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RESEARCH ARTICLE Usharani Chatragadda et.al / IJIPSR / 2 (7), 2014, 1321-1336

Department of Pharmacology ISSN (online) 2347-2154

Available online: www.ijipsr.com July Issue 1321

V PHARMACOLOGICAL EVALUATION ON GLUCOSE

LOWERING EFFICAY OF LEAVES OF PRUNUS PERSICA

1Usharani Chatragadda*,

2Nori Kodandaram,

3Vimochana Bowjanku

Department of pharmacology, Hindu College of Pharmacy, Guntur- 522002, Andhrapradesh,

INDIA

Corresponding Author:

CH. UshaRani

Department of pharmacology

Hindu College of Pharmacy, Guntur- 522002

Andhrapradesh, INDIA

Email: [email protected]

Phone: +91 9293105722

International Journal of Innovative

Pharmaceutical Sciences and Research www.ijipsr.com

Abstract

The aim of the present study was to evaluate the glucose lowering efficacy of methanolic and aqueous

extracts of leaves of Prunus persica using oral glucose loaded normoglycemic rat model, alloxan induced

diabetic rat model and inhibition of intestinal glucose absorption by everted gut sac model. Diabetes mellitus

[DM] is a group of heterogeneous disorders in which carbohydrate metabolism is reduced while that of

proteins and lipids are increased. Hyperglycaemia is a common end point for all types of DM and is an

important parameter to evaluate the efficacy of antidiabetic drugs. The results shows that in case of everted

gut sac model, both methanolic and aqueous extracts showed significant results at p<0.01 and p<0.001

respectively compared to control. In oral glucose loaded normoglycemic rat model administration of test

drug at different doses and at different dosing intervals were administered to evaluate the glucose lowering

efficacy. Acarbose was used as refrence standard in all models. The results revealed that there is suppression

of postprandial spike initially and later maintenance of blood glucose levels were noticed. In case of alloxan

induced diabetic rat model blood glucose levels were estimated on 7th and 10

th day after alloxan induction.

The results showed that the percentage reduction of blood glucose for methanolic extract 500mg/kg on 7th

day and 10th day was 49.58 and 69.27 respectively. In case of Aqueous extract the percentage reduction of

blood glucose on 7th and 10

th day was 42.83 and 55.26. Hence the present study clearly demonstrated the

antihyperglycemic activity of leaves of Prunus persica

Keywords: Prunus persica, Diabetes mellitus, Acarbose, Hyperglycemia, Alloxan.

mailto:[email protected]




INTRODUCTION

Diabetes mellitus [DM] is a group of heterogeneous disorders in which carbohydrate metabolism

is reduced while that of proteins and lipids are increased. Hyperglycaemia is a common end point

for all types of DM and is an important parameter to evaluate the efficacy of antidiabetic drugs.

The hallmarks of DM are three “polys”: an excessive urine production (polyuria), an excessive

thrist (polydipsia) and an excessive eating (polyphagia) [1].

Herbal drugs play a major role of all the officially recognized systems of Health in India such as

Ayurveda, Yoga, Unani, Siddha, Homeopathy and Naturopathy, except Allopathy. The World

Health Organization (WHO) estimates that 80 percent of the population of some Asian and

African countries presently use herbal medicine for some aspect of primary health care [2].

In diabetes, some herbal alternatives are proven to provide symptomatic relief and assist in the

prevention of the secondary complications of the disease. Some herbs have also been proven to

help in the regeneration of ß-cells and in overcoming resistance. In addition to maintaining

normal blood sugar level, some herbs are also reported to possess antioxidant activity and

cholesterol-lowering action [3].

Around 75 per cent of people have type 2 diabetes mellitus. The reasons for this are poor

nutrition, rising stress levels, an improper metabolism and lack of fitness [4]. In the United States

25.8 million people or 8.3% of the population have diabetes. Of these, 7.0 million have

undiagnosed diabetes. In 2010, about 1.9 million new cases of diabetes were diagnosed in

population over 20 years. It is said that if this trend continues, 1 in 3 Americans would be

diabetic by 2050 [5].

The peach, Prunus persica, is a deciduous tree, native to North-West China, in the region

comprised between the Tarim basin and the north slopes of the Kunlun Shan mountains, where it

was first domesticated and cultivated [6] It belongs to the Rosaceae family. It is highly useful in

treating inflammatory disorders [7]. Peach leaves are used in the treatment of constipation

[8]

Peach leaves possess good antioxidant properties [9].

Antioxidants are used as supportive therapy in the treatment of DM [[10] and hypoglycemic

plants have been shown to regulate the oxidative complications of DM [11] Many species of

prunus posses antidiabetic properties Prunus davidiana, Prunus amygdalus [12,13] Hence the

present study was carried out to evaluate the antidiabetic activity of Prunus persica.

http://en.wikipedia.org/wiki/World_Health_Organization

http://en.wikipedia.org/wiki/World_Health_Organization

http://timesofindia.speakingtree.in/topics/emotions/stress

http://en.wikipedia.org/wiki/Deciduous

http://en.wikipedia.org/wiki/Tree

http://en.wikipedia.org/wiki/Native_plant




MATERIALS AND METHODS

PLANT MATERIAL

Plant material used in this study consisted of the leaves of Prunus persica, collected and

authentified by Prof. V. Chelladurai, Ph.D., Research officer – Botany, Tirunelveli. A specimen

was deposited in the Hindu College of Pharmacy, Guntur.

PREPARATION OF EXTRACT

Preparation of methanolic extract:

The dried leaf powder was packed into soxhlet column and extracted successively with methanol

at a temperature not exceeding the boiling point of the solvent.

The extract was filtered using Whattman filter paper (No.1) and then concentrated in vacuum at

40 ℃ using Rotary evaporator.

The dried extract was labelled as Prunus persica methanolic leaf extract (MEPP) and preserved

in refrigerator for further use.

Preparation of aqueous extract:

The dried powdered material of leaves of Prunus persica , and then boiled in distilled water for

60min.

The decoction was taken and allowed to cool for 30min at room temperature (24 ± 5° C). This

decoction was filtered twice and the filtrate was then concentrated in vacuum at 60 ℃ using

Rotary evaporator.

The dried extract was labelled as Prunus persica aqueous leaf extract (AEPP) and preserved in

refrigerator for further use.

ANIMALS

Wistar male albino rats (150-180 g) were selected for the present study.

The animals had free access to standard rat pellet, with water supplied ad libitum under strict

hygienic conditions.

The experimental protocol was approved by IAEC (Institutional Animal Ethics Committee) of

HCOP (Hindu college of pharmacy).

The study followed all the rules of (CPCSEA) Committee for the Purpose of Control and

Supervision of Experiments on Animals. The protocol number is HCOP/IAEC/2013-14/02.




ANTIDIABETIC ACTIVITY

EXPERIMENTAL DESIGN

S.No Model Parameters Species, No. Of animals

and groups

1. Exvivo – Rat everted

sac model

Intestinal glucose

absorbtion

Wistar albino rats, 4 groups,

6 sacs at each interval

2.

Invivo – oral glucose

loaded normoglycemic

rat model

Blood glucose at

different intervals


6 animals in each group

3. Invivo – Alloxan

induced diabetic model Blood glucose


6 animals in each group

1. Effect of methanolic extract of prunus persica (MEPP) & aqueous extract of prunus

persica (AEPP) on intestinal glucose absorption by everted sac model

Overnight fasted rats were killed by light ether. The small intestine was washed out carefully

with tyrode solution using a syringe equipped with blunt end. The midportion of the small

intestine from each animal was used in order to minimize the transport variability of the

segments. Intestinal segments (5±0.2 cm) were then everted. The empty sacs was filled with

0.5ml of distlled water, then the sacs were placed inside the beakers containing 60 ml of the

tyrode solution which contain respective concentrations of test samples and maintained at a

temperature of 37±2 ◦C. The external medium was continuously bubbled with a gas mixture of

95% oxygen and 5% carbon dioxide during the whole experimental period [14] The prepared

sacs were placed in different groups of drug solutions

At respective time points (30 min, 60 min) the sacs were removed from the beakers and the

serosal fluid was drained through a small incision into a test tube.At each time interval 6 sacs are

prepared inorder to minimize the error and the average values are taken.

2. Effect of methanolic extract of prunus persica (MEPP) on blood glucose levels in oral

glucose loaded normoglycemic rat model

Fasted rats were divided into five groups of six animals each. Group I served as control and

received 3g/kg orally glucose, group II served as standard and received acarbose 50mg/kg orally

Group Dose

I control

II Acarbose 2mg/ml

III MEPP 2mg/ml

IV AEPP 2 mg/ml




30 min prior to glucose load, group III received MEPP 500 mg/kg orally at 0min glucose load,

group IV received MEPP 1g/kg orally 30 min prior to glucose load, group V received MEPP 1

g/kg orally 60 min prior to glucose load. Blood glucose was determined by small cut at the tip of

the tail with surgical blade and drop of blood was placed in glucometer to read the value at

repective time intervals of 0min,30 min, 60 min, 90min and 120 min respectively.

3. Effect of aqueous extract of prunus persica (AEPP) on blood glucose levels in oral

glucose loaded rats

Fasted rats were divided into four groups of six animals each. Group I served as control and

received 3g/kg oral glucose, group II served as standard and received acarbose 50mg/kg orally

30 min prior to glucose load, group III received AEPP 500 mg/kg orally at 0min glucose load,

group IV received AEPP 1 g/kg orally 60 min prior to glucose load. Blood glucose was

determined by small cut at the tip of the tail with surgical blade and drop of blood was placed in

glucometer to read the value at repective time intervals of 0min,30 min, 60 min, 90min and 120

min respectively.

4. Methanolic and aqueous extracts of prunus persica at different doses & at different

dosing intervals.

Fasted rats were divided into six groups of six animals each. Group I served as control and

received 3g/kg glucose, group II served as standard and received acarbose 50mg/kg 30 min prior

to glucose load, group III received MEPP 500 mg/kg orally at 0min glucose load, group IV

received AEPP 1 g/kg orally at 0 min glucose load, group V received MEPP 1g/kg orally 60 min

prior to glucose load and group VI received AEPP 1g/kg orally 60 min prior to glucose load.

Blood glucose was determined by small cut at the tip of the tail with surgical blade and drop of

blood was placed in glucometer to read the value at repective time intervals of 0min,30 min, 60

min, 90min and 120 min respectively

5. Effect of methanolic and aqueous extracts of prunus persica on blood glucose levels in

alloxan induced diabetic rat model.

Hyperglycemia was induced by single intraperitonial injection of 150 mg/kg body weight alloxan

monohydrate freshly dissolved in normal saline before use to overnight fasted rats. After 48 hrs

check the blood glucose levels of each rat and select those rats which show blood glucose more

than 300 mg/dl and were used for the experiment. The diabetic rats were divided into 4 groups




each containing 5 animals each. Group I served as control and received water daily for 10 days,

group II received acarbose 50 mg/kg orally daily twice a day and served as control, group III

received MEPP 500 mg/kg orally daily twice a day and group IV received AEPP 500 mg/kg

orally daily twice a day for 10 days. On 7th

day and on 10th

day the blood glucose was checked

with rite check glucometer respectively.

STATISTICAL ANALYSIS

The data expressed as the mean± SD. Data were analyzed by one-way ANOVA followed by

using Dunnetts T test. Instat® (Graph Pad software, U.S.A).

RESULTS

1. Effect of methanolic extract of prunus persica (MEPP) & aqueous extract of prunus

persica (AEPP) on intestinal glucose absorption by everted sac model

This study showed that the MEPP at a concentration of 2mg/ml inhibit the intestinal glucose

absorption. Acarbose was used as reference standard. For MEPP at 30 min and 60 min interval

the % of glucose absorption was 26.13 and 46.09 respectively. Compared to control the

methanolic extract showed significant results. Compared to methanolic extract aqueous extract

showed better results. Both methanolic and aqueous extracts showed significant results at p<0.01

and p<0.001 respectively compared to control.

Table: 1 Effect of methanolic extract of prunus persica (MEPP) & aqueous extract of

prunus persica (AEPP) on intestinal glucose absorption by everted sac model

n=6, All values are Mean ± sd ‘a’ significant at p<0.01 and ‘b’ significant at p<0.001 compared

to control

2. Effect of methanolic extract of Prunus persica (MEPP) on blood glucose levels in oral

glucose loaded normoglycemic rat model

The present study showed at 15min after glucose administration the peak of the blood glucose

level increased rapidly from the fasting value in case of the control group. MEPP 500mg/kg at

% of glucose absorption

30 min 60 min

Control 79.47±2.83 96.60±5.99

Acarbose 2mg/ml 37.48±2.73a 46.24±3.43

a

MEPP 2mg/ml 26.13±2.70a 46.09±1.05

AEPP 2mg/ml 21.01±2.83b 41.92±2.06

b




0min glucose load showed that there is significant decrease in the postprandial spike at 30min

and there is steady maintenance of blood level at 90 and 120min was observed. Suppression of

blood glucose for acarbose at 15 min was observed. In the case of MEPP 500mg/kg at 0min

glucose load there is significant reduction in postprandial spike at 30 min was observed. In the

case of MEPP 1g/kg 30min prior to glucose load significant reduction of postprandial spike is

observed at 15min compared to control. Significant p<0.001 MEPP 1g/kg at 30 min prior to

glucose load was observed compared to control group at 15 min time interval after glucose load.

Significant p<0.001 MEPP 1g/kg at 30 min prior to glucose load glucose load was observed

compared to acarbose.

Table 2: ffect of methanolic extract of prunus persica (MEPP) on blood glucose levels in

oral glucose loaded normoglycemic rat model

Values are mean ±sd .,n = 6 ‘a’ significant p<0.001 compared to control ‘b’ significant p<0.001

compared to acarbose

Group 0 min 15 min 30 min 60 min 90 min 120 min

control 107±

1.41

190.66±

4.17

133.5±

2.73

118±

3.03

96.33±

2.80

84.5±

1.76

Acarbose (50

mg/kg) 30 min

prior to glucose

load

74.5±

1.87

118±

3.28a

135.66±

5.39

140.83±

5.94

141.16±4

.16

143.16±

4.21

MEPP (500

mg/kg) with

glucose load (O

min)

97.33±

1.96

172.5±

2.25

114.16±

1.72a

92.33±

3.26

102.16±3

.43

100.5±

2.88

MEPP (1g/kg)-

30mins prior to

glucose load

84.16±

1.47

100.33±

1.63a,b

131.5±

2.07

142.66±

2.94

133.66±1

.21

140.5±

1.76

MEPP (1g/kg) -

60 min prior to

glucose load

85.83±

2.23

115.5±

2.25a

137.83±

3.43

142.33±

0.81

143.5±

3.01

105.16±

2.78




Effect of aqueous extract of prunus persica (AEPP) on blood glucose levels in oral glucose

loaded rats

The present study showed that the blood glucose levels in case of the control group reached

maxium peak value at 15 min time interval after glucose administration. AEPP 500mg/kg at

0min glucose load showed that there is significant decrease in the postprandial spike at 60min

and there is steady maintenance of blood level at 90 and 120min was observed. In case of AEPP

1g/kg 60min prior to glucose load suppression of glucose spike is observed at 15min time

interval after glucose load and steady maintenance of blood glucose was observed at 60 and

90min time interval after glucose load. The significant p<0.0001 for AEEP 1g/kg 60min prior to

glucose load compare to control was observed.

Table 3: Effect of queous extract of prunus persica (AEPP) on blood glucose levels in oral

glucose loaded rats.

Values are means ±sd., N = 6 ‘a’ significant p<0.001compared to acarbose and ‘b’ significant

p<0.0001 compared to control

Group 0 min 15 min 30 min 60 min 90 min 120

min

Control

107±

1.41

190.66±

4.17

133.5±

2.73

118±

3.03

96.33±

2.80

84.5±

1.76

Acarbose (50

mg/kg) 30

min prior to

glucose load

74.5±

1.87

118±

3.28b

135.66±

5.39

140.83±

5.94

141.16±

4.16

143.16±

4.21

AEPP (500

mg/kg) at 0

min glucose

load

105.5±

2.34

181.16±

1.47

137.33±

2.42

119±

2.82a

121±

2.28

117±

3.48

AEPP 1g/kg

60min prior to

glucose load

87.66±

2.42

125.83±

4.49b

144.16±

2.29

146.16±

2.92

147±

2.96

115.16±

1.32




4. Methanolic and aqueous extracts at different doses & at different dosing intervals.

In this present study methanolic and aqueous extracts were administered at different doses

and at different dosing intervals and the results revealed that the MEPP at 500mg/kg at omin

glucose load suppressed the postprandial spike at 30min time interval after the glucose load

but AEPP 500 mg/kg at 0min glucose load suppressed the postprandial spike at 60min time

interval after the glucose load. MEPP 1g/kg 60min prior to glucose load and AEPP 1g/kg

60min prior to glucose load suppressed the glucose spike at 15min time interval after the

glucose load. Compared to AEPP, MEPP showed better results. The significant p< 0.001 for

MEPP 1g/kg 60min prior to glucose load and AEPP 1g/kg 60 min prior to glucose load

compared to MEPP 500mg/kg at 0min glucose load was observed. The significant p<

0.0001 for MEPP 1g/kg 60min prior to glucose load and AEPP 1g/kg 60 min prior to

glucose load compared to control group was observed

Table 4: Effect of Methanolic and aqueous extracts at different doses & at different

dosing intervals.

Group 0 min 15 min 30 min 60 min 90 min 120 min

Control 107±

1.41

190.66±

4.17

133.5±

2.73

118±

3.03

96.33±

2.80

84.5±

1.76

Acarbose (50

mg/kg) 30 min

prior to glucose

load

74.5±

1.87

118±

3.28b

135.66±

5.39

140.83±

5.94

141.16±

4.16

143.16±

4.21

MEPP (500

mg/kg) at 0 min

glucose load

97.33±

1.96

172.5±

2.25

114.16±

1.72

92.33±

3.26

102.16±

3.43

100.5±

2.88

AEPP (500

mg/kg) at 0 min

glucose load

105.5±

2.34

181.16±

1.47

137.33±

2.42

119±

2.82

121±

2.28

117±

3.48

MEPP (1g/kg) -

60 min prior to

glucose load

85.83±

2.23

115.5±

2.25ab

137.83±

3.43

142.33±

0.81

143.5±

3.01

105.16±

2.78

AEPP 1g/kg 60

min prior to

glucose load

87.66±

2.42

125.83±

4.49ab

144.16±

2.29

146.16±

2.92

147±

2.96

115.16±

1.32

Values are means ±sd .,N = 6 ‘a’ significant p<0.001 compared to MEPP 500 mg/kg at 0 min

glucose load ‘b’ significant p<0.0001 compared to control




5. Effect of methanolic and aqueous extracts of Prunus persica on blood glucose levels in

alloxan induced diabetic rat model.

The present study showed that there is no decrease in the blood glucose levels in the case of the

control group and there is maintenance of steady blood glucose level was observed. Acarbose

was used used as reference standard. Blood glucose levels were estimated on 7th

and 10th

day

after alloxan induction. The results showed that the percentage reduction of blood glucose for

MEPP 500mg/kg on 7th

day and 10th

day was 49.58 and 69.27 respectively. In case of AEPP the

percentage reduction of blood glucose on 7th

and 10th

day was 42.83 and 55.26 respectively.

Compared to aqueous extract methanolic extract showed better results. The significant p<0.001

for MEPP 500mg/kg and AEPP 500mg/kg compared to control on 7th

day and significant

p<0.0001 for MEPP 500mg/kg and AEPP 500mg/kg compared to control on 10th

day.

Table 5: Effect of methanolic and aqueous extracts of Prunus persica on blood glucose

levels in alloxan induced diabetic rat model.

Values are means±sd N=6 566., ‘a’ significant at p< 0.001 compared to control at 7th

day and ‘b’

significant at p<0.0001 compared to control at 10th

day

Fig. 1: Effect of Methanolic Extract of Prunus Persica (MEPP) & Aqueous Extract of

Prunus Persica (AEPP) On Intestinal Glucose Absorption By Everted Sac Model

Group 0th

day on 7th

day on 10th

day

Control 592.83±4.62 592.83±4.62 589.83±4.92

Acarbose 50

mg/kg 518.16±8.54 274.5±11.11

a 164±11.74

b

MEPP 500

mg/kg 555±10.82 279.83±9.68

a 170.5±10.21

b

AEPP 500 mg/kg .16±11.49 410.67±11.31 309.12±11.03a




Fig. 2: Effect of Methanolic Extract of Prunus Persica (MEPP) On Blood Glucose Levels In

Oral Glucose Loaded Normoglycemic Rat Model

Fig. 3: Effect of Aqueous Extract Of Prunus Persica (AEPP) On Blood Glucose Levels In

Oral Glucose Loaded Rats

Fig. 4: Methanolic and aqueous extracts at different doses & at different dosing intervals




Fig. 5: Effect of methanolic and aqueous extracts of prunus persica on blood glucose levels

in alloxan induced diabetic rat model.

DISCUSSION

Diabetes mellitus is a metabolic disorder of multiple aetiologies that is characterized by chronic

hyperglycaemia with disturbed carbohydrate, fat and protein metabolism [15]. Many diverse

therapeutic strategies for the treatment of Type 2 diabetes are in use. The conventional available

therapies for diabetes include stimulation of endogenous insulin secretion, enhancement of the

action of insulin at the target tissues, oral hypoglycemic agents, such as biguanids and

sulfonylureas and the inhibition of degradation of dietary starch by glycosidases such as α-

amylase and α-glucosidase [16]. The results of the preliminary phytochemical screening of the

test extracts of leaves of Prunus persica revealed the presence of alkaloids, tannins, flavonoids,

sapponons and glycosides. In the present study Prunus persica leaf extract was evaluated for its

role in glucose transport across the rat everted gut sacs exvivo. It might possible that the active

compounds of Prunus persica leaf extract decreased the blood glucose levels by inhibiting the

absorption of glucose from the alimentary tract. Therefore it is most probable that active

phytochemicals in the leaves of Prunus persica prevented the glucose transit across the gut

membrane by reducing the glucose transporter protein activity which might have lead to wash of

glucose from the body. The inhibition of intestinal glucose uptake might be due to the presence

of the phytochemical constituent namely naringenin [17] GLUT 2, a transmembrane carrier

protein of GLUT family, located in basolateral membrane of small intestine, is very efficient

carrier of glucose at small intestine level [18].




Encouraging data are available concerning effects of flavonoids on inhibiting GLUT 2 and their

hypoglycemic effect,Kwon et al[19], Song et al[20], Johnston et al[21]. Although our findings

are promising, uncertainties remain. Some investigators suggested that flavonoids decreased

glucose uptake by a sodium-dependent pathway via the sodium-dependent glucose transporter 1

SGLT1.

The presence of flavonoids in the leaves of Prunus persica might inhibiting GLUT2, SGLT1

transporters and reducing the glucose absorption across the intestine. In the present study the

effect of methanolic (MEPP) and aqueous extracts (AEPP) of Prunus persica on blood glucose

levels in oral glucose loaded normoglycemic rat model at different doses and different dosing

intervals were investigated and the results suggests that the incretin hormones are released during

meals from gut endocrine cells.

They potentiate glucose-induced insulin secretion and may be responsible for up to 70% of

postprandial insulin secretion. The incretin hormones include glucagon-like peptide-1 (GLP-1)

and glucose-dependent insulin tropic polypeptide (GIP), contribute to insulin secretion from the

beginning of a meal and their effects are progressively amplified as plasma glucose

concentrations rise [22]

But the results shows that there is significant decrease in the postprandial spike initially and there

is steady maintenance of blood glucose level is observed indicating that the test extract might

interfering with glucose mediated insulin stimulation and delay in the luminal carbohydrate

metabolism might be the reason for maintenance of steady straight line.

Flavonoids are promising alternative for diabetes and its associated complications. However,

effective control of not only keeping the blood glucose level optimum will provide better results

but compromising continued damage of human islets as well as stabilizing the cellular

components is more essential for effective diabetic management [23].

Experimental studies conducted in alloxan induced diabetic rats revealed that orally administered

methanolic and aqueous extracts of Prunus persica leaves at doses of 500mg/kg twice daily

showed antidiabetic activity. The reductions in serum glucose concentration by Prunus persica

may be due to one or a combination of different mechanisms, including modulation of digestion,

glucose absorbtion, insulin sensitivity, or insulin secretion




CONCLUSION

In the present study, Prunus persica leaves showed significant antidiabetic activity in dose

dependent manner. The leaves of Prunus persica may be a new natural source for the treatment

of diabetes. When compared to the conventional drugs the herbal plant extracts shows the same

effect in a normal physiological way and with least side effects. Further research is needed to

find the exact mechanism of action and the chemical constuitents that are responsible for the

antidiabetic activity.

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http://www.sciencedirect.com/science/journal/22517294

http://www.sciencedirect.com/science/journal/22517294

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