OBJECTIVE: To observe the effect of amygdalin on liver fibrosis in a liver fibrosis mouse model, and the underlying mechanisms were partly dissected in vivo and in vitro. METHODS: Thirty-two male mice were randomly divided into 4 groups, including control, model, low- and high-dose amygdalin-treated groups, 8 mice in each group. Except the control group, mice in the other groups were injected intraperitoneally with 10% carbon tetrachloride (CCl₄)-olive oil solution 3 times a week for 6 weeks to induce liver fibrosis. At the first 3 weeks, amygdalin (1.35 and 2.7 mg/kg body weight) were administered by gavage once a day. Mice in the control group received equal quantities of subcutaneous olive oil and intragastric water from the fourth week. At the end of 6 weeks, liver tissue samples were harvested to detect the content of hydroxyproline (Hyp). Hematoxylin and eosin and Sirius red staining were used to observe the inflammation and fibrosis of liver tissue. The expressions of collagen I (Col-I), alpha-smooth muscle actin (α-SMA), CD31 and transforming growth factor β (TGF-β)/Smad signaling pathway were observed by immunohistochemistry, quantitative real-time polymerase chain reaction and Western blot, respectively. The activation models of hepatic stellate cells, JS-1 and LX-2 cells induced by TGF-β1 were used in vitro with or without different concentrations of amygdalin (0.1, 1, 10 µmol/L). LSECs. The effect of different concentrations of amygdalin on the expressions of liver sinusoidal endothelial cells (LSECs) dedifferentiation markers CD31 and CD44 were observed. RESULTS: High-dose of amygdalin significantly reduced the Hyp content and percentage of collagen positive area, and decreased the mRNA and protein expressions of Col-I, α-SMA, CD31 and p-Smad2/3 in liver tissues of mice compared to the model group (P<0.01). Amygdalin down-regulated the expressions of Col-I and α-SMA in JS-1 and LX-2 cells, and TGFβ R1, TGFβ R2 and p-Smad2/3 in LX-2 cells compared to the model group (P<0.05 or P<0.01). Moreover, 1 and 10 µmol/L amygdalin inhibited the mRNA and protein expressions of CD31 in LSECs and increased CD44 expression compared to the model group (P<0.05 or P<0.01). CONCLUSIONS Amygdalin can dramatically alleviate liver fibrosis induced by CCl₄ in mice and inhibit TGF-β/Smad signaling pathway, consequently suppressing HSCs activation and LSECs dedifferentiation to improve angiogenesis.
Rats ; Male ; Mice ; Animals ; Transforming Growth Factor beta/metabolism* ; Amygdalin/therapeutic use* ; Endothelial Cells/metabolism* ; Olive Oil/therapeutic use* ; Rats, Wistar ; Smad Proteins/metabolism* ; Liver Cirrhosis/metabolism* ; Liver ; Transforming Growth Factor beta1/metabolism* ; Signal Transduction ; Collagen Type I/metabolism* ; Carbon Tetrachloride ; Hepatic Stellate Cells

ABSTRACT Phenolic compounds are secondary metabolites of plants metabolism and can be found in olive oil. They exhibit antimicrobial activity towards both gram-positive and gram-negative bacteria. However, little is known about the antibacterial activity of the compounds towards periodontopathogens. The study aimed to investigate the potential of these compounds as antibacterial agents towards pathogens, specifically Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum. Phenolic compounds were extracted from extra virgin olive oil (EVOO) through liquid-liquid separation using methanol:water (70:30), and hexane. It was then prepared in various concentrations to determine its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against the periodontopathogens. The anti-adhesion activity was quantified using crystal violet staining while the effects on the morphology were examined through scanning electron microscopy (SEM). The MICs of the phenolic compounds on A. actinomycetemcomitans, P. gingivalis and F. nucleatum were 31.25 mg/mL, 62.5 mg/mL and 125 mg/mL, respectively. The MBCs of the phenolic compounds on A. actinomycetemcomitans and F. nucleatum were 62.5 mg/mL and 125 mg/mL, respectively suggesting this compound can eradicate these bacteria. There was no bactericidal effect on P. gingivalis. The adhesion of all the bacteria was interrupted by the compounds at the lowest concentration (1.95 mg/mL). SEM findings showed disruption of bacterial cell surfaces such as blebs and disintegration of cells after exposure to this extract. Phenolic compounds of olive oil exhibited antibacterial activity against the tested pathogens, with bactericidal effects on A. actinomycetemcomitans and F. nucleatum and bacteriostatic effects on P. gingivalis.
Anti-Bacterial Agents ; Phenols ; Periodontal Diseases ; Olive Oil

To investigate the therapeutic effect of Jingfang Granules on carbon tetrachloride(CCl_4)-induced liver fibrosis in mice and its mechanism. Forty-nine 8-week-old male C57 BL/6 J mice were randomly divided into a blank group, a CCl_4 group, a silybin group(positive control, 100 mg·kg~(-1))+CCl_4, a Jingfang high-dose(16 g·kg~(-1)) group, a Jingfang high-dose(16 g·kg~(-1))+CCl_4 group, a Jingfang medium-dose(8 g·kg~(-1))+CCl_4 group, and a Jingfang low-dose(4 g·kg~(-1))+CCl_4 group, with 7 mice in each group. The mice in the blank group and Jingfang high-dose group were intraperitoneally injected olive oil solution, and mice in other groups were intraperitoneally injected with 10% CCl_4 olive oil solution(5 mL·kg~(-1)) to induce liver fibrosis, twice a week with an interval of 3 d, for 8 weeks. At the same time, except for the blank group and CCl_4 group, which were given deionized water, the mice in other groups were given the corresponding dose of drugs by gavage once daily for 8 weeks with the gavage volume of 10 mL·kg~(-1). All mice were fasted and freely drank for 12 h after the last administration, and then the eyeballs were removed for blood collection. The liver and spleen were collected, and the organ index was calculated. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bile acid(TBA), and triglyceride(TG) in the serum of mice were detected by an automated analyzer. Tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and interleukin-1β(IL-1β) levels were detected by enzyme-linked immunosorbent assay(ELISA). Kits were used to detect the contents of superoxide dismutase(SOD), malondialdehyde(MDA), and glutathione(GSH) in the liver tissue. Pathological changes in the liver tissue were observed by hematoxylin-eosin(HE), Masson, and Sirius red staining. Western blot was used to detect protein expressions of transforming growth factor-β(TGF-β), α-smooth muscle actin(α-SMA) and Smad4 in the liver tissue. The results indicated that Jingfang Granules significantly reduced the organ index, levels of ALT, AST, TBA,TG, TNF-α, IL-6, and IL-1β in the serum, and the content of MDA in the liver tissue of mice with CCl_4-induced liver fibrosis. Jingfang Granules also significantly increased the content of SOD and GSH in the liver tissue. Meanwhile, Jingfang Granules down-regulated the protein levels of TGF-β, α-SMA, and Smad4. Furthermore, Jingfang Granules had no significant effect on the liver tissue morphology and the above indexes in the normal mice. In conclusion, Jingfang Granules has obvious therapeutic effect on CCl_4-induced liver fibrosis, and its mechanism may be related to reducing the expression of pro-inflammatory factors, anti-oxidation, and regulating TGF-β/Smad4 signaling pathway.
Mice ; Male ; Animals ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Olive Oil/therapeutic use* ; Carbon Tetrachloride/metabolism* ; Liver Cirrhosis/metabolism* ; Liver ; Superoxide Dismutase/metabolism* ; Transforming Growth Factor beta/metabolism*

BACKGROUND/OBJECTIVES: To compare five indices of adherence to the Mediterranean Diet (MD) among adults living in the Mediterranean region. SUBJECTS/METHODS: A total of 100 healthy Lebanese adults aged between 18 and 65 years. Face-to-face interviews to collect sociodemographic and medical information, to take anthropometric measurements, and to fill a validated, culturally adapted, food frequency questionnaire (FFQ). The score for each item was calculated following the recommendations for each corresponding index. The five MD indices were Mediterranean Diet Scale (MDScale), Mediterranean Food Pattern (MFP), MD Score (MDS), Short Mediterranean Diet Questionnaire (SMDQ), and the MedDiet score. RESULTS: Significant correlations were detected between items with P-values < 0.001. Minimal agreement was seen between MDScale and MedDiet score and maximal agreement between MDS and MedDiet score. Univariate and multivariate analyses showed that MDS and MedDiet scores had significant correlations with fiber and olive oil intake, main components of the MD. MDScale showed a significant correlation with waist-to-hip ratio and with total energy intake but none of the five indices was correlated to body mass index (BMI). CONCLUSIONS: The indices that showed the highest correlation with variables related to the MD are the MDScale and the MedDiet score; therefore, they can be used to assess our future study populations. Based on the current results, more than half of the study population was non-adherent to the MD and adherence to this diet did not appear to protect against being overweight (BMI ≥ 30).
Adult ; Body Mass Index ; Diet ; Diet, Mediterranean ; Energy Intake ; Humans ; Mediterranean Region ; Multivariate Analysis ; Olive Oil ; Overweight ; Waist-Hip Ratio

Intestinal barrier dysfunction always accompanies cirrhosis in patients with advanced liver disease and is an important contributor facilitating bacterial translocation (BT), which has been involved in the pathogenesis of cirrhosis and its complications. Several studies have demonstrated the protective effect of Vitamin D on intestinal barrier function. However, severe cholestasis leads to vitamin D depletion. This study was designed to test whether vitamin D therapy improves intestinal dysfunction in cirrhosis. Rats were subcutaneously injected with 50% sterile CCl₄ (a mixture of pure CCl₄ and olive oil, 0.3 mL/100 g) twice a week for 6 weeks. Next, 1,25(OH)₂D₃(0.5 µg/100 g) and the vehicle were administered simultaneously with CCl₄ to compare the extent of intestinal histologic damage, tight junction protein expression, intestinal barrier function, BT, intestinal proliferation, apoptosis, and enterocyte turnover. Intestinal heme oxygenase-1 (HO-1) expression and oxidative stress were also assessed. We found that vitamin D could maintain intestinal epithelial proliferation and turnover, inhibit intestinal epithelial apoptosis, alleviate structural damage, and prevent BT and intestinal barrier dysfunction. These were achieved partly through restoration of HO-1 and inhibition of oxidative stress. Taken together, our results suggest that vitamin D ameliorated intestinal epithelial turnover and improved the integrity and function of intestinal barrier in CCl₄-induced liver cirrhotic rats. HO-1 signaling activation was involved in these above beneficial effects.
Animals ; Apoptosis ; Bacterial Translocation ; Cholestasis ; Enterocytes ; Fibrosis ; Heme Oxygenase-1 ; Heme ; Humans ; Liver ; Liver Diseases ; Olive Oil ; Oxidative Stress ; Rats ; Tight Junctions ; Vitamin D ; Vitamins

An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca²⁺ and was slightly inhibited by Mn²⁺ and Zn²⁺ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.
Ascomycota/enzymology* ; Calcium ; Catalysis ; Corn Oil/metabolism* ; Detergents/chemistry* ; Enzyme Stability ; Fungal Proteins/chemistry* ; Glucans/chemistry* ; Hexanes/chemistry* ; Hydrogen-Ion Concentration ; Hydrolysis ; Industrial Microbiology ; Lipase/chemistry* ; Manganese/chemistry* ; Olive Oil/metabolism* ; Peanut Oil/metabolism* ; Sesame Oil/metabolism* ; Substrate Specificity ; Sunflower Oil/metabolism* ; Surface-Active Agents ; Temperature ; Zinc/chemistry*

PURPOSE: The aim of study was to determine the effects of carbohydrate, fat, protein, and fiber contents on glycemic responses in a single food item or meal. METHODS: Glycemic responses were measured in 30 healthy young adults (17 males and 13 females) with various test foods, including rice, egg whites, bean sprouts, olive oil, noodles, prune, broccoli, Korean dishes, Western dishes, and salad dishes, etc. Test foods were designed to contain various carbohydrate, fat, protein, and fiber contents in single or mixed foods or dishes. After 12 hours of fasting, participants consumed test foods, and the glycemic response was measured for a subsequent 120 min (0, 15, 30, 60, 90, and 120 min). Three hundred and fifty three glycemic responses from 62 foods were collected. The incremental area under the curve (AUC) was calculated for each test food for each subject to examine glycemic responses. Statistical analysis was conducted to identify which macronutrient (carbohydrate, fat, protein and fiber) affected the AUC using a mixed model. RESULTS: Carbohydrates (β= 37.18, p < 0.0001) significantly increased while fat (β= −32.70, p = 0.0054) and fiber (β= −32.01, p = 0.0486) significantly reduced the glycemic response. CONCLUSION: It can be concluded that the glycemic response of a meal can be modified depending on the fat and fiber contents of ingredient foods, even though carbohydrate content is maintained.
Area Under Curve ; Brassica ; Carbohydrates ; Egg White ; Fasting ; Glycemic Index ; Humans ; Male ; Meals ; Olive Oil ; Young Adult

BACKGROUND: The prevalence of allergic conditions is increasing in most countries. One possible explanation may be childhood nutrition. OBJECTIVE: The aim of the study was to investigate the relationship between consumption of pre-specified types of food in school-aged children and presence of respiratory and allergic symptoms. METHODS: A total of 191 students aged 9–11 years were recruited from 5 schools to geographically cover all of Malta. Data was collected between October 2011 and February 2012. This was part of a bigger study which included clinical and environmental tests besides standardized health questionnaires. For the purposes of this part of the study only the health questionnaires were used. These standardized health questionnaires based on the International Study of Asthma and Allergies in Childhood (ISAAC) were used to identify the presence of respiratory and allergic symptoms and to identify the types of foods and the frequency of consumption of various types of foods. RESULTS: We found that milk, meat, butter, olive oil, and yoghurt consumption had a negative association with allergic symptoms in children, whilst fish consumption had a detrimental effect. These relationships remained significant after correction for paternal level of education. CONCLUSION: The study highlights the fact that nutrition in early childhood may have a significant effect on the risk of allergic conditions. Our results, taken together with data in the literature, suggest that different types of fish might have had different effects. This is probably related to their different fatty acid constitution thus warranting further studies.
Asthma ; Butter ; Child ; Constitution and Bylaws ; Education ; Humans ; Hypersensitivity ; Malta ; Meat ; Milk ; Olive Oil ; Prevalence ; Yogurt

PURPOSE: This study is a comparative evaluation of the incidence of parenteral nutrition-associated liver disease (PNALD) when administering intravenous fat emulsions containing fish oil. METHODS: The medical records of patients who were in the neonatal intensive care unit at Severance Hospital from January, 2012 to December 2015, were reviewed retrospectively. Patients who were administered either soybean oil (SO) or SMOF (containing soybean oil, medium chain triglycerides, olive oil, and fish oil) more than 14 days were included. The patients were excluded if they were administered both agents or had underlying hepatic disease. An increase in bilirubin to 2 mg/dL was defined as PNALD. RESULTS: PNALD occurred in only 8 out of a total of 77 patients: 6 out of 31 (19.4%) in the SO group and 2 out of 46 (4.3%) in the SMOF group (P=0.055). The number of patients, whose lab values, such as direct bilirubin, total bilirubin, asparate aminotransferase (AST), alanine amino-transferase, gamma-glutamyl transpeptidase, C-reactive protein, serum triglyceride, and alkaline phosphate, exceeded the normal range, were similar in both groups. The gestational age, birth body weight, and APGAR score at 1 min and 5 min were significantly higher in the SO group and the PN duration was significantly long in the SMOF group. Considering only term infants, there were no significant differences in baseline characteristics and incidence of PNALD. The number of patients whose AST exceeded the normal range was significantly higher in the SO group (P=0.034). CONCLUSION: The incidence of PNALD was similar in both groups. On the other hand, considering the tendency, there was a high correlation between the type of lipid emulsion and an increased direct bilirubin level in the SO group.
Alanine ; Apgar Score ; Bilirubin ; Body Weight ; C-Reactive Protein ; Emulsions* ; Fat Emulsions, Intravenous ; Fish Oils ; gamma-Glutamyltransferase ; Gestational Age ; Hand ; Humans ; Incidence ; Infant ; Infant, Newborn* ; Intensive Care, Neonatal ; Liver Diseases* ; Liver* ; Medical Records ; Olive Oil ; Parenteral Nutrition ; Parturition ; Reference Values ; Retrospective Studies ; Soybean Oil ; Triglycerides

PURPOSE: Appropriate animal models of atherosclerotic plaque are crucial to investigating the pathophysiology of atherosclerosis, as well as for the evaluation of the efficacy and safety of vascular devices. We aimed to develop a novel animal model that would be suitable for the study of advanced atherosclerotic lesions in vivo. MATERIALS AND METHODS: Atherosclerotic plaque was induced in 24 iliac arteries from 12 rabbits by combining a high cholesterol diet, endothelial denudation, and injection into the vessel wall with either saline (n=5), olive oil (n=6), or inflammatory proteins [n=13, high-mobility group protein B1 (HMGB1) n=8 and tumor necrosis factor (TNF)-α n=5] using a Cricket™ Micro-infusion catheter. Optical coherence tomography (OCT) was performed to detect plaque characteristics after 4 weeks, and all tissues were harvested for histological evaluation. RESULTS: Advanced plaque was more frequently observed in the group injected with inflammatory proteins. Macrophage infiltration was present to a higher degree in the HMGB1 and TNF-α groups, compared to the oil or saline group (82.1±5.1% and 94.6±2.2% compared to 49.6±14.0% and 46.5±9.6%, p-value<0.001), using RAM11 antibody staining. On OCT, lipid rich plaques were more frequently detected in the inflammatory protein group [saline group: 2/5 (40%), oil group: 3/5 (50%), HMGB1 group: 6/8 (75%), and TNF-α group: 5/5 (100%)]. CONCLUSION: These data indicate that this rabbit model of atherosclerotic lesion formation via direct injection of pro-inflammatory proteins into the vessel wall is useful for in vivo studies investigating atherosclerosis.
Animals ; Cholesterol, Dietary/administration & dosage ; *Disease Models, Animal ; Endothelium/surgery ; HMGB1 Protein/*adverse effects ; Iliac Artery/diagnostic imaging/pathology/surgery ; Injections, Intra-Arterial ; Macrophages ; Male ; Olive Oil/adverse effects ; Plaque, Atherosclerotic/*chemically induced/diagnostic imaging/pathology ; Rabbits ; Sodium Chloride/adverse effects ; Tomography, Optical Coherence ; Tumor Necrosis Factor-alpha/*adverse effects