species feed on algae or diatoms, but a few are predatory, eating each other, while everyone else snacks on them. In other words, a larger population of predator mayflies in the Yakima could, over time, change a species' dynamics. But it's another predator that people wonder about.
The Yakama Nation—in a well-publicized effort to reintroduce indigenous salmon to the Yakima watershed—has reared 810,000 spring chinook at its Cle Elum Supplementation and Research Facility since 1997. Averaging 4 to 5 inches when released, the smolts are discharged in the spring from three acclimation points, two in the Yakima River and a third on the North Fork of the Teanaway River. What impact, if any, has the sudden presence of thousands of smolts had on insect populations in the river?
Dave Fast, research manager with Yakama Nation Fisheries, says, "What people don't realize is that the smolts are only in the system for two to three weeks. Since they have been reared on a vegetarian diet in the hatchery before release, they are slow to start eating insects. By the time they do, they are usually migrating out." Additionally, Fast notes that the biomass contributed by salmon spawn and salmon carcasses should lead to higher production and survival of insects, not less.
Because the Yakama's spring chinook salmon smolts are known as aggressive feeders, albeit they begin life as vegetarians, I asked Montana Fish, Wildlife & Parks senior biologist David Schmetterling what he thinks would happen to aquatic invertebrates if hundreds of thousands of smolts were introduced into systems like the Clark Fork or Bitterroot Rivers near Missoula. Schmetterling is a renowned research scientist and a frequent contributor to magazines and periodicals. He told me, "I am totally speculating, but I don't think there would be any deleterious consequences to aquatic invertebrates; in fact, on the contrary, maybe a benefit. Smolts could be a source of food for some invertebrates as many of the released smolts would die before leaving [such rivers] either naturally or as a result of shock or confusion. These smolts would also be a source of food for other scavengers [crayfish] as well as mink, herons, kingfishers, and, of course, other fish."
Aquatic entomologist and author Rick Hafele, when asked the same question, agreed with Schmetterling. "Typically, from my experience, if resident fish are growing larger, healthier, and gradually increasing in numbers . . . as they are in the Yakima, then there would have to be abundant food to support the increase in biomass, which includes invertebrates."
There is a silver lining in this playbook—the overall size and health of the trout being caught in the Yakima are exceptional. "Even though there are fewer small mayflies hatching during big flows in some stretches, there is abundant food and clean, cool water, which we have seen by the size and health of the fish the past few seasons," says Steve Joyce.
Some speculate the river's cutthroat trout may be faring even better than the rainbows. The cutts have expanded their range in the river and are more numerous than ever according to longtime Yakima anglers. They are now caught all the way down to Roza Dam; 15 years ago, cutts were almost unheard of in the canyon. Not only are the native cutts numerous now, they are also growing to impressive proportions—last year, an 8-pound cutthroat turned up during electroshocking surveys.
In addition to catching healthier and heavier wild trout, fly anglers are also adjusting their techniques and meeting with success by fishing more underneath than on top.
"No question the nymph game on the Yakima is productive because 80 percent of what trout eat lives underwater, not on top," says Joe Rotter. This upheaval in technique, notes Rotter, has gradually caught fire and today, nymphing is often the meat-and-potatoes technique for catching trout on the Yakima. "The standard nymph rig usually features an indicator with two flies like a Pat's Stone on top and a smaller mayfly or stonefly imitation on the bottom," explains Rotter.
Most anglers, he observes, equate nymph fishing with success. However, he also points out that when he adds up the time he often spends untangling nymph rigs, he will frequently suggest using one nymph or switching tactics to cast dries or streamers when hatches and conditions dictate.
Worth noting, too, is that the Yakima is also home to a variety of baitfish. Streamer patterns that imitate sculpins, shiners, and other minnows can be very effective, especially in the shoulder seasons leading to fall, winter, and spring. Fished on the swing with floating or 10- to 15-foot sinking-tip lines, streamers elicit violent strikes from big fish.
Salmon Homecoming
The construction of Roza Dam in the late 1930s was one of the many regional efforts to take water born of mountain snowpacks and use it for expanding agricultural development. In an era before terms like "ecosystem" and "environmental impact," visions were followed and potential consequences were not considered, if even they were understood. Strong runs of great sea-run fish were taken for granted in some cases and often undervalued at the same time. The resultant impact on regional migratory fish was catastrophic by almost all accounts. But times have changed and, to many, the current fish stocks are unacceptable. Compensatory actions are even mandated by law in some cases.
Today, some river systems are seeing measured success, but it comes at a steep price. The Yakama Nation Fisheries has invested heavily in an attempt to restore part of its tribal history and culture. In addition to the Yakama Nation's Cle Elum Supplementation and Research Facility, which focuses on spring chinook salmon, the Yakamas have also reintroduced a run of summer chinook, which are released on a lower stretch of the Naches River.
Coho and sockeye, notes research manager Dave Fast, were extirpated but are also making a comeback in the Yakima. "The long-term health of the watersheds is dependent on how coho recover because they migrate up tributaries, spawn, and die. Their carcasses provide a food source for steelhead, bull trout, and rainbows."
Fast says that adult sockeye are transported from Priest Rapids Dam in the summer and spawn above Lake Cle Elum. Historically, 200,000 sockeye used to return annually to four lakes—Bumping, Keechelus, Kachess, and Cle Elum. The 2014 season marked the first returns of the sockeye transplanted to Lake Cle Elum. While their returns are in smaller numbers, anglers have been reporting sightings of silvers in the fall during recent years.
For the river's trout anglers, burgeoning salmon stocks hold the promise of even better things to come: if these species take hold, the river's biomass will continue to increase.
The farthest upstream WDFW public launch is at Ensign, below Easton, adjacent to a ranch owned by the Mormon Church. Take exit 78 off Interstate 90 onto Golf Course Road. Private cabins and homes frame most of the 5-mile float down to Bullfrog, the site of a former state park now gone to seed. The first three weeks of June on this section can offer incredible Green Drake hatches. Anglers can also walk and wade upstream from Ensign's public access point.
Bullfrog to South Cle Elum comes next and is ripe with sweepers, logjams, and tight corners downriver to the confluence with the Cle Elum River; it's a treacherous float for drift boats or any novice in a pontoon boat. There are big fish to be had, no question. However, if you are determined to try this stretch, get advice from Jim Gallagher at Troutwater in Cle Elum first.
The 5-mile float from South Cle Elum to "State" (East Cle Elum) begins at a primitive public launch at the bridge in town. It requires skilled trailer maneuvering to back a drift boat or raft down the steep bank. The float presents no major obstacles and the take-out is at a state launch site off State Route 10 in East Cle Elum. From I-90, take exit 85 and follow old US Route 10 a short distance toward Blewett Pass on SR 970. Turn southeast off SR 970 at the Public Fishing sign. Trout hold in very specific places in this reach—off a drop below a riffle behind a rock. You need to be able to read the water. Expect bigger fish but not as many as you move downriver. Some anglers will forgo the float, choosing instead to walk and wade upstream to the
Cle Elum confluence.
Next comes State to Green Bridge (Thorp). Just below the launch, the river splits and staying right is always recommended. In June, after runoff, Green Drakes hatch, and in July, Golden Stoneflies steal the show. Long riffles, deep pockets, big boulders, and silky tailouts hold many 14- to 20-inch trout. Below Thorp, a short run of 2.5 miles to Diversion Dam sees little pressure. Expect consistent dry-fly fishing with stoneflies active from July through September and BWOs and Mahogany Duns in fall.
KOA Campground to Irene Rinehart Riverfront Park in Ellensburg is a relatively short float that offers some protection from the notorious winds that whip up in the Kittitas Valley. In spring and fall, the next section, the float from Irene to Ringer, is usually passable, but be careful: from KOA to the Ringer boat launch, narrow channels, whitewater drops, and sweepers can form hazards, depending on water levels. Before you float this stretch, call a local Ellensburg shop for updates on blockages
or portages.
In fact, regardless of where you float, it's wise to check in with any of the shops or guides before making your first drift of the season or after any high-water event because some stretches of the river are prone to changes and hazards come and go
every year.
The Canyon and More
Though diverse and productive, the upper sections of the Yakima don't bask in the fame of Yakima Canyon, which is in a league of its own in terms of both fishing and scenery. Yakima River trout populations reach their highest density here. The river runs deep, cold, and fish friendly; precipitous arid slopes rise dramatically to towering heights.
In mid-February, Skwala stoneflies begin to hatch in earnest. Nymphs migrate shoreward when the water reaches 38 to 40 degrees. Adults hatch when the water temperature rises into the mid- to upper 40s. Fish nymphs early in the day, then switch to an adult Skwala pattern in the afternoon. In April, March Browns often emerge about 1:30 p.m. It's a tough hatch to predict. For the past two years, it's been a stronger emergence in the farmlands and above.
Mention the month of May and Yakima regulars immediately think Mother's Day Caddis. Sunny days bring indescribable swarms of egg-laying caddisflies, which are good for trout gorging below the surface but not so good for you. Cloudy days are much more angler friendly because the adult caddisflies sit on the surface, making a size 14 or 16 X-Caddis deadly.
After June runoff, casting ants, beetles, and even small grasshopper patterns downstream and tight to the bank, drag free, can bring the big boys to the net. In July and August, canyon flows stabilize at around 4,000 cubic feet per second. The trout are fat and feisty. Shortwing Stoneflies (aka Summer Stones), along with those grasshoppers, are always on the menu, as are caddisflies after the sun moves off the water.
Things change dramatically in autumn, when regional irrigation slackens and the heavy flows that feed it subside. The river drops and gravel bars emerge again. When low flows stabilize in October, the river reawakens in dramatic fashion. A new crop of insects stirs surface action upriver, including renewed hatches of Baetis (watch for spinner falls in the evenings and imitate them with a tiny Red Quill Spinner).
Fall BWO hatches can be fleeting in the canyon but are especially strong in the farmlands and on the upper river. Anywhere on the river, if luck holds, you may find larger Mahogany Duns and possibly even Pale Evening Duns hatching alongside the little Blue-Winged Olives. On some sections, giant October Caddis appear. Of course, fall is also a fine time to pull big streamers for the largest trout.
Is the Yakima River changing? Are its fish and hatches and ecology in a state of flux? Probably. That's the nature of rivers—they are dynamic ecosystems and in the case of Washington's unofficial blue-ribbon trout stream, the changes are for the better.
Even while Yakima Canyon continues to get most of the good press, the river as a whole offers amazing and diverse prospects for anglers. And while changes wrought by years of surge flows, the reintroduction of salmon, and other catalysts may not be quantified by science, for anglers, conclusions are easy to come by: the trout themselves are bigger and better than ever, native cutthroat seem more numerous and more widely distributed, and hatches such as Skwala stoneflies and March Browns have increasingly become predictable major events in the farmlands and upriver stretches.
So, if you haven't fished the Yakima lately or at all, it's time to introduce yourself. Explore it head to toe, embrace all the seasons, learn the hatches all over again, and rejoice in the big, beautiful wild trout. If you've never cast flies on the Yak, ponder this river in the reverent tones usually reserved for waters like the Madison and Henrys Fork and Bighorn: the amazing Yakima is one of those places every angler needs to experience.


The land area of Colorado has roots in the Louisiana Purchase (1803), a Spanish land cession pursuant to the Adams-Onis Treaty (1819) the Annexation of Texas (1845) and a Mexican land cession pursuant to the Treaty of Guadalupe Hidalgo (1848). The present-day boundaries, in turn, are the result of a series of mid-nineteenth century legislative initiatives by the federal government.
As part of the Compromise of 1850, Texas, which entered the Union in 1845, traded land to the federal government in exchange for debt relief. Some of that land, along with other federal government holdings (from the 1848 Mexican cession) was organized into the New Mexico and Utah territories – their formation was also a part of the Compromise.
A few years hence, the Kansas-Nebraska Act of 1854 created the territories of Kansas and Nebraska. The Colorado territory was born in 1861 and included portions of the Kansas, Nebraska, New Mexico and Utah territories.The entire Colorado territory would become the State of Colorado 15 years later.
Three meridians (north-south lines) serve as the principal lines of reference for dividing the state into 6 mile square townships.1 The 6th Principal Meridian (longitude 97° 22′ 08″ West) resulted from survey work begun in 1855 and is the reference line for most of Colorado. The corresponding baseline (east-west lines) for the 6th P.M begins at the Missouri River and is marked by a monument on the west side on a bluff overlooking the river at 40° North latitude. The baseline extends west across Colorado. Baseline Road in Boulder follows the baseline.
The New Mexico Principal Meridian (longitude 106° 53′ 12″ West) is the reference line for the San Luis Valley. The corresponding baseline for the New Mexico Principal Meridian runs across New Mexico at 34° 15′ 35″ North latitude. The Ute Principal Meridian (longitude 108° 31′ 59″ West) is the reference line for portions of Mesa and Delta counties on the Western Slope.
A legal description of property locates its township(s) relative to the appropriate baseline (north or south) and meridian (east or west). Property within a township is further defined by section – sections being 1 square mile divisions of a township.
So, for example, the downtown Colorado Springs office for Unified Title Company is located in section 18 of a township that is the 66th west of the 6th P.M. and the 14th south of the baseline.
1. In Colorado, counties are a basic unit of government, but land is divided into survey townships pursuant to centuries old federal legislation – the Land Ordinance of 1785.


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