Therapeutic Potential of Citrus medica L. Peel Extract in Carrageenan Induced Inflammatory Pain in Rat

Research Article
Therapeutic Potential of Citrus medica L. Peel Extract in Carrageenan Induced Inflammatory Pain in Rat

S. Sood, S. Bansal, A. Muthuraman, N.S. Gill and M. Bali

Research Journal of Medicinal Plants, 2009, 3(4), 123-133.

Abstract

In this study, we planned to evaluate the antioxidative, anti-inflammatory and analgesic potential of Citrus medica peel extract. Antioxidant activity in different solvent systems was evaluated. Ethyl acetate extract of Citrus medica peel (EtCM) showed maximum 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydrogen peroxide radical scavenging activity in a dose dependent manner as compared to ascorbic acid. Further, anti-inflammatory and analgesic activities of EtCM (200, 300 and 400 mg kg^-1) were studied on carrageenan induced inflammatory pain in rats. Anti-inflammatory activity was assessed by measuring paw volume in rats. Analgesic activity was evaluated for its central and peripheral pharmacological actions by using hot plate, plantar, pin prick and mechanical allodynia tests in rats. EtCM (400 mg kg^-1) produced significant decrease in paw volume and pain as compared to diclofenac. Therefore, the Citrus medica peel extract may be used as a future antioxidant for the treatment of inflammation and pain.

ASCI-ID: 82-64

Halliwell, 1996, 2006). Natural antioxidants exhibit a wide range of pharmacological activities and have been shown to possess analgesic, antiulcer, anticancer, anti-inflammatory and anti-aging properties because oxidative stress produced by accumulation of free radicals is supposed to play an important role in the pathogenesis of these diseases (Mayne, 2003; Pinnell, 2003). More recently, it has become evident that phenolic compounds obtained from natural products may reduce oxidative stress by antioxidant action (Vinson et al., 1998). For example, various flavonoids, which are found naturally in fruits and vegetables have been demonstrated to exert antioxidant effect through a number of different mechanisms. Numerous vegetables, crops, spices and medicinal herbs have been tested in an effort to identify new and potentially useful antioxidants (Zheng and Wang, 2001; Nijveldt et al., 2001).

Citrus fruits and juices have long been recognized to contain secondary metabolites including antioxidant such as ascorbic acid, flavanones, phenolics and pectin that are important to human nutrition. Many researchers have found antioxidant in juice and edible parts of oranges of different origin and from different varieties (Miller and Rice-Evans, 1997; Gorinstein et al., 2001). Citrus processing byproducts represent a rich source of naturally occurring flavonoids. The citrus peels represent roughly half of the fruit mass and their extracts were found to have a good antioxidative potential (Zia-ur-Rehman, 2006; Anagnostopoulou et al., 2006). The peel extract was also found to contain the highest concentration of flavonoids as compared to juice or pulp part. Numerous work on citrus peels of different species, like Citrus sinensis, Citrus limon, Citrus paradisi, Citrus reticulata and Citrus junos particularly dealing with antioxidant effects and pharmacological actions are well documented in previously reported studies (Alicia et al., 2005; Parmar and Kar, 2008; Yi et al., 2008). Our previous studies also reported that Citrus decumana peel extract possessed ameliorative potential in stress induced peptic ulcer in rat (Sood et al., 2009). However, little information is available on the potential therapeutic effect of Citrus medica peels. Therefore, in the present investigation an attempt has been made to explore the anti-inflammatory and analgesic potential of the peel extract of Citrus medica.

MATERIALS AND METHODS

Chemicals
DPPH (1,1-diphenyl-2-picrylhydrazyl), hydrogen peroxide, ascorbic acid, carrageenan and diclofenac sodium were obtained from Sigma Chemical Co. All other chemicals and solvents used were of analytical grade.

Plant Material
The fruits of Citrus medica were collected in the month of January, 2008 from Northern region of India particularly Himachal Pradesh. The plant material was identified and authenticated in the P.G. Department of Horticulture, Khalsa College, Amritsar (Voucher No. HD-1109). The peels were removed manually and dried under shade at room temperature. The dried peels were grounded into a coarse powder in a mixer. The powder was sieved through a 1 mm metal sieve to achieve a standard size of particles. Further extraction process, analgesic and anti-inflammatory studies were carried out in the month of November, 2008.

Animals
Wistar rats of either sex (180-220 g) were obtained from Sanjay Biologicals, Amritsar. They were kept at standard laboratory diet, environmental temperature and humidity. A 12 h light-dark cycle was maintained throughout the experimental protocol. The experimental protocol was duly approved by Institutional Animal Ethics Committee (IAEC) and care of the animals was carried out as per the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environment and Forest, Government of India (Reg No. 874/ac/05/CPCSEA).

Extraction
The finely grounded and dried peel powder was subjected to maceration using non-polar and polar solvents. First maceration was done using hexane followed by chloroform, ethyl acetate and methanol. The extraction with each solvent was carried out three times for a period of 24 h at room temperature. The material was kept for a period of 24 h between each successive solvent for proper drying. The extracts were dried under vacuum on a rotary evaporator at 40°C and stored in a refrigerator to pursue further analysis.

Phytochemical Screening
The concentrated extracts were used for preliminary screening of phytochemicals such as alkaloids, tannins, saponins, flavonoids, steroids, terpenoids and phenolic acids using standard procedures of analysis (Harborne, 2007; Evans, 2002). Ethyl acetate and methanol extracts showed the presence of flavonoid and phenolic acid. The confirmation of these was carried out using shinoda test and ferric chloride test, respectively (Audu et al., 2007).

In vitro Studies
Radical Scavenging Activity Using 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) Method
The free radical scavenging activity of the extracts obtained was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method (Shyu and Hwang, 2002). A solution of 0.5 mmol DPPH in methanol was prepared. The 1.0 mL of this solution was mixed with 0.1 mL of extract solution (250-400 μg mL^-1), 2 mL acetate buffer (pH 5.5) and 1.9 mL methanol. The reaction mixture was left in the dark at room temperature for 30 min. The absorbance of the mixture was measured spectrophotometrically at 517 nm. For each concentration, mixture without DPPH was used as blank and mixture without extract was used as control. Ascorbic acid was used as reference. The scavenging ability was calculated as follows:

Radical Scavenging Activity Using Hydrogen Peroxide Method
Hydrogen peroxide scavenging activity was measured using the method of Hui-Yin et al. (2007) with some modifications. An aliquot of 2.4 mL of 0.1 M phosphate buffer (pH 7.4) and 1.0 mL of various concentrations (80, 100, 120, 140 μg mL^-1) of extract solution was mixed with 0.6 mL of 43 mmol hydrogen peroxide solution. The absorbance was measured at 230 nm after 10 min. For each concentration, a separate blank sample (without hydrogen peroxide) was used. Ascorbic acid was used as reference. The scavenging activity was calculated as follows:

Pharmacological Evaluation
Experimental Protocol for Anti-Inflammatory and Analgesic Activity
Six groups, each comprising of six rats, were employed in the present study.

•	Group I (Normal Control Group): Administration of 1% CMC, p.o. on the day of experiment

•	Group II (Disease Control Group): 0.1 mL of carrageenan (1%) was injected at the plantar surface of the right hind paw

•	Group III (Reference Standard Group): Administration of diclofenac sodium (12.5 mg kg^-1, p.o.) on the day of experiment

•	Group IV to VI (EtCM Low, Medium and High Dose): Administration of EtCM (200, 300 and 400 mg kg^-1) for 7 consecutive days

In group III to VI pretreatment with diclofenac and extracts was performed 1 h before carrageenan administration on the day of experiment. Further, analgesic activities were assessed at different time interval of 0, 2, 4, 6, 8 and 10 h.

Acute Inflammation Activity
Paw Edema Test
All groups of animal were employed for the paw edema test by using plethysmometer (Winter et al., 1962). To assess anti-inflammatory activity paw volume was measured in all the groups at different time interval of 0, 1, 3, 6 and 12 h.

Analgesic Activity
Hot Plate Test
Thermal nociceptive (conduct heat) threshold, as an index of thermal hyperalgesia was assessed by the hot plate test as described by Andreas and Rainer (2005). Eddy’s hot plate was pre-heated and maintained at temperature of 52.5±0.5°C. Rats were placed on the hot plate and pain withdrawal threshold was assessed with respect to hind paw licking. The response of paw licking/jumping was recorded in seconds. Cut-off time of 30 sec was maintained.

Plantar Test
Thermal nociceptive (radiant heat) threshold, as an index of thermal hyperalgesia was assessed in the right hind paw under the radiant heat lamp source as described by Hargreaves et al. (1988). The intensity of the radiant heat stimulus was maintained at 25±0.1°C. The response of paw withdrawal latency was noted in seconds. Cut-off time of 30 sec was maintained.

Pin Prick Test
Peripheral nociceptive (pin point stimuli) threshold as an index of mechanical hyperalgesia was assessed as described by Erichsen and Blackburn-Munro (2002). Surface of the injured hind paw was touched with the point of bent 18 gauge needle (at 90° angle) at an intensity suficient to produce a reflex withdrawal response in normal non-operated animals but at an intensity which was insuficient to penetrate the skin in all other group. The duration of the paw lifting response was recorded in seconds. Cut-off time of 20 sec was maintained.

Mechanical Allodynia Test
Peripheral nociceptive (tactile stimuli) threshold as an index of mechanical hyperalgesia was assessed as described by Field et al. (2000). The surface of the hind paw was lightly strocked with a cotton bud. Care was taken to perform this procedure to avoid recording general motor activity. At least three measurements were taken at each time point the mean of which represented the paw withdrawal threshold. The duration of the paw withdrawal response was recorded in seconds. Cut-off time of 15 sec was maintained.

Statistical Analysis
All the results were expressed as Standard Error of Means (SEM). The data was statistically analyzed by one way analysis of variance (ANOVA) followed by Tukey’s multiple range tests by using Sigmastat Version-2.0 Software. The p<0.05 was considered to be statistically significant.

RESULTS

In vitro Antioxidant Assay
Scavenging of 1,1-Diphenyl-2-Picryl Hydrazyl
Antioxidants react with DPPH, which is a nitrogen-centered radical with a characteristic absorption at 517 nm and convert it into 1,1-diphenyl-2-picryl hydrazine, due to its hydrogen donating ability at a very rapid rate. The various Citrus medica peel extracts showed concentration dependent antioxidant activity. Maximum antioxidant activity was found in the ethyl acetate extract followed by methanol, chloroform and hexane extract (Table 1). Ethyl acetate extract showed comparable in vitro anti-oxidative potential with that of standard ascorbic acid.

Scavenging of Hydrogen Peroxide
Hydrogen peroxide itself is not very reactive, but sometimes it can be toxic to cells because of rise in the hydroxyl radicals in the cells and attack on cellular energy systems. The hydrogen peroxide scavenging activity of various extracts is shown in Table 2. Concentration dependent antioxidant activity was observed for all the extracts. The maximum antioxidant activity was found in the ethyl acetate extract followed by methanol, chloroform and hexane. Ethyl acetate extract showed comparable in vitro anti-oxidative potential with that of standard ascorbic acid.

Pharmacological Evaluation
Effect of Ethyl Acetate Extract of Citrus medica (EtCM) on Paw Edema Test
EtCM showed dose dependent anti-inflammatory activity in carrageenan induced paw edema in rats. Low and medium doses i.e., 200 and 300 mg kg^-1 of the ethyl acetate extract had shown significant difference in the anti-inflammatory activity as compared to the normal control (*p<0.05) group as well as diclofenac (^†p<0.05) treated group whereas, insignificant difference when compared with disease control group. High dose (400 mg kg^-1, p.o.) treated group showed insignificant difference in the anti-inflammatory activity when compared to the normal control and standard groups but significant difference (**p<0.05) when compared with disease group. Hence, it was observed that the higher dose of EtCM significantly attenuated carrageenan induced ipsilateral right hind paw edema in rats (Fig. 1).

Table 1:	Effect of Citrus medica peel extracts on DPPH scavenging activity

Effects of Citrus medica peel extracts were determined by the 1,1-diphenyl-2-picrylhydrazyl scavenging activity in vitro. Data were expressed as standard error of mean for each group (n = 3)

Table 2:	Effect of Citrus medica peel extracts on hydrogen peroxide scavenging activity

Effects of Citrus medica peel extracts were determined by the hydrogen peroxide scavenging activity in vitro. Data were expressed as standard error of mean for each group (n = 3)


Fig. 1:	Time course of paw edema measured by the carrageenan induced increased paw volume in rats. Data were expressed as Mean±SEM, n = 6 rats per group. p<0.05 vs. normal control group, *p<0.05 vs. carrageenan treated group and ^†p<0.05 vs. diclofenac sodium control group


Fig. 2:	Time course of paw thermal hyperalgesia measured by the noxious conduct heat evoked paw licking reflex. Data were expressed as Mean±SEM, n = 6 rat per group. p<0.05 vs. normal control group, *p<0.05 vs. carrageenan treated group and ^†p<0.05 vs. diclofenac sodium control group

Analgesic Activity in Rat
Effect of Ethyl Acetate Extract of Citrus medica (EtCM) on Hot Plate Test
EtCM showed significant analgesic activity against hyperalgesia induced by thermal stimuli as shown in Fig. 2. The oral administration of EtCM showed a dose dependent increase in paw licking threshold. Low and medium doses showed no significant difference as compared to the disease control group but significant difference was observed as compared to normal control (*p<0.05) and diclofenac (^†p<0.05) treated group. Whereas, high dose showed significant difference in the analgesic activity when compared with disease (**p<0.05) group but insignificant difference when the comparison was done with that of normal control and standard group. Hence, it was observed that the EtCM significantly reduced the pain sensation in rat at higher dose (400 mg kg^-1, p.o.).

Effect of Ethyl Acetate Extract of Citrus medica (EtCM) on Plantar Test
EtCM showed significant analgesic activity against hyperalgesia induced by thermal stimuli as shown in Fig. 3. The oral administration of EtCM showed a dose dependent increase in ipsilateral right hind paw withdrawal threshold. Low and medium doses showed no significant difference as compared to the disease control group but significant difference was observed as compared to normal control (*p<0.05) and diclofenac (^†p<0.05) treated group. Whereas, high dose showed significant difference in the analgesic activity when compared with disease (**p<0.05) group but insignificant difference when the comparison was done with that of normal control and standard group. Hence, it was observed that the EtCM significantly reduced the pain sensation in rat at higher dose (400 mg kg^-1, p.o.).

Effect of Ethyl Acetate Extract of Citrus medica (EtCM) on Pin Prick Test
EtCM showed significant analgesic activity against hyperalgesia induced by mechanical stimuli as shown in Fig. 4. The oral administration of EtCM showed a dose dependent increase in paw lifting duration. Low and medium doses showed no significant difference as compared to the disease control group but significant difference was observed as compared to normal control (*p<0.05) and diclofenac (^†p<0.05) treated group. Whereas, high dose showed significant difference in the analgesic activity when compared with disease (**p<0.05) group but insignificant difference when the comparison was done with that of normal control and standard group. Hence, it was observed that the EtCM significantly reduced the pain sensation in rat at higher dose (400 mg kg^-1, p.o.).


Fig. 3:	Time course of paw thermal hyperalgesia measured by the noxious radiant heat evoked ipsilateral right hind paw withdrawal reflex. Data were expressed as Mean±SEM, n = 6 rat per group. p<0.05 vs. normal control groups, *p<0.05 vs. carrageenan treated group and ^†p<0.05 vs. diclofenac sodium control group


Fig. 4:	Time course of paw mechanical hyperalgesia measured by the pin prick evoked rise in paw lifting duration. Data were expressed as Mean±SEM, n = 6 rat per group. p<0.05 vs. normal control groups, *p<0.05 vs. carrageenan treated group and ^†p<0.05 vs. diclofenac sodium control group


Fig. 5:	Time course of paw mechanical allodynia measured by the dynamic cotton bud evoked paw withdrawal reflex. Data were expressed as Mean±SEM, n = 6 rat per group. p<0.05 vs. normal control groups, *p<0.05 vs. carrageenan treated group and ^†p<0.05 vs. diclofenac sodium control group

Effect of Ethyl Acetate Extract of Citrus medica (Etcm) on Mechanical Allodynia Test
EtCM showed significant analgesic activity against allodynia induced by mechanical stimuli as shown in Fig. 5. The oral administration of EtCM showed a dose dependent increase in paw withdrawal latency. Low and medium doses showed no significant difference as compared to the disease control group but significant difference was observed as compared to normal control (*p<0.05) and diclofenac (^†p<0.05) treated group. Whereas, high dose showed significant difference in the analgesic activity when compared with disease (**p<0.05) group but insignificant difference when the comparison was done with that of normal control and standard group. Hence, it was observed that the EtCM significantly reduced the pain sensation in rat at higher dose (400 mg kg^-1, p.o.).

DISCUSSION

In the present study, the peel extracts of Citrus medica in different solvents were evaluated for the presence of important phytoconstituents and for their in vitro antioxidant activity. Ethyl acetate and methanol extracts showed the presence of flavonoid and phenolic acid. Results also revealed that ethyl acetate extract of Citrus medica (EtCM) possessed highest in vitro antioxidant activity. Hence, EtCM was further evaluated for its in vivo anti-inflammatory and analgesic activity in rats. Carrageenan induced rat paw edema test has frequently been used to assess the anti-edematous effect of natural products. Carrageenan is a strong chemical for the release of inflammation mediators (histamine, kinnin, prostaglandin, leukotriene etc.) and proinflammatory cytokinins (tissue necrosis factor, interleukin etc.) (Di Martino et al., 1987; Di Rosa et al., 1971). Free radicals are known to play an important role in the pathogenesis of inflammation and algesia (Koblyakov, 2001; Winrow et al., 1993; Chung, 2004). Moreover, literature also reveals the antioxidant and anti-inflammatory activity of Citrus sinensis peel extract in both acute and chronic inflammatory models due to the presence of free phenolic constituents (Ramachandran et al., 2002). Red orange extract showed interesting antiinflammatory properties in human keratinocyte cells (Cardile et al., 2009). Antioxidant activity and phenolic composition of Citrus bergamia, Citrus limon and Citrus aurantium peel and seed extracts has also been studied. Methanolic extract of these species were found to contain free phenolic compounds (Bocco et al., 1998). Recently, peels of number of commercially available citrus species have also been evaluated for their phenolic and flavonoids content along with their antioxidative potential (Ghasemi et al., 2009). In our studies EtCM had shown statistically significant results in anti-inflammatory and analgesic tests in rats. As EtCM had also shown significant free radical scavenging activity in vitro, so this can be the possible reason for the reduction of inflammation and algesia in the carrageenan induced paw edema in rats. Moreover, phytochemical screening revealed the presence of flavonoids and phenolic acids in the EtCM so, these constituents may be responsible for the antioxidant, anti-inflammatory and analgesic activities (Shahidi and Wanasundara, 1992; Hanasaki et al., 1994). However, more elaborative studies are required to identify and characterize the active components and precise mechanism of anti-inflammatory and analgesic actions.

CONCLUSIONS

Ethyl acetate extract of Citrus medica peels (EtCM) reduced carrageenan induced paw edema and showed analgesic action in rats. Its free radical scavenging action may be responsible for the observed ameliorative effects. Therefore, this herbal candidate may act as a potent anti-inflammatory and analgesic agent in human subjects.

ACKNOWLEDGMENTS

Thanks to Professor A.C. Rana and all faculty members of Rayat Institute of Pharmacy for their encouragement and support. We are also grateful to Rayat and Bahra Educational and Research Trust for their unconditional helps to carryout this project.

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Italian Journal of Animal Science, 2015, 14(2), 3737. DOI: 10.4081/ijas.2015.3737

In Vitro Methane Production and Quality of Corn Silage Treated with Maleic Acid

Italian Journal of Animal Science, 2015, 14(4), 3994. DOI: 10.4081/ijas.2015.3994

Lactation Curve and Persistency of Anatolian Buffaloes

Italian Journal of Animal Science, 2015, 14(2), 3679. DOI: 10.4081/ijas.2015.3679

Administration of Aloe arborescens homogenate to cattle: interaction with rumen fermentation and gut absorption of aloin

Italian Journal of Animal Science, 2016, 15(2), 233. DOI: 10.1080/1828051X.2016.1157007

Testicular Growth and Comb and Wattles Development in Three Italian Chicken Genotypes Reared Under Free-Range Conditions

Italian Journal of Animal Science, 2015, 14(2), 3653. DOI: 10.4081/ijas.2015.3653

Efficacy of Supplemental Natural Zeolite in Broiler Chickens Subjected to Dietary Calcium Deficiency

Italian Journal of Animal Science, 2014, 13(2), 3141. DOI: 10.4081/ijas.2014.3141

Modulation of Plasma Antioxidant Activity in Weaned Piglets by Plant Polyphenols

Italian Journal of Animal Science, 2014, 13(2), 3242. DOI: 10.4081/ijas.2014.3242

Amino Acids Fortification of Low-Protein Diet for Broilers Under Tropical Climate: Ideal Essential Amino Acids Profile

Italian Journal of Animal Science, 2014, 13(2), 3166. DOI: 10.4081/ijas.2014.3166

Effects of Oregano Essential Oil Supplementation to Diets of Broiler Chicks with Delayed Feeding After Hatching. Morphological Development of Small Intestine Segments

Italian Journal of Animal Science, 2014, 13(2), 3172. DOI: 10.4081/ijas.2014.3172

Stress and reactivity in three Italian chicken breeds

Italian Journal of Animal Science, 2016, 15(2), 303. DOI: 10.1080/1828051X.2016.1185978

Effects of Saccharomyces Cerevisiae at Direct Addition or Pre-incubation on in Vitro Gas Production Kinetics and Degradability of Four Fibrous Feeds

Italian Journal of Animal Science, 2014, 13(2), 3075. DOI: 10.4081/ijas.2014.3075

Digestibility of Juncus acutus and its effects on ruminal cellulolytic bacteria

Italian Journal of Animal Science, 2016, 15(1), 69. DOI: 10.1080/1828051X.2016.1139327

Wool Quality Traits of Purebred and Crossbred Merino Lambs Orally Drenched with Spirulina (Arthrospira Platensisi)

Italian Journal of Animal Science, 2014, 13(2), 3174. DOI: 10.4081/ijas.2014.3174

Expression Profile of Heat Shock Protein 70 in Indigenous Huainan Partridge Chicken Exposed to Low Temperature

Italian Journal of Animal Science, 2014, 13(2), 2960. DOI: 10.4081/ijas.2014.2960

Genetic Variability at α s2-casein Gene in Girgentana Dairy Goat Breed

Italian Journal of Animal Science, 2014, 13(1), 2997. DOI: 10.4081/ijas.2014.2997

Comparison of Meat Quality and Fatty Acid Profile in Slow-Growing Chicken Genotypes Fed Diets Supplemented with Origanum Vulgare or Melissa Officinalis Leaves Under the Organic System

Italian Journal of Animal Science, 2013, 12(3), e64. DOI: 10.4081/ijas.2013.e64

Sexual Maturity and Performance of Pullets Fed Different Preparations and Concentrations of Brown Marine Algae (Sargassum Dentifebium) in Pre-Laying and Early Laying Periods

Italian Journal of Animal Science, 2014, 13(1), 3102. DOI: 10.4081/ijas.2014.3102

Effects of Birth Weight and Feeding System on Fattening Performance and Feeding Behaviour of Karayaka Male Lambs

Italian Journal of Animal Science, 2013, 12(4), e89. DOI: 10.4081/ijas.2013.e89

Glutamine and Glutamic Acid Supplementation Enhances Performance of Broiler Chickens Under the Hot and Humid Tropical Condition

Italian Journal of Animal Science, 2015, 14(1), 3263. DOI: 10.4081/ijas.2015.3263

Analysis of the Non-Genetic Factors Affecting the Growth of Segureño Sheep

Italian Journal of Animal Science, 2015, 14(1), 3683. DOI: 10.4081/ijas.2015.3683

Effects of supplemental feeding on performance of Kilis goats kept on pasture condition

Italian Journal of Animal Science, 2016, 15(1), 110. DOI: 10.1080/1828051X.2015.1132542

Effect of Faba Bean Vicia Faba Var. Minor) Inclusion in Starter and Growing Diet on Performance, Carcass and Meat Characteristics of Organic Slow-Growing Chickens

Italian Journal of Animal Science, 2013, 12(4), e76. DOI: 10.4081/ijas.2013.e76

Effects of Dietary Quercetin on Growth Performance, Serum Lipids Level and Body Composition of Tilapia (Oreochromis Niloticus)

Italian Journal of Animal Science, 2013, 12(4), e85. DOI: 10.4081/ijas.2013.e85

An Update on the Safety of Foods of Animal Origin and Feeds

Italian Journal of Animal Science, 2014, 13(4), 3571. DOI: 10.4081/ijas.2014.3571

Amino Acids Fortification of Low-protein Diet for Broilers Under Tropical Climate. 2. Nonessential Amino Acids and Increasing Essential Amino Acids

Italian Journal of Animal Science, 2014, 13(3), 3297. DOI: 10.4081/ijas.2014.3297

Estimation of Optimal Lysine in Quail Chicks During the Second and Third Weeks of Age

Italian Journal of Animal Science, 2013, 12(4), e84. DOI: 10.4081/ijas.2013.e84

Genetic Diversity of Kıvırcık Sheep Breed Reared in Different Regions and its Relationship with Other Sheep Breeds in Turkey

Italian Journal of Animal Science, 2014, 13(3), 3382. DOI: 10.4081/ijas.2014.3382

Pentachlorophenol Degradation by Janibacter sp., a New Actinobacterium Isolated from Saline Sediment of Arid Land

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/296472

New Perspectives on Antiacne Plant Drugs: Contribution to Modern Therapeutics

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/301304

Antioedematous and Analgesic Properties of Fertile Fronds of Drynaria quercifolia

ISRN Inflammation, 2014, 2014(), 1. DOI: 10.1155/2014/302089

Pharmacodynamics and Pharmacokinetics Evaluation of Ranitidine Microemulsion on Experimental Animals

Advances in Pharmaceutics, 2014, 2014(), 1. DOI: 10.1155/2014/304392

Effect of Differently Coloured Clothes on Fear and Stress Responses, Some Meat Quality Traits and Performance in Broilers

Italian Journal of Animal Science, 2014, 13(3), 3453. DOI: 10.4081/ijas.2014.3453

A spatial analysis of the ionospheric TEC anomalies prior to M7.0+ earthquakes during 2003–2014

Advances in Space Research, 2016, (), . DOI: 10.1016/j.asr.2016.06.040

New Advances of Preimplantation and Prenatal Genetic Screening and Noninvasive Testing as a Potential Predictor of Health Status of Babies

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/306505

Reputation-Based Secure Sensor Localization in Wireless Sensor Networks

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/308341

Immobilization of a Pleurotus ostreatus Laccase Mixture on Perlite and Its Application to Dye Decolourisation

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/308613

Genetic Diversity Assessment and Identification of New Sour Cherry Genotypes Using Intersimple Sequence Repeat Markers

International Journal of Biodiversity, 2014, 2014(), 1. DOI: 10.1155/2014/308398

In Vitro Cadmium-Induced Alterations in Growth and Oxidative Metabolism of Upland Cotton (Gossypium hirsutum L.)

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/309409

Efficacy of 2-Phenoxyethanol as an Anaesthetic for Adult Redline Torpedo Fish, Sahyadria denisonii (Day 1865)

International Journal of Zoology, 2014, 2014(), 1. DOI: 10.1155/2014/315029

The Effect of Maternal Healthcare on the Probability of Child Survival in Azerbaijan

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/317052

Modeling and Optimization of Beam Pumping System Based on Intelligent Computing for Energy Saving

Journal of Applied Mathematics, 2014, 2014(), 1. DOI: 10.1155/2014/317130

A stable reactive approach in dynamic flexible flow shop scheduling with unexpected disruptions: A case study

Computers & Industrial Engineering, 2016, 98(), 360. DOI: 10.1016/j.cie.2016.06.018

Inhibition of 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase (Ex Vivo) by Morus indica (Mulberry)

Chinese Journal of Biology, 2014, 2014(), 1. DOI: 10.1155/2014/318561

Antisecretory Action of the Extract of the Aerial Parts of Eremomastax speciosa (Acanthaceae) Occurs through Antihistaminic and Anticholinergic Pathways

Advances in Pharmacological Sciences, 2014, 2014(), 1. DOI: 10.1155/2014/323470

Antidepressant Effects of Mallotus oppositifolius in Acute Murine Models

ISRN Pharmacology, 2014, 2014(), 1. DOI: 10.1155/2014/324063

Automatic Extraction System for Common Artifacts in EEG Signals Based on Evolutionary Stone’s BSS Algorithm

Mathematical Problems in Engineering, 2014, 2014(), 1. DOI: 10.1155/2014/324750

A Review of Botanical Characteristics, Traditional Usage, Chemical Components, Pharmacological Activities, and Safety of Pereskia bleo (Kunth) DC

Evidence-Based Complementary and Alternative Medicine, 2014, 2014(), 1. DOI: 10.1155/2014/326107

Comprehensive Aspectual UML Approach to Support AspectJ

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/327808

Development and Evaluation of Novel Self-Nanoemulsifying Drug Delivery Systems Based on a Homolipid from Capra hircus and Its Admixtures with Melon Oil for the Delivery of Indomethacin

Journal of Pharmaceutics, 2014, 2014(), 1. DOI: 10.1155/2014/340486

Evaluation of common bean (Phaseolus vulgaris L.) genotypes for drought stress adaptation in Ethiopia

The Crop Journal, 2016, (), . DOI: 10.1016/j.cj.2016.06.007

Neuroantibody Biomarkers: Links and Challenges in Environmental Neurodegeneration and Autoimmunity

Autoimmune Diseases, 2014, 2014(), 1. DOI: 10.1155/2014/340875

An Evaluation of 3-Rhamnosylquercetin, a Glycosylated Form of Quercetin, against the Myotoxic and Edematogenic Effects of sPLA2 from Crotalus durissus terrificus

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/341270

Development and targeting of transcriptional regulatory network controlling FLU1 activation in Candida albicans for novel antifungals

Journal of Molecular Graphics and Modelling, 2016, (), . DOI: 10.1016/j.jmgm.2016.07.009

Viola canescens: Herbal Wealth to Be Conserved

Journal of Botany, 2014, 2014(), 1. DOI: 10.1155/2014/345451

An Effective Protocol for Micropropagation of Edible Bamboo Species (Bambusa tulda and Melocanna baccifera) through Nodal Culture

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/345794

An Improved Mathematical Model for Computing Power Output of Solar Photovoltaic Modules

International Journal of Photoenergy, 2014, 2014(), 1. DOI: 10.1155/2014/346704

Epidemiological aspects and risk factors for infection by Leishmania infantum chagasi in dogs from municipality of Petrolina, Northeastern Brazil

Veterinary Parasitology: Regional Studies and Reports, 2016, (), . DOI: 10.1016/j.vprsr.2016.07.001

A New Mathematical Inventory Model with Stochastic and Fuzzy Deterioration Rate under Inflation

Chinese Journal of Engineering, 2014, 2014(), 1. DOI: 10.1155/2014/347857

Waste Material Adsorbents for Zinc Removal from Wastewater: A Comprehensive Review

International Journal of Chemical Engineering, 2014, 2014(), 1. DOI: 10.1155/2014/347912

Genetic Transformation of Metroxylon sagu (Rottb.) Cultures via Agrobacterium-Mediated and Particle Bombardment

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/348140

The Therapeutic Potential of Medicinal Foods

Advances in Pharmacological Sciences, 2014, 2014(), 1. DOI: 10.1155/2014/354264

Bacteria in Nanoparticle Synthesis: Current Status and Future Prospects

International Scholarly Research Notices, 2014, 2014(), 1. DOI: 10.1155/2014/359316

Selection Index in the Study of Adaptability and Stability in Maize

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/360570

Haematological and Biochemical Parameters during the Laying Period in Common Pheasant Hens Housed in Enhanced Cages

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/364602

Jatropha gossypiifolia L. (Euphorbiaceae): A Review of Traditional Uses, Phytochemistry, Pharmacology, and Toxicology of This Medicinal Plant

Evidence-Based Complementary and Alternative Medicine, 2014, 2014(), 1. DOI: 10.1155/2014/369204

Trypanosomosis, Its Risk Factors, and Anaemia in Cattle Population of Dale Wabera District of Kellem Wollega Zone, Western Ethiopia

Journal of Veterinary Medicine, 2014, 2014(), 1. DOI: 10.1155/2014/374191

Microcystin Contamination in Sea Mussel Farms from the Italian Southern Adriatic Coast following Cyanobacterial Blooms in an Artificial Reservoir

Journal of Ecosystems, 2014, 2014(), 1. DOI: 10.1155/2014/374027

In Vitro Activity of Lawsonia inermis (Henna) on Some Pathogenic Fungi

Journal of Mycology, 2014, 2014(), 1. DOI: 10.1155/2014/375932

Function Formula Oriented Construction of Bayesian Inference Nets for Diagnosis of Cardiovascular Disease

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/376378

Identification of meat origin in food products–A review

Food Control, 2016, 68(), 379. DOI: 10.1016/j.foodcont.2016.04.013

Exploration of microwave dielectric and near infrared spectroscopy with multivariate data analysis for fat content determination in ground beef

Food Control, 2016, 68(), 260. DOI: 10.1016/j.foodcont.2016.03.031

Host-Range Dynamics of Cochliobolus lunatus: From a Biocontrol Agent to a Severe Environmental Threat

BioMed Research International, 2014, 2014(), 1. DOI: 10.1155/2014/378372

Spatial Variability of Physical Soil Quality Index of an Agricultural Field

Applied and Environmental Soil Science, 2014, 2014(), 1. DOI: 10.1155/2014/379012

Detection of Rift Valley Fever Virus Interepidemic Activity in Some Hotspot Areas of Kenya by Sentinel Animal Surveillance, 2009–2012

Veterinary Medicine International, 2014, 2014(), 1. DOI: 10.1155/2014/379010

The occurrence and effect of unit operations for dairy products processing on the fate of aflatoxin M1: A review

Food Control, 2016, 68(), 310. DOI: 10.1016/j.foodcont.2016.04.007

Effects of Brown Seaweed (Sargassum polycystum) Extracts on Kidney, Liver, and Pancreas of Type 2 Diabetic Rat Model

Evidence-Based Complementary and Alternative Medicine, 2014, 2014(), 1. DOI: 10.1155/2014/379407

Transient Downregulation of Nanog and Oct4 Induced by DETA/NO Exposure in Mouse Embryonic Stem Cells Leads to Mesodermal/Endodermal Lineage Differentiation

Stem Cells International, 2014, 2014(), 1. DOI: 10.1155/2014/379678

Incidence and Evaluation of Incidental Abnormal Bone Marrow Signal on Magnetic Resonance Imaging

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/380814

Utility Potential of Parthenium hysterophorus for Its Strategic Management

Advances in Agriculture, 2014, 2014(), 1. DOI: 10.1155/2014/381859

Calculating Super Efficiency of DMUs for Ranking Units in Data Envelopment Analysis Based on SBM Model

The Scientific World Journal, 2014, 2014(), 1. DOI: 10.1155/2014/382390

A Review and Classification of Computer-Based Manufacturing Scheduling Tools

Computers & Industrial Engineering, 2016, (), . DOI: 10.1016/j.cie.2016.07.020

In Vitro Antimicrobial Activity of Thymus vulgaris Essential Oil Against Major Oral Pathogens

Journal of Evidence-Based Complementary & Alternative Medicine, 2017, (), 215658721770077. DOI: 10.1177/2156587217700772

An Evidence-Based Study on Medicinal Plants for Hemorrhoids in Medieval Persia

Journal of Evidence-Based Complementary & Alternative Medicine, 2017, (), 215658721668859. DOI: 10.1177/2156587216688597

Extraction, characterization and biological activity of citrus flavonoids

Reviews in Chemical Engineering, 2018, 0(0), . DOI: 10.1515/revce-2017-0027

Citrus medica “Otroj”: Attenuates Oxidative Stress and Cardiac Dysrhythmia in Isoproterenol-Induced Cardiomyopathy in Rats

Nutrients, 2013, 5(12), 4269. DOI: 10.3390/nu5114269

Cytotoxicity, genotoxicity, and mutagenicity potential of ethanolic extract of Heracleum persicum Desf. ex Fischer

Journal of Food Safety, 2019, (), e12640. DOI: 10.1111/jfs.12640

Monpa, memory, and change: an ethnobotanical study of plant use in Mêdog County, South-east Tibet, China

Journal of Ethnobiology and Ethnomedicine, 2020, 16(1), . DOI: 10.1186/s13002-020-0355-7

Analgesic and anti-inflammatory effects and safety profile of Cucurbita maxima and Cucumis sativus seeds

Saudi Journal of Biological Sciences, 2021, (), . DOI: 10.1016/j.sjbs.2021.04.020