Formulations vary depending on the type of tree selected; Sexual dysfunction remains a major problem by the third-to-forth decade of male life and continues to increase in prevalence with each subsequent decade. I purchased Hard Steel from a local gas station. 1999). Naphthalene was used as internal standard. Compared with other markers of recent alcohol intake, such as blood and urinary methanol, 5HTOL/5HIAA remains elevated for a longer time (615 hours vs. 26 hours for methanol) after blood alcohol levels have returned to normal levels. SPG warns against counterfiet versions of Hard Steel Male Enhancement. 1999; Sillanaukee et al. We have extensively investigated the safety of magnesium stearate, which is also considered safe and non-toxic and has no conflict when consumed with Alcohol. Examples include caffeine, morphine, strychnine, and nicotine. 2000), there does not appear to be a gender effect with %CDT, a procedure that determines the, SA levels also rise in conditions other than heavy drinking. We concluded that extract of EbM relax the CC smooth muscle through multitargets in NO/cGMP/PDE 5 pathway and might bring into perspective the treatment strategy for those patients with erectile dysfunction. The relaxant responses are normalized as the percentage of relaxation of the contraction originally induced by PE. ; Robinson, D.R. One of the most commonly used and researched of the ginsengs is Panax ginseng, also called Asian or Korean ginseng. The isometric muscle transducer in turn was connected to a polygraph recorder (Gould TA240, Ohio, USA). Then, the EP-20 extract was chromatographed on a column of Cosmosil5-C18-AR II and eluted with a gradient of water-acetonitrite (9:1 to acetonitrite alone), to yield six fractions (EP-20-1 to EP-20-6) by time interval. Fatiguein people withmultiple sclerosis(MS). The first catch The involvement of NO/cGMP pathway in EP-20-induced relaxation effect, After co-incubation withL-NAME (3 104M), the EP-20-induced relaxation effects were partially blocked by 30% in PE-precontracted CC strips, Whereas in co-incubation with a guanlyl cyclase inhibitor (ODQ, 3 105M), the relaxation effects were also partially blocked. The discrepancy between biological responses to crude extracts and pure compounds are commonly observed in many herbal remedies on clinical aspect, for which more evidence is needed to prove or disprove it. Reapply the solvent to the glue and use a razor blade to lift the loosened super glue. (1999a) reported a positive relationship between alcohol intake and SA levels in serum. Effect ofL-NAME or guanylyl cyclase inhibitor on CRCs of EP-20 in CC strips. Nehra A, Moreland RB . in the ingredients panel. Catuaba Bark - is a natural remedy derived from the bark of trees found in the Brazilian rainforest. Phosphodiesterases (PDE) are key enzymes in the regulation of CC smooth muscle tone. In fact, the fact that EbM extract potentiates the commonly used PDE 5 inhibitors, such as sildenafil and vardenafil, provides a new treatment strategy for those patients with ED. InFigure 3, EP-20 (0.01mg/ml) was pretreated in PE-precontracted CC strips for 10min, followed by incubation with different concentration of NO donor (SNP). and is safe. Several steps for quality experiments including retention time, spiking of authentic standard, change of wavelength and change of the composition of mobile phase were used to examine the contents in EbM extracts.13To standardize and quantify the EbM extracts, icariin (C33H40O15, molecular weight: 676.65Da) was used as an internal standard with a chromatograph column (Acq-Tac, Waters, Kyoto, Japan) and a UV detector (G1314A, Agilent, Waldbronn, Germany).11,12Furthermore, theL-arginine level was determined and quantified by HPLC analysis with a chromatograph column (38023-11, Cosmosil, Kyoto, Japan) as described previously.14In brief, 10mgL-arginine (A5006, Sigma, ST Louis, USA) was dissolved in 0.1MHCl as stock solution. FDA requires all products containing FD&C Black & White to identify it on their labels. Maximize your sexual performance with a unique herbal blend developed and manufactured by physicians in the USA. Mop it up . 1983), myocardial infarction (Woollen and Turner 1965), thyrotoxicosis (Oberkotter et al. product, specifically because it stated it worked with alcohol. Its a mixture of pure stearic acid and palmitic acid, where the content of stearic acid is not less than 40.0% and the sum of the two acids is not less than 90.0%. Relaxation responses are expressed as percentage of residual tension of contraction originally induced by PE (1M). Enclosed is a Comprehensive Analytical and Advisory Service physical performance test and physicochemical test for components formulated in Hard Steel Original Formulated Male Enhancement. It is synthesized in the intestinal chromaffin cells or in the central or peripheral neurons and is found in high concentrations in many body tissues. Determination of cGMP level in rabbit CC strips, The CC strips were pretreated with EP-20 in the presence or absence of ODQ (3 105M) or SNP (1 106M) or phosphodiesterase 5 (PDE 5) inhibitor (sildenafil, 1 107M) for 20min, followed by freezing the CC strips into liquid nitrogen. According to ICH Q6B guidelines, impurities in biological products can be classified as process-related or product-related. The effects of alcohol on males are a bit more straightforward. In HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT, Silica is included to improve Digestive issues by increasing the absorption of the product total formulation. People with diabetes, high blood pressure, high cholesterol, or heart disease often take nitrates and should always seek a physician before taking any male enhancements. Heavydrinking male college students produced higher absolute values than their heavydrinking female counterparts, although 74 percent of the women versus 44 percent of the men had levels above the 99th percentile for abstainers. Mean red cell volume as a marker of alcohol intake. Furthermore, it is of note that crude extracts of EbM (EPW and EP-20) concentration- dependently relax the PE-contracted CC strips. package said. Best Male Enhancement Pill, Penis Enlargement Procedure, Hard Steel Works With Alcohol, Details On Kangaroo Pills And Testosterone . Gelatin Scientifically, isrich in protein, and has a unique amino acid profile that gives it many potential health benefits. Heart: Drinking a lot over a long time or too much on a single occasion can damage the heart, causing problems including: Cardiomyopathy - Stretching and drooping of heart muscle. Owing to the fact that therapeutic doses of PDE 5 inhibitors exhibit slight blood-pressure-lowering effects, the combination of PDE 5 inhibitors with any NO donor is absolutely contraindicated because of potentially life-threatening hypotension.3, Recently, it is interesting to find that the vasodilating effect of wine-derived phenolic compounds is associated with the inhibition of PDEs and, in particular, PDE 5. These issues have a bearing particularly for new markers for which fully automated test procedures in which we have developed. CDT levels appear to elevate following alcohol consumption of 6080 g/d for 2 or 3 weeks without having impact on the effects on erectile dysfunction capsules whether medically or non-medically as in the case of HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT. The blue ice lotus continued to enlarge and quickly smashed towards the man in black. 1980), whereas urine hex normalizes after 4 weeks of abstinence (Martines et al. The data suggested that EP-20 enhanced relaxation effect with SNP in rabbit CC strips. Early warning of rejection? Early warning of rejection? The mobile phase was filtered through a millipore 0.45m filter and degassed before use. In contrast, SNP (0.01mg/ml) was pretreated in PE-precontracted CC strips for 10min, followed by incubation with different concentration of EP-20. ; and Thompson, A.E. Identification:Mass spectrometry(ESI-Triple Quad, MALDI-TOF, Q-TOF),NMR(1D and 2D), FTIR, Raman and UV-Vis spectroscopy, N-terminal Edman sequencing, LC-MS/MS. It is a magnesium salt of fatty acids C16 to C18. Mean red cell volume as a marker of alcohol intake. This can help men with erectile dysfunction. Evernew's take on the popular backpacking stove, however, is practically featherweight, weighing just over one ounce. All lab(s) relevant USP, EP, and JP testing procedures are certified by its Analytical Team. [12]Other amino acids that contribute highly include:alanine(Ala) 811%;arginine(Arg) 89%;aspartic acid(Asp) 67%; andglutamic acid(Glu) 1012%.[11]. I had a turkey sandwich from Arbys, and 2 rum Flu (influenza). Effective (a, 0.1mg/ml) and ineffective (b, 0.01mg/ml) concentration of EP-20 were co-incubated without () or with PDE 5 inhibitor alone () or in combination (). 1994). Magnesium is an essential mineral, serving as a cofactor for hundreds of enzymatic reactions and is essential for the synthesis of carbohydrates, lipids, nucleic acids and proteins, as well as neuromuscular and cardiovascular function.2, The FDA has affirmed that stearic acid is GRAS (Generally Regarded As Safe) and can be added to foods in accordance with Good Manufacturing Practices (GMP).3HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT is produced in a GMP-certified manufacturer lab. Alcohol works two ways to kill bedbugs. Increased levels of the 5HTOL/5HIAA ratio have been reported in association with disulfiram treatment, calcium cyanamide therapy, and glyburide treatment. It was hard steel works with alcohol pitch black and terrifying. Data points represent means.e.m. In nature stearic acid occurs primarily as a mixed triglyceride, or fat, with other long-chain acids and as an ester of a fatty alcohol. Accelerated alcohol metabolism has been observed in regular drinkers. HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT uses stearates tested to U.S. Pharmacopeia monograph standards; known as pharmaceutical grade, the highest purity. Alkaloids are organic compounds from plants, some of which have psychoactive effects. Do not exceed 1 pill in 48 hours. After confirming the model validity, the responses of EbM extracts or its vehicle were obtained by adding agents cumulatively to the organ bath. (2010, 2011) evaluated the effects of an ethanol extract obtained from muira puama and identified promnesic (improving memory), anti-amnesic, and acetyl-cholinesterase inhibition properties in laboratory animals (mice) treated orally with the extract. Thats because it has compounds called terpenoids. Epimedium Brevicornum Base Ingredient in HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT Materials and Methods. HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT uses palm oil that already contains significant amounts of saturated fats providing abundant stearic acid. Therefore, 5HTOL has been regarded as a marker of recent alcohol consumption. When he travels for work, specifically when attending booking conferences for music tours with his business partner, he's constantly turning down offers for drinks. Getting hard might be difficult Yep, "whiskey dick" is a thing. However, further six purified extracts from EP-20, namely, EP-20-1 to EP-20-6, presented little relaxation effect as did the extracts of EPW or EP-20 (Figure 1b). It has been reported that nonginkgolide nonflavonoid fraction of ginkgo biloba extract (NGE) has the most potent relaxing effect on vascular smooth muscle. In our study, we found the increasedL-arginine level at least plays a non-negligible role for the action of EP-20, even though there was no functional evidence demonstrated. Soak a dry cloth in the mixture and soak the stain for five minutes. ; Bayliss, R.I.; and Whitfield, A.G. Pnax Ginseng The herbal remedies referred to as "ginseng" are derived from the roots of several plants. In addition, our study assessed the in vitro antioxidant activities of plants from the Brazilian Amazon (Byrsonima japurensis, Calycophyllum spruceanum, Maytenus guyanensis, Passiflora nitida and muira puama Ptychopetalum olacoides. A typical supplement with stearate excipients may have less than a tenth of one percent of a typical persons daily consumption of dietary stearates. The Food and Drug Administration (FDA) recommends that you dont consume more than 10-30 grams, or 2% of your daily food intake (500-1,500 grams), of silica per day. These results indicate that polyphenols-induced vasorelaxation may also be sustained by smooth muscle PDE inhibition by anthocyanins present in red wines and grapes. Upon ingestion, magnesium stearate is dissolved into magnesium ion and stearic and palmitic acids. Alcohol abusers exhibit a higher IgA response to acetaldehydemodified proteins. 1999). This last characteristic is what separates wines and beers from the so-called hard liquors of the world: vodka, whiskey, rum, scotch, and many more. In fact, the fact that EbM extract potentiates the commonly used PDE 5 inhibitors, such as sildenafil and vardenafil, provides a new treatment strategy for those patients with ED. Types of Hard Liquor. Construction of CRCs was based on degrees of relaxation of the PE-induced contractions. As it was difficult to use rat CC strips in organ bath for mechanism studies, rabbit's CC strips were commonly used as described previously. The results showed that there was a shift to left of the constructed CRC in EP-20 as compared to SNP alone (. Why should you spray your bed with alcohol? After overnight fasting with free access to water, rabbits were anesthetized and killed with overdoses of intramuscular injection of ketamine hydrochloride (50mg/kg) and xylazine (5mg/kg). Taking Panax ginseng by mouth seems to improve sexual function in adults with ED. Always talk to your doctor before taking any male enhancements. Therefore, our study is aimed to test the hypothesis that EbM extracts relaxed the corpus cavernosum (CC) smooth muscle through activation of multitargets on nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling pathway. Three possibilities concerning the potency of crude extract being stronger than that of pure compounds have been proposed. Whitehead, T.P. However, only PE-20 at a concentration of 0.3mg/ml increased the duration of EFS-induced relaxation in PE-precontracted CC strips (Figure 5b). For this reason, we concluded that muira puama extracts could be a potential treatment for Alzheimers disease in humans. When detected by a wavelength of 270nm, the retention time of icariin was 36.628min. There is consensus that sildenalfil, tadalafil and vardenafil are promising oral agents for the treatment of patients with erectile dysfunction (ED). no tachyphylaxis, desensitization or tissue fatigue was observed). The extracts were filtered through layers of gauze, the residues discarded and the filtrates were kept at 20C, followed by lyophilizing the samples. (n>12). To identify the involvement of NO/cGMP pathway in the relaxation actions of EbM extracts, the CC strips were incubated with a potent NO syntheses inhibitor, The CC strips were pretreated with EP-20 in the presence or absence of ODQ (3 10, All the changes in CC strips tension in response to drugs are expressed on percentile basis and all the values are expressed as means.e.m. Figueir, et al. Hard Steel is specially designed to work with alcohol. Although CDTect shows less sensitivity for females than for males (Allen et al. ASTM 1008 is the catch-all, beginning grade, and has from 0 to 0.10 percent carbon. EP-20 enhanced relaxation effect with SNP in rabbit CC strips. In this study, we have clearly demonstrated that EbM extract causes concentration-dependent relaxation of isolated rabbit CC smooth muscle through multitargets related toL-arginine/NO/cGMP pathway including the presence of Alcohol. Approved by the FDA; Protective coatings to preserve the efficacy of pharmaceuticals over time TiO2is an essential component of the protective coatings of pharmaceuticals, enhancing their safety, efficacy, and quality for long periods. Relaxation responses are expressed as percentage of residualtension of contraction originally induced by PE (1M). "These are people I want to impress and I want to like me. Adding a flow agent such as vegetable magnesium stearate is imperative for ensuring a homogenous blend of ingredients and a consistent dosage in each and every HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT capsule. The well-known pure compounds extracted from EbM are the glycosides icariin and noricariin.11, EbM was purchased from a local wholesale distributor (Taipei, Taiwan, Republic of China). After overnight fasting with free access to water, rabbits were anesthetized and killed with overdoses of intramuscular injection of ketamine hydrochloride (50mg/kg) and xylazine (5mg/kg). 1992; Patel et al. Boil 2 cups (.5 liters) of water, and carefully pour the water into the flask while it is still hot. This small tree is native to the Amazonian region of South America and has a plethora of uses among the indigenous peoples of the Amazon, including the following: adaptogen, alopecia (applied topically for hair loss), anorexia, antinociceptive, antioxidant, aphrodisiac, ataxia, beri beri (vitamin deficiency disease), debility, digestive problems, dysentery, fatigue, impotence, neurasthenia, nerve tonic, rheumatism, stimulant, tonic (adding bark decoction to a bath), paralysis, and tremors (Quattrocchi, 2012; Piato et al., 2010; Berdonces, 2009; Duke et al., 2009; Taylor, 2005; Rutter, 1990). Data points represent means.e.m. Hydrolyzed collagen contains 19 amino acids, predominantlyglycine(Gly) 2634%,proline(Pro) 1018%, andhydroxyproline(Hyp) 715%, which together represent around 50% of the total amino acid content. The EbM extracts were prepared as follows. Woollen, J.W., and Turner, P. Plasma Nacetylglucosaminidase and glucuronidase in health and disease. It was, however, more effective than many of them as well. Secondly, there are some active factors other than well-known pure compounds in the crude extract. Oliveira et al. Stearates are nutrients that represent a natural part of every type of fat, whether animal or vegetable, and are typically consumed in amounts of several thousand milligrams per day from common food sources. The percentage of relaxation in sildenafil, vardenafil and tadalafil alone group was 23.74.8, 26.04.9 and 16.16.6%, respectively; whereas that of each combination groups was 36.24.9, 40.73.4 and 34.22.7%, respectively (Figure 6a). Although CDTect shows less sensitivity for females than for males (Allen et al. All the changes in CC strips tension in response to drugs are expressed on percentile basis and all the values are expressed as means.e.m. and cokes. The well-known pure compounds extracted from EbM are the glycosides icariin and noricariin.11,12Icariin (molecular weight=626.65Da) as an internal standard was purchased from Nacalai Tesque (Kyoto, Japan). The ability to have erections is reliant on the complex interaction among neurological, psychological and vascular responses. Isolated rabbit CC strips were mounted in organ baths and isometric tension was recorded in the presence or absence of specific inhibitors related to NO/cGMP signaling such asL-NG-nitro-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo-[4,3-a] quinoxalin-1-one (ODQ, a guanylyl cyclase inhibitor) or phosphodiesterase 5 (PDE 5) inhibitors. when you need it to work while you are drunk. I did consume a couple of drinks with this Relaxation responses are expressed as percentage of residual tension of contraction originally induced by PE (1M). The CC strips were pretreated with EP-20 in the presence or absence of ODQ (3 105M) or SNP (1 106M) or sildenafil (1 107M) for 20min, followed by freezing the CC strips into liquid nitrogen. tract. 1999b). These disruptions can change mood and behavior, and make it harder to think clearly and move with coordination . Must be 18 to purchase. To characterize these impurities, we conducted six (6) unique analytical techniques for the isolation and characterization of process- and product-related impurities: Identification:Mass spectrometry(ESI-Triple Quad, MALDI-TOF, Q-TOF),NMR(1D and 2D), FTIR, Raman and UV-Vis spectroscopy, N-terminal Edman sequencing, LC-MS/MSde novosequencing, peptide mapping, disulfide bond mapping, amino acid analysis for the following: Gelatin, FD&C Black & White, Titanium Dioxide, Cellulose, Magnesium Stearate, Silica, Pnax Ginseng Pnax Ginseng, Gingko Biloba, Mvira Puama, Catuba Bark and Epimedium Brevicornum Derivative. ; Hall, J.H. Presence of proline restricts the conformation. It lasted about 20 minutes to an hour, but When dry and converted into powdery form as in the case of HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT, gelatin total inclusions consist of 9899% protein, but it is not a nutritionally complete protein since it is missingtryptophanand is deficient inisoleucine,threonine, andmethionine. To our knowledge, this is the first study to provide evidence concerning the herbal extracts on PDE 5 inhibitors for proerectile functions. Water extract (a, ), methanol extract (a, ) and EP-20 (a,) derived from water extract were prepared as described in methods. Hard Steel Original Formulated Male Enhancement statemens have not been evaluated by the Food and Drug Administration. Clinical studies show that SA is elevated in alcoholic subjects as compared with social drinkers, demonstrating sensitivity and specificity values, respectively, of 58 percent and 96 percent for women and 48 percent and 81 percent for men (Sillanaukee et al. There is no credible concern that the comparatively tiny amounts in dietary supplements may inhibit absorption of nutrients. 1995; Arndt et al. This component utilizes cookies, and can only be shown if they are accepted. That gives you your headache. After 9 days of abstinence, levels of whole bloodassociated acetaldehyde (WBAA) declined to control levels. Effect of EP-20 on electrical field-stimulated CC strips. Humans cannot digest cellulose, butit is important in the diet as fiber. Selfreport screening scales, on the other hand, generally use a diagnosis of alcohol dependence as the criterion against which they are evaluated. Cellulose Cellulose is a naturally Approved FDA compound. EVERNEW Titanium Alcohol Stove. The main active components of Panax ginseng are ginsenosides, which have been shown to have a variety of beneficial effects, including anti-inflammatory, antioxidant, and anticancer effects. Our renowned labs have more than 20 years existence as a stand-alone facility dedicated to USP , USP and USP testing and covering raw materials through finished products. This common thickening and gelling agent is an animal-based product that contains high levels of protein. They can thus add credibility to research dealing with alcohol treatment efficacy and can provide our clinicians with an additional source of objective information. Use a damp alcohol-soaked microfiber towel to clean any stainless steel in your home. First, it acts as a solvent, which means it eats away the bug's . (n>12). While the packaging is professional, they almost always are! Magnesium is absorbed primarily in the small intestine, and to a lesser extent, in the colon. Firstly, the activity of pure compound is not prominent as that of crude extract. If you are aware of any counterfeiter's, we advise you contact us immediately. 1992; Reintgen et al. Possible acids include muriatic (hydrochloric) acid (HCL), nitric acid (HNO3), or sulfuric acid (H2SO4). The relaxation responses of CC strips were validated by cumulative doses of sodium nitroprusside (SNP). ; and Thompson, A.E. In addition, it found those who took the extract had fewer signs of dementia than those who didnt. 1983), hypertension (Mansell et al. Put on the cap and give the flask a good shake for about a minute, then dump the water out. In conclusion, we can safely determine that Hard Steel Original Formulated Male Enhancement is affective while social, moderate and heavy drinkers consume alcohol. Although silicon is a natural part of some foods and sees wide commercial application, the U.S. Food and Drug Administration has set out strict regulations around using silica as an additive. Magnesium stearate is a combination of stearic acid and the essential mineral magnesium. Three common alloy steels of different carbon content were se-lected for this work: SAE 8620, 4140, and 5160. Total SA and/or lipidassociated SA levels are elevated in patients suffering from tumors, inflammatory conditions, diabetes, and cardiovascular diseases (Sillanaukee et al. This is important for gelation properties of gelatin. Process-related impurities include those that are derived from the manufacturing process (e.g., cell substrates, cell culture media components or downstream processing). For much of its history, moonshine was produced in secret to avoid high taxes or outright bans on alcoholic drinks. In HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT, Silica is included to improve Digestive issues by increasing the absorption of the product total formulation. The parts are then cleaned and heated in a furnace in an atmosphere of dissociated Ammonia (containing N and H) for 10 to 40 hours at 500-625 C (932 - 1157 F). Serum and urine activities of hex are increased in alcoholics and in healthy volunteers drinking > 60 g/d for at least 10 days (Hultberg et al. Your stomach . The pharmacology of the counteractive drugs has been investigated clinically and a combination of 50mg sildenafil with 5mg dihydro-ergotamine has been prescribed under the therapeutic concept to increase blood influx and or to decrease blood efflux in patients with ED. 1999. Alcohol consumption can alter the metabolism of serotonin by inducing a shift toward the formation of 5HTOL. ; Hall, J.H. plastic capsule container. 1998). The results showed that both EPW and EP-20 relaxed the PE-precontracted CC strips in a concentration-dependent manner. The carbon content of a steel is a major factor influencing the amount and appearance of phases and co n-stituents. With chronic ethanol exposure, and in a nonenzymatic reaction, acetaldehyde can form stable adducts with a number of compounds, including proteins such as albumin and hemoglobin (Collins 1988; Goldberg and Kapur 1994; Niemel 1999). 1987a); it is also an indicator of rejection after kidney transplantation (Wellwood et al. The high performance liquid chromatography (HPLC) system consisted of a chromatographic pump (PM-80, Bioanalytical System, West Lafayette, IN, USA), an injector (Rheodyne 7125, Cotati, CA, USA) equipped with a sample loop and an ultraviolet detector (Varian, Walnut Creek, CA, USA). Titanium Dioxide Single most sustaining ingredient preserving HARD STEEL ORIGINAL FORMULATED MALE ENHANCEMENT 2-Year Shelf Life. Steel is a solid solution of iron and carbon, plus other significant alloys. Hard Steel Original Formulated Male Enhancement, Clinical Analysis Reporting for: Hard Steel Original Formulated Male Enhancement Supercore Products Group, Inc. (SPG). Only a doctor can give you medical advice. The results showed that EP-20 enriched the content ofL-arginine in the process of purification and relaxed the CC smooth muscle precontracted with phenylephrine (PE, 1M) in a concentration-dependent manner. Then, a 0.2g frozen tissue was homogenized with 1ml 5% trichloroacetic acid (w/v), followed by determination of cGMP levels by commercially available EIA kits.16.