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      The Medical Thread.

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      RedPuppy
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      Re: The Medical Thread.
      Reply #46: Jun 07, 2014 09:18:07 pm
      I feel as though I need one too :D I've Got this sciatic nerve trouble since I bought down that F***ing chimney wall in my house, started to get pins & needles in my left foot with the hamstring & calf muscle very tight, especially in the morning, been digging out foundations,Dry mixing & running in blocks for the rear extension and not one sign of any nerve problem, only at bedtime & early dawn.

      You should see your GP, to determine the cause.

      You may have a disc pinching a nerve, or it could be piriformis, which can give similar symptoms.

      An osteopath may be of benefit to give your back a good cracking. They would be able to tell if your vertebra are out of alinement.

      £400 for a chiropractor is a lot of money, I pad paid £40.00 for an assessment and full body treatment (osteopath).
      andylfcynwa
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      Re: The Medical Thread.
      Reply #47: Jun 08, 2014 10:14:07 am
      You should see your GP, to determine the cause.

      You may have a disc pinching a nerve, or it could be piriformis, which can give similar symptoms.

      An osteopath may be of benefit to give your back a good cracking. They would be able to tell if your vertebra are out of alinement.

      £400 for a chiropractor is a lot of money, I pad paid £40.00 for an assessment and full body treatment (osteopath).

      I,m with you £400 seems a hell of a lot  you can go to bupa and have an assessment  for roughly a hundred quid .
      shabbadoo
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      Re: The Medical Thread.
      Reply #48: Jun 08, 2014 10:49:14 am
      I,m with you £400 seems a hell of a lot  you can go to bupa and have an assessment  for roughly a hundred quid .

       About £400 for 10 sessions for cracking my back.

      Initially paid £70 for a consultation & a scan of the spine then £35 per session.
      andylfcynwa
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      Re: The Medical Thread.
      Reply #49: Jun 08, 2014 01:38:33 pm
      Well thats not to bad then.
      Swab
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      Re: The Medical Thread.
      Reply #50: Jun 18, 2014 02:03:15 pm
      Off to hospital for surgery tomorrow.
      They're hoping I'll just be in for the day and home in the evening, but we'll see.

      A long road ahead, but it has to be done.
      This is what I get for trying to do a marathon  :laugh:
      CoutinhoRed
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      Re: The Medical Thread.
      Reply #51: Jul 08, 2014 09:20:58 am
      Booked in for 2 weeks time for hip replacement.
      I've decided to bite the bullet and get the f**king thing done so I can get back on the golf course properly.

      They're saying I could be home the same day, but I'll wait and see.

      The good news is that I'm still fit and strong.
      The bad news is there's a lot of muscle around the area, so it might be more painful for me that it would for others.
      Approx. 6 months rehab and with luck I'll be good to go.

      What has your rehab been like?
      waltonl4
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      Re: The Medical Thread.
      Reply #52: Jul 09, 2014 08:28:11 pm
      I feel as though I need one too :D I've Got this sciatic nerve trouble since I bought down that f**king chimney wall in my house, started to get pins & needles in my left foot with the hamstring & calf muscle very tight, especially in the morning, been digging out foundations,Dry mixing & running in blocks for the rear extension and not one sign of any nerve problem, only at bedtime & early dawn.

      physio mate the bringers of unbelievable pain but with it relief.Get youself a Swiss ball they are magic for back problems
      Swab
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      Re: The Medical Thread.
      Reply #53: Jul 09, 2014 08:46:19 pm

      Still on crutches for now, soon as I come off them I'm on the exercise bike.
      Physio is very gentle and not putting any stress on the joint, just small gentle movements that I do during the day as well.
      The worst thing is not being able to bend or move normally, but I'll get there.
      racerx34
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      Re: The Medical Thread.
      Reply #54: Sep 25, 2014 12:47:33 pm
      So, just because I was curious, I decided to eat quite a bit of "gluten free" junk food. You know the type.
      It's marketed as a saviour even though, deep down, you know it's sh*te.

      Safe to say I haven't felt this sh*t in over 6 months.
      Chronic tiredness. Massive brain fog.
      Why the F**k did I bother?

      Here two articles to confirm what I already knew.
      Gluten Free junk food is still junk food.

      http://articles.mercola.com/sites/articles/archive/2014/08/18/gluten-free-low-carb-paleo-diet.aspx

      http://glutendude.com/celiac-rant/stop-eating-gluten-free-foods/
      Roddenberry
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      Re: The Medical Thread.
      Reply #55: Nov 19, 2014 01:59:53 am
      I've spent this year off my prescription meds.  It probably wasn't a good idea, but some of the side effects have had permanent repercussions, though coming off them may have reversed the damage, it hasn't.  I'm going to see out the year, but I'm back on them after that. My weight issues and sleep problems have both been acerbated by being clean and frankly I am seriously struggling with the pain issues.  Those who tell you that pain tells them they're alive, they're dead wrong. It makes me feel half-dead, anti-social, angry and depressed. 

      My biggest problem is, by going back on the drugs, though physically I'll feel a sh*t tonne better, they make me, trying to word this right, a little fuzzy on the mental side.  My work has been of a higher quality this year, both individually and as a team contributor, I've spotted things I probably should have noticed before and helped improve efficiency.   

      Billy1
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      Re: The Medical Thread.
      Reply #56: Nov 20, 2014 06:18:32 am
      I understand what you are going through Stuart, my doctor once prescribed me pain killers and gave me 500 at a time to take 2 tablets 3 times a day. I thought I am not going to make myself reliant on painkillers and stopped taking them, I must say I don't need them and the hour or the day is not regulated by them. Stay strong and you yourself will know if you need extra medication, cheers.
      AlwaysTheKop
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      Re: The Medical Thread.
      Reply #57: Dec 12, 2014 03:49:13 pm
      I suffer from pretty bad anxiety and possibly depression and I just needed somewhere to vent so sorry...

      Basically all my life I've been an anxious person, through school and college, I'd skip full days if I had a presentation or a one on one meeting that day. I'd get to the point that just having a conversation with someone would end up in me looking like I'd taken a swim in a lake through nervous sweats. Then after college I got a job running a machine where I presses one button in the same spot for 8 hours a day, and it totally depressed me but I didn't want to disappoint my family because they are big on the whole your worthless if your not working thing but after last Christmas it got so bad that I could no longer physically or mentally go back to that job and I quit.

      Over the past 12 months since then have been the best I remember since being a child! My anxiety pretty much vanished and I achieved a lot of personal goals such as losing almost 40 pounds and leaving my house to be social, I used to be scared of going to the supermarket believe it or not...

      During this year I've been unemployed I've been doing voluntary work and enjoying life while applying for jobs for Universal Credit and this month I heard back from Saintsbury's and my new way of life involved no fear so I blitzed the interview and even did well in the group sessions during the induction which if you told me a year ago I'd be standing in front of a group of strangers speaking confidently I'd have laughed...

      But after the induction I spent two hours on the shop floor and all that anxiety I used to feel came rushing back and hit me like a ton of bricks and since then I haven't slept and my body has been aching!

      I really don't want to go back to the person I was over a year ago.... where I was afraid to leave the house other than to work... I'm disappointed in myself because I'm already thinking about quitting and I haven't even done a full day but my anxiety is crippling!

      Today all I've done for my day off is search the internet about anxiety and that is exactly what I used to do while working for those 5 years on my time off...

      I don't know if I'm ready to work and that maybe I need to finally get my anxiety/depression looked at by a professional, but I don't want to disappoint people and appear to be a lazy person, because I'm not, and anybody that knows me would say the same.

      I just don't want everything I've accomplished this year to go to waste... like the weight loss, the daily exercise (I've skipped swimming today) and just nkt feeling anxious every second of the day....

      /rant over.
      racerx34
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      Re: The Medical Thread.
      Reply #58: Dec 12, 2014 04:44:00 pm
      I suffer from pretty bad anxiety and possibly depression and I just needed somewhere to vent so sorry...

      Basically all my life I've been an anxious person, through school and college, I'd skip full days if I had a presentation or a one on one meeting that day. I'd get to the point that just having a conversation with someone would end up in me looking like I'd taken a swim in a lake through nervous sweats. Then after college I got a job running a machine where I presses one button in the same spot for 8 hours a day, and it totally depressed me but I didn't want to disappoint my family because they are big on the whole your worthless if your not working thing but after last Christmas it got so bad that I could no longer physically or mentally go back to that job and I quit.

      Over the past 12 months since then have been the best I remember since being a child! My anxiety pretty much vanished and I achieved a lot of personal goals such as losing almost 40 pounds and leaving my house to be social, I used to be scared of going to the supermarket believe it or not...

      During this year I've been unemployed I've been doing voluntary work and enjoying life while applying for jobs for Universal Credit and this month I heard back from Saintsbury's and my new way of life involved no fear so I blitzed the interview and even did well in the group sessions during the induction which if you told me a year ago I'd be standing in front of a group of strangers speaking confidently I'd have laughed...

      But after the induction I spent two hours on the shop floor and all that anxiety I used to feel came rushing back and hit me like a ton of bricks and since then I haven't slept and my body has been aching!

      I really don't want to go back to the person I was over a year ago.... where I was afraid to leave the house other than to work... I'm disappointed in myself because I'm already thinking about quitting and I haven't even done a full day but my anxiety is crippling!

      Today all I've done for my day off is search the internet about anxiety and that is exactly what I used to do while working for those 5 years on my time off...

      I don't know if I'm ready to work and that maybe I need to finally get my anxiety/depression looked at by a professional, but I don't want to disappoint people and appear to be a lazy person, because I'm not, and anybody that knows me would say the same.

      I just don't want everything I've accomplished this year to go to waste... like the weight loss, the daily exercise (I've skipped swimming today) and just nkt feeling anxious every second of the day....

      /rant over.

      You tried going for a run or taking up some form of exercise?
      AlwaysTheKop
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      Re: The Medical Thread.
      Reply #59: Dec 12, 2014 04:54:00 pm
      You tried going for a run or taking up some form of exercise?

      I've been exercising all year, I took up swimming and gym aswell as walking everywhere within an hour range instead of taking the bus and lost 3 stone since January, but knowing I have to go to work the next day zaps every single bit of energy I have and makes all that stuff, which I've been enjoying, seem like a chore now.
      Bier
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      Re: The Medical Thread.
      Reply #60: Dec 12, 2014 04:59:14 pm
      AlwaysTheKop, you should do what's best for yourself mate, don't worry about what others think or if you might dissapoint them, that's waisted energy. I know that's easier said than done in your situation, it's certainly not a feeling you can just shut down, but you should try not to factor that into your decision making. Also, get professional help. If anxiety plays such a big role in your life then maybe a professional can help you learn to deal with it in a better way.
      racerx34
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      Re: The Medical Thread.
      Reply #61: Dec 12, 2014 09:04:30 pm
      I've been exercising all year, I took up swimming and gym aswell as walking everywhere within an hour range instead of taking the bus and lost 3 stone since January, but knowing I have to go to work the next day zaps every single bit of energy I have and makes all that stuff, which I've been enjoying, seem like a chore now.

      I know the feeling. World's greatest irony that you have to be at your strongest when you are at your weakest.

      Try and look for positive things in what you are doing. Easier Saud than done, I know.

      Good luck.
      shabbadoo
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      Re: The Medical Thread.
      Reply #62: Dec 12, 2014 09:56:41 pm
      I suffer from pretty bad anxiety and possibly depression and I just needed somewhere to vent so sorry...

      Basically all my life I've been an anxious person, through school and college, I'd skip full days if I had a presentation or a one on one meeting that day. I'd get to the point that just having a conversation with someone would end up in me looking like I'd taken a swim in a lake through nervous sweats. Then after college I got a job running a machine where I presses one button in the same spot for 8 hours a day, and it totally depressed me but I didn't want to disappoint my family because they are big on the whole your worthless if your not working thing but after last Christmas it got so bad that I could no longer physically or mentally go back to that job and I quit.

      Over the past 12 months since then have been the best I remember since being a child! My anxiety pretty much vanished and I achieved a lot of personal goals such as losing almost 40 pounds and leaving my house to be social, I used to be scared of going to the supermarket believe it or not...

      During this year I've been unemployed I've been doing voluntary work and enjoying life while applying for jobs for Universal Credit and this month I heard back from Saintsbury's and my new way of life involved no fear so I blitzed the interview and even did well in the group sessions during the induction which if you told me a year ago I'd be standing in front of a group of strangers speaking confidently I'd have laughed...

      But after the induction I spent two hours on the shop floor and all that anxiety I used to feel came rushing back and hit me like a ton of bricks and since then I haven't slept and my body has been aching!

      I really don't want to go back to the person I was over a year ago.... where I was afraid to leave the house other than to work... I'm disappointed in myself because I'm already thinking about quitting and I haven't even done a full day but my anxiety is crippling!

      Today all I've done for my day off is search the internet about anxiety and that is exactly what I used to do while working for those 5 years on my time off...

      I don't know if I'm ready to work and that maybe I need to finally get my anxiety/depression looked at by a professional, but I don't want to disappoint people and appear to be a lazy person, because I'm not, and anybody that knows me would say the same.

      I just don't want everything I've accomplished this year to go to waste... like the weight loss, the daily exercise (I've skipped swimming today) and just nkt feeling anxious every second of the day....

      /rant over.

      Have you tried meditation?, I know it helps as I've suffered in the past from anxiety & panic attacks.

      BKLFC
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      Re: The Medical Thread.
      Reply #63: Jan 01, 2015 09:56:55 am
      Dear Cornish!
      happy new year! So did you get the op done. Here is some info which may interest you regarding your spleen for alternative medicine when all else doesn't seem to work.

      Abstract
      Specialized B cells residing in the splenic marginal zone (MZ) continuously survey the blood for antigens and are important for immunity to systemic infections. However, the cues that uniquely attract cells to the MZ have not been defined. Previous work demonstrated that mice deficient in cannabinoid receptor 2 (CB2) have decreased numbers of MZ B cells but it has been unclear whether CB2 regulates MZ B cell development or positioning. We show that MZ B cells are highly responsive to the CB2 ligand 2-arachidonylglycerol (2-AG) and that CB2 antagonism rapidly displaces small numbers of MZ B cells to the blood. Antagonism for longer durations depletes MZ B cells from the spleen. In mice deficient in sphingosine-1-phosphate receptor function, CB2 antagonism causes MZ B cell displacement into follicles. Moreover, CB2 overexpression is sufficient to position B cells to the splenic MZ. These findings establish a role for CB2 in guiding B cells to the MZ and in preventing their loss to the blood. As a consequence of their MZ B cell deficiency, CB2-deficient mice have reduced numbers of CD1d-high B cells. We show that CB2 deficiency results in diminished humoral responses to a CD1d-restricted systemic antigen.

      Source: Cannabinoid receptor 2 positions and retains margi... [J Exp Med. 2011] - PubMed - NCBI





      BKLFC
      • Forum Emlyn Hughes
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      Re: The Medical Thread.
      Reply #64: Jan 01, 2015 09:58:43 am
      A Cyclooxygenase Metabolite Of Anandamide Causes Inhibition Of Interleukin-2
      Abstract

      Arachidonyl ethanolamine, which is commonly known as anandamide, was the first endogenous compound to be identified that binds to the cannabinoid receptors. Anandamide mimics many of the physiological effects of Δ9-tetrahydrocannabinol (Δ9-THC), including hypothermia, antinociception, immobility, catalepsy, and immune modulation. In the present studies, we show that anandamide caused a concentration-dependent inhibition of interleukin-2 in primary splenocytes. The CB1 and CB2 antagonists, SR141716A [N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorphenyl)-4-methyl-H-pyrazole-3 carboxyamidehydrochlorid e] and SR144528 [N-[(1S)-endo-1,3,3,-trimethylbicyclo[2,2,1]heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide], when used in combination, did not antagonize the inhibition of interleukin-2 by anandamide. Additionally, neither UCM707 [N-(3-furanylmethyl)-5Z,8Z,11Z,14Z-eicosatetraenamide], the inhibitor of the putative anandamide membrane transporter (AMT), nor methyl arachidonoyl fluorophosphonate (MAFP), the inhibitor of fatty acid amidohydrolase (FAAH), were able to affect the inhibitory activity of anandamide upon interleukin-2. Interestingly, arachidonic acid caused a concentration-dependent inhibition of interleukin-2 secretion (IC50 = 10.3 μM), which was similar to that of structurally related anandamide (IC50 = 11.4 μM). The inhibition of interleukin-2 by anandamide and arachidonic acid was partially reversed by pretreatment with the nonspecific cyclooxygenase inhibitors, flurbiprofen and piroxicam. Moreover, NS398 [N-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide], a cyclooxygenase-2-specific inhibitor, also attenuated the inhibitory effects of anandamide and arachidonic acid upon interleukin-2 secretion. Finally, pretreatment with a peroxisome proliferator-activated receptor γ (PPARγ)-specific antagonist, T0070907 [2-chloro-5-nitro-N-4-pyridinyl-benzamide], partially antagonized anandamide-mediated suppression of IL-2 secretion. Collectively, the aforementioned studies suggest that inhibition of interleukin-2 secretion by anandamide is independent of CB1/CB2 and the AMT/FAAH system. Additionally, these studies also suggest that inhibition of interleukin-2 is mediated by a PPARγ, which is activated by a cyclooxygenase-2 metabolite of anandamide.

      Two cannabinoid receptors have been isolated and cloned to date, CB1 and CB2 (Matsuda et al., 1990; Munro et al., 1993). CB1 is widely distributed in the central nervous system but is also found in a number of other tissues (Matsuda et al., 1990). CB2 is the predominant cannabinoid receptor expressed in cells of the immune system, although CB1 is also detected in many immune cells at lower levels (Kaminski et al., 1992; Munro et al., 1993). Both receptors have seven transmembrane domains and are G-protein-coupled. CB1 and CB2 share only 44% homology, which increases to 68% when comparing only the transmembrane regions that contain the ligand-binding domains (Munro et al., 1993). Arachidonyl ethanolamine, also known as anandamide, was the first endogenous cannabinoid receptor ligand to be identified through radioligand binding analysis and was therefore termed an endocannabinoid (Devane et al., 1992).

      Recently published studies suggest that CB2 may be involved with a number of immunomodulatory effects, including inhibition of macrophage-dependent T cell activation, induction of the immunosuppressive cytokine, transforming growth factor β, in human peripheral blood lymphocytes stimulated with anti-CD3, inhibition of antigen processing by macrophages, and induction of cytokine and chemokine production in HL-60 cells (McCoy et al., 1999; Buckley et al., 2000; Derocq et al., 2000; Gardner et al., 2002). Although CB1 is typically expressed at much lower levels in immune cells than CB2, there is also evidence to suggest a role for CB1 in certain immunological effects, such as inhibition of tumor necrosis factor α and interleukin-12 in LPS-treated mice as well as induction of interleukin-6 in mouse astrocytes (Molina-Holgado et al., 1998; Smith et al., 2001). Anandamide has been shown to modulate a number of immunological responses, including inhibition of nitric oxide and cytokine production in macrophages as well as inhibition of tumor necrosis factor α and neutrophil recruitment in lipopolysaccharide-induced pulmonary inflammation in mice (Berdyshev et al., 1998; Chang et al., 2001). Whether these effects are in fact mediated through the cannabinoid receptors has yet to be rigorously examined, however.

      Anandamide can be synthesized by a variety of cell types, including neurons, RBL-2HE basophils, and J774 macrophages (Sugiura et al., 2002). It has been proposed that anandamide is rapidly inactivated by a reuptake system consisting of the anandamide membrane transporter (AMT), which transports anandamide into the cell where fatty acid amidohydrolase (FAAH) hydrolyzes anandamide into arachidonic acid and ethanolamine (Di Marzo et al., 1994). Although FAAH appears to be the primary enzyme responsible for the hydrolysis of anandamide, an acid amidase has recently been identified that is also capable of hydrolyzing anandamide (Ueda et al., 2001b). In addition to metabolism by FAAH and the acid amidase, anandamide can also be oxygenated by cyclooxygenase-2 into prostaglandin E2 ethanolamide and other cyclooxygenase products as well (Yu et al., 1997). Prostaglandin E2-ethanolamide appears to mediate physiological effects similar to those produced by prostaglandin E2 (Ross et al., 2002).

      Anandamide is well known to cause a number of immunomodulatory effects including modulation of cytokine production, but nothing to date has been published concerning the effects of anandamide upon IL-2 secretion. Moreover, previously published studies from this laboratory have shown that 2-arachidonyl glycerol, a compound that is closely related to anandamide structurally, causes inhibition of interleukin-2 production in activated T cells (Ouyang et al., 1998). The objective of the present studies was then to determine the effect of anandamide upon interleukin-2 secretion. Interleukin-2 is an autocrine/paracrine factor secreted by activated T cells, which promotes T cell proliferation. Interleukin-2 has virtually no basal level of expression but is rapidly secreted upon T cell stimulation such that it is a hallmark of T cell activation. The current studies demonstrate that anandamide causes a concentration-dependent inhibition of interleukin-2 secretion in primary splenocytes, which is independent of CB1/CB2 and involves the activation of peroxisome proliferator activated receptor γ (PPARγ) by a cyclooxygenase-2 metabolite of anandamide.

      Materials and Methods

      Reagents. Anandamide, SR141716A, and SR144528 were provided by the National Institute of Drug Abuse. UCM707, MAFP, piroxicam, NS398, and T0070907 were purchased from Cayman Chemical (Ann Arbor, MI). All other reagents were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise indicated.

      Animals and Cell Cultures. Female B6C3F1 mice, 6 weeks of age, were purchased from Charles River Breeding Laboratories (Portage, MI). On arrival, mice were randomized, transferred to plastic cages containing sawdust bedding (5 animals/cage), and quarantined for 1 week. Mice were given food (Purina Certified Laboratory Chow) and water ad libitum. Mice were not used for experimentation until their body weight was 17 to 20 g. Animal holding rooms were kept at 21-24°C and 40 to 60% relative humidity with a 12-h light/dark cycle. Spleens were isolated aseptically and made into single-cell suspensions (1 × 106 cells/ml). Cells were cultured in RPMI-1640 supplemented with 100 units/ml penicillin, 100 units/ml streptomycin, 50 μM 2-mercapoethanol, and 2% bovine calf serum.

      Interleukin-2 Protein Quantification. Splenocytes were cultured in triplicate (1 × 106 cells/ml) in 48-well culture plates (800 μl/well). The cells were pretreated with antagonist or vehicle (or not pretreated in the case of the anandamide and arachidonic acid concentration responses) for 30 min prior to treatment with either anandamide or arachidonic acid. Splenocytes were activated 30 min later with 40 nM PMA and 0.5 μM ionomycin. The supernatants were collected 24 h after stimulation, and interleukin-2 protein was quantified using a sandwich enzyme-linked immunosorbent assay (ELISA) method previously described (Kaplan et al., 2003). The interleukin-2 standard (mouse recombinant interleukin-2), purified rat anti-mouse interleukin-2 antibody, and biotinylated anti-mouse interleukin-2 antibody were purchased from BD PharMingen (San Diego, CA).

      Statistical Analysis. The mean ± S.E. was determined for each treatment group in the individual experiments. Homogeneous data were evaluated by a parametric analysis of variance (ANOVA). For those experiments with two factors, a two-way ANOVA was employed. Dunnett's two-tailed t test was used to compare treatment groups with the vehicle control when significant differences were observed (Dunnett, 1955). IC50 values were calculated from the average of four different concentration responses using Prism GraphPad software (GraphPad Software Inc., San Diego, CA).

      Results

      Anandamide Causes a Robust Inhibition of Interleukin-2 Secretion, Which Is Unaffected by Pretreatment with the CB1/CB2 Antagonists. Previous studies from our laboratory have shown that the endogenous cannabinoid, 2-AG, produced a concentration-dependent inhibition of interleukin-2 in activated T cells (Ouyang et al., 1998). In light of these findings, a similar analysis of anandamide was performed. In splenocytes activated with PMA and ionomycin, anandamide caused a concentration-dependent inhibition of interleukin-2 with an IC50 of 11.4 μM (Fig. 1). The subsequent experiments were designed to determine the mechanism by which anandamide causes inhibition of interleukin-2 with the initial studies focusing upon the cannabinoid receptors. Although CB2 is the predominant cannabinoid receptor expressed in immune cells, low levels of CB1 transcripts have been detected in many immune cells, including T cells (Galiegue et al., 1995). Therefore, the role of both CB1 and CB2 on the inhibition of interleukin-2 secretion by anandamide was evaluated. Pretreatment of primary splenocytes with the CB1 and CB2 antagonists, SR141716A and SR144528, used in combination (0.05/0.05, 0.5/0.5, and 5/5 μM), prior to treatment with anandamide (10 μM) did not attenuate anandamide-mediated inhibition of interleukin-2 secretion (Fig. 2). At the highest concentration used (5/5 μM), SR141716A and SR144528, when employed in combination in the absence of anandamide, caused inhibition of interleukin-2 secretion. Additionally, neither pretreatment with SR141716A nor SR144528 alone attenuated anandamide-mediated suppression of interleukin-2 secretion (data not shown).

      Arachidonic Acid Also Causes a Concentration-Dependent Inhibition of Interleukin-2 Secretion. Because it was unclear whether the inhibitory effect of anandamide upon interleukin-2 secretion was due to the parent molecule or a hydrolysis product of anandamide, the effect of arachidonic acid upon interleukin-2 secretion was also evaluated. Treatment of primary splenocytes with various concentrations of arachidonic acid (0.1-20 μM) caused a concentration-dependent inhibition of interleukin-2 secretion (Fig. 3). The magnitude of interleukin-2 inhibition produced by arachidonic acid was very similar to that observed with anandamide as evidenced by the calculated IC50 values: 11.4 and 10.3 μM for anandamide and arachidonic acid, respectively.

      Neither Inhibition of the AMT nor Inhibition of FAAH Attenuates Suppression of Interleukin-2 by Anandamide. Because arachidonic acid produced a concentration-responsive suppression of interleukin-2 secretion with a profile of inhibition that resembled that of anandamide, it seemed reasonable that inhibition of interleukin-2 by anandamide was mediated by arachidonic acid, which is probably formed from the hydrolysis of anandamide rather than by the parent molecule of anandamide itself. It has been postulated that the AMT may be coupled to FAAH, the chief enzyme responsible for anandamide hydrolysis, and that these two proteins represent a major mechanism for the uptake and catabolism of anandamide (Giuffrida et al., 2001). As such, the potential roles of the AMT and FAAH in anandamide-mediated interleukin-2 inhibition were studied. Pretreatment with the AMT inhibitor, UCM707, at various concentrations (1-20 μM), did not attenuate anandamide-mediated suppression of interleukin-2 secretion (Fig. 4). Prior to evaluation of the effect of the FAAH inhibitor, MAFP, upon anandamide-mediated interleukin-2 inhibition, the effect of MAFP upon interleukin-2 in the absence of anandamide was determined. MAFP caused a concentration-responsive inhibition of interleukin-2 secretion at concentrations of 5 μM and higher (data not shown). Because MAFP is also a phospholipase A2 inhibitor and previous studies from this laboratory have shown that phospholipase A2 inhibitors cause a decrease in interleukin-2 production, the inhibitory effect of MAFP upon interleukin-2 secretion was not unexpected (Ouyang and Kaminski, 1999). Based upon the afore-mentioned study, 1 μM MAFP was the maximum concentration used for subsequent experiments due to the absence of an effect upon interleukin-2 coupled with the fact that the concentration is sufficient to inhibit FAAH (IC50 = 2.5 nM). Pretreatment with a broad range of concentrations of MAFP (0.001-1 μM) did not attenuate the inhibition of IL-2 secretion by anandamide (Fig. 5).

      Suppression of Interleukin-2 Secretion by Anandamide and Arachidonic Acid Is Attenuated by the Nonspecific Cyclooxygenase Inhibitors Flurbiprofen and Piroxicam. Due to the similarity in the concentration responses of anandamide and arachidonic acid in the inhibition of interleukin-2, the role of the cyclooxygenase enzymes was investigated. Initial evaluation of concentration responses of flurbiprofen determined 50 μM to be the optimum concentration for inhibition of cyclooxygenase-1 and cyclooxygenase-2 in splenocytes (data not shown). The higher concentration of 100 μM flurbiprofen caused marked inhibition of interleukin-2 by itself, which is not surprising since flurbiprofen (in the range of 100 to 1000 μM) is known to inhibit the activation of nuclear factor κ of B cells, an important transcription factor for interleukin-2 transcription (Tegeder et al., 2001). Although 50 μM flurbiprofen caused inhibition of interleukin-2 secretion in the absence of anandamide or arachidonic acid, the inhibition was far less robust than that which was caused by 100 μM flurbiprofen and nonetheless still caused an almost complete reversal of anandamide-mediated interleukin-2 inhibition (Fig. 6A). Likewise, flurbiprofen pretreatment also significantly attenuated the suppression of interleukin-2 mediated by arachidonic acid (Fig. 6B). Similar to flurbiprofen, pretreatment of primary splenocytes with piroxicam partially attenuated the inhibitory activity of both anandamide as well as arachidonic acid upon interleukin-2 secretion (Fig. 7). Moreover, the effect of piroxicam upon anandamide-mediated suppression of interleukin-2 is concentration-dependent (Fig. 9A).

      NS398, a Cyclooxygenase-2-Specific Inhibitor, Partially Attenuated the Inhibition of IL-2 Secretion by Anandamide and Arachidonic Acid. To determine whether the inhibitory activity of anandamide and arachidonic acid upon interleukin-2 was mediated by a cyclooxygenase-1 or cyclooxygenase-2 metabolite, the ability of NS398, a cyclooxygenase-2-specific inhibitor, to attenuate suppression of interleukin-2 by anandamide and arachidonic acid was evaluated. Similar to flurbiprofen and piroxicam, NS398 also attenuated the inhibition of IL-2 secretion by anandamide and arachidonic acid (Fig. 8). In addition, the attenuation by NS398 of anandamide-mediated inhibition of IL-2 secretion is also concentration-dependent (Fig. 9B). Conversely, the cyclooxygenase-1 specific inhibitors, SC560 and FR122047, did not have an effect upon the suppression of interleukin-2 secretion by anandamide (data not shown).

      Suppression of Interleukin-2 Secretion by Anandamide Is Partially Antagonized by Pretreatment with T0070907, a PPARγ Antagonist. Because activated PPARγ has been shown to cause inhibition of interleukin-2 secretion and a number of cyclooxygenase products, such as PGD2 and 15-deoxy-Δ12,14 PGJ2, have been shown to be endogenous agonists of PPARγ; the ability of T0070907, a specific PPARγ antagonist, to antagonize inhibition of interleukin-2 secretion by anandamide was examined. Pretreatment of primary splenocytes with T0070907, at various concentrations (0.1-10 μM), caused a concentration-dependent antagonism of anandamide-mediated suppression of interleukin-2 secretion (Fig. 10). Although the highest concentration of T0070907 used (10 μM), caused inhibition of interleukin-2 in the absence of anandamide, T0070907 nevertheless still caused a significant reversal of the effects of anandamide upon interleukin-2 at this concentration.

      Discussion

      The present studies demonstrate that anandamide causes a concentration-dependent inhibition of interleukin-2 secretion. The current studies also show that suppression of interleukin-2 secretion mediated by anandamide is not attenuated by pretreatment with the cannabinoid receptor antagonists, SR141716A and SR144528, suggesting that it is independent of both CB1 and CB2. Interestingly, arachidonic acid also caused a concentration-responsive inhibition of interleukin-2, which is similar to that of anandamide. The similarity was also evident from the calculated IC50 values, which were 11.4 μM and 10.3 μM for anandamide and arachidonic acid, respectively. Although the suppression of interleukin-2 by arachidonic acid suggests that inhibition of interleukin-2 by anandamide may be mediated by a product of anandamide hydrolysis rather than the parent molecule itself, neither pretreatment with an AMT inhibitor nor a FAAH inhibitor attenuated suppression of interleukin-2 secretion by anandamide. Pretreatment with two different nonspecific cyclooxygenase inhibitors, flurbiprofen and piroxicam, partially attenuated inhibition of interleukin-2 secretion by both anandamide and arachidonic acid, suggesting a role for the cyclooxygenase enzymes. Likewise, NS398, a cyclooxygenase-2-specific inhibitor also attenuated the inhibition of interleukin-2 secretion by both anandamide and arachidonic acid, whereas the cyclooxygenase-1 specific inhibitors, SC560 and FR122047, had no effect (data not shown). In addition, T0070907, a PPARγ-specific antagonist, caused a concentration-dependent partial antagonism of anandamide-mediated inhibition of interleukin-2. Collectively, the aforementioned observations suggest that the inhibition of interleukin-2 secretion is partially mediated by PPARγ, which is activated by a cyclooxygenase-2 metabolite of anandamide.

      The concentrations of anandamide, which cause inhibition of interleukin-2 secretion, are in the low micromolar range. Although concentrations of anandamide have been detected in the nanomolar range in rat and human plasma, the local concentrations of anandamide at the various target sites have yet to be determined. Because anandamide is synthesized de novo from membrane components, it seems likely, however, that concentrations of anandamide might be quite high within the microcosms of the target regions. Measurements of anandamide are further confounded by the lability of the compound. Moreover, anandamide has generally been measured in normal healthy tissue and plasma samples, whereas these levels may differ substantially in inflamed or damaged tissues. Arachidonic acid levels, for instance, have been shown to increase into the upper micromolar range in damaged tissues (Patel et al., 2003). Furthermore, arachidonic acid mobilization has been linked to increased anandamide synthesis (Pestonjamasp and Burstein, 1998).

      Although there have been a number of effects of endocannabinoids attributed to CB1/CB2, there are also a growing number of reports of endocannabinoid activities that are independent of CB1/CB2 in keeping with the studies reported here (Smart et al., 2000, Ross et al., 2002). One potential mechanism for cannabinoid receptor-independent activity is the vanilloid receptor, VR1. VR1 is a ligand-gated cation channel that is activated by heat or capsaicin. Anandamide is a full agonist of VR1 albeit at concentrations 10 to 20 times higher than those at which it activates CB1 (Smart et al., 2000). It is notable that the VR1 agonist, capsaicin, does not cause inhibition of interleukin-2 secretion and that the VR1 antagonist, capsazepine, does not attenuate anandamide-mediated interleukin-2 inhibition (data not shown). The current study is unique because it not only demonstrates anandamide-mediated activity that is independent of CB1/CB2 and VR1, but also because it shows that the metabolism of anandamide does not always lead to the cessation of physiological effects but in certain circumstances may actually be responsible for the biological activity observed.

      There have been a number of reports that anandamide mediates arachidonic acid release from various cell types, including human peripheral blood mononuclear cells (Berdyshev et al., 1997; Di Marzo et al., 1997). Therefore, an alternative explanation for the observations of the current investigation is that anandamide causes release of arachidonic acid from the primary splenocytes. The released arachidonic acid is then subsequently metabolized into an eicosanoid, which mediates the inhibition of interleukin-2 secretion. Although the current studies do not negate the possibility that anandamide causes arachidonic acid release, the striking similarity in the concentration responses of anandamide and arachidonic acid suggests that the same moiety is responsible for the activity of both anandamide and arachidonic acid. Further studies will be needed to determine whether this is the case, however.

      The primary mechanism that has been identified for the hydrolysis of anandamide is through the enzyme FAAH. FAAH has been detected in a variety of different tissues with the highest levels found in the brain, liver, small intestine, and testes, whereas relatively low levels have been found in the spleen (Bisogno et al., 2002). Although FAAH seems to be the primary mechanism for the hydrolysis of anandamide, at least one other enzyme has been identified that hydrolyzes anandamide. Unlike FAAH, the other amidase has an optimal pH of 5, is not inhibited by MAFP, and is also reported to have a high level of activity in the spleen (Ueda et al., 2001a). Under the conditions used for the current studies, it is likely that anandamide is either hydrolyzed by the acid amidase, another amidase that has yet to be identified, or is directly metabolized by cyclooxygenase-2.

      Whereas the vast majority of research on anandamide metabolism has focused upon FAAH, there has also been considerable interest in the role of the cyclooxygenase enzymes. There are two isoforms of the cyclooxygenase enzyme, cyclooxygenase-1 and cyclooxygenase-2. Cyclooxygenase-1 is constitutively expressed in most cell types, whereas cyclooxygenase-2 expression is induced in response to a stimulus only in certain cell types (Parente and Perretti, 2003). Both cyclooxygenase enzymes have been found in most cells within the immune system, including T cells (Tilley et al., 2001). It has been reported that anandamide can bind directly to cyclooxygenase-2 and be metabolized into prostaglandin E2-ethanolamide and other products as well (Yu et al., 1997). The same studies have also demonstrated, however, that anandamide neither binds directly to nor is oxygenated by cyclooxygenase-1.

      There is evidence to support that a number of different prostanoids can cause inhibition of interleukin-2 secretion, including prostaglandin E2, 15-deoxy-Δ12,14 prostaglandin J2, and prostaglandin I2 (Marcinkiewicz and Chain, 1993; Yang et al., 2000; Harris et al., 2002). Of the aforementioned eicosanoids, 15-deoxy-Δ12,14 prostaglandin J2 (15d-PGJ2) is the most recent to be recognized to suppress interleukin-2 secretion. There has been growing interest in 15d-PGJ2 in a number of different research areas due to its identification as one of the most potent endogenous ligands of PPARγ. Structurally related to the hormone receptors, PPARs are also members of the nuclear receptor superfamily. Currently, there are three subtypes of PPARs that have been identified: PPARα, PPARδ, and PPARγ. Although the function of PPARδ remains unclear, PPARα and PPARγ have been best characterized for their roles in lipid metabolism and have only recently been discovered to also be involved with immunoregulation (Clark, 2002). Although both PPARα and PPARγ are expressed in T cells, only activated PPARγ causes an inhibition of IL-2 secretion. Physical association of activated PPARγ with the transcription factor, NFAT, appears to suppress NFAT binding to the IL-2 promoter, which is thought to be the mechanism for the decrease in interleukin-2 production by PPARγ agonists (Yang et al., 2000). Similarly, activated PPARγ has also been shown to cause decreased cytokine production in activated macrophages through direct protein-protein interaction with nuclear factor κB, a key transcription factor for a number of different cytokines (Ricote et al., 1998).

      15d-PGJ2 is produced through the sequential metabolism of arachidonic acid by cyclooxygenase and PGD synthase, followed by a series of nonenzymatic transformations (Zhang and Young, 2002). Whether 15d-PGJ2 is produced in vivo has been somewhat of a controversy; however, recent studies have identified elevated levels of 15d-PGJ2 in LPS-stimulated RAW264.7 macrophages (Shibata et al., 2002). Future studies will determine whether 15d-PGJ2 is produced in other cell types, but it is notable that both enzymes required for 15d-PGJ2 formation, cyclooxygenase and PGD synthase, are expressed in T cells. Although 15d-PGJ2 is one of the most potent activators of PPARγ, other cyclooxygenase products are also able to activate PPARγ. More studies will be needed to determine exactly which cyclooxygenase products are produced from anandamide metabolism and which are subsequently responsible for the observed inhibition of interleukin-2 production.

      Although several studies have emerged which suggest that under certain conditions activated PPARγ may cause apoptosis in activated T cells, there are also a number of studies suggesting that PPARγ activation may be protective against apoptosis or have no effect upon viability (Clark, 2002; Wang et al., 2002). Whereas the cause of the differential effects of activated PPARγ upon T cell viability is unclear, it is likely that the concentration of 15d-PGJ2, the kinetics of cell treatment, and/or the state/mode of activation may be factors. Under the specific conditions used in the present studies, there was no effect upon cellular viability. It should be noted, however, that anandamide and arachidonic acid are likely to be metabolized into a number of different products, some of which may be protective against apoptosis.

      Although in the majority of cases studied thus far the metabolism of anandamide has been associated with the cessation of its physiological activity, the present studies are significant because they show that metabolism of anandamide in certain cases may yield a biologically active product. It has been demonstrated here that the metabolism of anandamide by cyclooxygenase-2 yields a product that causes inhibition of interleukin-2 secretion through activation of PPARγ.

      Source, Graphs and Figures: A Cyclooxygenase Metabolite of Anandamide Causes Inhibition of Interleukin-2 Secretion in Murine Splenocytes
      BKLFC
      • Forum Emlyn Hughes
      • ****
      • Started Topic

      • 822 posts |
      Re: The Medical Thread.
      Reply #65: Jan 01, 2015 10:00:16 am
      Molecular Characterization Of A Peripheral Receptor For Cannabinoids
      Abstract

      The major active ingredient of marijuana, delta 9-tetrahydrocannabinol (delta 9-THC), has been used as a psychoactive agent for thousands of years. Marijuana, and delta 9-THC, also exert a wide range of other effects including analgesia, anti-inflammation, immunosuppression, anticonvulsion, alleviation of intraocular pressure in glaucoma, and attenuation of vomiting. The clinical application of cannabinoids has, however, been limited by their psychoactive effects, and this has led to interest in the biochemical bases of their action. Progress stemmed initially from the synthesis of potent derivatives of delta 9-THC, and more recently from the cloning of a gene encoding a G-protein-coupled receptor for cannabinoids. This receptor is expressed in the brain but not in the periphery, except for a low level in testes. It has been proposed that the nonpsychoactive effects of cannabinoids are either mediated centrally or through direct interaction with other, non-receptor proteins. Here we report the cloning of a receptor for cannabinoids that is not expressed in the brain but rather in macrophages in the marginal zone of spleen.

      Source: PMID:7689702 [PharmGKB]

      Please let us know how you are doing Cornish YNWA
      Mad4LFC
      • Forum Ian Callaghan
      • ****

      • 989 posts | 81 
      Re: The Medical Thread.in
      Reply #66: Jan 01, 2015 11:34:21 am
      I suffer from pretty bad anxiety and possibly depression and I just needed somewhere to vent so sorry...

      Basically all my life I've been an anxious person, through school and college, I'd skip full days if I had a presentation or a one on one meeting that day. I'd get to the point that just having a conversation with someone would end up in me looking like I'd taken a swim in a lake through nervous sweats. Then after college I got a job running a machine where I presses one button in the same spot for 8 hours a day, and it totally depressed me but I didn't want to disappoint my family because they are big on the whole your worthless if your not working thing but after last Christmas it got so bad that I could no longer physically or mentally go back to that job and I quit.

      Over the past 12 months since then have been the best I remember since being a child! My anxiety pretty much vanished and I achieved a lot of personal goals such as losing almost 40 pounds and leaving my house to be social, I used to be scared of going to the supermarket believe it or not...

      During this year I've been unemployed I've been doing voluntary work and enjoying life while applying for jobs for Universal Credit and this month I heard back from Saintsbury's and my new way of life involved no fear so I blitzed the interview and even did well in the group sessions during the induction which if you told me a year ago I'd be standing in front of a group of strangers speaking confidently I'd have laughed...

      But after the induction I spent two hours on the shop floor and all that anxiety I used to feel came rushing back and hit me like a ton of bricks and since then I haven't slept and my body has been aching!

      I really don't want to go back to the person I was over a year ago.... where I was afraid to leave the house other than to work... I'm disappointed in myself because I'm already thinking about quitting and I haven't even done a full day but my anxiety is crippling!

      Today all I've done for my day off is search the internet about anxiety and that is exactly what I used to do while working for those 5 years on my time off...

      I don't know if I'm ready to work and that maybe I need to finally get my anxiety/depression looked at by a professional, but I don't want to disappoint people and appear to be a lazy person, because I'm not, and anybody that knows me would say the same.

      I just don't want everything I've accomplished this year to go to waste... like the weight loss, the daily exercise (I've skipped swimming today) and just nkt feeling anxious every second of the day....

      /rant over.

      Mate i feel your pain and anxiety is a crippler when it attacks, in my job i went from giving evidence at the old bailey to being scared of going to the local shop and speaking to the sales staff due to one episode of anxiety. Thing is though as hard as it may seem there is only one cause of the anxiety and that is YOU nothing else. When you come to terms with this you can the take the next step and try to find a treatment that works for you.

      I have tried medication and for me that doesnt or should i say didnt work, it masks the anxiety when you in non social situations, but you will still get an attack when you put yourself is stressful situations. Obviously you can have therapy but like i said above this will just get you to understand that the cause for your anxiety is YOU.

      All i can say is what worked for me and that is a process called mindfulness, many an article to search for on the internet, also ( although she is as funny as cramp ) Ruby Wax's book is a very good read on the subject.

      good luck dont give up hope.
      Roddenberry
      • Forum Legend - Paisley
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      • 16,568 posts | 1876 
      Re: The Medical Thread.
      Reply #67: Jun 06, 2015 12:01:47 am
      Any one with some tips about dealing with a concussion?  Feel nauseous but not like I'm going to throw up, feel tired but not sleepy, stand up and the world moves around me.  In the last couple of hours, my neck's been sore as hell, nothing seems to ease it.  Typing this out was a pain as well, got a touch of light sensitivity and can't focus properly.
      HScRed1
      • LFC Reds Subscriber
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      • 20,172 posts | 4401 
      Re: The Medical Thread.
      Reply #68: Jun 06, 2015 12:04:51 am
      Any one with some tips about dealing with a concussion?  Feel nauseous but not like I'm going to throw up, feel tired but not sleepy, stand up and the world moves around me.  In the last couple of hours, my neck's been sore as hell, nothing seems to ease it.  Typing this out was a pain as well, got a touch of light sensitivity and can't focus properly.

      Get your self to A/E you silly fool.

      Your symptoms sound like a migraine FWIW.

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