Carob (Ceratonia siliqua L.), Pharmacological and Phytochemical Activities of Neglected Legume of the Mediterranean Basin, as Functional Food.

Carob (Ceratonia siliqua L.) has been widely cultivated in different parts of the world, particularly in the Mediterranean region, and the tree belongs to the family Leguminosae. Several studies have indicated that carobs and their products can improve human health and help prevent different specific chronic diseases. Carob can considered as functional food due to its high content in dietary fibers, low-fat content, and high content of minerals. Its fruit is a pod containing 10%-20% seeds, and the pods consist of sugars, proteins, crude fibers, minerals, vitamins, polyphenols, vitamins, and lipids. In many countries in the Middle east, carob is mainly used to prepare as a traditional drink and some kinds of confectioneries. The powders can be utilized to prepare carob juice concentrate. The systematic review of documents from clinical trials and scientific societies dedicated to traditional medicine in China has been carried out. The goal of this review article is a survey of chemical compounds, and pharmaceutical benefits of carob, especially by considering traditional medicinal sciences. Moreover, clinical trials research promotes studies to highlight and focus on the scope of application of traditional medicinal science in the growing system of medicine.

Effects of Carob Extract on the Intestinal Microbiome and Glucose Metabolism: A Systematic Review and Meta-Analysis.

Abstract: The legume tree known as carob (Ceratonia siliqua L.) is indigenous to the Mediterranean area and over the centuries its pods had been traditionally used mostly as animal feed. However, it has gained great attention in human nutrition due to the molecular compounds it contains, which could offer many potential health benefits: for example, carob is renowned for its high content of fiber, vitamins, and minerals. Moreover, in traditional medicine it is credited with the ability to control glucose metabolism and gut microbiome. Modern science has also extensively acknowledged the numerous health advantages deriving from its consumption, including its anti-diabetic, anti-inflammatory, and antioxidant properties. Due to its abundant contents of pectin, gums, and polyphenols (such as pinitol), carob has garnered significant attention as a well-researched plant with remarkable therapeutic properties. Notably, carob is extensively used in the production of semi-finished pastry products, particularly in ice cream and other creams (especially as a substitute for cocoa/chocolate): these applications indeed facilitate the exploration of its positive effects on glucose metabolism. Our study aimed at examining the effects of carob extract on intestinal microbiota and glucose metabolism. In this review, we conducted a thorough examination, comprising in vitro, in vivo, and clinical trials to appraise the consequences on human health of polyphenols and pectin from different carob species, including recently discovered ones with high polyphenol contents. Our goal was to learn more about the mechanisms through which carob extract can support a balanced gut flora and improve one's glucose metabolism. These results could influence the creation of novel functional foods and dietary supplements, to help with the management and prevention of chronic illnesses like diabetes and obesity.

Exploring Carob (Ceratonia siliqua L.): A Comprehensive Assessment of Its Characteristics, Ethnomedicinal Uses, Phytochemical Aspects, and Pharmacological Activities.

The carob tree (Ceratonia siliqua L.) is currently considered one of the most valuable fruit and forest trees in various fields and sectors of activity. It is a versatile plant, belonging to the Fabaceae family. It is widely used in traditional medicine to treat many diseases such as diabetes, hypertension, and gastrointestinal disorders, given that all its parts (leaves, flowers, pods, seeds, wood, bark, and roots) are useful and hold value in many areas. Its importance has increased significantly in recent years. Originating from the Middle East, it is recognized for its ecological and industrial significance. Previous studies conducted on Ceratonia siliqua L. have revealed the presence of several compounds, including polyphenols, flavonoids, carbohydrates, minerals, and proteins. The carob tree demonstrates antihypertensive, antidepressant, anti-obesity, and antihyperglycemic activities. This plant is known for its medicinal and therapeutic virtues. Moreover, it is particularly interesting to consider the pharmacological activities of the major phytochemical compounds present in the different extracts of this plant, such as phenolic acids, for example, coumaric and gallic acids, as well as flavonoids such as kaempferol and quercetin. Therefore, this review aims to analyze some aspects of this plant, especially the taxonomy, cytogeography, traditional uses, phytochemical constituents, and pharmacological activities of Ceratonia siliqua L., in addition to its biological properties.

Colour Stability, Fatty Acid Profile, and Lipid Oxidation in Meat Stored in Modified Atmosphere Packaging from Light Lambs Fed with Concentrate with Carob Pulp (Ceratonia siliqua L.).

There is a growing interest in using by-products rich in polyphenols, such as carob pulp (Cp, Ceratonia siliqua L.), as a dietary source of antioxidants for animals. This study assesses the effects of including Cp in lambs' diet and meat display time (0, 7, 9, and 11 days) in modified atmosphere packaging on meat colour, fatty acid (FA) composition, tocopherol levels, and lipid oxidation values in the Semimembranosus muscle of 40 light lambs. The lambs were fed with concentrates supplemented with increasing Cp levels (0, 150, and 300 g/kg) for 45 days before slaughter. Metmyoglobin (MMb) and malondialdehyde (MDA) contents increased linearly with display time (p < 0.05), regardless of diet (p > 0.05). At 11 days of display, MMb (28 ± 0.8%) and MDA (0.6 ± 0.1 mg MDA/kg of meat) contents remained within the acceptable limits. The α-tocopherol content was lower in the 30% Cp group and meat (p < 0.05). Total saturated and monounsaturated FA contents (934 ± 64 and 823 ± 65 mg/100 g of meat, respectively) did not differ significantly among the groups. However, the meat from lambs fed with 30% Cp showed reduced levels of branched-chain FAs, while polyunsaturated FAs increased (p < 0.05) compared to the control lambs. The inclusion of Cp in the lamb's diet, up to 30%, did not lead to meat deterioration and improved certain quality parameters, including a healthier FA profile. These findings highlight Cp's potential as an alternative antioxidant source in animal diets.

Polymorphisms at Candidate Genes for Fat Content and Fatty Acids Composition: Effects on Sheep Milk Production and Fatty Acid Profile Using Two Dietary Supplementations.

The nutritional value of sheep's milk and its derivatives is influenced by the lipid fraction, which is affected by diet and genetics. This study aimed to explore the genetic variations in the DGAT1 and SCD genes and assessed the impact of the DGAT1 genotype on milk quality in Valle del Belìce sheep, considering diet supplementation with carob pulp and barley grain. Among the potentially polymorphic sites, only DGAT1 g.127 C > A and SCD g.87 C > A showed variability. The DGAT1 genotype did not significantly impact milk yield and composition, except for higher urea content in the CA genotypes than in the CC ones. Carob pulp increased the milk fat content compared to barley grain. Genetic variation in DGAT1 was associated with changes in the milk fatty acid profile; specifically, the CA genotype exhibited higher levels of short-chain fatty acids and lower levels of polyunsaturated fatty acids compared to the CC genotype. Carob pulp supplementation increased saturated fatty acids and reduced unsaturated fractions, leading to milk with higher atherogenic and thrombogenic indices. No significant interaction was found between genotype and diet. This study provides insights into the genetic and dietary factors influencing sheep's milk composition. Further research is needed to understand the impact of these genetic variations on milk production and composition, as well as to determine optimal levels of carob pulp for improving fat percentage and promoting sustainable sheep breeding practices.

FTIR-ATR detection method for emerging C3-plants-derivated adulterants in honey: Beet, dates, and carob syrups.

The European Union Publications Office has recently presented a report on the European Union's coordinated action with the Joint Research Centre to determine certain fraudulent practices in the honey sector, in which it has been indicated that 74% of the samples analyzed, imported from China, and 93% of the samples analyzed, imported from Turkey, the two largest honey producers worldwide, presented at least one indicator of exogenous sugar or suspicion of being adulterated. This situation has revealed the critical state of the problem of honey adulteration worldwide and the need to develop analytical techniques for its detection. Even though the adulteration of honey is carried out in a general way with sweetened syrups derived from C4 plants, recent studies have indicated the emerging use of syrups derived from C3 plants for the adulteration of honey. This kind of adulteration makes it impossible to analyze its detection using official analysis techniques. In this work, we have developed a fast, simple, and economical method based on the Fourier transform infrared spectroscopy technique, with attenuated total reflectance, for the qualitative, quantitative, and simultaneous determination of beetroot, date, and carob syrups, derived from of C3 plants; whose available bibliography is very scarce and analytically not very conclusive for its use by the authorities. The proposed method has been based on the establishment of the spectral differences between honey and the mentioned syrups at eight different points in the spectral region between 1200 and 900 cm-1 of the mid-infrared, characteristic of the vibrational modes of carbohydrates in honey, which allows the pre-discrimination of the presence or absence of the syrups studied, and their subsequent quantification, with precision levels lower than 2.0% of the relative standard deviation and relative errors lower than 2.0% (m/m).

Evaluation of The Effects of Carob (Ceratonia siliqua L.) Fruits on the Puberty of Rats

Objective: This study was planned to determine the effects of carob use on puberty because of the observation of early puberty or pubertal variants due to the long-term use of carob in our clinic. Methods: Forty-eight Wistar albino rats, on postnatal day 21, were assigned into two groups female (n=24) and male (n=24). Groups were divided into four groups Control, and Carob-150, Carob-300, and Carob-600. Ceratonia siliqua L. extract was given to rats in a 0.5% carboxymethylcellulose (CMC) solution. CMC (0.5%) was given to the control, Ceratonia siliqua L. extract was given 150 mg/kg/day to the Carob-150, 300 mg/kg/day to the Carob-300, 600 mg/kg/day to the Carob-600 by oral gavage. The treatments were performed once daily until the first sign of puberty. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, total testosterone, leptin, glutathione, glutathione peroxidase (GPx), and malondialdehyde were measured by commercial rat-specific ELISA kits. Testis, uterus and ovarian tissue were examined histologically. Results: The median time of preputial separation in male rats was 38th, 31st, 31st, and 31st days in the Control, Carob-150, Carob-300, and Carob-600 groups, respectively (p=0.004). The median day of vaginal opening day was the 39th, 31st, 34th, and 31st days in the Control, Carob-150, Carob-300, and Carob-600 groups, respectively (p=0.059). FSH, LH, testosterone (male), estradiol (female) and leptin levels of the groups were similar. However, GPx levels were higher in male and female animals given C. siliqua extract compared to the Control (male p=0.001 and female p=0.008). Testicular and ovarian tissues were concordant with the pubertal period in all groups. As the dose of Ceratonia siliqua extract increased, it induced spermatogenesis and spermiogenesis, causing abnormal changes, such as ondulation in the basement membrane, capillary dilatation, and increased congestion in males. In females, edema in the medulla gradually increased with increased dosage, and granulosa cell connections were separated in Carob-300 and Carob-600 groups. Conclusion: This study demonstrated that C. siliqua caused early puberty and increased spermiogenesis and folliculogenesis. Antioxidant mechanisms were impaired with increasing dose, possibly leading to tissue damage at high doses.

Carob extract induces spermatogenesis in an infertile mouse model via upregulation of Prm1, Plzf, Bcl-6b, Dazl, Ngn3, Stra8, and Smc1b.

ETHNOPHARMACOLOGICAL RELEVANCE: Ethnopharmacological studies for drug discovery from natural compounds play an important role for developing current therapeutical platforms. Plants are a group of natural sources which have been served as the basis in the treatment of many diseases for centuries. In this regard, Ceratonia siliqua (carob) is one of the herbal medicine which is traditionally used for male infertility treatments. But so far the main mechanisms for effects of carob are unknown. Here, we intend to investigate the ability of carob extract to induce spermatogenesis in an azoospermia mouse model and determine the mechanisms that underlie its function. AIM OF THE STUDY: This is a pre-clinical animal model study to evaluate the effect of carob extract in spermatogenesis recovery. METHODS: We established an infertile mouse model with the intent to examine the ability of carob extract as a potential herbal medicine for restoration of male fertility. Sperm parameters, as well as gene expression dynamics and levels of spermatogenesis hormones, were evaluated 35 days after carob administration. RESULTS: Significant enhanced sperm parameters (P < 0.05) showed that the carob extract could induce spermatogenesis in the infertile mouse model. Our data suggested an anti-apototic and inducer role in the expressions of cell cycle regulating genes. Carob extract improved the spermatogenesis niche by considerable affecting Sertoli and Leydig cells (P < 0.05). The carob-treated mice were fertile and contributed to healthy offspring that matured. Our data confirmed that this extract triggered the hormonal system, the spermatogenesis-related gene expression network, and signaling pathways to induce and promote sperm production with notable level (P < 0.05). We found that the aqueous extract consisted of a polar and mainly well water-soluble substance. Carob extract might upregulate spermatogenesis hormones via its amino acid components, which were detected in the extract by liquid chromatography-mass spectrometry (LC-MS). CONCLUSION: Our results strongly suggest that carob extract might be a promising future treatment option for male infertility. This finding could pave the way for clinical trials in infertile men. This is the first study that has provided reliable, strong pre-clinical evidence for carob extract as an effective candidate for fertility recovery in cancer-related azoospermia.

Optimization and kinetic modeling of media composition for hyaluronic acid production from carob extract with Streptococcus zooepidemicus.

Hyaluronic acid (HA), a mucopolysaccharide belonging to the glycosaminoglycan family, consists of repeating disaccharide units and has been used directly or indirectly in numerous human health practices. This study focused on evaluating carob pods for microbial HA production and kinetic modeling of HA fermentation. Therefore, the optimal medium composition was determined using Plackett-Burman Design (PBD) for HA production from carob extract with Streptococcus zooepidemicus. Maximum HA production of shake flask fermentation was 2.6 g/L (1.25 × 106) in the optimum medium, comprising 10°Bx of carob pods extract, 0.5 g/L of MgSO4.7H2O, 10.0 g/L of casein, 2.5 g/L of KH2PO4, 2.0 g/L of NaCl, 1.5 g/L of K2HPO4, 0.002 g/L of FeSO4 and 10.0 g/L of beef extract. In the continuation of the study, the fermentation performed with the optimal medium composition was modeled using three different models including the logistic model for biomass production, the Luedeking-Piret model for HA production, and the modified Luedeking-Piret model for substrate consumption. Based on the results, the experimental HA production data agreed with the Luedeking-Piret model with an R2 of 0.989. Since the α value was 63-fold higher than the value of ß, the HA production is growth-associated. Consequently, carob extract can be evaluated as a promising carbon source for producing HA.

Effect of feeding carob (Ceratonia siliqua L.) pulp powder to broiler chicken on growth performance, intestinal microbiota, carcass traits, and meat quality.

In this study, the influence of a partial substitution of corn with carob (Ceratonia siliqua L.) pulp powder on broiler performance, intestinal microbiota, carcass traits, and meat quality, was investigated. Two dietary treatments were compared: the control group received a diet containing basically corn, soybean meal, and wheat bran for all the breeding periods while the experimental group received the same starter diet as the control group and grower and finisher diets containing 3% and 7% of carob pulp powder, respectively. Growth performance (weight gain, feed intake, and feed conversion ratio), carcass traits, lactic acid bacteria, and total coliforms were determined. Physicochemical parameters, proximate composition, fatty acid profiles, lipid oxidation index, and sensory characteristics of broiler thigh and breast muscles were determined. Dietary supplementation with carob pulp powder did not show an effect (P > 0.05) on growth performance and carcass traits except for slaughter yield and breast relative weight. An increase in sum lactic acid bacteria count and a decrease in total coliforms were observed in the experimental group. No effect (P > 0.05) of dietary carob was observed on pH, cooking losses, ash, mineral, and protein contents in breast and thigh muscles from broilers. However, the experimental diet decreased (P < 0.05) moisture and fat contents in thigh muscles but not in breast muscles. Both muscles from the experimental group of birds exhibited lower palmitic and oleic acids contents, and higher linoleic and linolenic acids levels than those from the control group. Whereas the polyunsaturated fatty acid contents in broiler breast and thigh meat was increased by dietary carob, the concentration of lipid oxidation products decreased in this group. The dietary intervention had no significant effect on the sensory profile of breast and thigh muscles. Thus, carob pulp powder can be applied to broiler diet at 7% as a nonconventional feed with no negative impact on growth performance and carcass traits and to increase the degree of unsaturation with no negative impact on oxidative stability or sensory traits.

Dietary carob fruit (Ceratonia siliqua L.) supplementation improves spermatogenesis, semen quality and embryonic death via antioxidant effect in aging broiler breeder roosters.

Rooster semen contains high content of long-chain polyunsaturated fatty acids; therefore, spermatozoa are very susceptible to lipid peroxidation. Carob fruit (CF) contains abundant amounts of flavonoid known as natural antioxidants and semen enhancers. The purpose of the study was to investigate the effect of dietary supplementation of CF on semen characters, blood parameters, immune response, testicular histology, and reproductive performance of aging broiler breeder roosters. Thirty-two 47-week-old Ross-308 breeder roosters were randomly assigned to either control or supplemented (CF, 0 or 1 g/kg diet) dietary treatment for 14 continuous weeks. Semen, blood and testis samples were collected at the end of trial. Furthermore, the rate of fertility and hatchability were evaluated using 440 natural mated hens. The sperm kinematic values and progression ratios, seminal antioxidant enzymes (superoxide dismutase and glutathione peroxidase) activity, and total antioxidant capacity of semen and blood were increased, and malondialdehyde concentration in semen and blood were decreased by dietary CF supplementation (P < 0.01). The treatment had no effect on the seminal sperm concentration, blood parameters and immune response to Newcastle disease virus vaccine. The testes weight, seminal tubes spermatozoids number, spermiogenesis index and repopulation index, were improved by dietary supplementation (P < 0.05). Fertility and hatchability rate of roosters in the CF group were insignificant. In conclusion, CF supplementation in aging roosters' diet enhanced the sperm motility, seminal and blood antioxidant capacity, and testicular spermatogenenic indexes.

Evaluation of the Potential of a Lectin Extracted from Polygonumpersicaria L. as a Biorational Agent against Sitophilusoryzae L.

Rice weevil, Sitophilus oryzae L. (Coleoptera: Curculionidae), is one of the most destructive stored-product pests that is resistant to a wide range of chemical insecticides. In the present study, we investigated whether a lectin extracted from Polygonum persicaria L. (PPA) can be used as a biorational agent to control such insect pests. Along with the lethal digestive assay, the sub-lethal insecticidal activities of PPA, including the effects on digestive, detoxifying, and antioxidant enzyme activities, were evaluated against S. oryzae adults. The effect of feeding a diet containing PPA and carob extract as a food attractant on the mortality of S. oryzae adults was also investigated. Feeding on the diet containing PPA resulted in a significant mortality of S. oryzae adults with a LC50 (Lethal Concentration to kill 50% of insects) of 3.68% (w/w). The activity of digestive enzymes, including α-amylase, α-glucosidase, TAG-lipase, trypsin, chymotrypsin, elastase, and carboxy- and aminopeptidase, were decreased by the sub-lethal concentration of PPA. Detoxifying and antioxidant enzymes, including esterase, superoxide dismutase, catalase, glutathione-S-transferase, ascorbate peroxidase, glucose 6-phosphate dehydrogenase, and malondialdehyde, were activated in adults affected by PPA. These findings indicated that PPA, in addition to causing digestive disorders, leads to oxidative stress in S. oryzae. The presence of carob extract had no effect on the PPA-induced mortality of the insect. According to the results of the present study, PPA has promising insecticidal efficiency against S. oryzae. In addition, the usage of PPA with a food attractant carob extract in bait traps can be recommended as a new biorational formulation in S. oryzae management.

Dietary Supplementation of Carob and Whey Modulates Gut Morphology, Hemato-Biochemical Indices, and Antioxidant Biomarkers in Rabbits.

This study aimed to evaluate the influence of the carob powder (CP) and sweet whey powder (WhP) inclusion into weaning feed on the gut morphology, hemato-biochemical parameters, and antioxidant biomarkers. The addition of 10 g/kg (basal diet +10 g/kg of CP, of WhP) or the mixture (5 g/kg of CP and 5 g/kg of WhP) in the rabbit's standard diet was assessed. A total of 40 weaned New Zealand White rabbits (4 weeks old) was distributed according to body weight (BW) into four treatments (n = 10) and the feed additives were provided for 7 weeks. Tissue samples and blood were obtained after slaughter. Final BW, daily weight gain, feed conversion ratio, intestinal morphology, and carcass dressing were positively affected by CP-WhP treatments compared with the control diet. Also, CP-WhP treatments significantly increased total proteins, calcium and iron levels, fecal cholesterol excretion, total antioxidants capacity, superoxide dismutase, and catalase in different tissues and significantly decreased total cholesterol, triglycerides, and glucose in blood serum. These changes were associated with a diminution of blood tumor necrosis factor alpha, lipid peroxidation, and carbonyl proteins in rabbit tissues. Both the additives separately and especially in the mix may enhance productive performance, protein profile, gut function, immunity, and antioxidant activity, with reducing lipid peroxidation, essential inflammatory mediator, and protein-carbonyl residues of growing rabbits. These findings suggest that CP-WhP dietary supplementation provides novel insights into a variety of bioactive compound mixtures with different beneficial modes of actions.

The Effects of Carob (Ceratonia siliqua L.) on Emotional Behavior Impairment and Metabolic Disorders Induced by Estrogen Deficiency in Rats.

Carob (Ceratonia siliqua L.) contains a wide variety of polyphenols with high antioxidant properties. In this study, we investigated the effects of aqueous extract of carob pods (AECP) on emotional behavior impairments and metabolic disorders in ovariectomized (OVX) rats. Female Wistar rats were assigned to three groups: group 1, control non-OVX rats; group 2, OVX rats; and group 3, OVX rats orally treated with AECP (500 mg/kg) for15 days after ovariectomy. Elevated plus-maze and open-field tests were performed on the 26th and 27th post-ovariectomy days, respectively. Afterwards, the rats were anesthetized and their serums were collected for biochemical analysis. We found that AECP improved emotional behavior impairments revealed by elevated plus-maze and open-field tests in OVX rats. Moreover, ovariectomy significantly increased triglyceride, lactate dehydrogenase, alanine aminotransferase, and aspartate aminotransferase levels in the serum. AECP administration significantly reversed ovariectomy-induced biochemical alterations. Thus, we suggest that the AECP may have an anxiolytic-like effect and prevent biochemical disorders associated with menopause or ovariectomy.

Optimization of lactic acid production using immobilized Lactobacillus Rhamnosus and carob pod waste from the Lebanese food industry.

The valorization of a solid carob waste from the Lebanese industry was investigated by optimizing the production of lactic acid using immobilized Lactobacillus rhamnosus in alginate beads and response surface methodology. The results showed that pH and alginate concentration had a significant effect on the production of lactic acid. The fermentation of non-enriched carob waste juice needed an additional nitrogen source to improve lactic acid production and yield. From extracts with 65 g/L sugars, the optimum conditions were found to be 2% for the concentration of alginate, 4% bacteria cells entrapped in beads, 80 rpm agitation speed and pH 6.4. Lactic acid concentration obtained under these conditions was 22 g/L with a yield of 76.9 g/g consumed sugar and a productivity of 1.22 g/L/h. The use of invertase pretreatment increased lactic acid concentration from 22 to 40 g/L, but reduced yield at 66.6%. Finally, cells immobilized in alginate beads could be used for at least five successive cycles.

ß-Mannanase production and kinetic modeling from carob extract by using recombinant Aspergillus sojae.

The main objectives of this study were to optimize ß-mannanase fermentation conditions by using Response Surface Methodology (RSM) and to model kinetically using the kinetic models. Based on the results, the optimum fermentation conditions were found to be initial sugar concentration of 10°Bx, whey concentration of 0.75% [w/v], and inoculum size of 8% (v/v). Under optimized conditions, ß-mannanase activity (P), sugar consumed (ΔS), maximum ß-mannanase production rate (QP ), and sugar utilization yield (SUY) were 687.89 U/mL, 47.38 g/L, 118.54 U mL-1 day-1 , and 69.73%, respectively. Kinetic models were employed to describe the optimum ß-mannanase fermentation process. The kinetic analysis of ß-mannanase fermentation showed that ß-mannanase fermentation is growth associated because the α value (U/mgX) is approximately 330-fold higher than the ß value (U/mgX·hr). Nevertheless, maintenance value (Z) was lower than γ value, thus showing that Aspergillus niger mainly utilizes the sugars for ß-mannanase production and fungal growth. Consequently, carob extract and whey powder could be used to be cost-effective carbon and organic nitrogen sources, respectively. It was clearly indicated that the suggested kinetic models can successfully describe the fungal growth, ß-mannanase production, and substrate consumption.

Can Carob-Fruit-Extract-Enriched Meat Improve the Lipoprotein Profile, VLDL-Oxidation, and LDL Receptor Levels Induced by an Atherogenic Diet in STZ-NAD-Diabetic Rats?

Carob fruit extract (CFE) has shown remarkable in vitro antioxidant properties and reduces postprandial hyperglycemia and hyperlipidemia in healthy animals. Development of functional meat products that contain bioactive components are presented as a great nutritional strategy. Until now, the effect of the consumption of restructured meat enriched with CFE in a murine model of diabetes has not been investigated. The objective of this study was to evaluate the effect on glycemia, lipemia, lipoprotein profile, Ldlr, arylesterase (AE), and very low-density lipoproteins (VLDL) and liver oxidation in streptozotocin-nicotinamide (STZ-NAD) growing Wistar diabetic rats fed restructured meat in the frame of a high cholesterol/high saturated-fat diet. In the present study, three groups (D, ED and DE) were fed cholesterol-enriched (1.4% cholesterol and 0.2% cholic acid) and high saturated-fat diets (50% of total energy from fats and 20.4% from saturated fatty acids). Rats were subjected to a STZ-NAD administration at the 3rd week. Group D did not receive CFE, while ED and DE rat groups received CFE before and after the diabetic induction, respectively. After eight weeks, D rats showed hyperglycemia and hypercholesterolemia, an increased amount cholesterol-enriched VLDL (ß-VLDL), IDL and LDL particles and triglyceride-enriched HDL. ED and DE partially blocked the hypercholesterolemic induction with respect to D group (p < 0.001) and improved glycemia, cholesterol levels, lipoprotein profile, Ldlr, plasma AE activity and liver oxidation (p < 0.001). Fecal fat, moisture and excretion were higher while dietary digestibility was lower in ED and DE vs. D counterparts (p < 0.001). In conclusion, CFE-enriched meat shows, for the first time, hypoglycemic and hypolipidemic effects in STZ-NAD animals fed high cholesterol/high saturated-fat diets. Likewise, it manages to reverse possible diabetes lipoprotein alterations if CFE-enriched meat is consumed before pathology development or improves said modifications if Type 2 Diabetes Mellitus is already established.

Effect of Wakame and Carob Pod Snacks on Non-Alcoholic Fatty Liver Disease.

Snacks combining different functional ingredients could represent a useful therapeutic strategy against NAFLD. The present study aimed to analyze the effect of two snack formulations based on carob and wakame flour in the treatment for NAFLD in rats. For this purpose, metabolic syndrome was induced in 50 adult rats by a high-fat high-fructose diet over eight weeks. After this period, rats were fed either normal calorie diets supplemented or not with snack A (1/50 wakame/carob pod) and snack B (1/5 wakame/carob pod) for four additional weeks. After sacrifice, liver composition and serum parameters were analyzed. Different pathways of triacylglycerol metabolism in liver were studied including fatty acid oxidation, fatty acid synthesis, triglyceride assembly and release, fatty acid uptake and glucose uptake. Oxidative stress was also measured. Snack treatment, and mainly B snack, reduced liver triacylglycerol levels by increasing fat oxidation. Moreover, this snack reduced oxidative stress. Therefore, this snack formulation could represent an interesting tool useful for fatty liver treatment.

Potential Usefulness of a Wakame/Carob Functional Snack for the Treatment of Several Aspects of Metabolic Syndrome: From In Vitro to In Vivo Studies.

Metabolic syndrome (MetS) greatly increases the risk of cardiovascular diseases and type 2 diabetes mellitus. The aim of this study was to evaluate the efficacy of functional snacks containing a combination of wakame (W) and carob pod (CP) flours in reducing markers associated with MetS. The mechanisms of action underlying these effects were also evaluated. In vitro approaches were carried out in mature 3T3-L1 adipocytes and RAW 264.7 macrophages treated with different doses of extracts from W, CP, or a combination of both. Furthermore, an in vivo experiment was conducted in rats with MetS treated with normal-caloric diets containing different snack formulations with combinations of 1/50 (snack A) or 1/5 of wakame/carob (snack B). In vitro experiments results indicated that both W and CP had delipidating effects, but only the latter induced anti-inflammatory and anti-hypertensive effects. As far as the in vivo study is concerned, snack B was ineffective and snack A showed an anti-hypertensive effect in rats with MetS. The present study shows for the first time the in vitro efficacy of a W and CP combination as an anti-inflammatory, delipidating, and anti-hypertensive tool, and its potential usefulness in treating MetS.

Antioxidant Activity and Protective Effect of Carob Honey in CCl4-induced Kidney and Liver Injury.

BACKGROUND/AIM: Various honey samples exhibited protective effect against drug and chemical induced toxicity. The study was designed to determine the antioxidant content and activity of carob honey and to investigate its hepato-renal protective effect in carbon tetrachloride (CCl4) induced kidney and liver injury in rats. MATERIAL AND METHODS: Phenolic, flavone and flavonol in carob honey were quantified. DPPH, ABTS•+, ferric reducting antioxidant power, and total antioxidant activity were used to evaluate the antioxidant activity. Rats were used for the experiment, and received either intraperitoneal injection of CCl4 (1 mL/kg.b.wt); honey (orally, 2 g/kg.b.wt) and CCl4; or honey. Liver and kidney function parameters were assessed. Oxidative parameters including lipid peroxidation (MDA), protein carbonyl formation (PCO), advanced protein oxidation products (AOPP), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), and ascorbic acid were measured in the kidney and liver tissues. RESULTS: CCl4 caused a significant elevation of liver enzymes, lactic acid dehydrogenase, blood glucose, uric acid, blood urea and serum creatinine as compared to the control group. Also, it significantly increased MDA, PCO and AOPP level, and markedly decreased GHS, ascorbic acid, CAT and GPx in the liver and kidney tissues. These changes were significantly ameliorated by carob honey before and after CCl4 administration. Honey alone did not cause significant changes as compared to the control group. CONCLUSION: The data showed for the first time that carob honey has high antioxidant content, antioxidant property, and protective effect against CCl4 induced kidney and liver toxicity by maintaining the activity of antioxidant defense system.