scholarly journals The Methanolic Extract fromMurraya koenigiiL. Inhibits Glutamate-Induced Pain and Involves ATP-Sensitive K+Channel as Antinociceptive Mechanism

2016 ◽  
Vol 2016 ◽  
pp. 1-6
Author(s):  
Nushrat Sharmin Ani ◽  
Sudip Chakraborty ◽  
Md. Moniruzzaman
Keyword(s):  
Acetic Acid ◽  
Potassium Channels ◽  
Methanolic Extract ◽  
Scientific Basis ◽  
K Channel ◽  
Dependent Manner ◽  
Glutamatergic System ◽  
Antinociceptive Effects ◽  
Wide Range ◽  
Antinociceptive Mechanisms

Murraya koenigiiL. is a perennial shrub, belonging to the family Rutaceae. Traditionally, the leaves of this plant are extensively used in treatment of a wide range of diseases and disorders including pain and inflammation. Although researchers have revealed the antinociceptive effects of this plant’s leaves during past few years, the mechanisms underlying these effects are still unknown. Therefore, the present study evaluated some antinociceptive mechanisms of the methanolic extract ofM. koenigii(MEMK) leaves along with its antinociceptive potential using several animal models. The antinociceptive effects of MEMK were evaluated using formalin-induced licking and acetic acid-induced writhing tests at the doses of 50, 100, and 200 mg/kg. In addition, we also justified the possible participations of glutamatergic system and ATP-sensitive potassium channels in the observed activities. Our results demonstrated that MEMK significantly (p<0.01) inhibited the pain thresholds induced by formalin and acetic acid in a dose-dependent manner. MEMK also significantly (p<0.01) suppressed glutamate-induced pain. Moreover, pretreatment with glibenclamide (an ATP-sensitive potassium channel blocker) at 10 mg/kg significantly (p<0.05) reversed the MEMK-mediated antinociception. These revealed that MEMK might have the potential to interact with glutamatergic system and the ATP-sensitive potassium channels to exhibit its antinociceptive activities. Therefore, our results strongly support the antinociceptive effects ofM. koenigiileaves and provide scientific basis of their analgesic uses in the traditional medicine.

scholarly journals Antinociceptive and Anti-Inflammatory Activities ofBridelia retusaMethanolic Fruit Extract in Experimental Animals

2014 ◽  
Vol 2014 ◽  
pp. 1-12 ◽  
Author(s):  
Tekeshwar Kumar ◽  
Vishal Jain
Keyword(s):  
Acetic Acid ◽  
Animal Models ◽  
Methanolic Extract ◽  
Significant Inhibition ◽  
Dependent Manner ◽  
Paw Oedema ◽  
Antinociceptive Effects ◽  
Anti Inflammatory ◽  
Wbc Count ◽  
Dose Dependent

Antinociceptive and anti-inflammatory potentials of methanolic extract ofBridelia retusafruit (BRME) were evaluated against different animal models in rodents. Antinociceptive effects of BRME were assessed in mice using the acetic acid-induced writhing and formalin test. Anti-inflammatory effects of BRME in three different doses, namely, 100, 200, and 400 mg/kg, were evaluated by utilizing different animal models representing various changes associated with inflammation, namely, carrageenan-induced paw oedema, histamine and serotonin-induced paw oedema, arachidonic acid-induced paw oedema, formalin-induced paw oedema, TPA-induced ear oedema, acetic acid-induced vascular permeability, total WBC count in paw fluid, and myeloperoxidase assay. Also BRME was phytochemically evaluated using chromatographic method. The BRME did not exhibit any signs of toxicity up to a dose of 2000 mg/kg. The extract showed statistical significant inhibition of induced nociception and inflammation in dose dependent manner. The higher dose of extract significantly inhibited pain and inflammation against control (P<0.001). HPLC results revealed the presence of gallic acid and ellagic acid as phytoconstituents in BRME and it was proven as anti-inflammatory agents. The present study scientifically demonstrated the antinociceptive and anti-inflammatory potential of fruit ofB. retusamethanolic extract. These effects may be attributed to the presence of polyphenolic phytoconstituents in the extract.

scholarly journals Antinociceptive Activities of the Methanolic Extract of the Stem Bark ofBoswellia dalzieliiHutch. (Burseraceae) in Rats Are NO/cGMP/ATP-Sensitive-K+Channel Activation Dependent

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Marius Mbiantcha ◽  
Alain Ngouonpe Wembe ◽  
Amadou Dawe ◽  
William Yousseu Nana ◽  
Gilbert Ateufack
Keyword(s):  
Methylene Blue ◽  
Neuropathic Pain ◽  
Methanolic Extract ◽  
Stem Bark ◽  
Opioid Antagonist ◽  
K Channel ◽  
Therapeutic Effects ◽  
Pain Models ◽  
Antinociceptive Effects ◽  
Oxide Synthase

Boswellia dalzielii (B. dalzielii)is traditionally used in the treatment of rheumatism, pain, and inflammation. The present investigation evaluates the property and possible mechanism of action of the methanolic extract ofB. dalzielii(BDME) on inflammatory and neuropathic pain models. Effects of BDME (250 and 500 mg/kg), orally administered, were verified in mechanical hypernociception induced by LPS or PGE2. Mechanical hyperalgesia, cold allodynia, and heat hyperalgesia were used in vincristine-induced neuropathic pain. NW-nitro-L-arginine methyl ester (inhibitor of nitric oxide synthase), glibenclamide (ATP-sensitive potassium channel blocker), methylene blue (cGMP blocker), or naloxone (opioid antagonist receptor) has been used to evaluate the therapeutic effects of BDME on PGE2-induced hyperalgesia. Chemical profile of BDME was determined by using HPLC-XESI-PDA/MS. BDME showed significant antinociceptive effects in inflammatory pain caused by LPS and PGE2. The extract also significantly inhibited neuropathic pain induced by vincristine. The antinociceptive property of BDME in PGE2model was significantly blocked by L-NAME, glibenclamide, methylene blue, or naloxone. The present work reveals the antinociceptive activities of BDME both in inflammatory and in neuropathic models of pain. This plant extract may be acting firstly by binding to opioid receptors and secondly by activating the NO/cGMP/ATP-sensitive-K+channel pathway.

scholarly journals Antinociceptive and Antioxidant Activities of Methanolic Extract of Leaves of Azadirachta Indica (Neem)

2019 ◽  
Author(s):  
Akash S Mali ◽  
Mahesh B Thorat ◽  
Dhairyasheel M. Ghadge ◽  
Kumodini A. Nikam ◽  
Shraddha Sawant ◽  
...  
Keyword(s):  
Methylene Blue ◽  
Acetic Acid ◽  
Azadirachta Indica ◽  
Methanolic Extract ◽  
Antinociceptive Activity ◽  
Opioid System ◽  
K Channel ◽  
Hot Plate ◽  
Nitric Oxide Radical ◽  
Tail Immersion

AbstractGraphical abstractBackgroundAzadirachta indica (Neem) is a communal plant of Meliaceae family called Neem Or Kadunimb in Maharashtra, India Neem stated anti-inflammatory through regulation of proinflammatory enzyme activities with COX and LOX enzyme. Previous studies show that Azadirachta Indica (neem) and its chief constituents play essential role in anticancer management via the modulation of different molecular pathways including NF-κB, p53, PI3K/Akt, Bcl-2, pTEN and VEGF. Many parts of the plant are traditionally used in the treatment of various pharmacological action, the analgesic activity of Neem Seed Oil has already reported but Neem Leaves.MethodsThe antinociceptive activity of Azadirachta Indica Leaves (AZIL) was examined using heat-induced-mechanical (hot-plate and tail-immersion test) and chemical-induced (acetic acid, formalin, glutamic acid, cinnamaldehyde) nociception models in mice at 50,100, and 200 mg/kg doses. ATP-sensitive K+ channel pathway, cyclic guanosine monophosphate (cGMP) pathway and involvement of opioid system was also tested using glibenclamide, methylene blue and naloxone/morphine respectively. The methanolic extract of leaves of A.Indica was assessed by using different in vitro antioxidant models of screening like scavenging of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, and hydroxyl radical.ResultsAZIL showed antinociceptive activity and antioxidant activity. In both hot plate and tail immersion tests AZIL significantly increases the latency to the thermal stimuli. In acetic acid-induced writhing test the extract repressed the number of abdominal writhing. Similarly, AZIL produced substantial dose-dependent inhibition of paw licking in both neurogenic and inflammatory pain induced by intraplanar injection of formalin. As well, AZIL also expressively withdrawn cinnamaldehyde-induced pain and the glutamate-induced pain in mice. It was also proved that pretreatment with naloxone significantly reversed the antinociception produced by AZIL in mechanical tests signifying the involvement of opioid system in its effect. Furthermore, administration of methylene blue, enhanced AZIL induced antinociception while glibenclamide, an ATP-sensitive K+ channel antagonist, could not converse antinociceptive activity induced by AZIL.ConclusionBased on the results of the present study it can be said that AZIL keeps significant antinociceptive activity which acts in both central and peripheral mechanisms.Ethnopharmacological relevanceAzadiracta Indica having family Melliases is one of the common medicinal plant in United states and south Asia including India, Pakistan, Bangladesh. Various parts of the plants used in treatment of many inflammatory conditions, skin diseases, tooth protection, antidiabetic in the form of oil and herbal porridge.

scholarly journals Anti-inflammatory and antinociceptive activities of methanolic extract from red seaweed Dichotomaria obtusata

2013 ◽  
Vol 49 (1) ◽  
pp. 65-74 ◽  
Author(s):  
Neivys García Delgado ◽  
Ana Iris Frías Vázquez ◽  
Hiran Cabrera Sánchez ◽  
Roberto Menéndez Soto del Valle ◽  
Yusvel Sierra Gómez ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Methanolic Extract ◽  
Therapeutic Potential ◽  
Intraperitoneal Administration ◽  
Moderate Activity ◽  
Dependent Manner ◽  
Ear Edema ◽  
Antinociceptive Effects ◽  
Anti Inflammatory ◽  
Phospholipase A2 Activity

The aim of the present work was to investigate the anti-inflammatory and antinociceptive effects of methanolic extract from D. obtusata using classic models in mice (croton oil-induced ear edema and acetic acid-induced writhing) and a phospholipase A2 activity test. Qualitative analysis of the chemical composition of seaweed was also determined by extraction with solvents of increasing polarity and precipitation and color tests. Results of qualitative chemical study showed the presence of lactonic and phenolic compounds, reduced carbohydrates, other sugars, flavonoids, fatty compounds, triterpenes and steroids. The extract inhibited mouse ear edema in a dose-dependent manner with an efficacy higher than 90% and a mean effective dose of 4.87µg/ear, while intraperitoneal administration presented a moderate activity. The extract did not inhibit phospholipase A2 activity. In the writhing test, the intraperitoneal administration of the extract showed a strong antinociceptive activity (80.2%), while the oral route showed a lower efficacy. In conclusion, this study demonstrated the anti-inflammatory and antinociceptive effects of methanol extract of D. obtusata in experimental models, suggesting its therapeutic potential in the treatment of peripheral painful and/or inflammatory pathologies.

scholarly journals Antinociceptive and Antioxidant Activities of Methanolic Extract of Leaves of Azadirachta Indica (Neem).

2019 ◽  
Author(s):  
Akash Mali ◽  
Mahesh B Thorat ◽  
Dhairyasheel M. Ghadge ◽  
Kumodini A. Nikam ◽  
Shraddha D. Sawant ◽  
...  
Keyword(s):  
Methylene Blue ◽  
Acetic Acid ◽  
Azadirachta Indica ◽  
Methanolic Extract ◽  
Antinociceptive Activity ◽  
Opioid System ◽  
K Channel ◽  
Hot Plate ◽  
Nitric Oxide Radical ◽  
Tail Immersion

Background: Azadirachta indica (Neem) is a communal plant of Meliaceae family called Neem Or Kadunimb in Maharashtra, India Neem stated anti-inflammatory through regulation of proinflammatory enzyme activities with COX and LOX enzyme. Previous studies show that Azadirachta Indica (neem) and its chief constituents play an essential role in anticancer management via the modulation of different molecular pathways including NF-&kappa;B, p53, PI3K/Akt, Bcl-2, pTEN, and VEGF. Many parts of the plant are traditionally used in the treatment of various pharmacological action, the analgesic activity of Neem Seed Oil has already reported but Neem Leaves. Methods: The antinociceptive activity of Azadirachta Indica Leaves (AZIL) was examined using heat-induced-mechanical (hot-plate and tail-immersion test) and chemical-induced (acetic acid, formalin, glutamic acid, cinnamaldehyde) nociception models in mice at 50,100, and 200 mg/kg doses. ATP-sensitive K+ channel pathway, cyclic guanosine monophosphate (cGMP) pathway and involvement of the opioid system were also tested using glibenclamide, methylene blue, and naloxone/morphine respectively. The methanolic extract of leaves of A.Indica was assessed by using different in vitro antioxidant models of screening like scavenging of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, and hydroxyl radical. Results: AZIL showed antinociceptive activity and antioxidant activity. In both hot plate and tail immersion tests AZIL significantly increases the latency to the thermal stimuli. In the acetic acid-induced writhing test the extract repressed the number of abdominal writhing. Similarly, AZIL produced substantial dose-dependent inhibition of paw licking in both neurogenic and inflammatory pain induced by intraplantar injection of formalin. As well, AZIL also expressively withdrawn cinnamaldehyde-induced pain and the glutamate-induced pain in mice. It was also proved that pretreatment with naloxone significantly reversed the antinociception produced by AZIL in mechanical tests signifying the involvement of the opioid system in its effect. Furthermore, administration of methylene blue, enhanced AZIL induced antinociception while glibenclamide, an ATP-sensitive K+ channel antagonist, could not converse antinociceptive activity induced by AZIL. Conclusion: Based on the results of the present study it can be said that AZIL keeps significant antinociceptive activity which acts in both central and peripheral mechanisms.

scholarly journals Antinociceptive and antioxidant activities of methanolic extract of leaves of Azadirachta indica in vivo

2019 ◽  
Author(s):  
Akash S Mali ◽  
Mahesh B Thorat ◽  
Dhairysheel M Ghadge ◽  
Kumodini A Nikam ◽  
Shraddha D Sawant ◽  
...  
Keyword(s):  
Methylene Blue ◽  
Acetic Acid ◽  
Azadirachta Indica ◽  
Methanolic Extract ◽  
Antinociceptive Activity ◽  
Opioid System ◽  
K Channel ◽  
Hot Plate ◽  
Nitric Oxide Radical ◽  
Tail Immersion

Background: Azadirachta indica (Neem) is a communal plant of Meliaceae family called Neem Or Kadunimb in Maharashtra, India Neem stated anti-inflammatory through regulation of proinflammatory enzyme activities with COX and LOX enzyme. Previous studies show that Azadirachta Indica (neem) and its chief constituents play essential role in anticancer management via the modulation of different molecular pathways including NF- κ B, p53, PI3K/Akt, Bcl-2, pTEN and VEGF. Many parts of the plant are traditionally used in the treatment of various pharmacological action, the analgesic activity of Neem Seed Oil has already reported but Neem Leaves. Methods: The antinociceptive activity of Azadirachta Indica Leaves (AZIL) was examined using heat-induced-mechanical (hot-plate and tail-immersion test) and chemical-induced (acetic acid, formalin, glutamic acid, cinnamaldehyde) nociception models in mice at 50,100, and 200 mg/kg doses. ATP-sensitive K+ channel pathway, cyclic guanosine monophosphate (cGMP) pathway and involvement of opioid system was also tested using glibenclamide, methylene blue and naloxone/morphine respectively. The methanolic extract of leaves of A.Indica was assessed by using different in vitro antioxidant models of screening like scavenging of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, and hydroxyl radical. Results: AZIL showed antinociceptive activity and antioxidant activity. In both hot plate and tail immersion tests AZIL significantly increases the latency to the thermal stimuli. In acetic acid-induced writhing test the extract repressed the number of abdominal writhing. Similarly, AZIL produced substantial dose-dependent inhibition of paw licking in both neurogenic and inflammatory pain induced by intraplanar injection of formalin. As well, AZIL also expressively withdrawn cinnamaldehyde-induced pain and the glutamate-induced pain in mice. It was also proved that pretreatment with naloxone significantly reversed the antinociception produced by AZIL in mechanical tests signifying the involvement of opioid system in its effect. Furthermore, administration of methylene blue, enhanced AZIL induced antinociception while glibenclamide, an ATP-sensitive K+ channel antagonist, could not converse antinociceptive activity induced by AZIL. Conclusion: Based on the results of the present study it can be said that AZIL keeps significant antinociceptive activity which acts in both central and peripheral mechanisms.

scholarly journals The Role of K+, Ca2+ channels and Endothelial Hyperpolarizing Factors in Vasorelaxation Induced by Tribulus terrestris

2021 ◽  
Vol 2 (1) ◽  
pp. 94-100
Author(s):  
Thamer M. Bashir ◽  
Omar A.M. Al-Habib
Keyword(s):  
Methanolic Extract ◽  
K Channel ◽  
Tribulus Terrestris ◽  
Channel Blockers ◽  
Dependent Manner ◽  
The Novel ◽  
Concentration Dependent Manner ◽  
Relaxant Effect ◽  
Calcium Ion Channels

The present study focused on the relaxant effect of themethanolic extract (ME) of Tribulus terristris on rats’ thoracic aortae and included the study of underlying vasorelaxation mechanisms. The methanolic extract produced concentration-dependent relaxation in rats’ aorta. The methanolic extract produced concentration-dependent relaxation in the aortic rings. The use of different K+ channel blockers (BaCl2, 4-AP, GLIB, and TEA) indicated that Kv, KATP, KIR, and KCa and L-type Ca channels played no role in the methanolic extractinduced relaxation. However, with respect to endothelium-derived hyperpolarizing factors, PGI2 and sGC produced a mild inhibition in the relaxation response to ME while NO produced no effect at all. Based on the novel results of the current study, it can be concluded that T. terrestris methanolic extract (ME) mediated relaxation in isolated rat aortic tissues in a concentration-dependent manner. Moreover, we discovered that ME-mediated relaxation is endothelium-dependent and that potassium and calcium ion channels play no role in this relaxation with a limited role of PGI2 and sGC.

scholarly journals Analgesic Activity of Methanolic Extract of Tubers of Arisaema tortuosum (Wall.) Schott. in Swiss Albino Mice

2018 ◽  
Vol 17 (1) ◽  
pp. 37-41
Author(s):  
Priyanka Chakraborty ◽  
Nripendra Nath Bala ◽  
Sudipta Das
Keyword(s):  
Acetic Acid ◽  
Analgesic Activity ◽  
Methanolic Extract ◽  
Dependent Manner ◽  
Hot Plate ◽  
Plate Method ◽  
Swiss Albino Mice ◽  
Chemically Induced ◽  
Pain Reaction ◽  
Albino Mice

The aim of the the present study was to investigate the analgesic activity of methanolic extract of Arisaema tortuosum (MEAT) using acetic acid-induced writhing and hot plate methods. The hot plate method is useful in elucidating centrally mediated antinociceptive responses, while acetic acid-induced writhing is the chemically induced pain of peripheral origin. The MEAT was used at doses of 50, 100, 200 and 400 mg/kg body weight on swiss albino mice. The percentage inhibition of the abdominal constriction reflex increased dose dependently in case of acetic acid-induced pain and in the hot plate method model the extract at the dose of 400 mg/kg significantly increased the pain reaction time (PRT). These studies conclude that A. tortuosum (Wall.) Schott. tuber possesses analgesic activity in a dose dependent manner. In case of acetic acid-induced pain, the extract at the dose of 400 mg/kg body wt. showed 41.19% inhibition of writhing reflex. In case of hot plate method, after 60 minutes the PRT increased to 7.47 ± 0.05 seconds for the extract at the dose of 400 mg/kg body wt.Dhaka Univ. J. Pharm. Sci. 17(1): 37-41, 2018 (June)

scholarly journals Antinociceptive and antioxidant activities of methanolic extract of leaves of Azadirachta indica in vivo

2019 ◽  
Author(s):  
Akash S Mali ◽  
Mahesh B Thorat ◽  
Dhairysheel M Ghadge ◽  
Kumodini A Nikam ◽  
Shraddha D Sawant ◽  
...  
Keyword(s):  
Methylene Blue ◽  
Acetic Acid ◽  
Azadirachta Indica ◽  
Methanolic Extract ◽  
Antinociceptive Activity ◽  
Opioid System ◽  
K Channel ◽  
Hot Plate ◽  
Nitric Oxide Radical ◽  
Tail Immersion

Background: Azadirachta indica (Neem) is a communal plant of Meliaceae family called Neem Or Kadunimb in Maharashtra, India Neem stated anti-inflammatory through regulation of proinflammatory enzyme activities with COX and LOX enzyme. Previous studies show that Azadirachta Indica (neem) and its chief constituents play essential role in anticancer management via the modulation of different molecular pathways including NF- κ B, p53, PI3K/Akt, Bcl-2, pTEN and VEGF. Many parts of the plant are traditionally used in the treatment of various pharmacological action, the analgesic activity of Neem Seed Oil has already reported but Neem Leaves. Methods: The antinociceptive activity of Azadirachta Indica Leaves (AZIL) was examined using heat-induced-mechanical (hot-plate and tail-immersion test) and chemical-induced (acetic acid, formalin, glutamic acid, cinnamaldehyde) nociception models in mice at 50,100, and 200 mg/kg doses. ATP-sensitive K+ channel pathway, cyclic guanosine monophosphate (cGMP) pathway and involvement of opioid system was also tested using glibenclamide, methylene blue and naloxone/morphine respectively. The methanolic extract of leaves of A.Indica was assessed by using different in vitro antioxidant models of screening like scavenging of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, and hydroxyl radical. Results: AZIL showed antinociceptive activity and antioxidant activity. In both hot plate and tail immersion tests AZIL significantly increases the latency to the thermal stimuli. In acetic acid-induced writhing test the extract repressed the number of abdominal writhing. Similarly, AZIL produced substantial dose-dependent inhibition of paw licking in both neurogenic and inflammatory pain induced by intraplanar injection of formalin. As well, AZIL also expressively withdrawn cinnamaldehyde-induced pain and the glutamate-induced pain in mice. It was also proved that pretreatment with naloxone significantly reversed the antinociception produced by AZIL in mechanical tests signifying the involvement of opioid system in its effect. Furthermore, administration of methylene blue, enhanced AZIL induced antinociception while glibenclamide, an ATP-sensitive K+ channel antagonist, could not converse antinociceptive activity induced by AZIL. Conclusion: Based on the results of the present study it can be said that AZIL keeps significant antinociceptive activity which acts in both central and peripheral mechanisms.

scholarly journals Antispasmodic activity of Symplocos paniculata is mediated through opening of ATP-dependent K+ channel

2016 ◽  
Vol 11 (2) ◽  
pp. 495 ◽  
Author(s):  
Khalid Hussain Janbaz ◽  
Saba Akram ◽  
Fatima Saqib ◽  
Mamoona Khalid
Keyword(s):  
Methanolic Extract ◽  
Therapeutic Potential ◽  
Scientific Basis ◽  
K Channel ◽  
Antispasmodic Activity ◽  
Relaxant Effect ◽  
Rabbit Jejunum ◽  
Low K

<p class="Abstract"><em>Symplocos paniculata</em> is a medicinal plant used by native healers to manage gastrointestinal ailments. The crude methanolic extract of <em>S. paniculata</em> was screened pharmacologically both in vitro and in vivo for the validation of its therapeutic potential. It suppressed the spontaneous activity of isolated rabbit jejunum preparations and also caused inhibition of the low K<sup>+ </sup>(20 mM)- induced spastic contractions in isolated rabbit jejunum preparations in a manner comparable to cromakalim. The relaxant effect was found to be blocked following glibenclamide exposure of the isolated tissue preparations similar to cromakalim, suggesting that observed response was likely to be mediated through opening of ATP dependent K<sup>+ </sup>channels. Following oral administration to mice provided protection against castor oil-induced diarrhea in a manner similar to loperamide. The plant material was found safe in toxicity study up to oral dose of 8 g/kg in mice.  Hence, present study provides a scientific basis for the vernacular use of <em>S. paniculata</em> in gastro-intestinal system.</p><p> </p>

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