Botanical Briefs: Neem Oil (Azadirachta indica).

Azadirachta indica, commonly known as neem, has many uses as a natural remedy. We review and discuss the pharmacologic, biologic, and medicinal properties of neem in disease management. We also report a rare clinical case of a 77-year-old man who presented with a hypopigmented rash on the lower back, bilateral flanks, and buttocks after 6 months of repeated application of neem oil to treat persistent arthritis and lower back pain.

Performance and gut health status of broilers fed diets supplemented with two graded levels of a monoglyceride blend.

The present study was undertaken to evaluate the efficacy of 2 graded levels (0.03 and 0.05% of diet) of a monoglyceride blend containing butyric, caprylic, and capric acids in broilers' diet for optimizing gut structure and animal growth performance. For this purpose, a total of 210, one-day-old male Ross 308 broiler chicks were randomly allocated to 3 experimental treatments using 7 replicates each and 10 birds/replicate. The treatment groups involved supplementation of blend of short and medium chain fatty acids at the level of 0, 0.03, and 0.05% of the diet for 42 d. The incorporation of mixes of monoglycerides into broilers' diet linearly improved BWG between d 0 and 21 (P = 0.034). At the end of trial, however, no significant changes were observed in performance indexes (BWG, FI, FCR). Jejunal morphometric parameters (villus height, crypt depth, and their ratio) remained unaltered with the monoglyceride supplementation on d 21. The results further showed that monoglycerides supplementation increased the goblet cell counts along the jejunal villi (P = 0.034) and crypt regions (P = 0.022), as well as it effectively modulated the mRNA abundances of tight junction protein (ZO-1, P = 0.033) and nutrient transporters (SGLT, PePT1; P = 0.005, 0.023, respectively) in the jejunum. Moreover, the downregulation in mRNA abundance of TNFα (P = 0.030) was observed with the monoglyceride supplementation. The SCFAs analysis of cecal contents showed no notable differences with monoglyceride blend supplementation when compared to the unsupplemented group. Collectively, high goblet cell numbers in the jejunum along with downregulation of the mRNA abundances of pro-inflammatory cytokines, upregulation of tight junction proteins, and nutrient transporters showed favorable responses of low doses of monoglycerides blend in broiler feeding. Further studies should be conducted in different rearing conditions to examine the effectiveness of such low levels of a monoglyceride blend in the modulation of gut structure, its functionality and animal performance.

In vitro antibacterial activity of nimbolide against Helicobacter pylori.

ETHNOPHARMACOLOGICAL RELEVANCE: Nimbolide is one of hundreds of phytochemicals that have been identified within the neem tree (Azadirachta indica A. Juss). As an evergreen tree native to the Indian subcontinent, components of the neem tree have been used for millennia in traditional medicine to treat dental, gastrointestinal, urinary tract, and blood-related ailments, ulcers, headaches, heartburn, and diabetes. In modern times, natural oils and extracts from the neem tree have been found to have activities against a variety of microorganisms, including human pathogens. AIM OF THE STUDY: Helicobacter pylori, a prevalent gastric pathogen, shows increasing levels of antibiotic resistance. Thus, there is an increasing demand for novel therapeutics to treat chronic infections. The in vitro activity of neem oil extract against H. pylori was previously characterized and found to be bactericidal. Given the numerous phytochemicals found in neem oil extract, the present study was designed to define and characterize specific compounds showing bactericidal activity against H. pylori. MATERIALS AND METHODS: Azadirachtin, gedunin, and nimbolide, which are all common in neem extracts, were tested for antimicrobial activity; the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for nine strains of H. pylori. The specific properties of nimbolide were further characterized against H. pylori strain G27. Bactericidal kinetics, reversibility, effectiveness at low pH, and activity under bacteriostatic conditions were examined. The hemolytic activity of nimbolide was also measured. Finally, neem oil extract and nimbolide effectiveness against H. pylori biofilms were examined in comparison to common antibiotics used to treat H. pylori infection. RESULTS: Nimbolide, but not azadirachtin or gedunin, was effective against H. pylori; MICs and MBCs against the nine tested strains ranged between 1.25-5 µg/mL and 2.5-10 µg/mL, respectively. Additionally, neem oil extract and nimbolide were both effective against H. pylori biofilms. Nimbolide exhibited no significant hemolytic activity at biologically relevant concentrations. The bactericidal activity of nimbolide was time- and dose-dependent, independent of active H. pylori growth, and synergistic with low pH. Furthermore, nimbolide-mediated H. pylori cell death was irreversible after exposure to high nimbolide concentrations (80 µg/mL, after 2 h of exposure time and 40 µg/mL after 8 h of exposure). CONCLUSIONS: Nimbolide has significant bactericidal activity against H. pylori, killing both free living bacterial cells as well as cells within a biofilm. Furthermore, the lack of hemolytic activity, synergistic activity at low pH and bactericidal properties even against bacteria in a state of growth arrest are all ideal pharmacological and biologically relevant properties for a potential new agent. This study underscores the potential of neem oil extract or nimbolide to be used as a future treatment for H. pylori infection.

New Pyridone-Based Derivatives as Cannabinoid Receptor Type 2 Agonists.

The activation of the human cannabinoid receptor type II (CB2R) is known to mediate analgesic and anti-inflammatory processes without the central adverse effects related to cannabinoid receptor type I (CB1R). In this work we describe the synthesis and evaluation of a novel series of N-aryl-2-pyridone-3-carboxamide derivatives tested as human cannabinoid receptor type II (CB2R) agonists. Different cycloalkanes linked to the N-aryl pyridone by an amide group displayed CB2R agonist activity as determined by intracellular [cAMP] levels. The most promising compound 8d exhibited a non-toxic profile and similar potency (EC50 = 112 nM) to endogenous agonists Anandamide (AEA) and 2-Arachidonoylglycerol (2-AG) providing new information for the development of small molecules activating CB2R. Molecular docking studies showed a binding pose consistent with two structurally different agonists WIN-55212-2 and AM12033 and suggested structural requirements on the pyridone substituents that can satisfy the orthosteric pocket and induce an agonist response. Our results provide additional evidence to support the 2-pyridone ring as a suitable scaffold for the design of CB2R agonists and represent a starting point for further optimization and development of novel compounds for the treatment of pain and inflammation.

Effects of α-glyceryl monolaurate on growth, immune function, volatile fatty acids, and gut microbiota in broiler chickens.

This study was conducted to determine the effects of dietary addition of α-glyceryl monolaurate (α-GML) on growth performance, immune function, volatile fatty acids production and cecal microbiota in broiler chickens. A total of 480 1-day-old yellow-feathered broilers were randomly assigned in equal numbers to 4 dietary treatments: basal diet (NCO) or supplementations with 30 mg/kg bacitracin (ANT), 500 mg/kg α-GML, or 1,000 mg/kg α-GML (GML2). And, each treatment contained 8 replicates with 15 chickens per replicate. After supplementation with α-GML, the total BW gain and average daily weight gain of broilers increased significantly (P < 0.05) compared with the broilers on the NCO diet. Moreover, compared with the NCO group, higher levels of immune globulin M and immune globulin Y were observed in both GML groups and the ANT group. Concentrations of acetate, propionate, butyrate, valerate, and isovalerate in GML2 were significantly higher (P < 0.05) than those in the NCO group on day 28. However, acetate, propionate, valerate, and isovalerate concentrations were reduced to significantly (P < 0.05) lower than those in the NCO group on day 56. The abundance and diversity of microbiota were found to be improved in broilers that were supplemented with GML, using operational taxonomic unit and diversity analyses. Furthermore, the GML treatments increased favorable microbiota, particularly acid-producing bacteria, on day 28 and, also, reduced opportunistic pathogens, such as Alistipes tidjanibacter and Bacteroides dorei by day 56. These results suggest that α-GML supplementation modulates cecal microbiota and broiler immunity and improves volatile fatty acid levels during the early growth stages of broilers.

Monocaprin eye drop formulation to combat antibiotic resistant gonococcal blindness.

Neisseria gonorrhoeae bacteria are acknowledged as an urgent threat to human health because this species has developed resistances to all of the antibiotics used clinically to treat its infections. N. gonorrhoeae causes the sexually transmitted disease gonorrhoea, but also causes blindness when the bacteria infect the eyes. Infants are particularly susceptible, acquiring the infection from their mothers at birth. We have shown that the monoglyceride monocaprin rapidly kills N. gonorrhoeae and other bacterial species and is non-irritating in ocular assays. Here we show that the physical and chemical properties of monocaprin make it ideal for use in a thickened eye drop formulation to combat eye infections. Monocaprin-containing formulations were assessed using analytical techniques and for antimicrobial activity in vitro and in ex vivo infections. Monocaprin-containing formulations retained activity after three years and are non-irritating, unlike preparations of povidone iodine in our assays. A recommended formulation for further development and investigation is 0.25% monocaprin in 1% HPMC with 1% polysorbate 20.

Inhibition of monoacylglycerol lipase reduces nicotine reward in the conditioned place preference test in male mice.

Nicotine, the primary psychoactive component in tobacco, plays a major role in the initiation and maintenance of tobacco dependence and addiction, a leading cause of preventable death worldwide. An essential need thus exists for more effective pharmacotherapies for nicotine-use cessation. Previous reports suggest that pharmacological and genetic blockade of CB1 receptors attenuate nicotine reinforcement and reward; while exogenous agonists enhanced these abuse-related behaviors. In this study, we utilized complementary genetic and pharmacologic approaches to test the hypothesis that increasing the levels of the endocannabinoid 2-arachindonoylglycerol (2-AG), will enhance nicotine reward by stimulating neuronal CB1 receptors. Contrary to our hypothesis, we found that inhibition of monoacylglycerol lipase (MAGL), the primary catabolic enzyme of 2-AG, attenuates nicotine conditioned place preference (CPP) in mice, through a non-CB1 receptor-mediated mechanism. MAGL inhibition did not alter palatable food reward or Lithium Chloride (LiCl) aversion. In support of our findings, repeated MAGL inhibition did not induce a reduction in CB1 brain receptor levels or hinder function. To explore the potential mechanism of action, we investigated if MAGL inhibition affected other fatty acid levels in our CPP paradigm. Indeed, MAGL inhibition caused a concomitant decrease in arachidonic acid (AA) levels in various brain regions of interest, suggesting an AA cascade-dependent mechanism. This idea is supported by dose-dependent attenuation of nicotine preference by the selective COX-2 inhibitors valdecoxib and LM-4131. Collectively, these findings, along with our reported studies on nicotine withdrawal, suggest that inhibition of MAGL represents a promising new target for the development of pharmacotherapies to treat nicotine dependence.

Trilactic glyceride regulates lipid metabolism and improves gut function in piglets.

Glycerol-lactate esters are energy supplements for exercise, but effects of trilactic glyceride (TLG) on intestinal function and hepatic metabolism are unknown. We found that dietary supplementation with 0.5% TLG to weanling piglets decreased plasma concentrations of low-density lipoprotein and gamma-glutamyl transferase but increased those of D-xylose and high-density lipoprotein. TLG supplementation enhanced mRNA levels for fatty acid synthase (FASN) and SLC27A2 in white adipose tissue; insulin receptor in duodenum; aquaporin-8 in ileum, jejunum and colon; aquaporin-10 in duodenum and ileum; nuclear factor like-2 in jejunum and colon; glutathione S-transferase and phosphoenolpyruvate carboxykinase-1 in intestines; and abundances of claudin-1 and occludin proteins. TLG supplementation decreased mRNA levels for: hepatic hormone-sensitive lipase E, lipoprotein lipase, FASN, insulin-like growth factor-binding protein-3, and SLC27A2; and intestinal lipoprotein lipase, FASN and NADPH oxidase. Furthermore, TLG supplementation enhanced abundances of genus Bifidobacterium, while reducing abundances of family Enterobacteriaceae in ileum, colon and cecum; jejunal caspase-3 protein and diarrhea rate. In conclusion, dietary supplementation with TLG modulated lipid metabolism and alleviated diarrhea by improving intestinal function and regulating intestinal microflora in piglets.

A comparison of the efficacy of two commercial acaricides (fipronil and amitraz) with Azadirachta indica (neem) on the brown dog tick (Rhipicephalus sanguineus) from canines in Trinidad.

The brown dog tick (Rhipicephalus sanguineus) is prevalent on canids in Trinidad. It is directly (by causing anaemia) and indirectly (by acting as a vector of tick-borne pathogens) responsible for morbidity and mortalities in the canine population. The most commonly used commercial acaricides available to pet owners in Trinidad are amitraz and fipronil. Often, these acaricides may be abused and misused in a desperate attempt to rid pets of ticks. The objective of this study was to compare the efficacy of amitraz and fipronil with the herbal alternative, neem (Azadirachta indica). Triplicate in vitro trials utilizing the Larval Packet Test (LPT) were conducted using three concentrations (low, recommended and high) of fipronil (0.025%, 0.05% and 0.1%), amitraz (0.01%, 0.02% and 1%), neem oil (10%, 20% and 40%) and neem leaf extract (0.25%, 0.5% and 2%) for each trial. Statistical analysis using the mixed-effect Poisson regression analysis indicated that there was a significant difference (p < .05) in the survival of ticks pre-treatment versus post-treatment with amitraz, fipronil and all controls when compared to the neem oil. Fipronil and amitraz caused ≥99% mortality for all concentrations used in this study. Mortalities for neem oil and neem leaf extract ranged from 72.7% to 82% and 38% to 95.3%, respectively, with the greatest percentage of mortalities occurring at the lower concentrations. Neem oil and neem leaf extract can be used as alternative acaricides, and however, they are less efficacious against the brown dog tick than amitraz and fipronil.

Preparation of Monoacylglycerol Derivatives from Indonesian Edible Oil and Their Antimicrobial Assay against Staphylococcus aureus and Escherichia coli.

In the present work, linoleic acid and oleic acid were isolated from Indonesian corn oil and palm oil and they were used to prepare monoacylglycerol derivatives as the antibacterial agent. Indonesian corn oil contains 57.74% linoleic acid, 19.88% palmitic acid, 11.84% oleic acid and 3.02% stearic acid. While Indonesian palm oil contains 44.72% oleic acid, 39.28% palmitic acid, 4.56% stearic acid and 1.54% myristic acid. The oleic acid was purified by using Urea Inclusion Complex (UIC) method and its purity was significantly increased from 44.72% to 94.71%. Meanwhile, with the UIC method, the purity of ethyl linoleate was increased from 57.74% to 72.14%. 1-Monolinolein and 2-monoolein compounds were synthesized via two-step process from the isolated linoleic acid and oleic acid, respectively. The preliminary antibacterial assay shows that the 1-monolinolein did not give any antibacterial activity against Staphylococcus aureus and Escherichia coli, while 2-monoolein showed weak antibacterial activity against Staphylococcus aureus.

Effects of graded levels of Azadirachta indica seed oil on growth performance and biochemical profiles of broiler chickens.

Because of speculated risk in generating antibiotic resistance in pathogenic microbiota, natural products from plant origin due to their diverse biological activities, have recently gained a great attention in animal nutrition. This study was designed to evaluate graded levels of neem seed oil on growth performance of broiler chickens. A total of 400-day-old chicks were randomly allocated to five experimental treatment groups. Experimental rations consisted of supplementing basal diet (R0- ) with 1 g antibiotic (R0 + ), 15, 20 and 25 g neem seed oil/kg of feed. Data were recorded on feed intake, weight gain, feed conversion ratio, and serum biochemical parameters. Result revealed that feeding broiler chicks with 25 g of neem oil/kg of feed resulted in a marked (P < 0.05) decreased in feed intake as compared to the other treatments in the starter phase. The average live body weight and the weight gain decreased with increasing level of neem oil in the ration. Supplementation of poultry feed with graded levels of neem oil has no marked (P > 0.05) effect on carcass yield and relative weight of organs except for liver weight which significantly (P < 0.05) increased with the highest dose of neem oil. Serum content in total proteins, total cholesterol, HDL-cholesterol, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatinine were not significantly (P > 0.05) affected by the graded levels of neem oil. LDL-cholesterol significantly (P < 0.05) decreased with diets supplemented 20 g of neem oil/kg while triglycerides significantly increased with the highest doses of oil (20 and 25 g/kg) as compared to the negative and positive control rations. In conclusion, feeding broilers with Azadirachta indica seed oil has no beneficial effect on growth performance but may lead to the production of low-cholesterol chicken meat as demand by health-conscious consumers.

Monobutyrin Reduces Liver Cholesterol and Improves Intestinal Barrier Function in Rats Fed High-Fat Diets.

Butyric acid has been shown to reduce high-fat diet-related metabolic disturbances and to improve intestinal barrier function due to its potent anti-inflammatory capacity. This study investigates whether a butyric acid ester, monobutyrin (MB) affects lipid profiles and gut barrier function in a dose-response manner in rats fed butter- or lard-based high-fat diets. Four-week-old male Wistar rats were fed butter-based diets containing 0, 0.25, 0.75 and 1.5 MB g/100 g (dry weight basis) or 0.5 glycerol g/100 g, and diets with lard (La) containing 0 and 0.5 MB g/100 g or a low-fat control diet for 3⁻4 weeks. Lipid profiles in blood and liver tissue, intestinal permeability and cecal short-chain fatty acids were examined. The results showed a dose-dependent decrease in liver total cholesterol for 1.5 MB (p < 0.05) and liver triglycerides for 0.75 MB (p < 0.05) and 1.5 MB (p = 0.08) groups compared to the high-fat control group. Furthermore, a lower excretion of mannitol in urine in the 1.5 MB group indicated improved intestinal barrier function. When MB was supplemented in the lard-based diet, serum total cholesterol levels decreased, and total amount of liver high-density lipoprotein-cholesterol increased. Thus, MB dietary supplementation can be effective in counteracting lipid metabolism disturbances and impaired gut barrier function induced by high-fat diets.

Inhibitory Effect of Aqueous and Ethanolic Extracts of Neem Parts on Fungal Rot Disease of Solanum tuberosum.

BACKGROUND AND OBJECTIVE: The processing and preservation of Irish potato tubers like many other crops has been affected by various pathogens like fungi (particularly storage rot) which causes a major constraint to Irish potato production in Nigeria resulting to enormous post-harvest spoilage. The inhibitory effect of aqueous and ethanolic extracts of neem leaves, stem bark and seeds on fungal rot disease of Solanum tuberosum (Irish potato) as an alternative treatment for fungal storage disease on Irish potato tubers cultivated in Nigeria has been determined in this study. MATERIALS AND METHODS: Neem parts were harvested from Wukari, Nigeria and were processed for the experiment. Isolation of fungi and sub-culturing of the isolates was carried out to obtain pure culture. Pathogenicity study was carried out and effect of the plant extracts on mycelia growth of the test fungi was studied using the food poisoning techniques. The qualitative and quantitative phytochemical studies of the neem parts were carried out using standard methods. RESULTS: Qualitative phytochemical screening of the selected parts of neem plant showed the presence of alkaloids, glycoside, flavonoids, carbohydrates, reducing sugar, steroids, tannins and saponins. The quantitative test revealed that the percentage quantity of phytochemicals in these plant parts ranged from 10.17-23.88%. Effect of the extracts on the test organisms was significant (p<0.05). Ethanolic extract of the seed (ESE) exhibited the highest inhibitory effect on Aspergillus niger (88.37%) followed by aqueous extract of stem bark (ABE) (87.21%), while aqueous seed extract (ASE), ethanolic leaves extract (ELE), aqueous leaves extract (ALE) and ethanolic stem bark extract (EBE) exhibited inhibition of 81.78, 77.52, 72.87 and 39.53%, respectively. The inhibitory effect of EBE was significantly (p<0.05) lower than that of other extracts, while there was no significant difference in the inhibitory effects of ESE, ASE and ABE compared to ketoconazole on Aspergillus niger. The ethanolic extracts of the seed and leaves had 100% inhibitory effect on Fusarium oxyporium with 87.60% inhibition by aqueous extract of the leaves. The seed and the leaves ethanolic extracts exhibited the highest inhibitory effects on Pythium spp. and Fusarium oxyporium, while the aqueous leaves extract showed the least inhibition on Fusarium oxyporium. CONCLUSION: The plant extracts in this study were found to be very effective in inhibiting fungal mycelia growth and hence can be potentially effective for preservation during storage of Irish potatoes to minimize post-harvest lost.

Characterization, anti-oxidative effect of grape seed powder and in silico affinity profiling of polyphenolic and extra-phenolic compounds for calpain inhibition.

Vitis vinifera grape is a highly cultivated crop and solid wastes generated by the wine industry are largely under exploited. Plentiful studies have intended analyzing the polyphenolic content of grape seeds but characterization of non phenolic compounds is rather scarce. The present study aimed at the selective extraction of lipid, phenolic and aqueous phases from grape seed powder (GSP) in order to establish their intimate composition, as well as their antioxidant and chelating properties underlying partly their biological effects. Major non phenolic compounds identified in the lipid phase were glyceryl-monostearate and 2-monostearin whereas fructofuranose and sucrose were the most abundant in the aqueous phase. Among the most abundant compounds detected in the various phases, the polyphenol quercetin exhibited the best affinity and free binding energy towards the active site of the calcium-dependent protease calpain. Polyphenols likely constitute the bioactive part of GSP that should be exploited as safe modulators of intracellular signaling which is likely at the basis of their health beneficial effects. Nevertheless other compounds as lipids or sugars should be valorized along with polyphenols to improve their bioavailability into highly protected organs as brain or eye.

Can exposure to neem oil affect the spermatogenesis of predator Ceraeochrysa claveri?

Novel biological control methods and integrated pest management strategies are basic requirements for the development of sustainable agriculture. As a result, there is a growing demand for research on the use of plant extracts and natural enemies such as the green lacewing, Ceraeochrysa claveri, as natural pest control methods. Studies have shown that although natural compounds such as neem oil (Azadirachta indica) are effective as pest control strategies, they also cause sublethal effects on nontarget insects, such as C. claveri. The aim of this study was to examine the effects of neem oil on C. claveri testes. C. claveri larvae were fed Diatraea saccharalis eggs, which were pretreated with 0.5%, 1%, and 2% neem oil. Testes were collected from larvae, pupae, and adults and analyzed using light and electron (transmission and scanning) microscopy. Changes in cellular stress and possible cell death were also determined by TUNEL assay and the marker HSP-70. The results showed that neem oil affects the organization and distribution of cysts in the testes and the normal sequence of cyst development, causing a delay in spermatogenesis in the testes of treated insects. Tests for cellular stress and DNA fragmentation indicated there was no cellular alteration in the treated groups. Although neem oil does not induce cell death or changes in HSP-70 expression, this biopesticide negatively impacts the process of spermatogenesis and could decrease the perpetuation of this species in the agroecosystem, indicating that the use of neem oil in association with green lacewings as a biological control should be carefully evaluated.

In vitro activity of neem (Azadirachta indica) oil extract against Helicobacter pylori.

ETHNOPHARMACOLOGICAL RELEVANCE: The neem tree (Azadirachta indica A.Juss), of the Meliaceae family, has been used in India for millennia in traditional medicine. Parts of the tree are used to treat problems with the gastrointestinal tract, urinary tract, and hair; to combat infections of smallpox and plasmodium; and to treat ulcers, diabetes, blood pressure, headache, and heartburn. Natural products and extracts from the tree have been reported to have antimicrobial, antifungal, and antiparasitic activities. AIM OF THE STUDY: Antibiotic resistance in the gastric pathogen Helicobacter pylori is increasing, and novel therapeutics to eradicate this bacterium are needed. Given the growing interest in the use of natural products as antimicrobials, this study was designed to examine the bactericidal effects of an extract of neem oil against H. pylori. MATERIALS AND METHODS: Neem oil was obtained from a commercial source and subjected to liquid-liquid extraction with diethyl ether and aqueous methanol; the methanol-soluble fraction was retained. The minimum inhibitory (MIC) and bactericidal (MBC) concentrations were determined against nine strains of H. pylori. Additionally, specific properties of the extract were characterized using H. pylori strain G27: bactericidal kinetics, reversibility, and effectiveness under growth arrest conditions and at low pH. The hemolytic activity of the extract was measured in vitro. RESULTS: The MIC and MBC of the extract against the H. pylori strains were between 25 and 51 µg/mL and 43-68 µg/mL, respectively. The bactericidal activity was time- and concentration-dependent, and at the highest concentrations (75-105 µg/mL), no detectable bacteria were present by 6 h. The activity of the extract was reversible, independent of H. pylori growth, and increased at low pH. The extract exhibited no appreciable hemolytic activity. CONCLUSIONS: Neem oil extract has significant bactericidal activity against H. pylori. The extract has several favorable pharmacological properties, including ability to kill non-growing bacteria, increased activity at low pH, and no hemolytic activity. The compound(s) present in the extract could potentially be used as a future treatment for H. pylori infection.

Medium-chain glycerides affect gut morphology, immune- and goblet cells in post-weaning piglets: In vitro fatty acid screening with Escherichia coli and in vivo consolidation with LPS challenge.

The influence of medium-chain glycerides on performance and gastrointestinal well-being in weaning piglets was assessed. First, caproic (C6), caprylic (C8) and capric (C10) acid activity against Escherichia coli was screened in vitro. Pig flora of the whole small intestine was used as inoculum. Seven in vitro incubations were done in duplicate at pH = 3 and 5: C10 (15 mM), C8 (12 mM), C6 (15, 12, 10 mM), a non-incubated-negative control and incubated negative control. Culture suspensions were plated on E. coli-selective agar. Controls showed bacterial growth. C6 and C8 showed no growth at both pH-values, where C10 showed growth at pH = 5. Secondly, an in vivo study was done with 80 weaned piglets over 42 days, housed in pens of eight animals (five pens/treatment), fed a basal diet containing broken rice/soya bean meal/fish meal and supplemented with C6 and C8 in medium-chain glyceride form (MCT6/8, 0.175%) or antibiotic growth promoter (AGP, 0.020%) (Kasetsart University, Thailand) serving as control. Feed intake, daily gain and feed-to-gain ratio did not differ between MCT6/8 and AGP. Per replicate, two random selected piglets were challenged intravenously with E. coli-lipopolysaccharide (LPS) or saline solution (S) at Days 21 and 28. All challenged animals were sacrificed; blood and digestive tract samples (jejunum/ileum) were collected at Day 35. LPS challenge consistently reduced villus height and crypt depth for MCT6/8 and AGP. However, LPS-challenged piglets supplemented with MCT6/8 restored villus height, where AGP did not. MCT6/8 piglets had higher serum IgA, more jejunal IgA-positive plasma cells and goblet cells than AGP. At the ileal level, results were similar, though less pronounced. The present study offers new insight in the benefits of MCT6/8 over AGP in the post-weaning period. There is in vitro anti-microbial action of C6 and C8 on E. coli. In vivo, MCT6/8 also has protective effects in the small intestine that may result in growth promotion.

Isolation and Identification of Benzochroman and Acylglycerols from Massa Medicata Fermentata and Their Inhibitory Effects on LPS-Stimulated Cytokine Production in Bone Marrow-Derived Dendritic Cells.

Massa Medicata Fermentata (MMF), known as Shenqu, is an important traditional Chinese medicine widely used to treat indigestion, vomiting, and diarrhea. In this study, a new benzochroman, 3(S)-3,4-dihydro-5,10-di-ß-d-glucopyranoside-2,2-dimethyl-2H-naphtho(2,3-b)pyran-3-ol (1), and five known galactosyl acylglycerols (2⁻6) were isolated from a methanol extract from MMF. In addition, their chemical structures were determined by chemical and spectroscopic methods, which were compared with the previously reported data. Furthermore, the effects of isolated compounds on lipopolysaccharide (LPS)-stimulated bone marrow-derived dendritic cells were investigated. Compounds 1⁻3 exhibited significant inhibitory effects on the LPS-induced production of IL-6 and IL-12 p40, with IC50 values ranging from 1.6 to 10.2 µM. Compounds 2 and 3 also exhibited strong inhibitory effects on the LPS-stimulated production of TNF-α with IC50 values of 12.0 and 11.2 µM, respectively. The results might provide a scientific basis for the development of the active components in MMF, as well as for novel anti-inflammatory agents.

In vitro skin penetration of petrolatum and soybean oil and effects of glyceryl monooleate.

OBJECTIVES: Petrolatum and soybean oil are common ingredients incorporated in topical skin formulations for skin protection and moisturization. However, the stratum corneum (SC) penetration kinetics of these two cosmetic ingredients has not been systematically studied. Glyceryl monooleate (GlyMOle) has been shown to enhance skin penetration of various compounds. It was hypothesized that GlyMOle could enhance skin penetration of petrolatum and soybean oil. This study aimed to examine the in vitro skin penetration of petrolatum and soybean oil in the presence or absence of GlyMOle. METHODS: Skin permeation experiments were conducted using the in vitro Franz diffusion cell model with split-thickness human skin and human epidermal membrane (HEM). The effect of permeant dose and the kinetics of permeant penetration were examined with and without GlyMOle in vitro. RESULTS: Petrolatum and soybean oil were found to permeate across HEM, and no effect of GlyMOle on skin permeation into the receptor chamber was observed. GlyMOle enhanced the penetration of petrolatum into the split-thickness skin at 50 µg dose (petrolatum:GlyMOle, 49 : 1, w/w). However, no effect of GlyMOle on petrolatum penetration was observed at 200 µg dose (of the same petrolatum:GlyMOle ratio), indicating a dose-dependent effect. GlyMOle at the level used in the study did not enhance the penetration of soybean oil with 50 and 200 µg doses at any timepoints. CONCLUSION: GlyMOle was a skin penetration enhancer for petrolatum under the in vitro conditions identified in this study.

2-Arachidonoylglycerol as a possible treatment for anorexia nervosa in animal model in mice.

We have investigated the effects of 0.001mg/kg 2-arachidonoylglycerol (2-AG) administered in combination with compounds present in the body alongside 2-AG like 2-palmitoylglycerol and 2-linoleylglycerol (also termed "entourage"), on cognitive function,food intake, and neurotransmitter levels in the hippocampus and hypothalamus of mice under diet restriction. Young female Sabra mice were treated with vehicle, 2-AG, 2-AG+entourage, 2-AG+entourage+5-(4-Chlorophenyl)-1-(2,4-dichloro-phenyl)- 4-methyl-N-(piperidin-1-yl)-1H-pyrazole-3-carboxamide (SR141716A, a CB1 antagonist) and SR141716A. The mice were fed for 2.5h a day for 14days. Cognitive function was evaluated by the eight arm maze test, and neurotransmitter (norepinephrine, dopamine, L-DOPA and serotonin) levels were measured in the hippocampus and hypothalamus by high-performance liquid chromatography-electrochemical detection. Food intake was increased by 2-AG and, to an even greater extent, by 2-AG+entourage. SR141716A reversed the effect of 2-AG+entourage. The administration of 2-AG+entourage improved cognitive function compared to the vehicle mice, and this improvement was blocked by SR141716A. 2-AG+entourage-treated mice showed an increase in norepinephrine (NE), dopamine and L-DOPA levels in the hippocampus. SR141716A normalized NE and L-DOPA levels. There were no significant changes in hypothalamic neurotransmitter levels. The use of very low doses of the endocannabinoid 2-AG+entourage can improve cognitive function by elevating norepinephrine and L-DOPA levels in the hippocampus, without cannabinomimetic side effects. These findings may have implications for cognitive enhancement in anorexia nervosa.