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Thyroid Carcinoma in Dogs
What are Thyroid Tumors?
Thyroid tumors originate in the thyroid gland located along the trachea. The thyroid gland is responsible for producing thyroid hormone which plays a role in regulating metabolic rate and other functions. Despite its important role in producing hormones, the majority of thyroid tumors are "non-functional," meaning they do not produce thyroid hormones. Occasionally functional tumors are identified and evaluation of thyroid hormone levels should be performed in any dog with a thyroid tumor. Thyroid tumors are locally invasive and have a moderate-high rate of spread to other places in the body (called metastasis).
Diagnostic Testing
Diagnostic tests are recommended to determine which sites in the body are involved. These tests include:
- Complete blood count (CBC)
- Serum biochemistry panel
- Urinalysis (U/A)
- Regional lymph node evaluation
- Thoracic radiographs
- +/- Abdominal sonogram (indicated for tumors on the caudal portion of the trunk or proximal hindlimbs)
Treatment for thyroid carcinomas is aimed both at controlling the tumor in the site where it originates and delaying or preventing metastasis. Controlling the tumor in the site where it originates is best accomplished with either wide surgical excision or the combination of surgery to remove obvious gross disease and radiation therapy to destroy residual microscopic disease. In one study, dogs with moveable thyroid tumors and no evidence of metastasis were treated with surgery alone with a median survival of >36 months. However, many thyroid tumors are "fixed" (deeply attached) and invasive into adjacent soft tissues. These tumors that are not freely moveable are unlikely to be removed with clean margins and the best option is a combination of radiation therapy and surgery. The recommendation of the North Carolina State University Animal Cancer Treatment Program involves pre-operative radiation therapy consisting of 19 treatments administered on a Mon-Fri schedule with a follow-up CT scan done 3 months after completion of radiation therapy to determine resectability. This is based on reports of the gradual reduction in tumor size with thyroid tumors and an attempt to optimize timing of surgery. These are difficult surgeries given the many nerves in the area and the thyroid's rich blood supply with the associated risk of significant hemorrhage. A board-certified surgeon is often recommended for these reasons.
In conjunction with surgery and radiation therapy, we recommend systemic chemotherapy (carboplatin, cisplatin and doxorubicin have reported activity) to delay or prevent metastasis. In our hospital, chemotherapy is given as an intravenous injection at 3-week intervals for a total of four-six treatments. Chemotherapy drugs work by targeting fast-growing cells; cancer cells divide faster and abnormally compared to normal cells. Because of this, chemotherapy preferentially destroys cancer and spares normal tissues. There is the potential for some normal tissues with rapid growth rates (intestinal lining, bone marrow, hair cells) to be transiently damaged by chemotherapy. However, our goal in treating cancer in animals is quality of life, so the dosages are lower than those used in people. We do not normally see significant toxicity with chemotherapy. There is always some risk when a drug is given for the first time, and the oncologist will discuss what types of side effects may develop and what to watch for. If the pet has no problems the first time they receive a drug, they should have no problems with subsequent treatments of that drug. If the pet experiences any side effects, we address them as needed and lower the dosage of the causative drug for future treatments to prevent recurrent problems.
We also recommend piroxicam, an oral Non-Steroidal Anti-Inflammatory Drug (NSAID) which has anti-inflammatory and analgesic properties as well as possible anti-tumor activity against carcinomas. This medication should be given with food and the pet owner instructed to watch for signs of gastrointestinal upset (vomiting, change in stool, etc.). If such signs develop, piroxicam should be discontinued for 3-5 days. Restarting of therapy can then be considered in conjunction with the stomach-protectant misoprostil.
If definitive treatment is not an option, palliative radiation therapy can be considered. Palliative radiation therapy consists of two consecutive twice-daily doses of radiation therapy and is aimed at alleviating pain, inflammation, and swelling associated with the tumor. We recommend that the treatment be repeated in one month if no disease or symptom progression is noted in hopes of extending the duration of response.
Selected References
- Klein MK, Powers BE, Withrow SJ, et.al. Treatment of thyroid carcinoma in dogs by surgical resection alone: 20 cases (1982-1989). J Am Vet Med Assoc; 1995;206:1007-1009.
- Theon AP, Marks SL, Feldman ES, et al. Prognostic factors and patterns of treatment failure in dogs with unresectable differentiated thyroid carcinomas treated with megavoltage irradiation. J Am Vet Med Assoc 2000;216:1775-1779.
- Brearley MJ, Hayes AM, Murphy S. Hypofractionated radiation therapy for invasive thyroid carcinoma in dogs: a retrospective analysis of survival. J Sm Anim Pract 1999;40:206-210.
- Fineman LS, Hamilton TA, de Gortari A, et al. Cisplatin chemotherapy for treatment of thyroid carcinoma in dogs: 13 cases. J Am Anim Hosp Assoc 1998;34:109-1112.


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YIELD: Makes about 7 cups
Stir sugar, lemon juice and peel in medium bowl. Add buttermilk; stir until sugar dissolves. Chill until cold, about 4 hours. Process mixture in ice cream maker according to manufacturer's instructions. Transfer to container with lid; freeze. (Can be made 3 days ahead. Keep frozen.)

subset: commoncrawl


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Candlewood Suites Omaha Airport
Candlewood Suites Omaha Airport
Quick Info
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Important Update[{"start": "1280203200000","end":"1303876800000", "desc": "The Candlewood Suites Omaha Airport is now taking reservations for 2011 Berkshire Hathaway Stockholder Annual Meeting."},{"start": "1280203200000","end":"1309492800000", "desc": "General Manager Aaron Jones would like to let all College World Series Fans know that the reservations are now open for 2011 games!"},{"start": "1280203200000","end":"1285819200000", "desc": "Dont forget to make your reservations for RiverCity Round Up a four day western festival featuring the Douglas Country Fair. Ak-Sar-Ben 4-H livestock show and rodeo. In addition, numerous events like the Heritage Parade, Trail rides, Giddy Up gulch and the nebraska beef councils \" Whats for Dinner\" Cooking stage provide hours of excitement and activities."}]
- On-site Guest Self-Laundry Facilities
- Housekeeping Full Service
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It's hard to leave your best buddy at home when you're traveling for extended periods, so we provide a pet friendly experience. Restrictions and fees apply.
Whether you are booking suites for a family reunion or suites for your company's training needs, Candlewood Suites hotels can accomodate your group.
Workout whenever you want: Candlewood Gym always has an open door and lots of aerobic and cardio equipment to make it easier to stick to your exercise routine, no sweat.
At Candlewood Suites® hotels, we want you to experience comfort in every sense.
Getting Here
Guest Reviews
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Omaha trip
June 19, 2015
Hotel was outstanding. Entire staff, especially CJ, and Erin was outstanding
Top shelf experience
ByCheesehead Chili
June 18, 2015
I stay at Omaha Candlewood every year when we come to Omaha, primarily because of the kitchenette in the room. We have a particular need for the full size fridge as we are competing in the ICS Nebraska State Chili Cook-off. Our experience is such, however, that we would stay at Candlewood even if we did not need the kitchen facilities. We've had nothing but wonderful experiences always. This hotel is extremely comfortable, and we feel secure there --- just like being at home.
Better Than Expected for a Candlewood
June 17, 2015
Complimentary shuttle to the CWS and downtown Omaha was awesome. Front desk staff was extremely friendly and accommodating. Size of suite and bathroom was exceptional, even for 4 people.
Beds were not overly comfortable. Pillows were flat out terrible. Probably would have needed 5 of their pillows stacked to get to an acceptable height that I'd normally get from 2.
Location isn't great, but the shuttle service makes it much more accessible to downtown Omaha than I originally expected.
Quiet and relaxing overnight stay
June 16, 2015
Customer service was top notch from the moment I walked in to the moment I left. Hotel had a nice atmosphere with a pleasant and clean feel to it. Room was more than adequate with a nice flat screen television and plenty of space to move around. Bathroom towels were very soft and comfortable. I will definitely be staying at Candlewood Suites on future trips.


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The application is directed to the treatment of one or more symptoms associated with drug withdrawal by administering desmethylselegiline.
1. In a method for obtaining a selegiline therapeutic effect in a patient undergoing drug withdrawal, the improvement which comprises administering to said patient R(−)desmethylselegiline employing a dosage regimen effective to treat one or more withdrawal symptoms associated with one or more drugs selected from the group consisting of addictive psychostimulants, opiates, narcotics, barbiturates, alcohol, and nicotine.
2. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with an addictive psychostimulant.
3. The improvement of claim 2, wherein said addictive psychostimulant is cocaine.
4. The improvement of claim 2, wherein said addictive psychostimulant is selected from the group consisting of: amphetamines; methamphetamines; dextroamphetamines; chlorphentermine, methylphenidate, pipradol, p-hydroxymorphedrine, fenfluramine, 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane, bupropion and pemoline.
5. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with an addictive opiate.
6. The improvement of claim 5, wherein said addictive opiate is selected from the group consisting of: opium; morphine; and heroin.
7. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with an addictive narcotic.
8. The improvement of claim 7, wherein said addictive narcotic is selected from the group consisting of: alphaprodine, anileridine, bezitramide, codeine, dihydrocodeine, diphenoxylate, fentanyl, hydrocodone, hydromorphone, isomethadone, levomethorphan, levorphanol, metazocine, methadone, metopon, oxycodone, oxymorphone, pethidine, phenazocine, piminodine, racemethorphan, racemorphon and thebaine.
9. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with an addictive barbiturate.
10. The improvement of claim 9, wherein said addictive barbiturate is selected from the group consisting of: allobarbital, amylbarbital, butabarbital, hexabarbital, mephobarbital, methohexital, pentobarbital, phenobarbital, phenethylbarbital, secobarbital, talbutal and thiopental.
11. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with alcohol.
12. The improvement of claim 1, wherein said R(−)desmethylselegiline is administered to treat one or more withdrawal symptoms associated with nicotine.
13. The improvement of claim 1, wherein said one or more withdrawal symptoms are selected from the group consisting of: drug craving; depression, irritability; anergia; amotivation; appetite changes; nausea; shaking; psychomotoric retardation; and irregular sleep patterns.
14. The method of any one of claims 1-13, wherein said R(−)desmethylselegiline is administered orally.
15. The method of any one of claims 1-13, wherein said R(−)desmethylselegiline is administered non-orally.
16. A pharmaceutical composition, comprising:
a) R(−)desmethylselegiline; and
b) a second drug useful in the treatment of the symptoms associated with drug withdrawal;
wherein said R(−)desmethylselegiline and said second drug are present in an amount such that one or more unit doses of said composition are effective to treat one or more withdrawal symptoms associated with one or more drugs selected from the group consisting of addictive psychostimulants, opiates, narcotics, barbiturates, alcohol, and nicotine.
17. The composition of claim 16, wherein said second drug is selected from the group consisting of naloxone, naltrexone or nalorphine.
18. The composition of claim 16, wherein said one or more withdrawal symptoms are selected from the group consisting of: drug craving; depression, irritability; anergia; amotivation; appetite changes; nausea; shaking; psychomotoric retardation; and irregular sleep patterns.
19. The improvement of claim 13, wherein the one or more withdrawal symptoms is drug craving.
20. The improvement of claim 13, wherein the one or more withdrawal symptoms is depression.
21. The improvement of claim 13, wherein the one or more withdrawal symptoms is irritability.
22. The improvement of claim 13, wherein the one or more withdrawal symptoms is psychomotoric retardation.
23. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with an addictive psychostimulant.
24. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with an addictive opiate.
25. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with an addictive narcotic.
26. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with an addictive barbiturate.
27. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with alcohol.
28. The composition of claim 16, wherein the one or more withdrawal symptoms are associated with nicotine.
29. The composition of claim 18, wherein the one or more withdrawal symptoms is drug craving.
30. The composition of claim 18, wherein the one or more withdrawal symptoms is depression.
31. The composition of claim 18, wherein the one or more withdrawal symptoms is irritability.
32. The composition of claim 18, wherein the one or more withdrawal symptoms is psychomotoric retardation.
33. The composition of claim 16, wherein the R(−)desmethylselegiline and the second drug are administered orally.
34. The composition of claim 16, wherein the R(−)desmethylselegiline and the second drug are administered non-orally.
The present application is a continuation-in-part of U.S. application Ser. No. 08/679,330 filed Jul. 12, 1996, which is a continuation-in-part of PCT/US96/01561, with an international filing date of Jan. 11, 1996, a continuation-in-part of U.S. Provisional Application No. 60/001,979, filed Jul. 31, 1995 and a continuation-in-part of U.S. application Ser. No. 08/372,139, filed Jan. 13, 1995.
The present invention pertains to methods and pharmaceutical compositions for using the selegiline metabolite R(−)desmethylselegiline (also referred to simply as "desmethylselegiline" or "R(−)DMS") either alone or in combination with its enantiomer, ent-desmethylselegiline (also referred to as "S(+)desmethylselegiline" or "S(+)DMS"). In particular, the present invention provides compositions and methods for using these agents in the treatment of selegiline-responsive diseases and conditions, particularly diseases or conditions involving neuronal degeneration or neuronal rescue).
Two distinct monoamine oxidase enzymes are known in the art: monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B). The cDNAs encoding these enzymes show different promoter regions and distinct exon portions, indicating they are encoded independently at different gene positions. In addition, analysis of the two proteins has shown differences in their respective amino acid sequences.
The first compound found to selectively inhibit MAO-B was R-(−)-N-methyl-N-(prop-2-ynyl)-2-aminophenylpropane, also known as L-(−)-deprenyl, R-(−)-deprenyl, or selegiline. Selegiline has the following structural formula:
Selegiline is the active ingredient of a human drug product and is known in the art as a component of a therapeutic package. In particular, see Physicians Desk Reference (1995) pp. 2430-2432 (1995 PDR), describing Eldepryl� Tablets, manufactured by Somerset Pharmaceutical, Inc. and marketed by Sandoz, the active ingredient of which is selegiline. For example, the 1995 PDR describes a 5 mg selegiline hydrochloride tablet and further describes the manner in which selegiline-containing therapeutic packages are supplied for commercial use or sale. In particular, the 1995 PDR discloses that 5.0 mg Eldepryl Tablets are sold in "NDC 39506-011-25 bottles of 60 tablets."
In commercial use, selegiline-containing therapeutic packages are labeled and otherwise indicated for use in Parkinsonian patients receiving levodopa/carbidopa therapy. The 1995 PDR cited above provides an example of the complete approved labeling that is employed in known therapeutic packages. Accordingly, known in the prior art are therapeutic packages providing one or more unit doses of selegiline as an active ingredient thereof, supplied in a finished pharmaceutical container that contains said unit doses, and further contains or comprises labeling directing the use of said package in the treatment of a human disease or condition as described above.
The selectivity of selegiline in the inhibition of MAO-B is important to its safety profile following oral administration. Inhibition of MAO-A may cause toxic side effects by interfering with the metabolism of tyramine. Tyramine is normally metabolized in the gastrointestinal tract by MAO-A but when MAO-A is inhibited, tyramine absorption is increased following consumption of tyramine-containing foods such as cheese, beer, herring, etc. This results in the release of catecholamines which can precipitate a hypertensive crisis, producing the "cheese effect." This effect is characterized by Goodman and Gilman as the most serious toxic effect associated with MAO-A inhibitors.
One of the metabolites of selegiline is its N-desmethyl analog. Structurally, this metabolite is the R(−) enantiomeric form of a secondary amine of the formula:
Heretofore, desmethylselegiline was not known to have pharmaceutically useful MAO-related effects, i.e., potent and selective inhibitory effects on MAO-B. In the course of determining the usefulness of desmethylselegiline for the purposes of the present invention, the MAO-related effects of desmethylselegiline were more completely characterized. This characterization has established that desmethylselegiline has exceedingly weak MAO-B inhibitory effects and no advantages in selectivity with respect to MAO-B compared to selegiline.
For example, the present characterization established that selegiline has an IC50 value against MAO-B in human platelets of 5�10−9 M whereas R(−)desmethylselegiline's IC50 value is 4�10−7 M, indicating the latter is approximately 80 times less potent as an MAO-B inhibitor than the former. Similar characteristics can be seen in the following data measuring inhibition of MAO-B and MAO-A in rat cortex mitochondrial-rich fractions:
As is apparent from the above table, selegiline is approximately 128 times more potent as an inhibitor of MAO-B relative to MAO-A, whereas desmethylselegiline is about 97 times more potent as an inhibitor of MAO-B relative to MAO-A. Accordingly, desmethylselegiline appears to have an approximately equal selectivity for MAO-B compared to MAO-A as selegiline, albeit with a substantially reduced potency.
Analogous results are obtained in rat brain tissue. Selegiline exhibits an IC50, for MAO-B of 0.11�10−7 M whereas desmethylselegiline's IC50 value is 7.3�10−7 M, indicating desmethylselegiline is approximately 70 times less potent as an MAO-B inhibitor than selegiline. Both compounds exhibit low potency in inhibiting MAO-A in rat brain tissue, 0.18�10−5 for selegiline, 7.0�10−5 for desmethylselegiline. Thus, in vitro R(−)desmethylselegiline is approximately 39 times less potent than selegiline in inhibiting MAO-A.
Based on its pharmacological profile as set forth above, R(−)desmethylselegiline as an MAO-B inhibitor provides no advantages in either potency or selectivity compared to selegiline. To the contrary, the above in vitro data suggest that use of desmethylselegiline as an MAO-B inhibitor requires on the order of 70 times the amount of selegiline.
The potency of R(−)desmethylselegiline as an MAO-B inhibitor in vivo has been reported by Heinonen, E. H., et al. ("Desmethylselegiline, a metabolite of selegiline, is an irreversible inhibitor of MAO-B in human subjects," referenced in Academic Dissertation "Selegiline in the Treatment of Parkinson's Disease," from Research Reports from the Department of Neurology, University of Turku, Turku, Finland, No. 33 (1995), pp. 59-61). According to Heinonen, desmethylselegiline in vivo has only about one-fifth the MAO-B inhibitory effect of selegiline, i.e., a dose of 10 mg of desmethylselegiline would be required for the same MAO-B effect as 1.8 mg of selegiline. In rats, Barbe reported R(−)desmethylselegiline to be an irreversible inhibitor of MAO-B, with a potency about 60 fold lower than selegiline in vitro and about 3 fold lower ex vivo (Barbe, H. O., J Neural Trans. (Suppl.):32:131 (1990)).
The various diseases and conditions for which selegiline is disclosed as being useful include: drug withdrawal (WO 92/21333, including withdrawal from psychostimulants, opiates, narcotics, and barbiturates); depression (U.S. Pat. No. 4,861,800); Alzheimer's disease and Parkinson's disease, particularly through the use of transdermal dosage forms, including ointments, creams and patches; macular degeneration (U.S. Pat. No. 5,242,950); age-dependent degeneracies, including renal function and cognitive function as evidenced by spatial learning ability (U.S. Pat. No. 5,151,449); pituitary-dependent Cushing's disease in humans and nonhumans (U.S. Pat. No. 5,192,808); immune system dysfunction in both humans (U.S. Pat. No. 5,387,615) and animals (U.S. Pat. No. 5,276,057); age-dependent weight loss in mammals (U.S. Pat. No. 5,225,446); and schizophrenia (U.S. Pat. No. 5,151,419). PCT published application WO 92/17169 discloses the use of selegiline in the treatment of neuromuscular and neurodegenerative disease and in the treatment of CNS injury due to hypoxia, hypoglycemia, ischemic stroke or trauma. In addition, the biochemical effects of selegiline on neuronal cells have been extensively studied (e.g., see Tatton, et al., "Selegiline Can Mediate Neuronal Rescue Rather than Neuronal Protection," Movement Disorders 8 (Supp. 1):S20-S30 (1993); Tatton, et al., "Rescue of Dying Neurons," J. Neurosci. Res. 30:666-672 (1991); and Tatton, et al., "(−)-Deprenyl Prevents Mitochondrial Depolarization and Reduces Cell Death in Trophically-Deprived Cells," 11th Int'l Symp. on Parkinson's Disease, Rome, Italy, Mar. 26-30, 1994. Although selegiline is reported as being effective in treating the foregoing conditions, neither the precise number or nature of its mechanism or mechanisms of action are known. However, there is evidence that selegiline may provide neuroprotection or neuronal rescue by reducing oxidative neuronal damage, increasing the amount of the enzyme superoxide dismutase, and/or reducing dopamine catabolism. For example, PCT Published Application WO 92/17169 reports that selegiline acts by directly maintaining, preventing loss of, and/or assisting in, the nerve function of animals.
Selegiline is known to be useful when administered to a subject through a wide variety of routes of administration and dosage forms. For example U.S. Pat. No. 4,812,481 (Degussa A G) discloses the use of concomitant selegiline-amantadine in oral, peroral, enteral, pulmonary, rectal, nasal, vaginal, lingual, intravenous, intraarterial, intracardial, intramuscular, intraperitoneal, intracutaneous, and subcutaneous formulations. U.S. Pat. No. 5,192,550 (Alza Corporation) describes a dosage form comprising an outer wall impermeable to selegiline but permeable to external fluids. This dosage form may have applicability for the oral, sublingual or buccal administration of selegiline. Similarly, U.S. Pat. No. 5,387,615 discloses a variety of selegiline compositions, including tablets, pills, capsules, powders, aerosols, suppositories, skin patches, parenterals, and oral liquids, including oil-aqueous suspensions, solutions, and emulsions. Also disclosed are selegiline-containing sustained release (long acting) formulations and devices.
Although a highly potent and selective MAO-B inhibitor, selegiline's practical use is circumscribed by its dose-dependent specificity for MAO-B, and the pharmacology of selegiline metabolites generated after administration.
The present invention is based upon the surprising discovery that both desmethylselegiline ("DMS" or "R(−)DMS") and its enantiomer (ent-desmethylselegiline, abbreviated as "ent-DMS" or "S(+)DMS") are useful in providing selegiline-like effects in subjects, notwithstanding dramatically reduced MAO-B inhibitory activity and an apparent lack of enhanced selectivity for MAO-B compared to selegiline. It has been discovered that desmethylselegiline, ent-desmethylselegiline and their isomeric mixtures provide a more advantageous way of obtaining selegiline-like therapeutic effects in selegiline-responsive diseases or conditions. This is particularly true for diseases or conditions characterized by neuronal degeneration, neuronal trauma or which are hypodopaminergic in nature, i.e. diseases or conditions characterized by reduced dopamine release and formation.
Thus, the present invention provides novel pharmaceutical compositions in which desmethylselegiline, either alone or in a racemic mixture with ent-desmethylselegiline is employed as the active ingredient and novel therapeutic methods involving the administration of desmethylselegiline. Specifically, the present invention provides:
(1) An improved method for obtaining selegiline-like therapeutic effects in a subject suffering from a selegiline-responsive disease or condition, which comprises: administering to said subject desmethylselegiline in a dosage regime effective to produce said selegiline-like therapeutic effect.
(2) A non-oral pharmaceutical composition comprising an amount of desmethylselegiline such that one or more unit doses of said composition administered on a periodic basis is effective to treat one or more selegiline-responsive diseases or conditions in a subject to whom said unit dose or unit doses are administered.
The pharmaceutical composition may contain desmethylselegiline in a substantially enantiomerically pure state or, alternatively, the composition may contain a racemic mixture of enantiomers that are together present in an amount sufficient for one or more unit doses of the composition to be effective in treating a selegiline-responsive disease or condition. The composition may be designed in such a way as to make it suitable for sublingual, buccal, parenteral or transdermal administration and may be adapted for effecting neuronal rescue or protection. The composition may also be adapted for restoring or improving immune system function in a human.
In addition, the present invention is directed to a therapeutic package for dispensing to, or for use in dispensing to, a patient being treated for a neuronal-protective or neuronal-regenerative selegiline-responsive disease or condition. The package contains one or more unit doses, each such unit dose comprising an amount of desmethylselegiline such that periodic administration is effective in treating the patient's selegiline-responsive disease or condition. The therapeutic package also comprises a finished pharmaceutical container containing the unit doses of desmethylselegiline and further containing or comprising labeling directing the use of the package in the treatment of the selegiline-responsive disease or condition. The unit doses may be adapted for oral administration, e.g. as tablets or capsules, or may be adapted for non-oral administration.
The invention is also directed to a method of dispensing desmethylselegiline to a patient being treated for a neuronal-protective or neuronal-regenerative selegiline-responsive disease or condition. The method comprises providing patients with a therapeutic package having one or more unit doses of desmethylselegiline in an amount such that periodic administration to the patient is effective in treating their selegiline-responsive disease or condition. The package also comprises a finished pharmaceutical container containing the desmethylselegiline and having labeling directing the use of the package in the treatment the selegiline-responsive disease or condition. The unit doses in the package may be adapted for either oral or non-oral.
As used herein the term "selegiline-responsive disease or condition" refers to any of the various diseases or conditions in mammals, including humans, for which selegiline is disclosed in the prior art as being useful. In particular, a "selegiline-responsive disease or condition" refers to the various diseases and conditions described above, e.g., Alzheimer's disease, cognitive dysfunction, neuronal rescue, and the like. The term also refers to the use of selegiline as an appetite suppressant. Similarly, the term "selegiline-like therapeutic effect" refers to one or more of the salutary effects reported as being exerted by selegiline in subjects being treated for a selegiline-responsive disease or condition.
The selegiline-responsive diseases or conditions related to neuronal degeneration or trauma which respond to the present methods include Parkinson's disease, Alzheimer's disease, depression, glaucoma, macular degeneration, ischemia, diabetic neuropathy, attention deficit disorder, post polio syndrome, multiple sclerosis, impotence, narcolepsy, chronic fatigue syndrome, alopecia, senile dementia, hypoxia, cognitive dysfunction, negative symptomatology of schizophrenia, amyotrophic lateral sclerosis, Tourette's syndrome, tardive dyskinesia, and toxic neurodegeneration.
The present invention also encompasses the restoration or improvement of immune system function by R(−)DMS or mixtures of R(−)DMS and S(+)DMS. Such improvement or restoration has been reported to occur when selegiline is administered to animals. The conditions or diseases treatable include age-dependent immune system dysfunction, AIDS, infectious diseases and immunological loss due to cancer chemotherapy.
In addition, the invention includes the control of selegiline-responsive withdrawal symptoms (e.g., drug craving, depression, irritability, anergia, amotivation, appetite changes, nausea, shaking, psychomotoric retardation and irregular sleep patterns) associated with a variety of drugs including: addictive psychostimulants (e.g. cocaine, amphetamines, methamphetamines, dextroamphetamines, chlorphentermine, methylphenidate, pipradol, p-hydroxymorphedrine, fenfluramine, 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane, bupropion and pemoline), addictive opiates (e.g. opium, morphine and heroin), addictive narcotics (e.g. alphaprodine, anileridine, bezitramide, codeine, dihydrocodeine, diphenoxylate, fentanyl, hydrocodone, hydromorphone, isomethadone, levomethorphan, levorphanol, metazocine, methadone, metopon, oxycodone, oxymorphone, pethidine, phenazocine, piminodine, racemethorphan, racemorphon and thebaine), addictive barbiturates (e.g. allobarbital, amylbarbital, butabarbital, hexabarbital, mephobarbital, methohexital, pentobarbital, phenobarbital, phenethylbarbital, secobarbital, talbutal and thiopental), alcohol and nicotine. The desmethylselegilne may be administered either orally or non-orally and may be used in conjunction with other drugs. Specifically included within the invention are pharmaceutical compositions containing desmethylselegiline together with a second drug useful in the treatment of drug addiction and in which the desmethylselegiline and second drug are present in an amount such that one or more unit doses of the composition are effective in the symptomatic treatment of drug withdrawal. Specific second drugs that can be used include naloxone, naltrexone or nalorphine.
Desmethylselegiline may be administered either by a route involving gastrointestinal absorption or by a route that does not rely upon gastrointestinal absorption. Depending upon the particular route employed, desmethylselegiline is administered in the form of the free base or as a physiologically acceptable non-toxic acid addition salt. Such salts include those derived from organic and inorganic acids such as, without limitation, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulphonic acid, acetic acid, tartaric acid, lactic acid, succinic acid, citric acid, malic acid, maleic acid, sorbic acid, aconitic acid, salicylic acid, phthalic acid, embonic acid, enanthic acid, and the like. The use of salts, especially the hydrochloride, is particularly desirable when the route of administration employs aqueous solutions, as for example parenteral administration; use of delivered desmethylselegiline in the form of the free base is especially useful for transdermal administration. Accordingly, reference herein to the administration of DMS or ent-DMS or to mixtures thereof encompasses both the free base and acid addition salt forms.
The optimal daily dose of desmethylselegiline, or of a combination of R(−)DMS and S(+)DMS, useful for the purposes of the present invention is determined by methods known in the art, e.g., based on the severity of the disease or condition being treated, the condition of the subject to whom treatment is being given, the desired degree of therapeutic response, and the concomitant therapies being administered to the patient or animal. Ordinarily, however, the attending physician or veterinarian will administer an initial dose of at least about 0.015 mg/kg, calculated on the basis of the free secondary amine, with progressively higher, doses being employed depending upon the route of administration and the subsequent response to the therapy. Typically the daily dose will be about 0.5 mg/kg and may extend to about 1.0 mg/kg of the patient's body weight depending on the route of administration. Again, all such doses should be calculated on the basis of the free secondary amine. These guidelines further require that the actual dose be carefully titrated by the attending physician or veterinarian depending on the age, weight, clinical condition, and observed response of the individual patient or animal.
The daily dose can be administered in a single or multiple dosage regimen. Either oral or non-oral dosage forms may be used and may permit, for example, a continuous release of relatively small amounts of the active ingredient from a single dosage unit, such as a transdermal patch, over the course of one or more days. This is particularly desirable in the treatment of chronic conditions such as Parkinson's disease, Alzheimer's disease, and depression. Alternatively, it can be desirable in conditions such as ischemia or neural damage to administer one or more discrete doses by a more direct systemic route such as intravenously or by inhalation. In still other instances such as glaucoma and macular degeneration, localized administration, such as via the intraocular route, can be indicated.
Pharmaceutical compositions containing desmethylselegiline and/or ent-desmethylselegiline can be prepared according to conventional techniques. For example, preparations for parenteral routes of administration for desmethylselegiline, e.g., intramuscular, intravenous and intraarterial routes, can employ sterile isotonic saline solutions. Sterile buffered solutions can also be employed for intraocular administration.
Transdermal dosage unit forms of desmethylselegiline and/or ent-desmethylselegiline can be prepared utilizing a variety of previously described techniques (see e.g., U.S. Pat. Nos. 4,861,800; 4,868,218; 5,128,145; 5,190,763; and 5,242,950; and EP-A 404807, EP-A 509761, and EP-A 593807). For example, a monolithic patch structure can be utilized in which desmethylselegiline is directly incorporated into the adhesive and this mixture is cast onto a backing sheet. Alternatively desmethylselegiline, and/or ent-desmethylselegiline, can be incorporated as an acid addition salt into a multilayer patch which effects a conversion of the salt to the free base, as described for example in EP-A 593807.
Desmethylselegiline and/or ent-desmethylselegiline can also be administered by a device employing a lyotropic liquid crystalline composition in which, for example, 5 to 15% of desmethylselegiline is combined with a mixture of liquid and solid polyethylene glycols, a polymer, and a nonionic surfactant, optionally with the addition of propylene glycol and an emulsifying agent. For further details on the preparation of such transdermal preparations, reference can be made to EP-A 5509761.
Since the term "ent-desmethylselegiline" refers to the S(+) isomeric form of desmethylselegiline, reference above to mixtures of desmethylselegiline and ent-desmethylselegiline includes both racemic and non-racemic mixtures of optical isomers.
Subjects treatable by the present preparations and methods include both human and non-human subjects for which selegiline-like therapeutic effects are known to be useful. Accordingly, the compositions and methods above provide especially useful therapies for mammals, especially domesticated mammals. Thus, the present methods and compositions are used in treating selegiline-responsive diseases or conditions in canine and feline species.
Successful use of the compositions and methods above requires employment of an effective amount of desmethylselegiline, or mixtures of desmethylselegiline and ent-desmethylselegiline. Although both desmethylselegiline and ent-desmethylselegiline are dramatically less potent than selegiline as inhibitors of MAO, employment of these agents, or a mixture of these agents, for neuroprotection does not require a commensurately increased dosage to obtain a selegiline-like therapeutic response. Surprisingly, dosages necessary to attain a selegiline-like therapeutic appear to be on the same order as the known doses of selegiline. Accordingly, because both desmethylselegiline and ent-desmethylselegiline exhibit a much lower inhibition of MAO-A at such dosages, desmethylselegiline and ent-desmethylselegiline provide a substantially wider margin of safety with respect to MAO-A associated toxicity compared to selegiline. In particular, the risk of the adverse effects of MAO-A inhibition, e.g., hypertensive crisis, are minimized due to the 40-70 fold reduced potency for MAO-A inhibition.
As described above and notwithstanding its demonstrably inferior inhibitory properties with respect to MAO-B inhibition, desmethylselegiline and its enantiomer appear to be at least as effective as selegiline in treating certain selegiline-responsive conditions, e.g., conditions resulting from neuronal degeneration or neuronal trauma. Although the oral route of administration will generally be most convenient, drug may be administered by the parenteral, topical, transdermal, intraocular, buccal, sublingual, intranasal, inhalation, vaginal, rectal or other routes as well.
As noted above, the present invention encompasses the additional discovery that desmethylselegiline can be employed either alone or in mixture with ent-desmethylselegiline. Desmethylselegiline, its enantiomer and mixtures thereof are conveniently prepared by methods known in the art, as described in Example 1.
FIG. 1: HPLC Chromatogram of Purified R(−)DMS (Microsorb MV Cyano Column). The purity of a preparation of R(−)DMS was determined by HPLC on a Microsorb MV Cyano column and results are shown in FIG. 1. The column had dimensions of 4.6 mm�15 cm and was developed at a flow rate of 1.0 ml/min using a mobile phase containing 90% 0.01 M H3PO4 (pH 3.5) and 10% acetonitrile. The column was run at a temperature of 40� C. and effluent was monitored at a wavelength of 215 nm. The chromatogram shows one major peak appearing at a time of 6.08 minutes and having 99.5% of the total light-absorbing material eluted from the column. No other peak had greater than 0.24%.
FIG. 2: HPLC Elution Profile of R(−)DMS (Zorbax Mac-Mod C18 Column). The same preparation that was analyzed in the experiments discussed in FIG. 1 was also analyzed for purity by HPLC on a Zorbax Mac-Mod SB-C18 column (4.6 mm�75 mm). Effluent was monitored at 215 nm and results can be seen in FIG. 2. Greater than 99.6% of the light-absorbing material appeared in the single large peak eluting at a time of between 2 and 3 minutes.
FIG. 3: Mass Spectrum of R(−)DMS. A mass spectrum was obtained for purified R(−)DMS and results are shown in FIG. 3. The spectrum is consistent with a molecule having a molecular weight of 209.72 and a molecular formula of C12H15N.HCl.
FIG. 4: Infrared Spectrum (KBr) of Purified R(−)DMS. Infrared spectroscopy was performed on a preparation of R(−)DMS and results are shown in FIG. 4. The solvent used was CDCl3.
FIG. 5: NMR Spectrum of Purified R(−)DMS. A preparation of purified R(−)DMS was dissolved in CDCl3 and 1H NMR spectroscopy was performed at 300 MHZ. Results are shown in FIG. 5.
FIG. 6: HPLC Chromatogram of S(+)DMS. The purity of a preparation of S(+)DMS was examined by reverse phase HPLC on a 4.6 mm�75 mm Zorbax Mac-Mod SB-C18 column. The elution profile, monitored at 215 nm, is shown in FIG. 6. One major peak appears in the profile at a time of about 3 minutes and contains greater than 99% of the total light-absorbing material that eluted from the column.
FIG. 7: Mass Spectrum of Purified S(+)DMS. Mass spectroscopy was performed on the same preparation examined in FIG. 6. The spectrum is shown in FIG. 7 and is consistent with the structure of S(+)DMS.
FIG. 8: Infrared Spectrum (KBr) of Purified S(+)DMS. The preparation of S(+)DMS discussed in connection with FIGS. 6 and 7 was examined by infrared spectroscopy and results are shown in FIG. 8.
FIG. 9: Effect of Selegiline on Neuron Survival. Mesencephalic cultures were prepared from embryonic 14 day rats. Cultures were used at about 1.5 million cells per plate and were maintained either in growth medium alone (control cultures) or in growth medium supplemented with selegiline. On day 1, 8 and 15, cells were immunostained for the presence of tyrosine hydroxylase ("TH"). Striped bars represent results obtained for cultures maintained in the presence of 50 μM selegiline and open bars represent results for control cultures. In all cases, results are expressed as a percentage of TH positive cells present in control cultures on day 1. The abbreviation "DIV" refers to "days in vitro." Asterisks or stars above bars both in FIG. 9 and the figures discussed below indicate a result that differs from controls in an amount that is statistically significant, i.e. P<0.05.
FIG. 10: [3H]-Dopamine Uptake in Mesencephalic Cells. Cells, cultured as described above for FIG. 9, were tested for their uptake of labeled dopamine and results are shown in FIG. 10. Striped bars represent uptake in cells maintained in the presence of 50 μM selegiline and open bars represent uptake in control cultures.
FIG. 11: Effect of Selegiline on Glutamate Receptor Dependent Neuronal Cell Death. Rat embryonic mesencephalic cells were cultured as described above. After allowing cultures to stabilize, the culture medium was changed daily for a period of 4 days to induce glutamate receptor-dependent cell death. Depending on the culture, medium contained either 0.5, 5.0 or 50 μM selegiline. After the final medium change, cultured cells were immunostained for the presence of tyrosine hydroxylase. From left to right, bars represent results for controls, 0.5, 5.0 and 50 μM selegiline.
FIG. 12: Effect of Selegiline on Dopamine Uptake in Neuronal Cultures. Rat mesencephatic cells were cultured and medium was changed on a daily basis as discussed for FIG. 11. Uptake of tritiated dopamine by cells was measured and results are shown in the figure. From left to right, bars are in the same order as for FIG. 11.
FIG. 13: Effect of R(−)Desmethylselegiline on Glutamate Receptor Dependent Neuronal Cell Death. Rat embryonic mesencephalic cultures were prepared as described above except that R(−)DMS was used instead of selegiline. On day 9, the number of TH positive cells in cultures was determined. Results are expressed as a percentage of control. From left to right, bars show results for controls, 0.5, 5 and 50 μM R(−)DMS.
FIG. 14: Effect of R(−)Desmethylselegiline on Dopamine Uptake in Neuronal Cultures. Cell cultures were prepared as described above for FIG. 13 and then tested for uptake of tritiated dopamine. Results for controls and for cells maintained in the presence of 0.5 μM, 5 μM and 50 μM desmethylselegiline are shown from left to right in the figure.
FIG. 15: Comparison of Dopamine Uptake in Mesencephalic Cells. Incubated in the Presence of Different Monoamine Oxidase Inhibitors. Rat embryonic mesencephalic cells were prepared as described for FIGS. 11-14 and incubated in the presence of a variety of monoamine oxidase inhibitors. The inhibitors examined were selegiline; R(−) desmethylselegiline; pargyline; and clorgyline, all at concentrations of 0.5, 5 and 50 μM. In addition, cells were incubated in the presence of the glutamate receptor blocker MK-801 at a concentration of 10 μM. Cultures were tested for uptake of tritiated dopamine.
FIG. 16: Relative Effectiveness of R(−) and S(+)DMS in Maintaining [3H]-Dopamine Uptake by Cultured Mesencephalic Cells (NMDA Model). Preparations of R(−) and S(+)DMS were assayed for their effect on [3H]-dopamine uptake by cultured rat mesencephalic cells exposed to the toxin N-methyl-D-aspartate (NMDA). Results were expressed as a percentage of the uptake seen in control cultures not exposed to NMDA and are shown in FIG. 16. From the left, the bars represent: cells incubated with medium alone; medium +5 μM deprenyl; medium +0.5 μM R(−)DMS; medium +5 μM R(−)DMS; medium +50 μM R(−)DMS; medium +0.5 μM S(+)DMS; medium +5 μM S(+)DMS; and medium +50 μM S(+)DMS. All of the cell cultures shown in the figure were exposed to 100 μM NMDA. Statistical significance was determined by ANOVA followed by Dunnett's test. One star above a bar indicates a percentage uptake that differs significantly from control uptake at the 0.05 confidence level. Two stars indicate a result that differs at the 0.01 confidence level.
FIG. 17: Relative Effectiveness of R(−) and S(+)DMS on Survival of Cultured Mesencephalic Cells (NMDA Model). Rat mesencephalic cell cultures were exposed to 100 μM NMDA and incubated as described above in connection with FIG. 16. The effect of DMS enantiomers on the survival TH positive cells is shown in FIG. 17. The bars are in the same order as for FIG. 16 and results are expressed as a percentage of control. One star indicates p<0.05 and two stars indicates p<0.01 when results are compared to those obtained for cells exposed to NMDA and then incubated in unsupplemented medium.
FIG. 18: Inhibition of Neuronal Dopamine Re-Uptake by Deprenyl and the Two Enantiomers of Desmethylselegiline. An in vitro nerve terminal preparation (synaptosome preparation) was prepared using fresh rat neostriatal tissue. This was examined for its ability to take up tritiated dopamine in buffer alone or in buffer supplemented with various concentrations of selegiline, R(−)desmethylselegiline or S(+)desmethylselegiline. Uptake in the presence of each MAO inhibitor, expressed as a percent inhibition vs. log concentration of inhibitor is shown in FIG. 18. As indicated, the plot was used to determine the IC50 for each test agent.
FIG. 19: Determination of IC50 Values for Inhibition of Dopamine Re-Uptake. The experiment of FIG. 18 was repeated in a concentration range designed to more accurately provide an IC50 value and results are shown in FIG. 19. Using the log C vs. probit graphs, as shown in the figure, the IC50 for S(+)DMS was determined to be about 1 1 μM; for selegiline, about 46 μM; and for R(−)DMS about 54 μM.
FIG. 20: In Vivo MAO-B Inhibition in Guinea Pig Hippocampus. Various doses of selegiline, R(−)desmethylselegiline, and S(+)desmethylselegiline were administered daily to guinea pigs for a period of 5 days. Animals were then sacrificed and the MAO-B activity in the hippocampus portion of the brain was determined. Results were expressed as a percent inhibition relative to hippocampus MAO-B activity in control animals and are shown in FIG. 20. The plots were used to estimate the ID50 dosage for each agent. The ID50 for selegiline was about 0.008 mg/kg; for R(−)DMS, it was about 0.2 mg/kg; and for S(+)DMS, it was about 0.5 mg/kg.
The surprising utility of desmethylselegiline and ent-desmethylselegiline in treating selegiline-responsive diseases or conditions is attributable in part to their powerful action in preventing loss of dopaminergic neurons by promoting repair and recovery. Because desmethylselegiline prevents loss and facilitates recovery of nerve cell function, it is of value in a wide variety of neurodegenerative and neuromuscular diseases. In this regard, desmethylselegiline is at least as potent as selegiline, and in one model of neuroprotection, appeared to be substantially more potent. This is described more empirically in Examples 4 to 9 below.
Preparation of Desmethylselegiline and Ent-desmethylselegiline
A. Desmethylselegiline
Desmethylselegiline (designated below as "R(−)DMS") is prepared by methods known in the art. For example, desmethylselegiline is a known chemical intermediate for the preparation of selegiline as described in U.S. Pat. No. 4,925,878. Desmethylselegiline can be prepared by treating a solution of R(−)-2-aminophenylpropane (levoamphetamine):
in an inert organic solvent such as toluene with an equimolar amount of a reactive propargyl halide such as propargyl bromide, Br—CH2—C≡CH, at slightly elevated temperatures (70�-90� C.). Optionally the reaction can be conducted in the presence of an acid acceptor such as potassium carbonate. The reaction mixture is then extracted with aqueous acid, for example 5% hydrochloric acid, and the extracts are rendered alkaline. The nonaqueous layer which forms is separated, for example by extraction with benzene, dried, and distilled under reduced pressure.
Alternatively the propargylation can be conducted in a two-phase system of a water—immiscible solvent and aqueous alkali, utilizing a salt of R(+)-2-aminophenylpropane with a weak acid such as the tartrate, analogously to the preparation of selegiline as described in U.S. Pat. No. 4,564,706.
B. Ent-Desmethylselegiline
Ent-desmethylselegiline (designated below as "S(+)DMS") is conveniently prepared from the enantiomeric S(+)-2-aminophenylpropane (dextroamphetamine), i.e.,
C. Mixtures of Enantiomers
Mixtures of enantiomeric forms of desmethylselegiline, including racemic desmethylselegiline, are conveniently prepared from enantiomeric mixtures, including racemic mixtures of the above aminophenylpropane starting material.
D. Conversion into Acid Addition Salts
N-(prop-2-ynyl)-2-aminophenylpropane in either optically active or racemic form can be converted to a physiologically acceptable non-toxic acid addition salt by conventional techniques such as treatment with a mineral acid. For example, hydrogen chloride in isopropanol is employed in the preparation of desmethylselegiline hydrochloride. Either the free base or salt can be further purified, again by conventional techniques such as recrystallization or chromatography.
Characteristics of Substantially Pure R(−)DMS
A preparation of substantially pure R(−)DMS has the appearance of a white crystalline solid with a melting point of 162-163� C. and an optical rotation of [α]D 23� C.=−15.2+/−2.0 when measured at a concentration of 1.0 M using water as solvent. R(−)DMS appeared to be 99.5% pure when analyzed by HPLC on a Microsorb MV Cyano column (see chromatogram in FIG. 1) and 99.6% pure when analyzed by HPLC on a Zorbax Mac-Mod SB-C 18 column, (see chromatogram in FIG. 2). No single impurity is present at a concentration greater than or equal to 0.5%. Heavy metals are present at a concentration of less than 10 ppm and amphetamine hydrochloride at a concentration of less than 0.03%. The last solvents used for dissolving the preparation, ethyl acetate and ethanol are both present at a concentration of less than 0.1%. A mass spectrum performed on the preparation (see FIG. 3) is consistent with a compound having a molecular weight of 209.72 and a formula of C12H15N.HCl. Infrared and NMR spectra are shown in FIGS. 4 and 5 respectively. These are also consistent with the known structure of R-(−)-DMS.
Characteristics of Substantially Pure S(+)DMS
A preparation of substantially pure S(+)DMS has the appearance of a white powder with a melting point of approximately 160.04� C. and a specific rotation of +15.1 degrees when measured at 22� C. in water, at a concentration of 1.0 M. When examined by reverse phase HPLC on a Zorbax Mac-Mod SB-C18 column the preparation appears to be about 99.9% pure (FIG. 6). Amphetamine hydrochloride is present at a concentration of less than 0.13% (w/w). A mass spectrum is performed on the preparation and is consistent with a compound having a molecular weight of 209.72 and a molecular formula of C12H15N.HCl(see FIG. 7). Infrared spectroscopy is performed and also provides results consistent with the structure of S(+)DMS (see FIG. 8).
Neuronal Survival as Measured Using Tyrosine Hydroxylase
The effect of desmethylselegiline on neuron survival can be correlated to tyrosine hydroxylase, the rate limiting enzyme in dopamine biosynthesis. Assays are performed by determining the number of tyrosine hydroxylase positive cells in cultured E-14 embryonic mesencephalic cells over a period of 7 to 14 days. Protection in this system has been seen with a variety of trophic factors including BDNF, GDNF, EGF, and β-FGF.
A. Test Methods
Timed pregnant Sprague-Dawley rats are used to establish neuronal cultures from embryonic rat brain on the 14th day of gestation. Mesencephalon is dissected out without the membrane coverings and collected in Ca++ and Mg++ free balanced salt solution at 4� C. Tissue fragments are dissociated in chemically defined medium by mild trituration with a small bore pasteur pipette. Cell suspension is plated in polyornithine-coated 35 mm Falcon plastic dishes (0.1 mg/ml, Sigma) at a density of 1.5�106 cells/dish. Cultures are maintained at 37� C. in an atmosphere of 10% CO2:90% air and 100% relative humidity, and fed twice weekly with chemically defined medium consisting of MEM/F12 (1:1, Gibco), glucose (33 mM), HEPES (15 mM), NaHCO3 (44.6 mM), transferrin (100 mg/ml), insulin (25 mg/ml), putrescine (60 nM), sodium selenite (30 nM), progesterone (20 nM), and glutamine (2 mM). Control cells receive no further additions. The medium used for other cells also includes test substance, e.g. selegiline, at one or more concentrations.
Cultures are fixed in 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) for 30 minutes at room temperature, permeabilized with 0.2% Triton X-100 for 30 minutes and incubated with an antibody against tyrosine hydroxylase (1:1000; Eugene Tech) for 48 hours at 4� C. in the presence of a blocking serum. They are then stained using a peroxidase-coupled avidin-biotin staining kit (Vectastain ABC kit; Vector Labs) with 3′,3′-diaminobenzidine as a chromagen.
The number of dopaminergic neurons in cultures is determined by counting the cells positively immunostained with TH antibodies. 100 fields (0.5 mm�0.5 mm) in two transverse strips across the diameter of the dish, representing 2.5% of the total area, are counted using a Nikon inverted microscope at 200�magnification.
B. Results
Using the procedures described above, the following results were obtained:
Neuronal Survival as Measured Using Dopamine Uptake
In addition to determining the number of TH positive cells in culture (see Example 4), the protective effect of desmethylselegiline on neuronal cells also can be determined by directly measuring dopamine uptake. The amount of uptake by the cultured brain cells corresponds to axonal growth.
A. Test Methods
Cell cultures, established in the manner discussed above, are incubated with [3H]dopamine (0.5 mCi/ml; 37 Ci/mmol; New England Nuclear) for 15 minutes in the presence of ascorbic acid (0.2 mg/ml) in PBS (pH 7.3), supplemented with 0.9 mM CaCl2 and 0.5 mM MgCl2 at 37� C. After two rinses and a 5 minute incubation with fresh buffer, [3H]dopamine accumulated within the cells is released by incubating the cultures with 95% ethanol for 30 minutes at 37� C. Preparations are then added to 10 ml Ecoscint (National Diagnostics) and counted in a scintillation spectrometer. Nonspecific uptake values are obtained by blocking dopaminergic neuronal uptake with 10 mM mazindol.
B. Results
Using the above procedure, the results shown in Table 3 were obtained.
C. Conclusions from Examples 4 and 5
The results described in Examples 4 and 5 indicate that desmethylselegiline is substantially more potent superior to selegiline as a neuroprotective agent. This is true notwithstanding the fact that desmethylselegiline in much less potent than selegiline as an inhibitor of MAO-B.
Neuroprotective Action of Desmethylselegiline Enantiomers in Cultured Dopamine-Containing Mesencephalic Neurons In Vitro
The survival of mesencephalic, dopamine-containing neuronal cultures of rat brain tissue was used in these experiments to examine neuroprotective properties of selegiline and R(−)desmethylselegiline. The number of TH positive neurons is directly proportional to the survival of dopaminergic neurons and 3H-dopamine uptake is a measure of axonal growth in these neurons.
A. Effect of Selegiline on the Survival of Dopaminergic Neurons
Mesencephalic cultures prepared from embryonic day 14 rats were treated with 0.5, 5 or 50 μM selegiline for 15 days, beginning on the day of plating. (For a more detailed discussion of the culturing of cells and other methods used in these experiments see Mytilineou et al., J. Neurochem. 61:1470-1478 (1993)). Survival and growth of dopamine neurons was evaluated by tyrosine hydroxylase (TH) immunocytochemistry and [3H]dopamine uptake and results are shown in FIGS. 9 and 10.
B. Effect of Selegiline on Glutamate Receptor Dependent Cell Death
The neuroprotective effect of selegiline was also examined using an experimental paradigm that causes neuronal cell death that can be blocked by inhibition of glutamate receptors. In these experiments, cells were plated and allowed to stabilize for several days. The growth medium of the cells was then changed on a daily basis to induce cell death that can be prevented by blocking glutamate receptors, e.g. using MK-801. After 4 days of daily medium changes, cultures were stained for tyrosine hydroxylase and assayed for uptake of tritiated dopamine. The results shown in FIGS. 11 and 12 further support the conclusion that selegiline promotes the survival of dopaminergic neurons.
C. Effect of Desmethylselegiline on the Survival of Dopamine Neurons
Using the glutamate receptor dependent model of neuron death, an even more potent protection of dopaminergic neurons was provided when desmethylselegiline was used in place of selegiline. Even at the lowest dose tested (0.5 μM), desmethylselegiline caused a significant reduction in the loss of TH positive neurons (FIG. 13) and a significant increase in dopamine uptake (FIG. 14) relative to control cultures in which medium was used without supplementation with either selegiline or desmethylselegiline.
D. Comparison with Other MAO Inhibitors
Using the glutamate receptor dependent model of neurotoxicity, the effects of selegiline and desmethylselegiline were compared with two other MAO inhibitors, pargyline and clorgyline (FIG. 15). In agreement with previous results, measurement of dopamine uptake indicated neuron protection by 50 μM deprenyl and 5 and 50 μM desmethylselegiline. Pargyline did not appear to offer any protection at the concentrations used, while clorgyline protected at 50 μM. As expected, protection was also obtained by the NMDA receptor blocker MK-801 (10 μM).
E. Effect of DMS Enantiomers on 3H-Dopamine Uptake
The data summarized in Table 4 suggests that both (R−)DMS and S(+)DMS are effective as neuroprotectants in mesencephalic dopamine-containing neurons in culture.
These results were obtained using the medium change model of cell death. Compared to untreated control cells, there was 40% and 134% more axonal growth and terminal axonal survival after treatment with R(−)DMS and S(+)DMS, respectively. In this study, S(+)DMS showed greater potency as a neuroprotectant than R(−)DMS.
Comparison of the Neuroprotective Effect of R(−)DMS and S(+)DMS
The neuroprotective effect of R(−)DMS and S(+)DMS on cultured rat mesencephalic cells was examined using two models of neuronal cell death. In the first model, cells were exposed to 100 μM N-methyl-D-aspartate (NMDA), an agent which causes cell death by binding to glutamate receptors. Cells exposed to NMDA were incubated in the presence of either medium alone; medium supplemented with 50 μM deprenyl; medium with 0.5, 5, or 50 μM R(−)DMS; or medium containing 0.5, 5 or 50 μM S(+)DMS. The effect of these treatments on [3H]-dopamine uptake and the survival of TH positive cells was determined and results are shown in Tables 5-8 and FIGS. 16 and 17. It. can be seen that both forms of DMS had a neuroprotective effect, with S(+)DMS being the most effective treatment to a statistically significant degree as determined by tritiated dopamine uptake. Experiments examining the neuroprotective effect of DMS enantiomers were also performed using the medium change model of cell death described previously (see Example 6). As can be seen in Tables 9-12, both the R(−) and S(+) enantiomers significantly enhanced [3H]-dopamine uptake and the survival of TH positive cells. In this model, the relative potency of both enantiomers appears to be equal to treatment with 50 μM selegiline.
Desmethylselegiline and Ent-Desmethylselegiline as Inhibitors of Dopamine Re-Uptake
The biological actions of the brain neurotransmitter dopamine are terminated at the synapse by a high-affinity, sodium and energy-dependent transport system (neuronal re-uptake) present within the limiting membrane of the presynaptic dopamine-containing nerve terminal. Inhibition of this transport mechanism would extend the actions of dopamine at the synapse and therefore enhance dopamine synaptic transmission.
A. Method of Testing
The R(−) and S(+) enantiomers of desmethylselegiline (DMS) were tested for their ability to inhibit the dopamine re-uptake system and compared to selegiline. Inhibitory activity in this assay is indicative of agents of value in the treatment of diseases which require enhanced synaptic dopamine activity. Presently this would include Parkinson's disease, Alzheimer's disease and attention deficit hyperactivity disorder (ADHD).
The assay system used was essentially that described by Fang et al (Neuropharmacology 33:763-768 (1994)). An in vitro nerve-terminal preparation (symptosome-preparation) was obtained from fresh rat neostriatal brain tissue. Transport by dopamine nerve-terminals was estimated by measuring the uptake of tritiated dopamine.
B. Results
As seen in the data presented in Table 13, selegiline, R(−)DMS and S(+)DMS all inhibited dopamine re-uptake by dopamine-containing nerve terminals. Selegiline and R(−) DMS were approximately equipotent. In contrast, S(+)DMS was 4-5 times more potent than either selegiline or R(−)DMS.
Relative potency can be expressed in terms of the concentration required to inhibit doparnine re-uptake by 50% (IC50). The IC50 values were determined graphically (see FIG. 18) and are shown below in Table 14.
The experiment described above was repeated in a concentration range designed to more accurately describe IC50 values and results are shown in FIG. 19. ID50 values determined based upon the graph are shown in Table 15.
C. Conclusions
The results demonstrate that, at the appropriate concentration, selegiline and each of the enantiomers of DMS inhibit transport of released dopamine at the neuronal synapse and enhance the relative activity of this neurotransmitter at the synapse. In this regard, S(+)DMS is more potent than selegiline which, in turn, is more potent than R(−)DMS. Of the agents tested, S(+)DMS is the most preferred with regard to the treatment of hypodopaminergic conditions such as ADHD.
Actions of the R(−) and S(+) enantiomers of Desmethylselegiline (DMS) on Human Platelet MAO-B and Guinea Pig Brain MAO-B and MAO-A Activity
Human platelet MAO is comprised exclusively of the type-B isoform of the enzyme. In the present study, the in vitro and in vivo inhibition of this enzyme by the two enantiomers of DMS was determined and compared with inhibition due to selegiline. The present study also examined the two enantiomers of DMS for inhibitory activity with respect to the MAO-A and MAO-B in guinea pig hippocampal tissue. Guinea pig brain tissue is an excellent animal model for studying brain dopamine metabolism, the enzyme kinetics of the multiple forms of MAO and the inhibitory properties of novel agents that interact with these enzymes. The multiple forms of MAO in this animal species show similar kinetic properties to those found in human brain tissue. Finally, the test agents were administered to guinea pigs and the extent to which they might act as inhibitors of brain MAO in vivo was assessed.
A. Method of Testing
In vitro: The test system utilized the in vitro conversion of specific substrates of MAO-A (14C-serotonin) and MAO-B (14C-phenylethylamine) by human platelets and/or guinea pig hippocampal homogenates. The rate of conversion of each substrate was measured in the presence of S(+)DMS, R(−)DMS or selegiline and compared to the isozyme activity in the absence of these agents. A percent inhibition was calculated from these values. Potency was evaluated by comparing the concentration of each agent which caused a 50% inhibition (IC50 value).
In vivo: R(−)DMS, S(+)DMS or selegiline was administered in vivo subcutaneously (sc), once a day for 5 days prior to sacrifice, preparation of enzyme hippocampal homogenates, and the in vitro assay of MAO-A and MAO-B activity. These experiments were performed to demonstrate that the DMS enantiomers were capable of entering brain tissue and inhibiting MAO activity.
B. Results
MAO-B Inhibitory Activity In Vitro
Results for MAO-B inhibition are shown in Tables 16 and 17. IC50 values for MAO-B inhibition and potency as compared to selegiline is shown in Table 18.
As observed, R(−)DMS was 20-35 times more potent than S(+)DMS as an MAO-B inhibitor and both enantiomers were less potent than selegiline.
MAO-A Inhibitory Activity In Vitro
Results obtained from experiments examining the inhibition of MAO-A in guinea pig hippocampus are summarized in Table 19. The IC50 values for the two enantiomers of DMS and for selegiline are shown in Table 20.
R(−)DMS was twice as potent as S(+)DMS as an MAO-A inhibitor and both were 20-40 times less potent than selegiline. Moreover, each of these agents were 2-3 orders of magnitude, i.e., 100 to 1000 times, less potent as inhibitors of MAO-A than inhibitors of MAO-B in hippocampal brain tissue. Therefore, selegiline and each enantiomer of DMS can be classified as selective MAO-B inhibitors in brain tissue.
Results of In Vivo Experiments
Each enantiomer of DMS was administered in vivo by subcutaneous injection once a day for five consecutive days, and inhibition of brain MAO-B activity was then determined. In preliminary studies, selegiline was found to have an ID50 of 0.03 mg/kg and both R(−)DMS and S(+)DMS were determined to be about 10 times less potent. More recent studies, performed on a larger group of animals, indicates that R(−)DMS is actually about 25 times less potent than selegiline as an inhibitor of MAO-B and that S(+)DMS is about 50 times less potent. Results are shown in FIG. 20 and ID50 values are summarized in Table 21.
This experiment demonstrates that the enantiomers of DMS penetrate the blood brain-barrier and inhibit brain MAO-B after in vivo administration. It also demonstrates that the potency differences as an MAO-B inhibitor observed in vitro between each of the DMS enantiomers and selegiline are substantially reduced under in vivo conditions.
In experiments examining the effect of 5 s.c. treatments on MAO-A activity in guinea pig cortex (hippocampus), it was found that selegiline administration at a dose of 1.0 mg/kg resulted in a 36.1% inhibition of activity. R(−)DMS resulted in an inhibition of 29.8% when administered at a dose of 3.0 mg/kg. S(+)DMS administration did not cause any observable inhibition at the highest dose tested (10 mg/kg) indicating that it has significantly less cross reactivity potential.
C. Conclusions
In vitro, R(−)DMS and S(+)DMS both exhibit activity as MAO-B and MAO-A inhibitors. Each enantiomer was selective for MAO-B. S(+)DMS was less potent than R(−)DMS and both enantiomers of DMS were less potent than selegiline in inhibiting both MAO-A and MAO-B.
In vivo, both enantiomers demonstrated activity in inhibiting MAO-B, indicating that these enantiomers are able to pass through the blood-brain barrier. The ability of these agents to inhibit MAO-B suggests that these agents may be of value as therapeutics for hypodopaminergic diseases such as ADHD and dementia.
In Vivo Neuroprotection by the Enantiomers of Desmethylselegiline
The ability of the enantiomers of DMS to prevent neurological deterioration was examined by administering the agents to the wobbler mouse, an animal model of motor neuron diseases, particularly amyotrophic lateral sclerosis (ALS). Wobbler mice exhibit progressively worsening forelimb weakness, gait disturbances, and flexion contractions of the forelimb muscles.
A. Test Method
A 0.1 mg/kg dose of R(−)DMS, S(+)DMS or placebo was administered to wobbler mice by daily intra-peritoneal injection for a period of 30 days in a randomized, doubleblind study. At the end of this time mice were examined for grip strength, running time, resting locomotive activity and graded for semi-quantitative paw posture abnormalities, and semi-quantitative walking abnormalities. The investigators who prepared and administered the test drugs to the animals were different than those who analyzed behavioral changes.
Assays and grading were performed essentially as described in Mitsumoto et al., Ann. Neurol. 36:142-148 (1994). Grip strength of the front paws of a mouse was determined by allowing the animal to grasp a wire with both paws. The wire is connected to a gram dynamometer and traction is applied to the tail of the mouse until the animal is forced to release the wire. The reading on the dynamometer at the point of release is taken as a measure of grip strength.
Running time is defined as the shortest time necessary to traverse a specified distance, e.g. 2.5 feet and the best time of several trials is recorded.
Paw posture abnormalities are graded on a scale based upon the degree of contraction and walking abnormalities are graded on a scale ranging from normal walking to an inability to support the body using the paws.
Locomotive activity is determined by transferring animals to an examination area in which the floor is covered with a square grid. Activity is measured by the number of squares traversed by a mouse in a set time interval, e.g., 9 minutes.
B. Results
At the beginning of the study, none of the groups were different in any variables, indicating that the three groups were comparative at the baseline. Weight gain was identical in all three groups, suggesting that no major side effects occurred in any animals. Table 22 summarizes differences that were observed in the mean grip strength of the test animals:
Grip strength dropped markedly at the end of the first week in all animals. At the end of the study, grip strength was the least in control animals. The variability in grip strength in the treated animal groups prevented a meaningful statistical analysis of this data, however, at a dose of 0.1 mg/kg, the mean grip strength measured in the DMS-treated animals was greater than for the controls. These results suggest that the dose may have been too low, and that a higher dose study should be performed.
Running time, resting locomotive activity, semiquantitative paw posture abnormality grading, and semi-quantitative walking abnormality grading were also tested. None of these tests, however, showed any difference among the three groups tested.
Immune System Restoration by R(−)DMS and S(+)DMS
There is an age-related decline in immunological function that occurs in animals and humans which makes older individuals more susceptible to infectious disease and cancer. U.S. Pat. Nos. 5,276,057 and 5,387,615 suggest that selegiline is useful in the treatment of immune system dysfunction. The present experiments were undertaken to determine whether R(−)DMS and S(+) are also useful in the treatment of such dysfunction. It should be recognized that an ability to bolster a patient's normal immunological defenses would be beneficial in the treatment of a wide variety of acute and chronic diseases including cancer, AIDS, and both bacterial and viral infections.
A. Test Procedure
The present experiments utilized a rat model to examine the ability of R(−)DMS and S(+)DMS to restore immunological function. Rats were divided into the following experimental groups:
1) young rats (3 months old, no treatment);
2) old rats (18-20 months old, no treatment);
3) old rats injected with saline;
4) old rats treated with selegiline at a dosage of 0.25 mg/kg body weight;
5) old rats treated with selegiline at a dosage of 1.0 mg/kg body weight;
6) old rats treated with R(−)DMS at a dosage of 0.025 mg/kg body weight;
7) old rats treated with R(−)DMS at a dosage of 0.25 mg/kg body weight;
8) old rats treated with R(−)DMS at a dosage of 1.0 mg/kg body weight;
9) old rats treated with S(+)DMS at a dosage of 1.0 mg/kg body weight.
Rats were administered saline or test agent ip, daily for 60 days. They were then maintained for an additional "wash out" period of 10 days during which time no treatment was given. At the end of this time, animals were sacrificed and their spleens were removed. The spleen cells were then assayed for a variety of factors which are indicative of immune system function. Specifically, standard tests were employed to determine the following:
1) in vitro production of γ-interferon by concanavalin A-stimulated spleen cells;
2) in vitro concanavalin A-induced production of interleukin-2;
3) percentage of IgM positive spleen cells (IgM is a marker of B lymphocytes);
4) percentage of CD5 positive spleen cells (CD5 is a marker of T lymphocytes).
B. Results
The effect of administration of selegiline, R(−)DMS and S(+)DMS on concanavalin A-induced interferon production by rat spleen cells is shown in Tables 23 and 24. Table 23 shows that there is a sharp decline in cellular interferon production that occurs with age. Administration of selegiline, R(−)DMS and S(+)DMS all led to a restoration of γ-interferon levels with the most dramatic increases occurring at dosages of 1.0 mg/kg body weight.
Table 24 shows the extent to which R(−)DMS, S(+)DMS and selegiline are capable of restoring γ-interferon production in the spleen cells of old rats. Interferon-γ is a cytokine associated with T cells that inhibit viral replication and regulate a variety of immunological functions. It influences the class of antibodies produced by B-cells, upregulates class I and class II MHC complex antigens and increases the efficiency of macrophage-mediated killing of intracellular parasites.
Histological immunofluorescence studies show a dramatic loss of innervation in rat spleens with age. When rats are treated with R(−)DMS, there is a significant increase in innervation in the spleens of animals and this increase occurs in a dose-response manner. S(+)DMS did not show any effect on histological examination, despite a modest increase in interferon-γ production. IL-2 production was not enhanced by treatment with R(−)DMS or S(+)DMS, suggesting that the effects of these agents may be limited to IFN-γ production.
C. Conclusions
The results obtained with respect to histological examination, the production of interferon, and the percentage of IgM positive spleen cells support the conclusion that the DMS enantiomers are capable of at least partially restoring the age-dependent loss of immune system function. The results observed with respect to IFN-γ are particularly important. In both humans and animals, IFN-γ production is associated with the ability to successfully recover from infection with viruses and other pathogens. In addition, it appears that R(−)DMS and S(+)DMS will have a therapeutically beneficial effect for diseases and conditions mediated by weakened host immunity. This would include AIDS, response to vaccines, infectious diseases and adverse immunological effects caused by cancer chemotherapy.
Examples of Dosage Forms
The two ingredients are thoroughly mixed, cast on a film backing sheet (e.g., Scotchpak� 9723 polyester) and dried. The backing sheet is cut into patches a fluoropolymer release liner (e.g., Scotchpak� 1022) is applied, and the patch is hermetically sealed in a foil pouch. One patch is applied daily to supply 1-10 mg of desmethylselegiline per 24 hours in the treatment of conditions in a human produced by neuronal degeneration or neuronal trauma.
B. Ophthalmic Solution
Desmethylselegiline (0.1 g) as the hydrochloride, 1.9 g of boric acid, and 0.004 g of phenyl mercuric nitrate are dissolved in sterile water qs 100 ml. The mixture is sterilized and sealed. It can be used ophthalmologically in the treatment of conditions produced by neuronal degeneration or neuronal trauma, as for example glaucomatous optic neuropathy and macular degeneration.
C. Intravenous Solution
A 1% solution is prepared by dissolving 1 g of desmethylselegiline as the HCl in sufficient 0.9% isotonic saline solution to provide a final volume of 100 ml. The solution is buffered to pH 4 with citric acid, sealed, and sterilized to provide a 1% solution suitable for intravenous administration in the treatment of conditions produced by neuronal degeneration or neuronal trauma.
D. Oral Dosage Form
Tablets and capsules containing desmethylselegiline are prepared from the following ingredients (mg/unit dose):
with an approximately 1:1 ratio of citric acid and sodium citrate.
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Brick Oven Plans: Table of Contents
The Pompeii Oven is a set of plans to build an Italian Brick Oven. It is comprised of a number of basic components, which are outlined here to give you quick access to that page. Click on these links to jump directly to the directions for each step of the process of building your brick oven.
Oven Overview
Insulating Hearth
Cooking Surface
Oven Dome
Vent Arch
Dome Insulation
Enclosure, including the Igloo and Gabled House, and Indoor ovens.
Click on Materials List to see what you need to buy and what tools you need.


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The Hatewatch blog is managed by the staff of the Intelligence Project of the Southern Poverty Law Center, an Alabama-based civil rights organization.
News Roundup for July 21, 2011
White supremacist Mark Anthony Stroman was executed Wednesday evening for his post-9/11 revenge spree. That spree left two South Asian men dead and one Bangladeshi man wounded in acts of revenge for the 9/11 terrorist attacks.
Police in Colorado Springs, Colo., are investigating the second possible anti-gay hate crime in a month. Two homeless men were reportedly stabbed as their assailants made anti-gay slurs. This follows an earlier incident in which at least one man was beaten because he was thought to be gay.
Two New Mexico men have pled guilty to federal hate crime charges for their part in the branding of a Navajo man. Paul Beebe, 27, and Jesse Sanford, 25, admitted to drawing sexually and racially offensive pictures on 22-year-old Vincent Kee, and then branding him with a swastika. Kee reportedly has fetal alcohol syndrome and was considered an easy target. Beebe and Sanford will reportedly receive up to 8 ½ years in prison for the crime.
Longtime Militiaman Drew Rayner Facing Trial for Mail Fraud
NOTE: HateWatch has learned that Drew Allen Rayner has prominent ties to the antigovernment "sovereign citizen" movement. In April 2010, Rayner was one several sovereign citizens who signed a letter sent to all 50 governors demanding they vacate their offices, or else. The Guardians for the free Republics (GFR), a group which included the vanguard of the sovereign citizen movement, organized the letter campaign. Another sovereign citizen who signed the letters was James Timothy Turner, a sovereign guru who heads the Republic for the united States of America (RUSA) – one of the largest and most ambitious sovereign groups active in the United States.
A major figure in the antigovernment militia movement for the past two decades has been indicted for mail fraud in Mississippi. Prosecutors allege that Drew Allen Rayner, 72, gave a fake promissory note to the town of Poplarville to pay a court-ordered fine.
Rayner, who is set to go to trial on Oct. 3, faces up to 55 years in prison and a $500,000 fine.
U.S. Senator Catches Anti-Gay Testifier Misrepresenting Study
A representative of one of the nation's largest organizations opposed to same-sex marriage learned, as the U.S. Senate and the entire nation watched, to be careful what studies you cite – because someone might actually read them.
U.S. Sen. Al Franken of Minnesota caught Thomas Minnery, vice president of government and public policy for Focus on the Family, blatantly mischaracterizing a government study on the benefits to children of being raised by two married parents as referring to opposite-sex parents, when the report itself drew no such distinction. It was the latest in a relentless campaign of misrepresentations, half-truths and outright lies in recent years that have defined anti-gay Christian right organizations intent on opposing equal rights and common dignity for LGBT people.
Judge Ignores 'Martian law,' Tosses 'Sovereign Citizen' Into Slammer
This Georgia judge had heard just about enough nonsense from a "sovereign citizen" charged in a scheme to steal a house.
Rockdale County Superior Court Judge David Irwin repeatedly advised Akeem Kwame – a self-described "Moorish national" charged with mortgage fraud for falsely claiming ownership of a house in Conyers, Ga. – to get a lawyer or accept a public defender.
And Kwame repeatedly rejected that advice. It was time for trial.
News Round Up for July 20, 2011
One of the prime suspects in the murder of civil rights activists in the 1960s died on Wednesday. Richard Willis, a former Philadelphia, Miss., police officer, was long seen as part of the group of Klansmen involved in the killing of civil rights activists James Chaney, Andrew Goodman and Michael Schwerner in 1964. The incident spawned a massive FBI investigation and the indictment of nearly 18 men on federal conspiracy charges.
Four Texas men connected to a white supremacist prison gang have been arrested for a brutal home invasion. Jason Skolaski, 34, Shannon Michael Kasper, 20, Ned Alan Smith, 42, and his 15-year old son broke into the house of a San Jacinto County couple, stole $300, and then cut off the finger of one of the residents with a pair of bolt cutters before fleeing. The three adult suspects are believed to be part of the white supremacist Aryan Brotherhood of Texas.
A New Jersey woman was arrested for illegally occupying a foreclosed house and committing forgery. Jolanda S. Bordley-Jackson EL, 41, is accused of moving into a foreclosed home in Mullica Township, filing a fraudulent deed claiming ownership of the house, and putting the utilities in her name while changing the locks of the house. Police say the woman is a member of the Moorish Science Temple of America, some of whose members hold the beliefs of the sovereign citizen movement.
April Gaede Claims Daughters' Interviews are a Money-Making Scam
April Gaede responded to her twin daughters Lynx and Lamb announcing they had left the movement with a theory: They're scamming the Jews, just as she taught them. "The girls are using the Jews [sic] media to make bank as usual. If they had done anything else they would not have gotten any attention," Gaede said on Facebook earlier this week. "For a money making scheme I guess I can't blame them too much."
The message came a day after her two daughters granted an exclusive interview to The Daily on Sunday in which they announced they had renounced the racist world in which they were raised in favor of "a place of love and light." "We should have been pushed toward something a little more mainstream and easier for us to handle than being front-men for a belief system that we didn't completely understand," Lynx said in the interview.
Given their role as what one publication called the "new face of hate," the announcement sent shock waves through the racist right and drew out many white nationalist and nativist figures eager to respond. Some offered a defense of the Gaede twins, but overwhelmingly the trend was to comfort their mother, April Gaede. Even William Pierce, an author and chief ideologue behind the once powerful neo-Nazi National Alliance, made concessions to practice his beliefs, one commenter said. "Pierce had a Jew publish his books too… Can't blame [Lynx and Lamb] too much," a user named Per Olsson said. (Pierce is most infamous for publishing The Turner Diaries, a race war novel that inspired Oklahoma City bomber Timothy McVeigh.) ( continue to full post… )
Internet Messages Detail 'Nazi Jihadist's' Radicalization
A Pennsylvania man who was indicted Thursday for allegedly using the Internet to encourage Islamic fundamentalists to launch terror attacks against U.S. targets has a long history of extremism and joined a neo-Nazi group in 2003.
Emerson Begolly, 22, of New Bethlehem, Pa., was about 14 when he first contacted the National Socialist Movement (NSM), America's largest neo-Nazi group, Hatewatch has learned.
Begolly was arrested on January 4 after he bit two FBI agents who approached him to serve search warrants as he sat alone in the front seat of a car parked at a Burger King. When the agents opened the door and identified themselves, he screamed and reached for his pocket, then bit the agents' hands, drawing blood. He was found to have a 9 mm Makarov handgun with the safety off and two fully loaded magazines in his pocket.
News Roundup for July 19, 2011
A former member of the white supremacist Aryan Circle prison gang pleaded guilty for his part in a 2008 murder. George Buford Scott pleaded to a single assault charge in return for testimony against a fellow gang member, Billy Wayne Haynes, in the murder of Rey Valdez in 2008. Scott was released for time served on a two-year sentence, while Haynes received life in prison.
Authorities in Asheville, N.C. are looking for suspects in a possible hate crime. Luke Hankins claims that a group of teenagers began to shout gay slurs at him and followed him to his car before one teen hit him, causing three factures in his face. Police are treating the incident a hate crime; however, state law does not include sexual orientation as a consideration for a hate crime.
A Washington couple connected to the sovereign citizen movement has been charged for fraudulently attempting to collect $320,600 in tax refunds. Prosecutors accuse Raymond Leo and Ute Christine Jarlik-Bell of requesting large refunds from the IRS through fraudulent means. Previously, the couple had participated in the "Sovereign Assemblies" movement that promoted sham business accounts to conceal income from the IRS.
Two Arkansas men charged with a hate crime for a cross burning pleaded guilty today. Tony Branscum, 25, and James Bradley Branscum, 23, confessed that they planned to burn a cross in the yard of an African-American man, and carried it out with a third defendant. The pair face up to 10 years in prison for the crime.
Neo-Nazi Twins Drop Racism for Love and Marijuana
Lynx and Lamb Gaede were twin pop sensations for all the wrong reasons. Their songs dealt with overt white nationalist themes, and even their band – Prussian Blue – was a reference to the distinctive color of Zyclon B residue in the Nazi gas chambers. In one of the most infamous photos from their childhood, the twins posed wearing white T-shirts with a smiley face adorned with Hitler's toothbrush mustache and neatly parted, jet-black hair.
That's all over, the girls say now. In an exclusive interview with The Daily, Lynx and Lamb – once described as the "new face of hate" – say they've abandoned the racism hoisted on them by their mother April Gaede and instead settled into "a place of love and light."
"I'm glad we were in the band," Lynx said. "But I think we should have been pushed toward something a little more mainstream and easier for us to handle than being front-men for a belief system that we didn't completely understand." Lamb put their transformation more bluntly. "I'm not a white nationalist anymore." ( continue to full post… )
News Roundup for July 18, 2011
A Texas white supremacist responsible for an anti-Muslim shooting spree faces execution on Wednesday. Mark Stroman, 41, is currently on death row for a series of revenge attacks in the days after the 9/11 terror attacks that killed two South Asian men and wounded another. The surviving victim, a Bangladeshi Muslim, is asking for Storman's sentence to be commuted to life in prison.
One of the defendants convicted for the murder of an Arizona man and his daughter has been sentenced to life in prison. Albert Gaxiola, along with two others, entered the home of Junior Flores pretending to be Border Patrol agents and shot Flores, his wife (who survived), and their daughter Brisenia. Two of the defendants have received the death penalty, but the jury was unable to reach a unanimous decision on a death sentence for Gaxiola.
A member of the Hutaree Militia has been deemed unfit for trial due to mental problems. Jacob Ward, 34, has been indicted, along with nine others, for an antigovernment plot to kill police officers. Judge Victoria Roberts stated that Ward suffers from a mental defect, and is unable to understand the charges he faces.
The hunt for a militia leader has been temporarily halted after the suspect disappeared into the Montana woods. David Burgert, 47, a former Marine and the leader of the Project 7 militia was released in 2010 after eight years in prison for illegal firearms possession. Last June, Burgert led law enforcement on a chase, before shooting at the officers and fleeing into the woods. Local authorities plan to continue the search when new leads arise.
Police in Cherry Hill, N.J., have upgraded charges over a series of vandalisms against local Christian sites to a hate crime. In the course of a week, six cases of vandalism have taken place against Christian shrines, including one instance in which more than 30 statues were damaged at a church. Police are still searching for the suspect.


text: Ballistic Publishing - Call for Entries The OutpostSome time ago I started to make a small series of a fantasy-like environment settings and this is one of them : a small isolated outpost, situated on a solitary mountain peak. Copyright (C) Ognian Bonev 2007 All Rights Reserved No unauthorised duplication or distribution without explicit permission from the copyright holder. Artist Name: Ognian Bonev Artist Country: US Entered for: Website: http://www.northflame.comSoftware used: Autodesk - 3ds MaxAdobe - Photoshop About us | Contact us | Privacy Policy Copyright © 2003 Ballistic Media Pty. Ltd.


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i think it was a good move for the nuggets to try to squeeze everything they can for carmelo but i do think the knicks made the mistake of doing the trade during the current nba season when they shouldve waited until he was a free agent
The reason why Melo didn't wait until after the season was because of the problems looming with next season. With contract changes, Melo was playing it safe. I think the Knicks made a good move, because I think they'll be in title contention a few years down the road.
Pretty good deal


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How to make difficult decisions will be explored at Lifetree Café 7 p.m. CT on Monday, March 4.
The program, titled "Making Life's Toughest Decisions," features a screening of the award-winning short film "The Last Race."
"The Last Race" portrays a family suddenly faced with a decision about whether to donate the organs of a loved one.
Admission to the 60-minute event is free. Snacks and beverages are available. Lifetree Café is located at 1602 U.S. Highway 98 in Mexico Beach across from El Governor Motel.
Lifetree Café is a place where people gather for conversation about life and faith in a casual coffeehouse-type setting. Questions about Lifetree may be directed to Gary Grubb at 334-806-5667 or email@example.com.


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Dirtbags Win Epic With North Carolina, Lose In Regional Final To End Season
Ino Patron finished with six hits in four games to lead the Dirtbags.
McNeil, Four Other Dirtbags Garner All-Big West Recognition
Junior Jeff McNeil earns first team honors.
Dirtbags Take Series With 5-4 Win Over UC Riverside
Senior Juan Avila led the Dirtbags with three hits Saturday.
Long Beach State Loses Heartbreaker in 2-1 Pitcher's Duel
Jeff McNeil was 3-for-4 with two doubles in the loss.
Dirtbags Complete Sweep of UC Davis With 7-2 Win
Junior Michael Hill finishes series with six hits.
Long Beach State (2014): All-Big West Second Team selection ... led the Dirtbags with 25 appearances on the mound with 24 of those coming in relief ... posted a 5-5 record with a 1.77 ERA and held opposing batters to a .238 batting average ... recorded five saves, which ranked second on the team ... made his only start on Sunday against Vanderbilt (2/16), allowing four runs over 5.2 innings ... made the move back to the bullpen and allowed just one run over his next 10 appearances, which totaled 15.1 innings ... allowed only one hit over two innings to Wright State (2/22) ... pitched a perfect two-thirds of an inning against USC while striking out one to pick up his first win (2/25) ... allowed a run in the ninth at Wichita State for his second loss (2/28) ... got the win at Arizona (3/4) after a scoreless inning of relief ... did not allow a hit over 2.1 innings and struck out two to get a win over Indiana (3/14) ... another 1.2 innings of scoreless relief led to another win over Indiana (3/16) ... did not allow a hit and struck out a pair in a scoreless two-inning relief appearance at Cal State Fullerton (3/22) ... held UCLA scoreless for an innings (3/25) ... limited CSUN to one hit over two scoreless innings (3/28) ... charged with a loss after allowing CSUN to score once (3/30) ... struck out two at USC but allowed a run on two hits for his fourth loss ... an unearned run against UCLA (4/4) charged him with his final loss of the season ... pitched a perfect inning at UC Davis (4/12) and another scoreless inning the next day ... held UCSB scoreless for three innings out of the bullpen (4/19) ... did not allow a hit over 2.1 innings against Cal Poly and struck out three to complete a shutout and collect his second save ... had another three-inning appearance against Cal State Fullerton (5/4), allowing one run on four hits ... struck out three in 3.1 innings at Hawaii (5/10) for his longest relief appearance of the year and third save ... picked up his fifth win with a scoreless appearance of 1.2 innings against UC Irvine (5/22) ... struck out two in the ninth against UC Irvine to complete a Dirtbag sweep and earn his fourth save ... pitched 2.1 perfect innings against North Carolina in the elimination game to get the his fifth save and help the Dirtbags advance.
(2013):Made 22 appearances on the mound - all out of the bullpen ... posted a 2.33 ERA with a 3-0 record in relief ... also came through with one save ... earned All-Big West Honorable Mention for middle relief ... did not allow a run in his first appearance of the year - one inning at Vanderbilt (2/15) ... tossed a perfect inning of relief against Valparaiso with a strikeout (2/22) ... had three consecutive hitless outings from Mar. 3-17 ... was perfect in an inning against Seattle (3/3) ... recorded one out at Arizona State (3/9) ... struck out a batter in a flawless inning against Wichita State (3/17) ... struck out the side in his only inning of work against San Diego State (4/9) to earn his first win ... threw six scoreless innings of relief over five outings from Mar. 28-Apr. 19 ... allowed just one hit and a walk in a two-inning effort against UC Davis (4/12) ... had another perfect inning against the Aggies (4/14) ... struck out two in a perfect inning of relief at UC Santa Barbara (4/19) ... did not surrender a hit or a walk in 1.1 innings against Cal Poly (4/26) ... pitched two scoreless innings of relief with a strikeout against Cal Poly (4/28) to record his only save ... did not allow a run in two appearances at Cal State Fullerton (5/3, 5/5) ... didn't allow a run in eight consecutive appearances from Apr.26-May 18 ... worked 13.1 innings of scoreless relief over those eight appearances ... went 2.1 innings against Pepperdine (5/7) to earn his second win of the season ... won his third game with a three-inning relief effort against UC Riverside (5/18) - his longest effort of the year.
(2012):Redshirted during the 2012 season.
High School:Earned two letters with Wilson High School in Long Beach ... A first-team All-Moore League selection as a senior.
Personal: Born in Long Beach ... Parents are Robert and Donna Provencher ... Majoring in communications.
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