KR101282055B1 - Anti diabetic fermented food comprising fermented noni and method of manufacturing the same - Google Patents

Abstract

The present invention relates to a fermented food using a noni fermented product and a method for producing the same, and more particularly, to a method for preparing an anti-diabetic fermented food comprising inoculating fermented with Chungkookjang powder on a noni medium. Antidiabetic Noni fermented food according to the present invention can provide antidiabetic effect through the following and multi-disease symptoms, intake of blood sugar, glycated glycated pigment and blood lipids, improved insulin sensitivity.

Description

Anti-diabetic fermented foods using noni fermented products as an active ingredient and a method of manufacturing the same {ANTI DIABETIC FERMENTED FOOD COMPRISING FERMENTED NONI AND METHOD OF MANUFACTURING THE SAME}

The present invention relates to an anti-diabetic fermented food comprising the noni fermented product and a method for producing the same.

Korean dietary culture is becoming more westernized, and metabolic syndrome is increasing. One of its representative diseases is diabetes. Diabetes is a disorder of the pituitary gland and hepatic brain, which is responsible for the control of diuretic function, and because of the fact that it does not produce the required amount of insulin in the body, or because insulin does not function properly in the cell, it does not absorb enough sugar into the body, resulting in high blood sugar It is a disease. Type 2 diabetes mellitus is insulin-independent diabetes. Insulin resistance does not occur due to insulin resistance or is caused by a relatively lack of insulin. 90% of all diabetics belong to type 2 diabetes. Diabetes mellitus can cause various complications such as hyperinsulinemia, hypertension, and atherosclerosis because glucose uptake in the body does not normally occur, causing glucose metabolism, lipid metabolism, and protein metabolism. Diabetes mellitus is a series of diseases with various causes and etiologies characterized by the occurrence of acute and chronic complications due to such metabolic abnormalities, and thus, there is no development of a drug that can be fundamentally treated.

In order to prevent or treat the above diabetes, insulin or oral hypoglycemic agents are administered. However, because of the economic burden and the risk of side effects, in recent years, diet and natural therapy using pharmacological substances such as natural plants are used. This is being tried. Natural products reported to be effective in treating diabetes include moxibustion extracts (Korean Patent No. 464815), Apanaic acid derivative compounds isolated from Jangung Beetle Mushrooms (Korean Patent No. 457690), and extracts of hawthorn trees (Korean Patent No. 441787). , Polysaccharides isolated from Korean Angelica gigas (International Publication No. 2001-60386), bile of ruminant animals (US Patent No. 6645355) and the like.

On the other hand, Noni is known to have an anti-cancer effect and high blood pressure, such as the present inventors have been found to be able to produce a more effective anti-diabetic composition by fermenting Cheonggukjang powder under Noni medium.

An object of the present invention is to provide a non-diabetic health functional food by preparing a noni fermented product prepared by inoculating Chonggukjang powder in noni medium.

In one embodiment, there is provided a method for producing an anti-diabetic fermented food comprising the step of inoculating the fermented Chungkukjang powder in Noni medium, wherein the cheonggukjang powder of the embodiment is 1 part by weight relative to the total weight ratio It provides a method for producing a fermented food, the step of fermenting the embodiment provides a method for producing an anti-diabetic fermented food, characterized in that the fermentation at a temperature of 20 to 30 ℃, before the step of fermentation of the embodiment It provides a method for producing an anti-diabetic fermented food, characterized in that it is pre-fermented at a temperature of 75 to 90 ℃, after the step of fermentation of the above embodiment anti-diabetic, characterized in that the fermentation at a temperature of 75 to 90 ℃ It provides a method for preparing a fermented food, the 75 to 90 ℃ of prior to the step of fermentation of the embodiment Pre-fermentation in the Figure, and after the fermentation step provides a method for producing an anti-diabetic fermented food, characterized in that the post-fermentation at a temperature of 75 to 90 ℃, the step of fermenting the embodiment of the relative humidity 55 to 65 It provides a method for producing an anti-diabetic fermented food, characterized in that the fermentation under% conditions, the step of fermentation of the embodiment provides a method for producing an anti-diabetic fermented food, characterized in that it is carried out for 40 days.

In one embodiment, there is provided an anti-diabetic fermented food comprising a noni fermented product inoculated with Nogyeojangjang powder in Noni medium, wherein the chonggukjang powder of the embodiment provides an anti-diabetic fermented food, characterized in that 1 parts by weight relative to the total weight ratio. do.

In one embodiment, there is provided an anti-diabetic fermentation composition comprising a noni fermented product inoculated with Chungkookjang powder in noni medium, and provides an anti-diabetic pharmaceutical composition comprising a noni fermentation inoculated with nongyo medium powder.

In the present invention, a feed supplemented with 0.4% noni fermented product to KKAy / TaJcl, which is genetically induced type 2 diabetes, was supplied as a diet and analyzed by analyzing feed and drinking water intake, blood sugar, glycated hemoglobin, insulin content, and blood lipid concentration. Diabetic activity was verified. In particular, HbA1c (Hemoglobin A1c), a glycated hemoglobin pigment value, is a glycemic control state that occurs during the red blood cell survival period of about 3 months, and thus has a glycemic control state for about 3 months. Often appears above%. In addition, as a significant indicator of the progression of diabetes, every 2% increase in HbA1c increases the risk of developing retinopathy, one of the complications of diabetes, and decreases β cell function. Therefore, glycated hemoglobin measurement was used as a major indicator of antidiabetic activity. In addition, the anti-diabetic activity of noni fermentation was measured to determine the antidiabetic activity and the mechanism of action by measuring glucose uptake and PPAR-gamma transcriptional activity.

Noni fermented food according to the present invention can provide an antidiabetic effect by reducing the following and polylactic symptoms upon ingestion, reducing blood sugar levels, glycated blood pigments and blood lipids, and improving insulin sensitivity.

1 is a graph showing the amount of blood glucose change in mice according to the diet.
Figure 2 is a graph showing the glycated blood pigment value of the mouse according to the diet (green tea fermented product).
Figure 3 is a graph showing the glycated hemoglobin pigment value of the mouse according to the diet (rice bran fermentation).
Figure 4 is a graph showing the blood glucose concentration of the mouse according to the diet.
5 is a graph showing the insulin concentration of the mouse according to the diet.
Figure 6 is a graph measuring the Homeostasis assessment-IR of the mouse according to the diet.
7 is a graph measuring the Quicki of the mouse according to the diet.
8 is a graph showing PPAR-gamma transcriptional activity.
9 is a graph showing 2-NBDG absorption.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

Example

Example  1. Noni Fermented  Produce

Cheonggukjang powder (Binmax Korea, Muan) was added to Noni (Korea, Seoul, Ilsan Sanghoe) powder at 1% ratio, and 40% of the total weight was supplied, followed by primary fermentation at 82.5 ± 2.5 ℃ for 4 hours. It was. Since the main fermentation was carried out for 40 days at a temperature of 25 ℃, 60% humidity. Dried at 82.5 ± 2.5 ° C for 2 hours, and then hot-air dried at 87.5 ± 2.5 ° C for 2 hours to prepare a final product having a moisture content of 3% or less.

Example  2. Anti-diabetic  Animal experiment

2-1. Diet of Laboratory Animals

KKA ^y / TaJcl (5 weeks old, male), a type 2 diabetes model, was purchased from G-BIO and purified for 1 week, and then classified into 4 groups of 7 by egg mass according to body weight. The control group was fed AIN-93G FORMULA general feed, and the positive control group, noni raw material, and noni fermented foods were fed to the diet by adding 0.4% of each sample to the general feed. At this time, Banaba extract, a commercial anti-diabetic dietary supplement, was used as a positive control group. The feeding room temperature was 23 ± 2 ℃, the humidity was 50 ± 5%, and the contrast period was set at 12 hour intervals.

2-2. Dietary efficiency

The body weight was measured at a constant time every 10 days, and the dietary intake and water intake were measured as the difference between the intake and the remaining amount, and the feed efficiency ratio (FER) was calculated by dividing the weight intake by the dietary intake.

In terms of weight change, fermented noni had the highest weight change rate, similar to the control and positive controls, and the noni intake group showed the lowest weight change rate. The food intake was significantly lower in the positive and fermented Noni intake groups than in the control group, and the drinking water intake was significantly lower in the fermented Noni and positive controls than in the control group. I could see. This symptom relief was especially noticeable in fermented noni intake group. In the case of FER calculated based on dietary intake and weight change, fermented noni, positive control, control, and noni intake group were higher. When diabetes occurs, dietary efficiency is known to be lowered due to degenerative changes in the body's metabolism. In the non-fermented group, the dietary efficiency is significantly higher than that of the control group, and the symptoms of diabetes are alleviated like the diet and drinking water intake results. It is showing.

Initial weight (g) Final weight (g) Weight change
(g / day) Intake
(g / day) Dietary efficiency (%)
Drinking Water Intake
(mL / day) Control group 27.29 ± 1.32 45.58 ± 2.35 0.20 + 0.02 7.11 ± 0.26 ^b 2.53 ± 0.24 22.28 ± 2.61 ^a Positive control group 26.61 ± 1.51 42.49 ± 9.16 0.20 ± 0.01 6.57 ± 0.27 ^c 2.94 ± 0.25 16.54 ± 3.50 ^b Noni 27.23 ± 1.17 43.76 ± 4.15 0.19 ± 0.02 7.53 ± 0.60 ^a 2.46 ± 0.26 20.67 ± 3.94 ^ab Noni Ferment 26.48 ± 0.77 45.71 ± 2.64 0.22 ± 0.01 6.85 ± 0.81 ^b 3.08 ± 0.28 13.36 ± 3.61 ^b

2-3. Blood glucose change measurement

Blood glucose was measured using a blood glucose strip (ACCU-CHEK Sensor, Roche, Basel, Switzerland) by taking whole blood from the tail vein at a constant time for 90 days every 10 days without fasting time.

KKA ^y / TaJcl is a type 2 diabetes disease model that introduces the Ay gene, which is associated with obesity, hyperglycemia and hair color, and is known to show more severe obesity and hyperglycemia at an early age (7-8 weeks of age) than KK mice. The blood glucose level starts to appear from about 40 g. In the control group, blood glucose began to increase rapidly from the 20th day when the body weight increased to 40 g or more, and high blood glucose of 400 mg / dL or more was maintained after 30 days. However, in the fermented noni group, the blood sugar level of 211.60-252.20 mg / dL was maintained without increasing blood glucose levels during the 90 days after the end of the experiment from the 10th day when the body weight increased to 30-40 g. appear. In particular, lower blood sugar level was maintained than the positive control group which consumed the anti-diabetic-related dietary supplement. On the other hand, in the noni intake group, blood sugar was significantly decreased compared to the control group after 50 days, but the antidiabetic activity of noni was enhanced through solid fermentation, considering that the fermentation in the noni fermentation group was lower than the control group. 1).

2-4. Glycated blood Color value  Measure

After completion of the experiment, 4 uL of whole blood was collected through the orbital vein of KKA ^y / TaJcl, which was fasted for 16 hours, and then injected into an easy A1c cartridge, which was measured using an easy A1c analyzer (Korea, Seoul, Asan Pharmaceutical).

HbA1c, a part of hemoglobin in erythrocytes, slowly transforms into glycated hemoglobin by non-enzymatic binding to glucose during the survival of blood cells. Since the survival time of the blood cells is about 90-120 days, the measurement of glycated hemoglobin is used as a basic indicator for integrating changes in blood glucose over a long period of time. As a result, the glycated hemoglobin values of the positive control group and the noni intake group were 8.96 ± 0.83 and 9.45 ± 0.32%, respectively, and there was no significant difference from the control group of 9.72 ± 0.95%. It was 7.62 ± 0.81%, which was significantly lower than the control group, indicating that blood glucose control was well controlled during the 90-day experiment. This is consistent with the results of the measurement of the change in blood sugar over 90 days, in which the blood sugar was maintained at a constant level without significant change during the experiment (FIG. 2).

In order to determine the degree of glycemic control ability of fermented noni, the glycated hemoglobin pigment value of mice ingested the rice bran fermented product was added to rice bran (Korean, Seoul, Ilshin Sanghoe) and rice bran instead of nonni.

As a result, the glycated blood pigment value of the mice fed the rice bran fermentation was maintained at a level similar to that of the control group, indicating that there was no glycemic control ability (FIG. 3). That is, in case of rice bran, the antidiabetic activity did not appear even when fermented with Cheonggukjang powder.

2-5. Blood insulin and Glucose  Content measurement

After 16 hours of fasting, the blood collected through the orbital vein of the mouse was centrifuged at 3000 rpm for 10 minutes to separate serum. Serum insulin levels were measured using a mouse insulin ELISA kit (TMB type) manufactured by SHIBAYAGI.

After separating the serum, 5 uL of the supernatant was added, 1 ml of tinder reagent was added, and the mixture was left at room temperature for 18 minutes, and then absorbance was measured at 505 nm to calculate blood glucose concentration using a glucose standard curve.

The results showed that the blood glucose concentrations in the positive control group, noni and noni fermentation group were significantly lower than those in the control group. Especially, the noni fermentation group showed a glucose concentration of 154 mg / dL, which was lower than the control group. 64% decrease. The serum insulin content was highest in the control group at 236.33 uU / mL, and the nonni intake group, the positive control group and the noni fermentation group were significantly lower than the control group. In particular, the insulin concentration of the noni fermentation group was 130.70 uU / mL was detected about 45% lower than the control group. In the control group, hyperglycemia was observed despite high insulin content, indicating that hyperinsulinemia was observed.In the non-fermented group, the blood sugar level remained constant during the experimental period even though the insulin content was lower than that of the control group. Noni fermented product was found to be effective in improving hyperinsulinemia (FIG. 4).

2-6. HOMA - IR  And QUICKI index

Fasting blood glucose and insulin levels were measured for Homeostasis assessment (IR) and Quicki, which are indicators of insulin resistance and sensitivity. HOMA-IR is an index indicating insulin resistance, and the larger the value, the higher the insulin resistance. Quicki is an index indicating insulin sensitivity, and the larger the value, the higher the insulin sensitivity. The positive control group, the noni intake group and the noni fermentation group were all lower in HOMA-IR and higher Quicki than the control group, which decreased insulin resistance and increased sensitivity, especially in the noni fermentation group. In view of this, it is thought that the noni fermented product has a hypoglycemic effect by improving insulin sensitivity and lowering insulin resistance (FIGS. 6 and 7).

2-7. Blood Lipid Level Measurement

After 16 hours of fasting, the blood collected through the orbital vein of the mouse was centrifuged at 3000 rpm for 10 minutes to separate serum and the lipid content was measured. Triglyceride is Triglycerides reagents (Bayer, USA), Total Cholesterol is Cholesterol reagent (Bayer, USA), HDL-Cholesterol is Direct HDL-Cholesterol (Bayer, USA), LDL- Cholesterol (LDL-cholesterol) was measured using an ADVIA 1650 (Bayer, USA) instrument using LDL-Cholesterol (Bayer, USA) Kit.

The most common lipid metabolism abnormality in patients with type 2 diabetes is hyperlipidemia, particularly hypertriglyceridemia, with a frequency of 20-60%, which has a significant effect on the development of atherosclerotic disease, which is the leading cause of death in diabetics. As a result, the triglyceride level of the control group was high as 235.60 ± 46.73 mg / dL, while the other experimental groups showed a low level, especially the noni fermented group was 148.40 ± 17.07 mg / dL. Triglycerides decreased by about 37%. On the other hand, higher serum glucose metabolism caused by hyperglycemia may increase the content of VLDL and LDL in the serum, which may increase the incidence of atherosclerotic coronary artery disease or cerebrovascular disease. By reducing the cholesterol accumulated in the action to suppress the occurrence of atherosclerosis. As a result, total cholesterol was not significantly different among experimental groups. However, HDL-cholesterol was higher in noni and noni fermentation groups than in control group, and LDL-cholesterol was higher in noni fermentation group than in control group. Significantly lower detection. This suggests that the Noni fermented group is effective in improving abnormal lipid metabolism caused by diabetes (Table 2).

Total cholesterol (mg / dL) HDL-
Cholesterol (mg / dL) LDL-
Cholesterol (mg / dL) Triglycerides (mg / dL) Control group 141.25 ± 16.69 116.00 ± 6.12 ^b 9.60 ± 2.06 235.60 ± 46.73 ^a Positive control group 144.80 ± 12.54 119.33 ± 9.48 ^b 9.75 ± 3.90 221.50 ± 31.05 ^a Noni 137.80 ± 13.53 133.40 ± 11.20 ^a 9.40 ± 2.15 ^a 169.00 ± 17.02 ^ab Noni Fermentation 137.00 ± 12.47 130.40 ± 10.13 ^a 7.80 ± 1.17 ^b 148.40 ± 17.07 ^b

Example  3. Noni Fermentation  Extract Anti-diabetic  Active measurement

3-1. Noni Fermentation  Preparation of extract

100 mL of water or 70% ethanol was added to 10 g of noni fermentation, followed by extraction with stirring at 25 ° C. for 24 hours. After centrifugation at 3000 rpm for 10 minutes, the supernatant was collected and filtered with filter paper (whatman No. 2). The filtrate was concentrated under reduced pressure, lyophilized and stored at -20 ° C and used for the experiment.

3-2. PPAR - gamma  Transcriptional activity

PPAR-gamma is a receptor in the cell nucleus of many tissues. When stimulated, these receptors produce enzymes that regulate lipid and glucose metabolism proteins and induce the proliferation of cells. This receptor is important because rosiglitazone, a typical diabetes drug used in clinical practice, has been developed for PPAR-gamma. PPAR-gamma angonist is effective in reducing blood sugar levels in diabetic patients. In the present invention, PPAR-gamma, its cofactor RXR, and binding DNA sequence PPRE were expressed intracellularly for luciferase assay to study transcription factor activity of HEK 293 cells.

As a result of measuring PPAR-gamma activity on water and 70% ethanol extract of Noni fermentation, Noni fermented water extract did not increase PPAR-gamma transcriptional activity. However, Noni fermented ethanol extract was confirmed to increase the transcriptional activity of PPAR-gamma in a concentration-dependent manner (Fig. 8). This suggests that noni fermented products have anti-diabetic activity by acting as a PPAR-gamma angonist, increasing insulin sensitivity and decreasing insulin resistance. In addition, even though the sample used in this experiment is a non-fermented ethanol crude extract, considering that the PPAR-gamma transcriptional activity is increased to a level similar to that of rosiglitazone, an antidiabetic drug, it has sufficient efficacy to be developed as an anti-diabetic health functional food.

3-3. C2C12 Myocyte  After differentiation induction glucose uptake  Measure

The main cause of insulin resistance in insulin-independent diabetes is known to be the reduction of non-oxidative glucose utilization in skeletal muscle. The glucose uptake was measured using C2C12 cells to measure the increase in glucose utilization by noni fermentation.

First, C2C12 myoblast cell lines were incubated for 48 hours in DMEM supplement 10% FBS medium, and then differentiated for 6-8 days until functional myotube in 1% horse serum medium. It was transferred to low glucose medium and starving for 4 hours, and the samples were incubated for 24 hours after treatment by concentration (50, 100, 200, 400 ug / mL). After incubation for 1 hour by adding 50 uM 2- (N- (7-nitrobens-2-oxa-1,3-diazol-4-yl) amino) -2-deoxyglucose, the medium was removed and washed with PBS. After lysis of cells with Lysis buffer, lysate was measured under the condition of EX / EM = 485/535 using a fluorescence micro reader.

As a result, the Noni fermented water extract did not increase glucose uptake, but the Noni fermented 70% ethanol extract was found to increase glucose uptake in a concentration-dependent manner (Fig. 9).

In conclusion, the antidiabetic activity of noni and noni fermentation was measured using KKA ^y / TaJcl, a type 2 diabetes disease model. Noni showed weak antidiabetic activity while noni fermented product showed excellent antidiabetic activity. Noni fermented KKA ^y / TaJcl not only maintained blood glucose of 211.60-252.20 mg / dL during 90 days of breeding, but also had a glycated hemoglobin level of 7.62% and a dietary anti-diabetic dietary supplement. It showed significantly lower blood glucose control ability than the anti-diabetic related dietary supplement foods, which was much lower than the group ingested. The antidiabetic activity of noni fermentation was found to be due to the lower insulin resistance by acting as a PPAR gamma angonist and showed similar efficacy to rosiglitazone, a diabetes treatment. In view of this, noni fermentation is highly commercially available as an anti-diabetic dietary supplement.

Compositions comprising the substances according to the invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories or sterile injectable solutions according to conventional methods. Can be used. More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to commonly used diluents such as water and liquid paraffin . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The extract of the present invention may vary depending on the age, sex, and weight of the patient, but in general, an amount of 0.01 to 500 mg / kg, preferably 0.1 to 100 mg / kg, may be administered once or several times a day. Can be. In addition, the dosage of the extract can be increased or decreased depending on the route of administration, the degree of disease, sex, weight, age and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

While the present invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know. In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Accordingly, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention be construed as including all embodiments falling within the scope of the appended claims.

Claims (14)

Method of producing a fermented food for the prevention and improvement of diabetes comprising the step of inoculating the fermented Chungkookjang powder to noni medium. The method of claim 1,
The Cheonggukjang powder is a method for producing a fermentation food for preventing and improving diabetes, characterized in that 1 part by weight relative to the total weight ratio. The method of claim 1,
The fermentation step is a method for producing a fermentation food for preventing and improving diabetes, characterized in that the fermentation at a temperature of 20 to 30 ℃. The method of claim 3,
Method for producing a fermentation food for the prevention and improvement of diabetes, characterized in that pre-fermentation at a temperature of 75 to 90 ℃ before the step of fermentation. The method of claim 3,
Method for producing a fermentation food for the prevention and improvement of diabetes, characterized in that after fermentation at a temperature of 75 to 90 ℃ after the step of fermentation. The method of claim 3,
Prefermentation at a temperature of 75 to 90 ℃ before the step of fermentation,
Method for producing a fermentation food for the prevention and improvement of diabetes, characterized in that after fermentation at a temperature of 75 to 90 ℃ after the step of fermentation. The method of claim 3,
The fermentation step is a method for producing a fermentation food for the prevention and improvement of diabetes, characterized in that the fermentation under conditions of 55 to 65% relative humidity. The method of claim 3,
The fermentation is a method for producing a fermentation food for the prevention and improvement of diabetes, characterized in that is carried out for 40 days. The method of claim 1,
The diabetes is a method of producing a fermented food for the prevention and improvement of diabetes, characterized in that the type 2 diabetes. Fermented food for the prevention and improvement of diabetes, including the Noni fermented product inoculated with Nogyeong medium powder. The method of claim 10,
The Cheonggukjang powder is a fermented food for the prevention and improvement of diabetes, characterized in that 1 part by weight relative to the total weight ratio. The method of claim 10,
The diabetes is a fermented food for the prevention and improvement of diabetes, characterized in that the type 2 diabetes. Fermented food composition for the prevention and improvement of diabetes, including the noni fermented product inoculated with Chungkookjang powder in noni medium. A pharmaceutical composition for the prevention and treatment of diabetes, comprising a noni fermented product inoculated with Chungkookjang powder on a noni medium.

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