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Proceedings, 2020, IECN 2020

1st International Electronic Conference on Nutrients - Nutritional and Microbiota Effects on Chronic Disease

Online | 02-15 November 2020

Volume Editors:
Maria Luz Fernandez, University of Connecticut, USA
Maria Dolores del Castillo, Instituto de Investigación en Ciencias de la Alimentación (CIAL) (CSIC-UAM), Spain

Number of Papers: 31
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Cover Story (view full-size image): This volume provides scientific evidence on the effects of nutrients and bioactive compounds on microbiota and how they are related to health and chronic disease. In addition, the existing [...] Read more.
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225 KiB  
Proceeding Paper
Vitamin D Supplementation Is Associated with Disease Activity in Systemic Lupus Erythematosus Patients
by María Correa-Rodríguez, Gabriela Pocovi-Gerardino, Irene Medina-Martínez, Sara Del Olmo-Romero, Norberto Ortego-Centeno and Blanca Rueda-Medina
Proceedings 2020, 61(1), 1; https://doi.org/10.3390/IECN2020-07013 - 30 Oct 2020
Viewed by 973
Abstract
Systemic lupus erythematosus (SLE) is a chronic disease characterized by inflammatory response and abnormal autoimmune disease. Vitamin D is essential in phosphorus-calcium metabolism, has immunosuppressive properties, and is considered a therapeutic option. Controversy exists about the role of this vitamin in the pathogenesis [...] Read more.
Systemic lupus erythematosus (SLE) is a chronic disease characterized by inflammatory response and abnormal autoimmune disease. Vitamin D is essential in phosphorus-calcium metabolism, has immunosuppressive properties, and is considered a therapeutic option. Controversy exists about the role of this vitamin in the pathogenesis of SLE. Thus, the aim of this study was to investigate the influence of the dietary intake of vitamin D and its supplementation in a cohort of patients with SLE. A cross-sectional study including a total 285 patients with SLE was conducted (248 females and 26 males; mean age 46.99 ± 12.89 years). The SLE disease activity index (SLEDAI-2K) and the SLICC/ACR damage index (SDI) were used to assess disease activity and disease-related damage, respectively. Levels of C-reactive protein (CRP; mg/dL), homocysteine (Hcy; mol/L), anti-double stranded DNA antibodies (anti-dsDNA) (IU/mL), complement C3 (mg/dL), and complement C4 (mg/dL), among other biochemical markers, were measured. The dietary intake of vitamin D and the intake of vitamin D supplement were obtained via a 24-h patient diary. A share of 57.1% of the patients took vitamin D supplements and the average of dietary vitamin D was 2.08 ± 2.94 μg/day. Note that 98.2% of patients did not reach the recommended dietary intakes for vitamin D intake. Multivariate regression analysis revealed that clinical and laboratory variables are not significantly affected by vitamin D intake levels after adjusting for age, gender, energy intake, and medical treatment (immunosuppressants, corticosteroids, and antimalarials). Patients with SLE who took vitamin D supplements had significantly higher serum complement C3 levels compared to patients who did not take them after adjusting for covariates (110.28 ± 30.93 vs. 107.38 ± 24.18; p = 0.018). Our findings suggest a potential impact of supplementation of vitamin D on the activity of SLE. Future longitudinal research on SLE patients, including intervention trials, are required to validate these preliminary data. Full article
283 KiB  
Proceeding Paper
Elateriospermum tapos Supplementation in Dams Ameliorating Obesity Development and Stress Hormone Level among Adult Male Offspring
by Azrina Zainal Abidin, Santhra Segaran Balan, Kokila Vani Perumal, Nurul Husna Shafie, Maizaton Atmadini Abdullah, Azmiza Syawani Jasni, Khairul Kamilah Abdul Kadir and Hasnah Bahari
Proceedings 2020, 61(1), 2; https://doi.org/10.3390/IECN2020-07006 - 30 Oct 2020
Cited by 4 | Viewed by 1150
Abstract
: Maternal obesity is a metabolic disorder described by chronic inflammation and an increase of stress hormones, influencing non-communicable diseases in offspring. Elateriospermum tapos has the potential as an antioxidant and inhibitor of lipids (pancreatic lipase) and carbohydrates (α-amylase and α-glucosidase) which are [...] Read more.
: Maternal obesity is a metabolic disorder described by chronic inflammation and an increase of stress hormones, influencing non-communicable diseases in offspring. Elateriospermum tapos has the potential as an antioxidant and inhibitor of lipids (pancreatic lipase) and carbohydrates (α-amylase and α-glucosidase) which are beneficial to combat obesity and diabetes. This study aimed to investigate the effect of E. tapos supplementation in obese rats prior to pregnancy on the dam and male offspring body weight and the level of stress hormones—adrenocorticotropic hormone (ACTH) and corticosterone (CORT). Thirty female Sprague Dawley rats were used in this study. Six rats were assigned to a normal diet (DND) group fed with a standard chow diet. The remaining rats were fed with a high-fat and cafeteria diet (HFCD) to generate obesity for five weeks. The obese rats were further divided into four groups (n = 6/group); dams negative control group (DNC, normal saline), dams positive control group (DPC, 200 mg/kg body weight of orlistat), dams treatment 1 group (DTX1, 200 mg/kg BW of E. tapos seed) and dams treatment 2 group (DTX2, 200 mg/kg BW of E. tapos shell). The treatment was given for six weeks daily before mating. At weaning, male offspring were designated into six groups according to their dam’s group (n = 6/group) and continued with the cafeteria diet except for the control group. The offspring were culled at 12 weeks of age for blood sample collections. DTX2 and their male adult offspring showed significantly lower body weight compared to DNC and their male offspring. Male offspring from DTX2 also showed significantly low ACTH and CORT levels compared to male offspring from the DNC group and a comparable level with the DND group. E. tapos shell supplementation was effective in reducing the development of obesity and suppressed stress through the regulation of ACTH and CORT release in male adult offspring. Full article
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<p>Effect of <span class="html-italic">E. tapos</span> supplement on the corticosterone and ACTH plasma level of 12-weeks-old adult offspring. Abbreviations: letter (O) indicates offspring. ND: normal diet; NC: negative control; PC: positive control; TX1: treatment 1 (<span class="html-italic">E. tapos</span> seed); and TX2: treatment 2 (<span class="html-italic">E. tapos</span> shell). Data are expressed as mean±SEM and were analyzed by one-way ANOVA, followed by post-hoc LSD. Significance level set at <span class="html-italic">p</span> &lt; 0.05. a: <span class="html-italic">p</span> &lt; 0.05 versus FNC; b: <span class="html-italic">p</span> &lt; 0.05 versus FND; c: <span class="html-italic">p</span> &lt; 0.05 versus FPC. n = 6/group.</p>
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248 KiB  
Proceeding Paper
Elacteriospermum tapos Ameliorates Maternal Obesity Effect on Serum Leptin Changes in Male Offspring
by Santhra Segaran Balan, Azrina Zainal Abidin, Kokila Vani Perumal, Nurul Husna Shafie, Maizaton Atmadini Abdullah, Azmiza Syawani Jasni, Khairul Kamilah Abdul Kadir and Hasnah Bahari
Proceedings 2020, 61(1), 3; https://doi.org/10.3390/IECN2020-07007 - 30 Oct 2020
Cited by 1 | Viewed by 1351
Abstract
The purpose of this study is to investigate the effect of Elateriospermum tapos aqueous extract supplementation on serum leptin of male offspring at weaning. A total of 30 female Sprague Dawley rats were assigned to two groups, where the control group (CG) [...] Read more.
The purpose of this study is to investigate the effect of Elateriospermum tapos aqueous extract supplementation on serum leptin of male offspring at weaning. A total of 30 female Sprague Dawley rats were assigned to two groups, where the control group (CG) consisted of six rats and the remaining rats had obesity-induced over five weeks with a high-fat diet pellet and cafeteria food. After five weeks, the obese group was further divided into four groups, a negative control group (NG), positive control group (PG) (orlistat 200 mg/kg), treatment 1 (TX1) (200 mg/kg BW of E. tapos seed) and treatment 2 (TX2) (200 mg/kg BW of E. tapos shell) for 6 weeks. After six weeks, all rats were mated and continued with their respective diet till weaning. One male pup from each dam culled at weaning (postnatal day 21 (PND21)). The results show that body weight in male offspring (M) from negative group dams (NG) was significantly heavier as compared to other pup groups. Total adipose tissue weight in MTX1 and MTX2 of the male offspring was also significantly lower compared to MNG. In mums, serum leptin of NG was significantly higher as compared to the CG group, whereas both treatment groups showed a significant reduction in serum leptin compared to the NG group. In pups, the MTX2 group showed a more substantial reduction in body weight and serum leptin compared to other pups from other mother’s groups. In conclusion, E. tapos aqueous extract supplementation has a greater effect on ameliorating maternal obesity effects on male offspring by lowering body weight, inhibit fat deposition, and reducing serum leptin. Full article
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<p>Impacts of <span class="html-italic">E. tapos</span> on serum leptin of dams. Abbreviations: CG; control group, NG, negative control, PG, positive group, TX1, treatment with seed, TX2, treatment with shell. Data are expressed as mean± SEM and were analyzed by one-way ANOVA, followed by post-hoc LSD.</p>
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<p>Impacts of <span class="html-italic">E. tapos</span> on serum leptin of male pups (PND21). Abbreviations: the first letter indicate male offspring (M); the second letter indicates the groups. MCG; offspring from control group, MNG, offspring negative control, MPG, offspring positive group, MTX1, offspring from dams treated with seed group and MTX2, offspring from dams treated with shell group. Data are expressed as mean± SEM and were analyzed by one-way ANOVA, followed by post-hoc LSD.</p>
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601 KiB  
Proceeding Paper
Anti-Proliferative and Apoptotic Induction Effect of Elateriospermum Extract on Human Lung Cancer Cell Line A549
by Siroshini K. Thiagarajan, Kokila Vani Perumal, Nurul Husna Shafie, Khairul Kamilah Abdul Kadir and Hasnah Bahari
Proceedings 2020, 61(1), 4; https://doi.org/10.3390/IECN2020-06985 - 30 Oct 2020
Cited by 2 | Viewed by 1368
Abstract
Natural products derived from plants are used to treat cancer due to fewer side effects compared to standard treatments available for cancer. The second highest cancer that causes death worldwide is lung cancer. Therefore, this study aimed to determine the cytotoxic activity of [...] Read more.
Natural products derived from plants are used to treat cancer due to fewer side effects compared to standard treatments available for cancer. The second highest cancer that causes death worldwide is lung cancer. Therefore, this study aimed to determine the cytotoxic activity of hot and cold aqueous extract of E. tapos seed and shell on human cancer cell line A549 as well as the apoptosis mechanism. The apoptosis mechanisms were evaluated by cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction assay; MTT assay), Hoechst 33358 staining and detection of reactive oxygen species (ROS) activity. The apoptosis inducing activity was analyzed in set of morphological and biochemical features. Hot aqueous shell extract (SHH) showed an anti-proliferative effect at the IC50 of 49.8 ± 0.06 µg/mL correlated with apoptosis induction by increasing the ROS activity by significant (p < 0.05) increase of 3.25 folds compared to control. Results suggest that SHH poses an anti-proliferative effect on account of apoptosis through ROS mediated mitochondria mutilation. Full article
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<p>Cell apoptosis morphology observation by Hoechst staining. SDH—Seed Hot Aqueous, SDC—Seed Cold Aqueous, SHH—Shell Hot Aqueous, SHC—Shell Cold Aqueous, Cisplatin, Control—Untreated cells.</p>
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<p>Fold increment of reactive oxygen species (ROS) released from treated A549 cells. Error bar indicates the standard error of the mean of three independent experiments. SDH—Seed Hot Aqueous, SDC—Seed Cold Aqueous, SHH—Shell Hot Aqueous, SHC—Shell Cold Aqueous, Cisplatin—Positive control, Control—Untreated cells. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 vs. control.</p>
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228 KiB  
Proceeding Paper
Assessment of Negative Factors Affecting the Intestinal Microbiota in People with Excessive Body Mass Compared to People with Normal Body Mass
by Karolina Osowiecka, Natalia Pokorna and Damian Skrypnik
Proceedings 2020, 61(1), 5; https://doi.org/10.3390/IECN2020-07010 - 30 Oct 2020
Viewed by 996
Abstract
Rationale: Intestinal microbiota plays a significant role in the human body. A range of negative factors may lead to dysbiosis, which results in many diseases (e.g., cancer) and metabolic disorders. It was shown that people with a body mass index (BMI) of ≥25 [...] Read more.
Rationale: Intestinal microbiota plays a significant role in the human body. A range of negative factors may lead to dysbiosis, which results in many diseases (e.g., cancer) and metabolic disorders. It was shown that people with a body mass index (BMI) of ≥25 kg/m2 present diminished microbial diversity. Additional negative factors such as stress, the use of non-steroidal anti-inflammatory drugs (NSAID) and proton pomp inhibitors (PPI) may result in greater dysbiosis compared to people with normal body mass. The aim of the study was to compare the incidence of negative factors affecting the intestinal microbiota in people with excessive body mass vs. people with normal body mass. Methods: The study involved volunteers aged 18–65 years: 582 people with normal BMI (18.5–24.99 kg/m2) and 538 people with BMI ≥ 25 kg/m2. The study was conducted using the author’s survey by the Computer-Assisted Web Interviewing method. The survey included the questions on sociodemographic features, level of physical activity, frequency of smoking, and stress. Frequency of NSAID and PPI use was also investigated. BMI was used to assess nutritional status. For statistics the Fisher test was implemented. p-Value < 0.05 was considered significant. Results: Women predominated in the study (p < 0.0001). People with normal BMI had a significantly higher level of physical activity (p < 0.0001) and smoked less (p = 0.0356). People with excessive body mass were less likely to report illness (p = 0.0004), but more often they took PPI (p = 0.0337). Conclusions: People with BMI ≥ 25 kg/m2 present more factors which may result in dysbiosis and lead to dysbiosis-related health problems in future compared to people with normal BMI. Full article
446 KiB  
Proceeding Paper
Dose-Dependent Cytotoxicity of the Origanum vulgare and Carvacrol on Triple Negative Breast Cancer Cell Line
by Brian Enrique Rojo-Ruvalcaba, Teresa Arcelia García-Cobián, Sara Pascoe-González, Tannia Isabel Campos-Bayardo, Luz María Guzmán-García, María Cristina Gil-Gálvez, Zyanya Escobar-Millán, Eduardo Huerta-García and Trinidad García-Iglesias
Proceedings 2020, 61(1), 6; https://doi.org/10.3390/IECN2020-07000 - 30 Oct 2020
Cited by 4 | Viewed by 2043
Abstract
Uncontrolled proliferation and death resistance are two hallmarks of cancer cells. Breast cancer (BC) has the highest incidence and mortality in women worldwide and national; subtype triple negative (TN) is the most aggressive type because it is not susceptible to conventional therapy. Since [...] Read more.
Uncontrolled proliferation and death resistance are two hallmarks of cancer cells. Breast cancer (BC) has the highest incidence and mortality in women worldwide and national; subtype triple negative (TN) is the most aggressive type because it is not susceptible to conventional therapy. Since 2004, the World Health Organization (WHO) approved use of alternative treatments as adjuvants based on evidence of its benefits. Since then, different natural alternatives to treat cancer have been studied, including Origanum vulgare (Ov) and carvacrol (Crv), one of its main compounds. However, cytotoxic potential of these products has not been studied in this subtype of BC. The objective of this study was to evaluate the cytotoxic effect of Ov and Crv on TN BC cell line (HCC-70). Lethal dose 50 was determined on control cell line HaCaT through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays with Ov and Crv stimulus in different doses and concentrations; the found dose was used on the HCC-70 cell line. The results were evaluated by ANOVA, finding that Ov reduced the proliferation by 94.05% ± 0.11 (p < 0.001) and Crv by 93.43% ± 0.21 (p < 0.001) compared to untreated cells. This suggests that both Ov and Crv present a powerful cytotoxic effect against the HCC-70 cell line and are promising options that should receive further study. Full article
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<p>Photographs of the carvacrol (Crv) solution (<b>A</b>) and <span class="html-italic">Origanum vulgare</span> (<span class="html-italic">Ov)</span> infusion (<b>B</b>). Both stimuli were prepared the same day that were used.</p>
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<p>(<b>A</b>) HaCaT cell culture at day 1. (<b>B</b>) HaCaT cell culture upon reaching &gt;90% confluence; in addition to cell proliferation it is appreciated how the cells are adhered to the base of the culture flask. (<b>C</b>) HaCaT cell culture after the recovery process by trypsinization; a more rounded shape is observed in cultured cells after the adherent cell recovery process. (<b>D</b>) Viable cells appear white and those that are not are stained blue. Practically no non-viable cells can be seen. Photographs of cell cultures were taken by observing them under an inverted microscope while the photograph of Neubauer’s chamber was taken by observing it under a microscope.</p>
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<p>Graphic representation of the proliferation index of HaCaT cells exposed to Crv. Significance * <span class="html-italic">p</span> &lt; 0.0001. Data are expressed as mean of three replicates.</p>
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<p>Graphic representation of the proliferation index of HaCaT cells exposed to the <span class="html-italic">Ov</span> infusion. Significance * <span class="html-italic">p</span> &lt; 0.0001. Data are expressed as mean of three replicates.</p>
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<p>Graphic representation of the proliferation index of triple negative breast cancer (TN BC) cells exposed to <span class="html-italic">Ov</span> infusion and Crv. Significance * <span class="html-italic">p</span> &lt; 0.0001 Data are expressed as mean of three replicates.</p>
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1072 KiB  
Proceeding Paper
Evaluation and Determinants of Secondary Metabolites and its Antioxidant Activities of Various Fractions from Albizia myriophylla Bark
by Goh Yong Meng, Razif Abas, Nurmawati Syakroni, Nur Izah Abdul Razak, Nurul Huda Mohd. Nor, Siti Fadziyah Mohamad Asri, Safuraa Salihan, Nurul Hayati Mohamad Zainal, Suryati Mohd. Thani and Azmah Saat
Proceedings 2020, 61(1), 7; https://doi.org/10.3390/IECN2020-07004 - 30 Oct 2020
Cited by 1 | Viewed by 1022
Abstract
Albizia myriophylla (ABZ) is a plant used in Thailand and Peninsular Malaysia for the treatment of diabetes mellitus type II. The antioxidant activities of ABZ have been suggested as one of the mechanisms for the observed beneficial effects. Hence, this study examined the [...] Read more.
Albizia myriophylla (ABZ) is a plant used in Thailand and Peninsular Malaysia for the treatment of diabetes mellitus type II. The antioxidant activities of ABZ have been suggested as one of the mechanisms for the observed beneficial effects. Hence, this study examined the phenolic, flavonoid, and saponin contents and antioxidant activity from methanol extract (ME) and its derived fractions hexane (HE), chloroform (CE), ethyl acetate (EAE), butanol (BE), and aqueous fraction of the bark of ABZ (AE). Amongst the extracts, EAE showed the highest total phenolic content of about 0.77 mg of gallic acid equivalent/g of extract (mg GAE/mg). However, the highest flavonoid content was detected in HE at 1.04 µg retinol equivalent ((RE)/g extract), while the saponin content was highest in CE at 1.1 µg diosgenin equivalent ((DE)/g extract). In 1,1-diphenyl-2-picrylhidrazyl (DPPH) radical scavenging test, EAE at 100 µg/mL had the highest percentage of inhibition of about 72.55%. 2-2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS radical scavenging test) showed BE had the highest percentage of inhibition at 100 µg/mL at 82.91%. Ferric reducing antioxidant power elucidated BE as having the highest percentage of inhibition which was 86.04% followed closely by ME at 85.90%. Thus, the different extracts of ABZ displayed various antioxidant capacity with probable free radical scavenging activity, which may be useful for the treatment of chronic inflammatory related metabolic diseases, such as diabetes and obesity. Full article
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<p>Extraction procedure of bark of <span class="html-italic">Albizia myriophylla</span>. Methods adapted from Kaur et al.</p>
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<p>The phenolic contents in 250 µg of aqueous fraction (AE), methanol extract (ME), hexane (HE), ethyl acetate extract (EAE), chloroform extract (CE), and butanol extract (BE) of <span class="html-italic">Albizi myriophylla </span>(ABZ) bark extract. GAE, gallic acid equivalent. Values were expressed as mean ± SEM. Differing alphabets denote significant difference between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The flavonoid levels in 250 µg of AE, ME, HE, EAE, CE, and BE of ABZ bark extract. RE, retinol equivalent. Values were expressed as mean ± SEM. Differing alphabets denotes significant difference between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The saponin levels in 250 µg of AE, ME, HE, EAE, CE, and BE of ABZ bark extract. DE, diosgenin equivalent. Values were expressed as mean ± SEM. Differing alphabets denotes significant difference between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The percentage of inhibition in 2-2′-azinobis 3-ethyl-benzothiazoline -6-sulfonic acid (ABTS) antioxidant test in 20 µg/mL, 40 µg/mL, 60 µg/mL, 80 µg/mL, and 100 µg/mL of AE, ME, HE, EAE, BE, and CE of ABZ bark extracts. Differing alphabets denotes significant difference between groups (<span class="html-italic">p</span> &lt; 0.05). Results showed no significant difference between the extracts.</p>
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<p>The percentage of inhibition in 1,1-diphenyl-2-picrylhidrazyl (DPPH) antioxidant test in 20 µg/mL, 40 µg/mL, 60 µg/mL, 80 µg/mL, and 100 µg/mL of AE, ME, HE, EAE, BE, and CE of ABZ bark extracts. Differing alphabets denotes significant difference between groups (<span class="html-italic">p</span> &lt; 0.05). Results showed no significant difference.</p>
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<p>The percentage of inhibition in the ferric reducing antioxidant power (FRAP) antioxidant test in 20 µg/mL, 40 µg/mL, 60 µg/mL, 80 µg/mL and 100 µg/mL of AE, ME, HE, EAE, BE, and CE of ABZ bark extracts. Differing alphabets denotes significant difference between groups (<span class="html-italic">p</span> &lt; 0.05). Results showed no significant difference.</p>
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1243 KiB  
Proceeding Paper
Gut Microbiota Is Linked to Physical Health Improvements Resulting from Energy-Restricted Diet and Exercise: A Randomized Controlled Trial in Healthy Adults
by Shakuntla Gondalia, Matthew Cooke, Stephen Keenan and Regina Belski
Proceedings 2020, 61(1), 8; https://doi.org/10.3390/IECN2020-06992 - 30 Oct 2020
Cited by 1 | Viewed by 1447
Abstract
Animal studies have demonstrated that energy-restricted diets and exercise affect the gut microbiome and are positively linked to physical health; however, less is known about the impacts of various patterns of dietary restriction combined with exercise on the gut microbiota and associated health [...] Read more.
Animal studies have demonstrated that energy-restricted diets and exercise affect the gut microbiome and are positively linked to physical health; however, less is known about the impacts of various patterns of dietary restriction combined with exercise on the gut microbiota and associated health outcomes in humans. This study aimed to determine if an energy-restricted diet combined with resistance training altered the gut microbiome, and whether any changes were associated with differences in body composition, dietary intake, or biomarkers of metabolic health. Twenty-six healthy males and females, aged 19–36 years with BMIs of 22–35 kg/m2, were enrolled in a 2-arm parallel, randomized controlled trial and followed either a 5:2 intermittent fasting (IFT, n = 13) or continuous energy restriction (CERT, n = 13) diet combined with supervised resistance training for 12 weeks. Both treatments resulted in decreased body weight and increased lean body mass. Shifts in the abundance of, Faecalibacterium prausnitzii, a high butyrate producer, was positively associated with changes in lean body mass (IFTp = 0.05, CERTp = 0.01) in both the groups. Moreover, in the CERT group, changes in Coprococcus genus were negatively associated with energy (p = 0.009) and fat intake (p= 0.03) and positively associated with body fat (p = 0.02). Overall, the findings indicate that using resistance training paired with either intermittent or continuous energy restriction, result in similar changes in bacterial diversity and shifts in relative abundance of bacterial taxa. The shift in specific bacterial taxa were positively associated with measures of physical health providing further support to the proposed relationship between energy consumption, exercise, gut microbiota, and physical health. Full article
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<p>Effect of dietary treatment on fecal bacterial α and β-diversity. (<b>A</b>) α-diversity analysis of microbial composition in two treatment groups at pre (baseline) and post (12 weeks; post-intervention) treatment. Microbial evenness, diversity and richness were determined based on, Shannon Diversity Index, InSimpson index and Observed species. (<b>B</b>) Non-Metric Multidimensional Scaling (NMDS) plots based on Bray-Curtis Distance metrics of IFT and CERT dietary treatment showing pre and post treatment changes. Adonis function was employed to evaluate compare shift in microbiome composition compare to baseline with significance at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Shift in the relative abundance of Bacterial Taxa. Mean fold change (log2 transformed) from baseline of Operational Taxonomic Units (OTUs) that showed significant effect (two-way repeated measure ANOVA, where <span class="html-italic">p</span> &lt; 0.05 considered significant. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Treatment-response relationships were evaluated on significantly changed OTUs, physical measures, blood cholesterol and dietary component (i.e., baseline to week 12) using spearman’s correlation, data represented are as correlation coefficient, where * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01.</p>
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224 KiB  
Proceeding Paper
Tropical Seaweeds Improve Cardiovascular and Metabolic Health of Diet-Induced Obese and Hypertensive Rats
by Ryan du Preez, Sunil K. Panchal and Lindsay Brown
Proceedings 2020, 61(1), 9; https://doi.org/10.3390/IECN2020-07011 - 30 Oct 2020
Viewed by 1079
Abstract
Seaweeds have been an important part of the diet of coastal populations in Asia possibly for millennia but only a few scattered coastal communities in Europe and the Americas have maintained these traditions. Our studies have investigated the potential of two tropical seaweeds [...] Read more.
Seaweeds have been an important part of the diet of coastal populations in Asia possibly for millennia but only a few scattered coastal communities in Europe and the Americas have maintained these traditions. Our studies have investigated the potential of two tropical seaweeds grown commercially in Asia, Sarconema and Caulerpa spp., as functional foods for the reversal of metabolic syndrome and possible mechanisms. Sarconema spp. are a source of carrageenans used as thickening and gelling agents in foods, while Caulerpa spp. are consumed in Southeast Asia as low-energy foods with high contents of vitamins and minerals. For our studies, male Wistar rats were divided into groups in a 16-week protocol: corn starch diet-fed rats (C); C rats supplemented with 5% dried seaweed for the last 8 weeks; high-carbohydrate, high-fat diet-fed rats (H); and H rats supplemented with 5% dried seaweed for the last 8 weeks. H rats developed obesity, hypertension, dyslipidaemia, glucose intolerance, fatty liver and increased left ventricular collagen deposition, infiltration of inflammatory cells and plasma liver enzyme activities. Seaweed supplementation decreased body weight, abdominal and liver fat, systolic blood pressure, plasma lipid concentrations, plasma activities of liver enzymes and collagen deposition. Furthermore, seaweed supplementation modulated gut microbiota. Possible mechanisms for improved cardiovascular and metabolic health include a reduced infiltration of inflammatory cells into organs as well as an increased intake of fibre modulating gut microbiota composition. Full article
623 KiB  
Proceeding Paper
Bacterial Taxa Associated with High Adherence to Mediterranean Diet in a Spanish Population
by Carles Rosés, Amanda Cuevas-Sierra, Salvador Quintana, José I. Riezu-Boj, J. Alfredo Martínez, Fermín I. Milagro and Anna Barceló
Proceedings 2020, 61(1), 10; https://doi.org/10.3390/IECN2020-07001 - 30 Oct 2020
Viewed by 1190
Abstract
The Mediterranean diet (MD) is recognised as one of the healthiest diets worldwide and is associated with the prevention of cardiovascular and metabolic diseases, among others. Dietary habits are considered one of the strongest modulators of the gut microbiota, which seems to play [...] Read more.
The Mediterranean diet (MD) is recognised as one of the healthiest diets worldwide and is associated with the prevention of cardiovascular and metabolic diseases, among others. Dietary habits are considered one of the strongest modulators of the gut microbiota, which seems to play a significant role in the health and disease of the host. The purpose of the present study was to evaluate interactive associations between gut microbiota composition and habitual dietary intake in 360 Spanish adults of the Obekit cohort (normal weight, overweight and obese subjects). Dietary intake and adherence to the MD tests together with faecal samples were collected from each subject. Faecal 16S rRNA sequencing was performed and checked against the dietary habits. MetagenomeSeq was the statistical tool applied to analyse at the species taxonomic level. Results from this study confirm that a strong adherence to the MD increases the population of some beneficial bacteria, improving microbiota status towards a healthier pattern. Bifidobacterium animalis is the species with the strongest association with the MD. One of the highlights is the positive association between several SCFA-producing bacteria and high adherence to the MD. In conclusion, this study shows that MD, fibre, legumes, vegetables, fruit and nuts intakes are associated with an increase in butyrate-producing taxa such as Roseburia faecis, Ruminococcus bromii and Oscillospira (Flavonifractor) plautii. Full article
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<p>Bacterial species that were significantly more abundant in the group with high adherence to the Mediterranean diet (MD) (FDR &lt; 0.05) by the metagenomeSeq test. Red boxes represent subjects with a higher adherence to the MD and blue boxes represent low adherence.</p>
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438 KiB  
Proceeding Paper
Intensive Training and Sex Influence Intestinal Microbiota Composition: A Preclinical Approach
by Patricia Ruiz-Iglesias, Malén Massot-Cladera, Sheila Estruel-Amades, Francisco J. PérezCano and Margarida Castell
Proceedings 2020, 61(1), 11; https://doi.org/10.3390/IECN2020-06989 - 30 Oct 2020
Cited by 1 | Viewed by 819
Abstract
Lifestyle, including regular physical activity and dietary habits, influences microbiota composition. Although some studies have demonstrated changes in microbiota composition due to moderate or high intensity training in athletes, the relationship between physical activity and changes in the intestinal bacteria is still a [...] Read more.
Lifestyle, including regular physical activity and dietary habits, influences microbiota composition. Although some studies have demonstrated changes in microbiota composition due to moderate or high intensity training in athletes, the relationship between physical activity and changes in the intestinal bacteria is still a matter of discussion. In addition, as most studies are performed on males, the role of sexual dimorphism deserves to be explored. Therefore, the aim of this preliminary study was to ascertain the influence of an intensive training and the rat’s sex on caecal microbiota composition. For this purpose, female and male 4-week-old Wistar rats were submitted to a 4-week running training program. At the end, caecal samples were collected immediately after performing an exhaustion test to characterize the microbiota composition by 16S rRNA sequencing technique. Parallel groups of female and male sedentary (SED) rats (age matched) were included. The results showed that young female rats had a higher ability to run than males but no sex- or training-associated changes were observed on microbiota diversity and richness indexes among groups. However, the Actinobacteria, Bifidobacteriaceae and Bifidobacterium spp. proportions were significantly higher in male than in female rats when comparing SED groups (p < 0.05), whose proportions in males were decreased by the training program (p < 0.05 vs. SED). On the contrary, female SED rats showed a higher proportion of Odoribacteraceae (belonging to Bacteroidetes), Clostridiaceae and Eubacteriaceae (both Firmicutes) than in the respective male group (p < 0.05), although Eubacteriaceae proportion decreased by running (p < 0.05 vs. SED). However, training increased the proportion of the Paraeggerthella genus (Actinobacteria) in female rats with respect to its sedentary counterpart (p < 0.05). Overall, caecal microbiota composition is modified by intensive training in young rats but there are also sex-based differences that should be considered for interventional studies. Full article
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<p>Experimental design of the intensive training program. Groups: female runners (F-RUN, n = 6), female sedentary (F-SED, n = 5), male runners (M-RUN, n = 6), male sedentary (M-SED, n = 5). At the end of the training RUN animals were subjected to a final exhaustion test (ET).</p>
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<p>Relative abundance of the family <span class="html-italic">Bifidobacteriaceae</span> in caecal content at the end of the exhaustion test differentiating between sexes and exercise condition. Groups: female runners (F-RUN), female sedentary (F-SED), male runners (M-RUN), male sedentary (M-SED). Results are expressed as mean ± SEM (n = 3/group). Statistical significance: * <span class="html-italic">p</span> &lt; 0.05 male vs. female with same exercise condition, <sup>Ψ</sup> <span class="html-italic">p</span> &lt; 0.05 RUN vs. SED with same sex condition.</p>
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<p>Venn diagrams showing the diversity in all families differentiating between exercise condition (<b>A, B</b>) and sexes (<b>C, D</b>). Results derived from n = 3/group. Groups: female runners (F-RUN), female sedentary (F-SED), male runners (M-RUN), male sedentary (M-SED).</p>
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261 KiB  
Proceeding Paper
Dynamic Multi-Stage Gastrointestinal Digestion Model Assessment of Microbial Fermentation Products of Collagen Hydrolysates
by Christina E. Larder, Michèle M. Iskandar and Stan Kubow
Proceedings 2020, 61(1), 12; https://doi.org/10.3390/IECN2020-06998 - 30 Oct 2020
Viewed by 1202
Abstract
Proteins, peptides and amino acids (AAs) that bypass upper gastrointestinal (GI) digestion can be fermented in the colonic regions. This could lead to microbial production of health promoting short-chain fatty acids (SCFAs). Nitrogenous compounds can also be fermented to generate potentially harmful branched [...] Read more.
Proteins, peptides and amino acids (AAs) that bypass upper gastrointestinal (GI) digestion can be fermented in the colonic regions. This could lead to microbial production of health promoting short-chain fatty acids (SCFAs). Nitrogenous compounds can also be fermented to generate potentially harmful branched chain fatty acids (BCFAs). As collagen hydrolysate (CH) supplements contain a high peptide content, we evaluated whether peptides that undergo intestinal CH digestion and microbial fermentation can generate SCFAs and BCFAs. Two bovine-sourced CH formulations (CH-GL and CH-OPT) underwent digestive processes and microbial fermentation for 24 h in a dynamic GI digestion model containing human fecal matter. After 24 h, CH-OPT showed a significant (p < 0.05) increase in SCFAs (propionic, butyric and valeric acids) in the ascending colonic vessel with no changes observed with CH-GL. Only CH-OPT showed a significant (p < 0.05) increase in BCFAs, also noted in the ascending colon. No significant (p < 0.05) changes to SCFAs and BCFAs were observed in the transverse and descending colons for both CHs. These findings demonstrate that CHs can induce microbial production of SCFAs and BCFAs although this appears to depend on the CH tested. More studies are needed to determine the physiological significance of these microbial metabolites from intake of CH supplements. Full article
346 KiB  
Proceeding Paper
Biopotential of Sea Cucumbers (Echinodermata) and Tunicates (Chordata) from the Western Coast of Portugal for the Prevention and Treatment of Chronic Illnesses
by Alessio Carletti, Carlos Cardoso, Diana Juliao, Jorge L. Arteaga, Paula Chainho, Maria Ana Dionísio, Sabrina Sales, Maria J. Gaudêncio, Inês Ferreira, Cláudia Afonso, Helena Lourenço, M. Leonor Cancela, Narcisa M. Bandarra and Paulo J. Gavaia
Proceedings 2020, 61(1), 13; https://doi.org/10.3390/IECN2020-06994 - 30 Oct 2020
Cited by 1 | Viewed by 1853
Abstract
In the present work, we aimed to explore the potential of two groups of marine invertebrates—sea cucumbers (Echinodermata) and ascidians (Chordata)—as sources of anti-inflammatory, anti-oxidant, and osteogenic compounds with potential to be used as pharmaceuticals and nutraceuticals for the treatment and prevention of [...] Read more.
In the present work, we aimed to explore the potential of two groups of marine invertebrates—sea cucumbers (Echinodermata) and ascidians (Chordata)—as sources of anti-inflammatory, anti-oxidant, and osteogenic compounds with potential to be used as pharmaceuticals and nutraceuticals for the treatment and prevention of chronic diseases. 24 extracts (ethanol, water, and ethyl acetate) from 4 species of sea cucumbers and 4 species of tunicates were produced and screened in vitro for their anti-inflammatory and anti-oxidant activities and in vivo for osteogenic activity through an assay using zebrafish larvae. Our results showed that ethanolic extracts presented anti-oxidant and anti-inflammatory activity, which revealed to be stronger in the ascidians. The osteogenic activity, which provides evidence of the bioactive potential of these organisms in preventing chronic disorders causing low bone density, was found to be strong in one species of ascidians and 3 of holothurians. This study demonstrates the high potential of extracts from these marine organisms for using as nutraceuticals in the prevention of chronic bone disorders. Full article
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<p>Osteogenic activity of water, ethanol, and ethyl acetate extracts from sea cucumbers and ascidians in zebrafish larvae. Both <span class="html-italic">HA</span> and <span class="html-italic">OpA</span>/<span class="html-italic">HA</span> are reported as percentage on the average of the control group (water, ethanol 0.1% <span class="html-italic">v</span>/<span class="html-italic">v</span> or DMSO 0.1% <span class="html-italic">v</span>/<span class="html-italic">v</span> respectively). The averages <span class="html-italic">HA</span> and <span class="html-italic">OpA</span>/<span class="html-italic">HA</span> of the control group for each extract tested is represented by the axis <span class="html-italic">y</span> =100 (%). Statistical differences among the means are tested through One-way ANOVA (between control and each condition). <span class="html-italic">p</span> values are indicated as follow: &lt;0.0332 (*), &lt;0.0021 (**), &lt;0.0002 (***), &lt;0.0001 (****).</p>
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Proceeding Paper
Effect of Common Bean Consumption on the Gut Associated Microbiome in an In Vivo Screening Model for Breast Cancer
by Henry J. Thompson, John N. McGinley, Elizabeth S. Neil and Tiffany L. Weir
Proceedings 2020, 61(1), 14; https://doi.org/10.3390/IECN2020-07008 - 30 Oct 2020
Cited by 2 | Viewed by 1652
Abstract
Population data indicate that consumption of common bean reduces breast cancer risk, an effect replicated in a well characterized rodent model of mammary carcinogenesis. However, low molecular weight fractions of bean failed to inhibit growth of established breast cancer cell lines. Given that [...] Read more.
Population data indicate that consumption of common bean reduces breast cancer risk, an effect replicated in a well characterized rodent model of mammary carcinogenesis. However, low molecular weight fractions of bean failed to inhibit growth of established breast cancer cell lines. Given that cell culture screens for anticancer activity are designed to detect a decrease in cell number accumulation, we reasoned that it might be possible to create a cell number accumulation assay for screening foods such as common bean in vivo by using an oncogene driven model for breast cancer. A benefit of this approach is that it permits simultaneous detection of systemic effects in the host and their potential mediation by the gut microbiome. We report an in vivo mammary cell accumulation assay driven by the polyoma middle T antigen (PyMT) oncogene and show that bean feeding reduced the accumulation of cells in developing mammary pathologies. As a candidate mediator, we report the impact of bean consumption on the gut associated microbiome. Differences were observed between common bean and the control diet in microbial phylogenetic diversity, beta diversity, abundance of various taxa, and predicted functional activity. Full article
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<p>Representative mammary glands wholemounts from PyMT (pP) female mice over a 22 day time course. Panel (<b>A</b>), 21 days of age (DOA); panel (<b>B</b>), 36 DOA; panel (<b>C</b>) 43 DOA; panel (<b>D</b>) is the linear regression for increase in mammary pathology area over time, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Representative mammary gland whole mounts from PyMT female mice fed control or bean containing purified diet for two weeks. (<b>A</b>) Control diet; (<b>B</b>) bean diet; location of mammary gland nipples 1–5 are labeled in each panel.</p>
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<p>Effect of bean feeding on PyMT driven mammary carcinogenesis, two sample t-test <span class="html-italic">p</span> &lt; 0.01. (<b>A</b>) Mammary pathology expansion, <span class="html-italic">p</span> &lt; 0.01; (<b>B</b>) levels of phospho-AMPK mammary pathologies.</p>
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<p>Endocannabinoid cancer inhibition pathway. Proteins that were assessed and that are nodes in this pathway are circumscribed in red. Pink shape-fill indicates activation and green shape-fill indicates inactivation.</p>
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<p>Effect of bean feeding on phylogenetic diversity. Alpha diversity, <span class="html-italic">p</span> = 0.009, bean <span class="html-italic">n</span> = 5, CTRL <span class="html-italic">n</span> = 5.</p>
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<p>Effect of bean on beta diversity within the gut.</p>
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<p>Predicted functional differences of cecal microbial populations in control and bean fed mice. (<b>A</b>) FASYN initial pathway, <span class="html-italic">p</span> = 0.424; (<b>B</b>) FAO pathway, <span class="html-italic">p</span> = 1.69 × 10<sup>−4</sup>; (<b>C</b>) PHOSLIPSYN pathway, <span class="html-italic">p</span> = 2.14 × 10<sup>−5</sup>; (<b>D</b>) FASYN-ELONG pathway, <span class="html-italic">p</span> = 9.92 × 10<sup>−6</sup>; CTRL <span class="html-italic">n</span> = 5, bean <span class="html-italic">n</span> = 5.</p>
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322 KiB  
Proceeding Paper
Amygdalin as a Plant-Based Bioactive Constituent: A Mini-Review on Intervention with Gut Microbiota, Anticancer Mechanisms, Bioavailability, and Microencapsulation
by Hassan Barakat
Proceedings 2020, 61(1), 15; https://doi.org/10.3390/IECN2020-06984 - 30 Oct 2020
Cited by 7 | Viewed by 1783
Abstract
Amygdalin—a plant-based bioactive constituent particularly abundant in bitter almond has been identified as featuring the cyanogenic glycoside chemical organic compound which was originally intended to be a medication for cancer treatment once hydrolyzed to hydrogen cyanide (HCN). Unfortunately, studies have revealed that HCN [...] Read more.
Amygdalin—a plant-based bioactive constituent particularly abundant in bitter almond has been identified as featuring the cyanogenic glycoside chemical organic compound which was originally intended to be a medication for cancer treatment once hydrolyzed to hydrogen cyanide (HCN). Unfortunately, studies have revealed that HCN can similarly affect normal cells, rendering it harming the human body. Both in vivo and in vitro investigations are extremely controversial and make its use unsafe. An updated substantial review on the source, structure, intervention with gut microbiota, anticancer therapy, bioavailability, and microencapsulation of amygdalin was summarized. Amygdalin provided anti-tumor, anti-fibrotic, anti-atherosclerosis, anti-inflammatory, immunomodulatory, analgesic, ameliorating digestive and reproductive systems, and enhancing neurodegeneration as well as myocardial hypertrophy. Studies confirmed the toxicity of amygdalin produced by its HCN after oral ingestion. However, the intravenous route of administration was much less toxic than the oral route and can be avoided with an oral dosage ranging from 0.6 to 1 g daily. The diversity of gut consortium is a key factor in inducing toxicity by amygdalin. Indeed, there is no guaranteed way to point out the microbial consortium for each person and provide a safe oral dosage. Recently, the encapsulating of amygdalin using alginate–chitosaninanoparticles (ACNPs) as transporter was investigated. As an active drug delivery mechanism for regulated and constant release of amygdalin with its enhanced cytotoxic effect on cancerous cells, biocompatible and biodegradable ACNPs can be applied while protectinginormal cells and tissues. In conclusion, still unproven and conflicting facts give way to a broad avenue of researchifor a compound that could potentially be the next stage of cancer therapy. Full article
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<p>(<b>A</b>) Chemical structure of amygdalin, (<b>B</b>) Chemical structure of laetrile.</p>
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1791 KiB  
Proceeding Paper
Chronic Dietary Zinc Deficiency Alters Gut Microbiota Composition and Function
by Omry Koren and Elad Tako
Proceedings 2020, 61(1), 16; https://doi.org/10.3390/IECN2020-06993 - 30 Oct 2020
Cited by 11 | Viewed by 2990
Abstract
Zinc (Zn) deficiency is a prevalent micronutrient insufficiency. Although the gut is a vital organ for Zn utilization, and Zn deficiency is associated with impaired intestinal permeability and a global decrease in gastrointestinal health, alterations in the gut microbial ecology of the host [...] Read more.
Zinc (Zn) deficiency is a prevalent micronutrient insufficiency. Although the gut is a vital organ for Zn utilization, and Zn deficiency is associated with impaired intestinal permeability and a global decrease in gastrointestinal health, alterations in the gut microbial ecology of the host under conditions of Zn deficiency have yet to be studied. By conducting a series of long-term in vivo (Gallus gallus) feeding trials, we aimed to characterize distinct cecal microbiota shifts induced by chronic dietary Zn depletion in the context of complete diets based on Zn-biofortified food crops that are relevant to target populations, and in geographical regions where dietary Zn deficiency is a major health concern. We demonstrate that Zn deficiency induces significant taxonomic alterations and decreases overall species richness and diversity, establishing a microbial profile resembling that of various other pathological states. Through metagenomic analysis, we show that the predicted Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways responsible for macro- and micronutrient uptake are significantly depleted under Zn deficiency; along with concomitant decreases in beneficial short-chain fatty acids, such depletions may further preclude optimal host Zn availability. We also identify several candidate microbes that may play a significant role in modulating the bioavailability and utilization of dietary Zn during prolonged deficiency. Our results are the first to characterize a unique and dysbiotic cecal microbiota during Zn deficiency, and they provide evidence for such microbial perturbations as potential effectors of the Zn-deficient phenotype. Full article
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<p>Microbial diversity of the cecal microbiome. (<b>A</b>) Measures of α-diversity using the Chao1 index [<a href="#B36-proceedings-61-00016" class="html-bibr">36</a>], and (<b>B</b>) total number of observed species. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, analysis of variance (ANOVA); <span class="html-italic">n</span> = 10 in Zn(+), <span class="html-italic">n</span> = 9 in Zn(−). (C) Measure of β-diversity using weighted UniFrac distances separated by the first three principal components (PCs). Each dot represents one animal, and the colors represent the different treatment groups.</p>
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<p>Phylum- and genera-level cecal microbiota shifts due to dietary Zn depletion. (<b>A</b>) Phylum-level changes between the Zn(+) and Zn(−) groups as measured at the end of the study (day 28). Only those phyla with abundance &gt; 1% are shown. (<b>B</b>) Increased Firmicutes to Bacteroidetes and Proteobacteria ratios in the Zn(+) group (* <span class="html-italic">p</span> &lt; 0.05, NS = not statistically significant). (<b>C</b>) Inverse correlation between Proteobacteria abundance and bodyweight. (<b>D</b>) Genus-level changes in the Zn(+) and Zn(−) group as measured at the end of the study (day 28). Only genera that were significantly different between groups are shown.</p>
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<p>Heat map describing a set of Spearman correlations, independently of treatment group, between the relative abundance of different operational taxonomic units (OTUs) and select biological indicators of Zn status. The color indicator ranges from a perfect negative correlation (−1, blue) to a perfect positive correlation (1, red) (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, ANOVA).</p>
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<p>The functional capacity of the cecal microbiota is perturbed under conditions of Zn deficiency. (<b>A</b>) Fold-change depletion of these pathways in the Zn(−) group (all <span class="html-italic">p</span> &lt; 0.01, Student’s <span class="html-italic">t</span>–test). (<b>B</b>) Relative abundance of differentially expressed Kyoto Encyclopedia of Genes and Genomes (KEGG) microbial metabolic pathways in cecal microbiota. Treatment groups are indicated by the different colors (all <span class="html-italic">p</span> &lt; 0.05, ANOVA).</p>
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<p>Microbial diversity of the cecal microbiome. (<b>a</b>) Measure of α-diversity using the Chao1 index and (<b>b</b>) α-diversity using the Observed Species Index. (<b>c</b>) Measure of β-diversity using the unweighted UniFrac distances separated by the first three principal components (PCoA). Each dot represents one animal, and the colors represent the different treatment groups.</p>
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<p>Compositional changes of gut microbiota in response to a biofortified diet. (<b>a</b>) Phylum-level changes in the control wheat-based diet (CZn) and Zn-biofortified wheat-based diet (BZn) groups as measured at the end of the study (day 42). Only phyla with abundance ≥1% are displayed. (<b>b</b>) Genus-level changes in the CZn and BZn groups as measured at the end of the study (day 42). Only genera with abundance ≥5% are displayed.</p>
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<p>The linear discriminant analysis effect size (LEfSe) method to identify the most differentially enriched taxa in the BZn and CZn groups. (<b>a</b>) Taxonomic cladogram obtained using LEfSe analysis of the 16S rRNA sequences. Treatment groups are indicated by the different colors, where the brightness of each dot is proportional to its effect size. (<b>b</b>) Computed linear discriminant analysis (LDA) scores of the relative abundance difference between the CZn and BZn groups. Negative LDA scores (red) are enriched in the CZn group, while positive LDA scores (green) are enriched in the BZn group.</p>
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<p>The functional capacity of the gut microbiota is altered with a Zn-biofortified diet: relative abundance of differentially enriched KEGG microbial metabolic pathways in the microbiota. Treatment groups are indicated by the different colors, and <span class="html-italic">p</span>-values are displayed on the <span class="html-italic">y</span>-axis.</p>
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<p>Intestinal functionality, short-chain fatty acid production, and cecal microbial composition following consumption of experimental diets. (<b>a</b>) Chicken intestinal goblet cell number and diameter (µm). (<b>b</b>) Chicken intestinal villus length and width (µm). (<b>c</b>) Cecal short-chain fatty acid (SCFA) composition. Bars represent the mean ± SEM of nine biological replicates. Relative abundance of microbial populations at the levels of (<b>d</b>) phyla and (<b>e</b>) families and genera. Asterisks denote significant differences for * <span class="html-italic">p</span> &lt; 0.05 and *** <span class="html-italic">p</span> ≤ 0.001 as determined by Student’s <span class="html-italic">t</span>-test. AU: arbitrary units.</p>
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<p>Microbial diversity and metabolic capacity following consumption of experimental diets. (<b>a</b>) Microbial α-diversity of chicken ceca using Faith’s phylogenetic diversity (PD). (<b>b</b>) Microbial β-diversity of chicken ceca using unweighted UniFrac distances separated by three principal components (PCs). Each dot represents either a ‘Control’ (green) or ‘Biofortified’ (red) chicken. (<b>c</b>) Computed linear discriminant analysis (LDA) scores of differences in relative microbial abundance and metabolic capacity, respectively. Positive LDA scores (green) are enriched in ‘Control’ and negative LDA scores (red) are enriched in ‘Biofortified’. Asterisks denote significant differences for * <span class="html-italic">p</span> &lt; 0.05 as determined by a Kruskal–Wallis test.</p>
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Proceeding Paper
Gut Microbiota Regulates the Interplay between Diet and Genetics to Influence Glucose Tolerance
by Jeralyn J. Franson, Julianne H. Grose, Kaitlyn Williams Larson and Laura C. Bridgewater
Proceedings 2020, 61(1), 17; https://doi.org/10.3390/IECN2020-07005 - 30 Oct 2020
Viewed by 943
Abstract
Per-arnt-sim (PAS) kinase is a nutrient sensing serine/threonine kinase whose absence protects against triglyceride accumulation, insulin resistance, a decreased metabolic rate and increased weight gain in response to a high fat diet, using phenotypes associated with the gut microbiome. Herein we further explored [...] Read more.
Per-arnt-sim (PAS) kinase is a nutrient sensing serine/threonine kinase whose absence protects against triglyceride accumulation, insulin resistance, a decreased metabolic rate and increased weight gain in response to a high fat diet, using phenotypes associated with the gut microbiome. Herein we further explored the metabolic effects of PAS kinase-deficiency(PASK−/−) on a high fat high sugar (HFHS) diet, including contributions from an altered microbiome. PASK−/− mice were not protected from weight gain on the HFHS diet but were resistant to liver triglyceride accumulation. Microbiome analysis of both WT and PASK−/− mice revealed a forked shift with two discrete clusters of HFHS-fed mice emerging, which displayed increased beta and decreased alpha diversity compared with the normal chow diet (NCD). A “lower” cluster associated with both increased weight gain and glucose intolerance contained elevated levels of Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria and Defferibacteres. Lower cluster PASK−/− mice also influenced glucose tolerance and Claudin-1 expression, a protein associated with leaky gut. These results suggest PAS kinase-deficiency can protect mice against the deleterious effects of liver triglyceride accumulation, leaky gut and glucose intolerance in response to diet; however, microbiome imbalance can override protection. In addition, these results support a healthy diet and suggest microbial culprits associated with metabolic disease. Full article
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<p>Diet, influenced by genotype, alters body and tissue weight of WT and Per-Arnt-Sim kinase (PASK)-deficient mice. (A) Weekly body weight of male and female mice displays weight gain in response to HFHS diet, influenced by genotype. (B) No significant difference in total weight gain of male and female mice. Relative tissue weights of male (C) and female (D) gonadal fat pad (GFP) and retroperitoneal fat pad (RFP) at dissection expressed as tissue weight/final body weight. (E) Male liver weight is influenced by diet, whereas female liver weight (F) is influenced by both diet and genotype, with PAS kinase-deficiency protecting against hepatic weight loss in response to HFHS diet. (G) PAS kinase-deficiency protects male mice against diet-induced hepatic triglyceride accumulation. All data is expressed as mean ± SE. All groups <span class="html-italic">n</span> = 13–19 with the exception of male liver TG (<span class="html-italic">n</span> = 4–8). Factorial ANOVA (JMP Pro version 14) statistical analysis was performed on male and female mice individually because statistical differences were observed for both sexes at all time points. Significant differences obtained through post hoc LSMeans Dunnett analysis using WT NCD as control or between other groups by post hoc Tukey HSD are shown; <span class="html-italic">p</span>-value less than or equal to 0.06 are shown as diet only (*) or interaction between diet and genotype (NCD with PASK <b><sup><span>$</span></sup></b>, HFHS with WT <b><sup>†</sup></b>, HFHS with PASK <b><sup>‡</sup></b>).</p>
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<p>The high fat high sugar (HFHS) diet affects both glucose tolerance testing (GTT) and insulin tolerance testing (ITT) responses. (<b>A</b>) Blood glucose levels during IP glucose tolerance testing (GTT) at 15 weeks and 23 weeks on the HFHS diet. (<b>B</b>) Area under the curve (AUC) from (<b>A</b>). (<b>C</b>) Blood glucose levels during IP insulin tolerance testing (ITT) at 16 weeks and 24 weeks on the HFHS diet. (<b>D</b>) Area under the curve (AUC) for (<b>C</b>). All data was analyzed by one factor ANOVA within genotype (HFHS versus NCD) followed by post hoc Tukey analysis and by Factorial ANOVA of the entire data set with post hoc Dunnetts analysis to investigate interactions between diet and genotype in JMP Pro version 14. “<span>$</span>” indicates differences between the NCD versus the HFHS diet in WT mice, and “‡” indicated differences between the two diets in PASK<sup>−/−</sup> mice, “†” indicates a significant difference for the PASK<sup>−/−</sup>-HFHS cohort compared to all others, * indicates differences due to diet (all mice on NCD versus all mice on HFHS diet). <span class="html-italic">p</span>-value &lt; 0.06. All groups <span class="html-italic">n</span> = 7.</p>
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<p>Diet and genotype induced changes in protein levels. (A) Colon claudin-1 levels are decreased in WT but not PASK-deficient mice on a HFHS diet. N = 5–7 per group. (B) Muscle Akt levels are decreased in PASK<sup>−/−</sup> mice on a NCD. N = 3–5 per group, (C) Muscle pAkt(ser473) levels are decreased in PASK<sup>−/−</sup> mice. N = 4–5 per group. Statistical analysis was performed using factorial ANOVA in JMP Pro version 14 with post hoc Dunnet analysis (* indicates <span class="html-italic">p</span>-value &lt; 0.05). Representative immunoblots are shown.</p>
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<p>Diet, influenced by genotype, alters gut microbial diversity. (A) Comparison of unweighted UniFrac distances between NCD (blue) and HFHS (red). (B) Comparison of unweighted UniFrac distances between WT (blue) and PASK<sup>−/</sup>- (red). (C) Comparison of alpha and beta diversity between NCD and HFHS. (D) Comparison of alpha and beta diversity by diet and genotype. (<b>E</b>) Relative abundance at phyla level in week 15 samples and week 22 samples. Alpha diversity is expressed as mean plus SEM, distances to NCD with Kruskal-Wallis follow-up. Beta diversity is expressed as mean plus SEM with PERMANOVA comparison.</p>
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<p>Gut microbial composition influences weight gain on a HFHS diet. (A) Unweighted UniFrac PCoA plot of week 15 samples and week 22 samples coded by final body weight tertile per group, with oval outline of the HFHS lower cluster (orange) and upper cluster (yellow). (B) Comparison of unweighted UniFrac distances by final body weight tertile of HFHS males at 15 and 22 weeks. (C) Final body weights of HFHS mice divided by unweighted UniFrac PCoA cluster compared to the WT-NCD. (D) Heatmap of week 22 male HFHS samples with the upper or lower cluster indicated on the x-axis. ANOVA from JMP Pro version14 software was used to determine significance followed by post hoc Tukey analysis (* indicates <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>Gut microbial composition influences glucose response. (<b>A</b>) Week 15, 19 and 23 glucose tolerance test (GTT). (<b>B</b>) Area under the curve (AUC) of the GTT shown in (<b>A</b>) (<span class="html-italic">n</span> = 8–14). (<b>C</b>) Comparison of unweighted UniFrac distances between week-19 AUC tertiles (<span class="html-italic">n</span> = 15–28). (<b>D</b>) Week 16 and 24 insulin tolerance test (ITT) (<span class="html-italic">n</span> = 10–13). ANOVA in JMP Pro version 14 was used to analyze significance with post hoc Tukey analysis to obtain the <span class="html-italic">p</span>-value (* indicates <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>An interplay between genotype and microbial composition influences weight and glucose response. (<b>A</b>) Comparison of weight gain, (<b>B</b>) triglyceride accumulation and (<b>C</b>) claudin-1 expression on the high fat high sugar diet by unweighted UniFrac PCoA cluster and genotype. Representative western blots for claudin-1 are shown. (<b>D</b>) Glucose tolerance(GTT) tests at week 15, 19 and 22. (<b>E</b>) Area under the curve (AUC) of the GTT’s shown in (<b>D</b>). Factorial ANOVA was performed on all samples, including individual GTT time points (symbols †, ‡, <span>$</span> for <span class="html-italic">p</span>-value &lt; 0.051) as well as the AUC (* is used for <span class="html-italic">p</span>-value &lt; 0.051) using JMP Pro version 14 and significant time points suggested an interaction between diet, genotype and upper or lower cluster. Sample number is provided (<span class="html-italic">n</span>).</p>
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1219 KiB  
Proceeding Paper
Influence of Sex and Diet on the Gastrointestinal Tract in a Mice Model with Partial Deficiency for TGF-β3
by Paula Gallego, Ana Bagüés, Elia Escasany, Yolanda López-Tofiño, Antonio Márquez, Gema Medina-Gomez, José Antonio Uranga and Raquel Abalo
Proceedings 2020, 61(1), 18; https://doi.org/10.3390/IECN2020-06983 - 30 Oct 2020
Cited by 1 | Viewed by 1472
Abstract
Functional and structural alterations of the gastrointestinal (GI) tract were evaluated in mice with partial deficiency of TGF-β3 (heterozygous; HZ) and the influence of sex and diet in them. No signs of inflammatory bowel disease (IBD) were detected. Wild type (WT) females presented [...] Read more.
Functional and structural alterations of the gastrointestinal (GI) tract were evaluated in mice with partial deficiency of TGF-β3 (heterozygous; HZ) and the influence of sex and diet in them. No signs of inflammatory bowel disease (IBD) were detected. Wild type (WT) females presented lower body weight and delayed GI transit compared to males. The HZ genotype modified the latency of marked feces expulsion, in a sex and diet dependent manner, without GI macroscopic structural alterations. A high fat diet (HFD) counteracted TGF-β3 heterozygosity functional effect in males. Sex, diet, and TGF-β3 may alter GI tract motility and structure, with a possible impact on the obesity-associated IBD. Full article
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<p>Number of feces excreted by the mice belonging to the different experimental groups. Male and female mice of wild type (WT) or mutant (heterozygous; HZ) genotypes were used, which were exposed to a control (CD) or a high fat (HFD) diet for 3 months. The number of feces excreted per hour is shown. The difference due to gender in the WT-CD groups is shown in (<b>A</b>). The effect of diet and genotype on the number of feces excreted by males and females are shown in (<b>B</b>) and (<b>C</b>), respectively. Values are represented by the mean ± SEM (standard error of the mean). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 vs. WT-CD (two-way ANOVA followed by Sidak’s multiple comparisons test).</p>
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<p>Wet weight of the feces excreted by the mice of the different experimental groups. Male and female mice of wild (WT) or mutant (HZ) genotypes were used, which were exposed to a control (CD) or a high fat (HFD) diet for 3 months. The wet weight of feces excreted per hour is shown. The difference due to gender in the WT-CD groups is shown in (<b>A</b>). The effect of diet and genotype on wet weight of feces excreted by males and females are shown in (<b>B</b>) and (<b>C</b>), respectively. Values are represented by the mean ± SEM (two-way ANOVA followed by Sidak’s multiple comparisons test).</p>
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<p>Dry weight of the feces excreted by the mice belonging to the different experimental groups. Male and female mice of wild (WT) or mutant (HZ) genotypes were used, which were exposed to a control (CD) or a high fat (HFD) diet for 3 months. The dry weight of the feces per hour is shown. The difference due to gender in the WT-CD groups is shown in (<b>A</b>). The effect of diet and genotype on dry weight of feces excreted by males and females are shown in (<b>B</b>) and (<b>C</b>), respectively. Values are represented by the mean ± SEM (two-way ANOVA followed by Sidak’s multiple comparisons test).</p>
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<p>Fecal moisture expressed as percentage of water content in the feces excreted by the mice of the different experimental groups. Male and female mice of wild (WT) or mutant (HZ) genotypes were used, which were exposed to a control (CD) or a high fat (HFD) diet for 3 months. The percentage of water in the excreted feces per hour is shown. The difference due to gender in the WT-CD groups is shown in (<b>A</b>). The effect of diet and genotype on moisture of feces excreted by males and females are shown in (<b>B</b>) and (<b>C</b>), respectively. Values are represented by the mean ± SEM. *** <span class="html-italic">p</span> &lt; 0.001 vs. WT-CD and # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 vs. HZ-CD (two-way ANOVA followed by Sidak’s multiple comparisons test).</p>
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<p>Stool marking index in mice of the different experimental groups. Male and female mice of wild (WT) or mutant (HZ) genotypes were used, which were exposed to a control (CD) or a high fat (HFD) diet for 3 months. The stool marking index per hour is shown. The difference due to gender in the WT-CD groups is shown in (<b>A</b>). The effect of diet and genotype on stool marking index in feces excreted by males and females are shown in (<b>B</b>) and (<b>C</b>), respectively. Values are represented by the mean ± SEM. <span class="html-italic">p</span> &lt; 0.05 vs. WT-CD ### <span class="html-italic">p</span> &lt; 0.001 vs. HZ-CD (two-way ANOVA followed by Sidak’s multiple comparisons test).</p>
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<p>Submucosa thickness in distal colon. Wild type (WT) and mutant (HZ) mice were exposed to a control (CD) or a high fat (HFD) diet for 3 months. Histological samples of WT (left: <b>A</b>, <b>C</b>, <b>E,</b> and <b>G</b>) or HZ (right: <b>B</b>, <b>D</b>, <b>F,</b> and <b>H</b>) mice stained with Van Gieson´s stain. Male (<b>A</b>–<b>D</b>) and female (<b>E</b>–<b>H</b>) animals. Arrow shows submucosa tissue stained in red. Bar 50 μm.</p>
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837 KiB  
Proceeding Paper
Simulated Gastrointestinal Digestion Influences the In Vitro Hypolipidemic Properties of Coffee Pulp, a Potential Ingredient for the Prevention of Non-Alcoholic Fatty Liver Disease
by Cheyenne Braojos, Miguel Rebollo-Hernanz, Vanesa Benitez, Silvia Cañas, Yolanda Aguilera, Silvia M. Arribas and Maria A. Martin-Cabrejas
Proceedings 2020, 61(1), 19; https://doi.org/10.3390/IECN2020-06997 - 30 Oct 2020
Cited by 2 | Viewed by 1411
Abstract
Approximately 90% of the coffee cherry is discarded as waste during coffee bean processing. Coffee pulp has been validated as a potential safe ingredient and is a potential source of nutrients and health-promoting compounds that could be used as nutraceuticals to manage some [...] Read more.
Approximately 90% of the coffee cherry is discarded as waste during coffee bean processing. Coffee pulp has been validated as a potential safe ingredient and is a potential source of nutrients and health-promoting compounds that could be used as nutraceuticals to manage some chronic diseases. Metabolic disorders associated with dysregulated energy and cellular processes, such as obesity and hyperlipidemia, contribute to non-alcoholic fatty liver disease (NAFLD). In this sense, the main objective of this study was to evaluate the impact of an in vitro simulated digestion on the hypolipidemic properties of coffee pulp flour and the biological activity of the digested fractions of its flour and extract in HepG2 cells. The hypolipidemic properties of coffee pulp flour were tested by measuring the capacities of the residual fraction of each digestion to bind cholesterol and bile salts and to inhibit the lipase activity after simulated gastric, intestinal, and colonic in vitro digestion. The results exhibited that coffee pulp residual fraction had up to 58% (p < 0.05) more capacity to bind cholesterol, 1.9-fold (p < 0.05) higher bile salts binding capacity, and 1.5-fold (p < 0.05) higher ability to reduce the lipase activity than control residues. Likewise, the digested fractions of coffee pulp flour and extract (50–250 µg/mL) significantly (p < 0.05) alleviated the accumulation of fat (14–35%), triglycerides (5–27%), and cholesterol (9–48%) triggered by the stimulation of HepG2 cells with palmitic acid (500 µM) to simulate NAFLD. In conclusion, simulated gastrointestinal and colonic digestion improves coffee pulp hypolipidemic properties, enhancing its biological activity in cell culture models. Therefore, this coffee by-product could be an interesting potential ingredient to be used to prevent hyperlipidemia and regulate lipid metabolism. Full article
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<p>Hypolipidemic properties of coffee pulp digested residual fractions. (<b>A</b>) Cholesterol-binding capacity of coffee pulp digested residual fractions at pH 2.0; (<b>B</b>) Cholesterol-binding capacity of coffee pulp digested residual fractions at pH 7.0; (<b>C</b>) Bile salts binding capacity of coffee pulp digested residual fractions. The results are expressed as mean ± SD. Bars with different letters significantly (<span class="html-italic">p</span> &lt; 0.05) differ according to ANOVA and Tukey’s multiple range test. G: Gastric digestion residual fraction; GI: Intestinal residual fraction; C: Colonic residual fraction; RAW: Raw flour.</p>
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<p>Illustrative representation of HepG2 morphologies after incubation in different conditions (<b>A</b>) Cells cultured in DMEM-BSA 1%; (<b>B</b>) Cells cultured in DMEM-BSA 1% with PA (500 μM); (<b>C</b>) cells cultured in DMEM-BSA 1% with PA (500 μM) with treatment (Intestinal CPE, 100 μg/mL).</p>
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578 KiB  
Proceeding Paper
Regulation of Lipid and Glucose Metabolism in Hepatocytes by Phytochemicals from Coffee By-Products and Prevention of Non-Alcoholic Fatty Liver Disease In Vitro
by Miguel Rebollo-Hernanz, Yolanda Aguilera, Maria A. Martin-Cabrejas and Elvira Gonzalez de Mejia
Proceedings 2020, 61(1), 20; https://doi.org/10.3390/IECN2020-06996 - 25 Nov 2020
Cited by 3 | Viewed by 1510
Abstract
This study aimed to assess the effect of the primary phytochemicals from coffee by-products and two aqueous extracts from the coffee husk and silverskin on lipid and glucose metabolism regulation in hepatocytes using an in vitro model of non-alcoholic fatty liver disease. Coffee [...] Read more.
This study aimed to assess the effect of the primary phytochemicals from coffee by-products and two aqueous extracts from the coffee husk and silverskin on lipid and glucose metabolism regulation in hepatocytes using an in vitro model of non-alcoholic fatty liver disease. Coffee husk and silverskin were used to prepare two aqueous extracts (CHE and CSE, respectively) using water. The phytochemical composition was determined using UPLC-MS/MS analysis. HepG2 cells were co-treated with 10–50 µmol L‒1 of either pure caffeine, chlorogenic acid, caffeic acid, protocatechuic acid, or gallic acid, and kaempferol, CHE, or CSE (20–100 µg mL‒1) in the presence or absence of palmitic acid (PA, 500 µmol L‒1). Different biomarkers of cell metabolism were assessed 24 h after the co-treatment in cell supernatants and lysates using chemical, biochemical, and immunochemical techniques. Phytochemicals from coffee by-products decreased PA-triggered lipid accumulation (16–94%, p < 0.05) by reducing fatty acid synthase activity and stimulating lipolysis (8–83%, p < 0.05). CHE, CSE, and therein-bioactive compounds promoted glucose uptake (13–45%) via the increase in the phosphorylation of the insulin receptor (1.9- to 2.7-fold), protein kinase B (AKT) (1.4- to 3.1-fold), AMPKα (1.6- to 2.4-fold), and PTEN (2.0- to 4.2-fold). In conclusion, our results proved that phytochemicals from coffee by-products, mainly caffeine and chlorogenic acid, could regulate hepatic lipid and glucose metabolism. Overall, our results generate new insights into the use of coffee by-products as a sustainable food ingredient to encounter non-alcoholic fatty liver disease. Full article
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<p>Venn diagram illustrating the overlap of differentially phosphorylated protein in adipocytes by coffee silverskin (CSE) and coffee husk (CHE) aqueous phenolic extracts (<b>A</b>), circos plot illustrating the biological processes modulated by CSE and CHE indicating which are shared among treatments (<b>B</b>), KEGG pathways associated with the differentially phosphorylated proteins (<b>C</b>), and protein–protein interaction networks built with Metascape from the differentially phosphorylated protein in hepatocytes by CSE (<b>D</b>) and CHE (<b>E</b>).</p>
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238 KiB  
Proceeding Paper
Dietary Consumption of Polyphenols in University Students—Relationship with Their Health-Related Habits
by Maria J Rodríguez-Lagunas, Francisco J Pérez-Cano, Filipa Vicente, Paula Pereira and Margarida Castell
Proceedings 2020, 61(1), 21; https://doi.org/10.3390/IECN2020-06991 - 30 Oct 2020
Cited by 1 | Viewed by 1066
Abstract
Polyphenols are compounds derived from plants found in foods with potential health benefits due to their antioxidant properties. The aim of this study was to establish the consumption of polyphenols in university students and the relationship between the amount of polyphenols consumed and [...] Read more.
Polyphenols are compounds derived from plants found in foods with potential health benefits due to their antioxidant properties. The aim of this study was to establish the consumption of polyphenols in university students and the relationship between the amount of polyphenols consumed and their health-related habits. For this, 270 university students answered a validated food consumption frequency questionnaire (FFQ), a physical activity questionnaire (IPAQ), and a health and lifestyle questionnaire. From the FFQ, the daily consumption of classes and subclasses of polyphenols was calculated. Then, the population was classified in terciles according to the polyphenol consumption, and the physical activity, smoking habits, and the prevalence of allergy and obesity in each tercile was established. The consumption of polyphenols was about 1.5 g/day, being flavonoids and phenolic acids the most consumed. The main sources of polyphenol consumption were cocoa, coffee, and to a lesser extent, fruits. There were no significant differences in consumption according to sex, BMI, health status, or physical activity. The smoking habit was related to a high consumption of polyphenols, mainly those that came from coffee and cocoa. Students who smoke had a higher consumption of polyphenols with respect to nonsmokers, especially those derived from coffee. Full article
496 KiB  
Proceeding Paper
Lyophilized Extract of Hibiscus sabdariffa L. Induces Cytotoxicity in Breast Cancer Cell Lines
by Zyanya Escobar Millan, Maritza Roxana García-Garcia, Juan Gómez Leyva, Ricardo Figueroa Martinez, Jorge Alfredo Vargas Vital, Eduardo Huerta García and Trinidad García Iglesias
Proceedings 2020, 61(1), 22; https://doi.org/10.3390/IECN2020-07002 - 30 Oct 2020
Cited by 1 | Viewed by 1683
Abstract
Some types of cancer are resistant to conventional treatments, such as triple-negative (TN) breast cancer (BC), which, due to its molecular characteristics, does not respond effectively. In recent years, there has been an increase in the use of medicinal plants for prevention and [...] Read more.
Some types of cancer are resistant to conventional treatments, such as triple-negative (TN) breast cancer (BC), which, due to its molecular characteristics, does not respond effectively. In recent years, there has been an increase in the use of medicinal plants for prevention and as an alternative in adjuvant therapy for cancer. Hibiscus sabdariffa L. (H.S.) is a plant which has anti-inflammatory and antioxidants properties. The objective of this study is to evaluate the cytotoxicity effect of the lyophilized extract of H.S. in the TN BC cell line (HCC-70) exposed to different concentrations (0.10, 0.25, 0.5 and 1.0 mg/mL). The results were evaluated by ANOVA, finding that H.S. reduces the proliferation on TN BC cell line HCC-70 exposed at 1.0 mg/mL by 62%. The use of natural extracts as coadjuvant therapy for cancer is an alternative with great potential for future studies and provides a precedent in the area of natural medicine. Full article
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<p>(<b>A</b>) Photographs of stock solution of HSE and (<b>B</b>) dilution of the HSE stock solution in culture medium to obtain the desired concentrations respectively (0.10 mg/mL, 0.25 mg/mL, 0.5 mg/mL, and 1.0 mg/mL).</p>
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<p>Cell line BC HCC-70. (<b>A</b>) Cell culture after the recovery process by trypsinization, a more rounded shape is observed in cultured cells after the adherent cell recovery process; these were observed through an inverted microscope. (<b>B</b>) Viable cells appear white and those that are not are stained blue, practically no non-viable cells can be seen; the photograph of Neubauer’s chamber was taken by observing it under a microscope.</p>
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<p>Cells of the HCC-70 cell line corresponding to TN BC exposed to different concentrations of H.S. (HCC70 0.10 mg/mL, HCC70 0.25 mg/mL, HCC70 0.5 mg/mL and HCC70 1.0 mg/mL. As negative control (HCC70−), we used etoposide and in the positive control (HCC70+) no H.S. was added.</p>
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Proceeding Paper
Effect of Pulse Consumption on Obesity and the Metagenome
by Tiffany L. Weir, John N. McGinley, Elizabeth S. Neil and Henry J. Thompson
Proceedings 2020, 61(1), 23; https://doi.org/10.3390/IECN2020-07009 - 30 Oct 2020
Viewed by 1238
Abstract
Grain legumes, which are commonly referred to as pulses, are staple foods in many parts of the world, but are infrequently consumed in most economically developed countries where the obesity pandemic is prominent. However, even in low pulse consuming countries such as the [...] Read more.
Grain legumes, which are commonly referred to as pulses, are staple foods in many parts of the world, but are infrequently consumed in most economically developed countries where the obesity pandemic is prominent. However, even in low pulse consuming countries such as the United States, there are sub-groups of individuals who consume large amounts of pulses. Systematic reviews of population studies indicate that pulse consumers have a lower risk for developing obesity. To determine whether these population-based findings could be modeled in preclinical systems in which such findings can be deconstructed, we used rat and mouse models of dietary induced obesity and reported that lipid accumulation was inhibited. In this study, we examined the relationship between inhibition of fat accumulation and changes in the gut associated microbiome in male C57BL/6 mice fed either a high fat diet with casein as the protein source or that diet formulation in which one of four pulses (chickpea, common bean, dry pea, or lentil) was substituted to provide 70% dietary protein with the remainder provided by casein. The seeds of each pulse were soaked, cooked, and then freeze-dried and milled; the resulting powder was used for diet formulation. Mice were ad libitum fed over the 17-week duration of the feeding trial. Cecal content was obtained at necropsy and immediately snap frozen in liquid nitrogen. Extracted genomic DNA was processed for 16s rRNA sequencing on an Illumina system. Significant differences were observed between each pulse and the high fat control diet in microbial phylogenetic diversity (p < 0.001) and accumulation of lipid in adipose depots (p < 0.01). Differences among pulses were also observed in both metrics. Microbiome alpha and beta diversity metrics, differences in abundance for each detected taxon among treatment groups, and their relationships to changes in lipid accumulation in adipose storage depots are reported. Full article
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<p>Principal components analysis of the effect of feeding pulse diets on adipose tissue mass.</p>
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<p>Principal components analysis of the effect of feeding pulse diets on the abundance of microorganism (level of genus).</p>
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<p>Regression analyses evaluating the relationship among principal components for adipose depot mass and microbial abundance (cecal luminal content). All regressions were statistically significant (<span class="html-italic">p</span> &lt; 0.01), except for the regression analysis shown in panel D; lower confidence limit: LCL; upper confidence limit: UPL; lower prediction limit: LPL; upper prediction limit: UPL.</p>
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<p>Effect of pulse consumption on microbial abundance (species level) using CLC Genomics Taxonomic Profiler.</p>
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<p>Effect of pulse consumption on alpha diversity.</p>
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<p>Effect of pulse consumption on beta diversity/.</p>
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<p>Effect of pulse consumption predicted function of the microbiome.</p>
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658 KiB  
Proceeding Paper
Identification of Antihypertensive Tripeptides in the Culture Medium of Lactobacillus helveticus D75 and D76 Strains
by Kirill Kuvakin, Timur Vakhitov, Stanislav Sitkin, Evgeniya Roshchina, Veronika Lisitskaya, Olga Ivanchenko, Yaroslav Dubrovskii and Elena Demyanova
Proceedings 2020, 61(1), 24; https://doi.org/10.3390/IECN2020-07014 - 30 Oct 2020
Viewed by 3782
Abstract
The aim of the current work was to search for bioactive peptides Ile-Pro-Pro (IPP) and Val-Pro-Pro (VPP) with antihypertensive action in the culture medium (CM) of probiotic Lactobacillus helveticus D75 and D76 strains when cultured in milk. The CM composition was analyzed using [...] Read more.
The aim of the current work was to search for bioactive peptides Ile-Pro-Pro (IPP) and Val-Pro-Pro (VPP) with antihypertensive action in the culture medium (CM) of probiotic Lactobacillus helveticus D75 and D76 strains when cultured in milk. The CM composition was analyzed using HPLC-UV, HPLC-MS, and MALDI Fourier transformation ion-cyclotron resonance mass spectrometry (MALDI FT-ICR MS, Varian). Direct analysis of CM using FT-ICR failed to detect peptides in the original CM due to the high concentration of concomitant components, but after preliminary fractionation of CM by HPLC, both peptides were detected. The masses (m/z) of characteristic ions were 312.19 (VPP) and 326.21 (IPP). Peptides were quantified using a single reference method with HPLC and time-of-flight mass spectrometry. The concentrations of VPP and IPP in the CM of the D75 strain, respectively, reached 18.0 µg/mL and 25.4 µg/mL, in the CM of the D76 strain they were lower (12.7 µg/mL and 16.2 µg/mL), and in the mixed CM (both strains) the concentrations reached intermediate values of 15.7 µg/mL and 24.0 µg/mL. The data obtained indicate the existence of an active complex of extracellular enzymes in L. helveticus D75 and D76, which provide the production of IPP and VPP peptides with antihypertensive effect. The results also confirm the symbiotic properties of D75 and D76 strains (synergism and syntrophy). Functional foods based on milk fermented by probiotic L. helveticus D75 and D76 strains, which contains IPP and VPP peptides in effective doses (>3.0 mg/day), can be considered a safe and effective non-pharmacological intervention in patients with mild hypertension. Full article
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<p>HPLC of a mixture of Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) standards. The column was applied to 20 mL of a solution of IPP and VPP with a concentration of 75 mg/mL each. The column was a Phenomenex Luna C18. Eluent of 0.1% TFC with an acetonitrile gradient from 3% (0–1 min) to 50% (23 min), t = 45 °C, V = 1 mL/min, registration by optical density at a wavelength of 206 nm.</p>
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<p>MALDI-mass spectrometric analysis of fractions of culture medium (CM) ultrafiltrates of the <span class="html-italic">L. helveticus</span> D75 strain. The mass spectra were obtained using a Varian ion-cyclotron resonance (ICR) mass spectrometer. The relative intensity is plotted on the ordinate, and the mass-to-charge ratio (m/z) is plotted on the abscissa. (<b>a</b>,<b>b</b>) Ultrafiltrates of culture medium (UCM) fractions: 24 h of growth of the <span class="html-italic">L. helveticu</span>s D75 strain: (<b>a</b>) fractions of 6.5–7.5 min; (<b>b</b>) fractions of 7.5–8.5 min.</p>
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<p>HPLC-MS of CM ultrafiltrates on an Intensity Solo 2C 18 column in a gradient system of 0.1% formic acid–acetonitrile (<a href="#proceedings-61-00024-t001" class="html-table">Table 1</a>). Relative intensity is plotted on the ordinate, and time in minutes is plotted on the abscissa. (<b>a</b>–<b>d</b>) Registration of total ion current: (<b>a</b>) <span class="html-italic">L. helveticus</span> D75, (<b>b</b>) <span class="html-italic">L. helveticus</span> D76, (<b>c</b>) mixed culture of D75 and D76 strains, and (<b>d</b>) a mixture of synthetic peptides of IPP and VPP in concentrations of 10 micrograms/mL. In the insets to the figures, registration of chromatographic separation by ion current of characteristic ions with m/z = 326.21 (IPP) – (a-1, b-1, c-1, d-1) and 312.19 (VPP) – (a-2, b-2, c-2, d-2).</p>
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<p>HPLC-MS of CM ultrafiltrates on an Intensity Solo 2C 18 column in a gradient system of 0.1% formic acid–acetonitrile (<a href="#proceedings-61-00024-t001" class="html-table">Table 1</a>). Relative intensity is plotted on the ordinate, and time in minutes is plotted on the abscissa. (<b>a</b>–<b>d</b>) Registration of total ion current: (<b>a</b>) <span class="html-italic">L. helveticus</span> D75, (<b>b</b>) <span class="html-italic">L. helveticus</span> D76, (<b>c</b>) mixed culture of D75 and D76 strains, and (<b>d</b>) a mixture of synthetic peptides of IPP and VPP in concentrations of 10 micrograms/mL. In the insets to the figures, registration of chromatographic separation by ion current of characteristic ions with m/z = 326.21 (IPP) – (a-1, b-1, c-1, d-1) and 312.19 (VPP) – (a-2, b-2, c-2, d-2).</p>
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549 KiB  
Proceeding Paper
Aging Effects on Gut Microbiota in SAMP8 Mice
by Lluïsa Miró, Miquel Moretó, Concepció Amat, Javier Polo and Anna Pérez-Bosque
Proceedings 2020, 61(1), 25; https://doi.org/10.3390/IECN2020-06995 - 30 Oct 2020
Cited by 3 | Viewed by 1176
Abstract
We have studied the effects of aging on the fecal microbiota composition in the senescence-accelerated prone mice SAMP8 strain. We compared animals two, four, and six months old. Feces were collected at the end of each period and a genomic study was carried [...] Read more.
We have studied the effects of aging on the fecal microbiota composition in the senescence-accelerated prone mice SAMP8 strain. We compared animals two, four, and six months old. Feces were collected at the end of each period and a genomic study was carried out on fecal DNA using the Illumina MiSeq analyzer. The Shannon diversity index showed similar values along this period and the number of species was neither affected by aging. The phylum Verrucobacteria went up with age, showing a seven-fold increase at six months, compared to two-month old mice. At the family level, changes observed between two and six months of age involved significant increases in Bacteroidaceae (q < 0.001) and strong reductions in Lactobacillaceae (q < 0.0001) and Prevotellaceae (q < 0.05); at the genus level, there was a significant reduction in probiotic Lactobacillus. At the species level, we observed an age-related reduction in Lactobacillus hayakitensis, a species involved in mucosal immune homeostasis, and in Blautia hansenii, which provides protection against Clostridium difficile infection. Interestingly, aging increases Parabacteroides goldsteiini, which is involved in the regulation of the TLR4 pathway. These results support the view that aging results in the proliferation of bacterial species that are associated with the immune deterioration of the gut mucosa. Full article
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<p>Microbial Shannon’s index (<b>A</b>), number of species (<b>B</b>). Results are expressed as mean ± SEM (<span class="html-italic">n</span> = 11 mice).</p>
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<p>Effects of aging on fecal bacterial composition (phylum level) at 2M (<b>A</b>), 4M (<b>B</b>), and 6M (<b>C</b>). Results are expressed as percent of the total population at phylum level (mean) from 11 mice.</p>
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<p>Effects of aging on fecal bacterial composition at family level. Results are expressed as percent of the total population at family level (mean ± SEM; <span class="html-italic">n</span> = 10–11 mice). Statistical differences were considered significant at <span class="html-italic">q</span> &lt; 0.05 (corrected <span class="html-italic">p</span> values).</p>
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243 KiB  
Proceeding Paper
Association between Intake of Fermented Dairy Products and Diet Quality, Health Beliefs in a Representative Sample of Polish Population
by Katarzyna E. Przybyłowicz, Jakub Morze, Anna Danielewicz, Katarzyna Staniewska, Aneta Dąbrowska, Maria Baranowska, Małgorzata Darewicz, Justyna Żulewska and Bogusław Staniewski
Proceedings 2020, 61(1), 26; https://doi.org/10.3390/IECN2020-06988 - 30 Oct 2020
Cited by 1 | Viewed by 1267
Abstract
This study aimed to evaluate the association between diet quality, perception of benefits consumption, and fermented dairy product intake in a representative sample of the Polish population. The study was carried out in February 2020. 2009 men and women were randomly sampled from [...] Read more.
This study aimed to evaluate the association between diet quality, perception of benefits consumption, and fermented dairy product intake in a representative sample of the Polish population. The study was carried out in February 2020. 2009 men and women were randomly sampled from the representative Polish population stratified for two age groups (19–30 and 66–75 years). Dairy product intake was evaluated using a qualitative frequency questionnaire. Diet quality was assessed by calculating the Mediterranean Diet Adherence Screener (MEDAS) score. Perceived health benefits of dairy product consumption were assessed with a literature-based questionnaire. The Health Concern Scale was used to measure participants’ attitudes toward health. Median intake of fermented dairy products was 0.8 portion/day (IQR: 0.4–1.6). Intake of fermented dairy products was associated with higher MEDAS. We observed that people with the highest intake of fermented dairy product consumed more oils, vegetables, wine, legumes, fish and seafood, sweets and pastries, nuts, and higher preference for white meat and were more likely to report perceived benefits to maintain body weight, reduce cardiovascular risk, and improve immune and dental health. Moreover, a high intake of fermented dairy products was positively related to paying more attention to health. Our study identified patterns of health behaviors associated with frequent consumption of fermented products. We observed that the intake of fermented dairy products is associated with better diet quality, consumer self-consciousness, and a greater attitude to own health. Full article
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Proceeding Paper
Effect of Docosahexaenoic Acid Supplementation on Microbiota in Obese ChiLdrEn: A Pilot Study. (The DAMOCLE Study)
by Matilde Amatruda, Sara Vizzuso, Valentina Fabiano, Elisa Borghi, Gian VIncenzo Zuccotti and Elvira Verduci
Proceedings 2020, 61(1), 27; https://doi.org/10.3390/IECN2020-07012 - 30 Oct 2020
Cited by 1 | Viewed by 1377
Abstract
Abstract: Obesity is an inflammatory condition associated with metabolic alterations including insulin resistance. Recent researches suggested that gut microbiota plays a role in its pathogenesis. Obesity has been associated with lower bacterial diversity and higher Firmicutes/Bacteroidetes ratio (F/R ratio) compared to normal‐weight [...] Read more.
Abstract: Obesity is an inflammatory condition associated with metabolic alterations including insulin resistance. Recent researches suggested that gut microbiota plays a role in its pathogenesis. Obesity has been associated with lower bacterial diversity and higher Firmicutes/Bacteroidetes ratio (F/R ratio) compared to normal‐weight condition. The objective of our study was to determine the effect of 4 months algae DHA supplementation, combined with dietary and habits education, on gut microbiota composition and biochemical parameters of 12 caucasian obese children. Anthropometric measures, metabolic profile and gut microbiota analysis trough stool samples were assessed at baseline (t0), after 4 months DHA supplementation plus diet-lifestyle intervention (t1), and lastly after additional 4 months of dietary dietary-lifestyle intervention without DHA supplementation (t2). No significant improvement in dietary habits nor in metabolic profile was found after the intervention, while a significant lowering of F/R ratio was observed from t0 to t1 and even more evident at t2 control. Full article
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<p>Gut microbiota analysis in obese children baseline (t0), after 4 months DHA supplementation combined with diet–lifestyle intervention (t1), and after an additional 4 months of dietary–lifestyle intervention without DHA supplementation (t2). (<b>A</b>) Alpha-diversity (observed species metric is reported) showed no significant differences in gut microbiota biodiversity; (<b>B</b>) Principal Coordinate Analysis (PCoA) according to Weighted UniFrac distance: t0 samples (red) significantly (<span class="html-italic">p</span> &lt; 0.05) separate from t1 samples (green); (<b>C</b>) Firmicutes/Bacteroidetes (F/B) ratio significantly decreases (<span class="html-italic">p</span> &lt; 0.05) at t1 (blue) compared with t0 (red).</p>
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242 KiB  
Proceeding Paper
Dietary Fibre, Gut Microbiota Dysbiosis and Type 2 Diabetes
by Omorogieva Ojo, Qianqian Feng, Osarhumwese Osaretin Ojo and Xiaohua Wang
Proceedings 2020, 61(1), 28; https://doi.org/10.3390/IECN2020-06986 - 30 Oct 2020
Cited by 27 | Viewed by 1177
Abstract
Background: Diabetes prevalence is on the increase globally and its impact on those with the condition in terms of acute and chronic complications can be profound. People with type 2 diabetes constitute the majority of those with the condition and the risk factors [...] Read more.
Background: Diabetes prevalence is on the increase globally and its impact on those with the condition in terms of acute and chronic complications can be profound. People with type 2 diabetes constitute the majority of those with the condition and the risk factors include obesity, lifestyle and gut microbiota dysbiosis. Poor dietary intake has been reported to influence the community of the gut microbiome. Therefore, a higher intake of dietary fibre may alter the environment in the gut and promote microbial growth and proliferation. Aim: This is a systematic review and meta-analysis which examined the effect of dietary fibre on gut microbiota in patients with type 2 diabetes. Method: This review was conducted in line with the PRISMA framework. Databases were searched for relevant articles which were screened based on inclusion and exclusion criteria. Results: Nine articles which met the inclusion criteria were selected for the systematic review and meta-analysis. High dietary fibre intake significantly improved (p < 0.05) the abundance of Bifidobacterium, total short-chain fatty acids (SCFAs) and HbA1c. Discussion: The promotion of SCFA producers in terms of greater diversity and abundance by dietary fibre may have resulted in improvement in glycated haemoglobin, partly due to increased GLP–1 production. Conclusion: High consumption of dietary fibre has a significant (p < 0.05) effect on Bifidobacterium, total SCFAs and HbA1c, but not (p > 0.05) on propionic, butyric and acetic acid, fasting blood glucose and the homeostatic model assessment of insulin resistance HOMAR–IR. Full article
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<p>The effect of dietary fibre on Bifidobacterium (%).</p>
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<p>The effect of dietary fibre on total short-chain fatty acids.</p>
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<p>The effect of dietary fibre on glycated haemoglobin (%).</p>
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222 KiB  
Proceeding Paper
Clinical Applications of Ketogenic Diet-Induced Ketosis in Neurodegenerative and Metabolism-Related Pathologies
by Raffaele Pilla
Proceedings 2020, 61(1), 29; https://doi.org/10.3390/IECN2020-06982 - 30 Oct 2020
Cited by 1 | Viewed by 1590
Abstract
Metabolic-based therapies such as nutritional ketosis have been proven effective for seizure disorders and various acute and chronic neurological pathologies. In a healthy brain, glucose is the primary metabolic fuel for cells. However, neurodegenerative disorders, such as Alzheimer’s disease (AD), Parkinson’s disease (PD), [...] Read more.
Metabolic-based therapies such as nutritional ketosis have been proven effective for seizure disorders and various acute and chronic neurological pathologies. In a healthy brain, glucose is the primary metabolic fuel for cells. However, neurodegenerative disorders, such as Alzheimer’s disease (AD), Parkinson’s disease (PD), seizure disorders, and traumatic brain injury (TBI) are associated with impaired glucose transport and metabolism and with mitochondrial dysfunction leading to energy deficit. Therapeutic ketosis can be considered as a form of metabolic therapy by providing alternative energy substrates. In addition, ketosis leads to metabolic adaptations that improve brain metabolism, restore mitochondrial ATP production, decrease reactive oxygen species production, reduce inflammation, and increase the activity of neurotrophic factors. Moreover, the synaptic activity between neurons is also stabilized through the increase of Szent-Györgyi–Krebs cycle intermediates, antioxidant effects, increased GABA-to-glutamate ratio, and activation of ATP-sensitive potassium channels. Full article
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Proceeding Paper
Cocoa and Cocoa Fibre Intake Modulate Reactive Oxygen Species and Immunoglobulin Production in Rats Submitted to Acute Running Exercise
by Patricia Ruiz-Iglesias, Raquel Gómez-Bris, Malén Massot-Cladera, María J. Rodríguez-Lagunas, Francisco J. Pérez-Cano and Margarida Castell
Proceedings 2020, 61(1), 30; https://doi.org/10.3390/IECN2020-06990 - 30 Oct 2020
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Abstract
Acute high-intensity exercise can impair the immune system, and lead to oxidative stress. Cocoa intake might help in protecting against oxidative damage and impaired immune functioning. The aim of this study was to establish the effect of cocoa and cocoa fibre on the [...] Read more.
Acute high-intensity exercise can impair the immune system, and lead to oxidative stress. Cocoa intake might help in protecting against oxidative damage and impaired immune functioning. The aim of this study was to establish the effect of cocoa and cocoa fibre on the oxidative status and the immunoglobulin (Ig) production of rats following a bout of acute exercise on a treadmill. The production of reactive oxygen species (ROS) by macrophages and the concentration of serum and mucosal Ig was assessed 16 h after the running session. Exercise increased ROS production and decreased the serum IgG concentration and the salivary gland IgM content. A cocoa fibre-enriched diet prevented the increased ROS production and the reduction in salivary IgM induced by exercise, although it decreased the IgA content in serum and the salivary glands. Overall, cocoa, by means of its fibre content, can partially prevent the alterations in ROS and Ig production induced by a single session of intensive running exercise. Full article
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<p>Experimental design. R = running session; ET = exhaustion test.</p>
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<p>Reactive oxygen species (ROS) production by peritoneal macrophages at the end of the study (percentage of increase from 0 to 120 min). The sedentary groups are represented by bars without any pattern and the runner groups by a striped pattern. REF = reference diet; CF = 5% cocoa fibre enriched diet; C10 = 10% cocoa enriched diet. Data are expressed as mean ± SEM (<span class="html-italic">n</span> = 8). <span class="html-italic">* p</span> &lt; 0.05 vs. the sedentary group in the same dietary group; # &lt; 0.05 vs. the same exercise condition in the REF diet.</p>
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<p>Immunoglobulin concentrations (µg/mL) in serum at the end of the study. The sedentary groups are represented by bars without any pattern and the runner groups by a striped pattern. REF = reference diet; CF = 5% cocoa fibre enriched diet; C10 = 10% cocoa enriched diet. Data are expressed as mean ± SEM (<span class="html-italic">n</span> = 8). <span class="html-italic">* p</span> &lt; 0.05 vs. the sedentary group in the same dietary group; # &lt; 0.05 vs. the same exercise condition in the REF diet.</p>
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<p>Submaxillary salivary gland IgM and IgA (<b>a</b>), and gut wash IgA (<b>b</b>) concentration at the end of the study. The sedentary groups are represented by bars without any pattern and the runner groups by a striped pattern. REF = reference diet; CF = 5% cocoa fibre enriched diet; C10 = 10% cocoa enriched diet. Data are expressed as mean ± SEM (<span class="html-italic">n</span> = 8). <span class="html-italic">* p</span> &lt; 0.05 vs. the sedentary group in the same dietary group; # &lt; 0.05 vs. the same exercise condition in the REF diet; λ &lt; 0.05 vs. the same exercise condition in the CF diet.</p>
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375 KiB  
Proceeding Paper
In Vitro Bioaccessibility of Citrus Pomace Compounds Possessing Health Promoting Properties with Potential to Reduce the Risk of Diabetes
by Adriana Maite Fernández-Fernández, Eduardo Dellacassa, Tiziana Nardin, Roberto Larcher, Adriana Gámbaro, Alejandra Medrano-Fernandez and María Dolores del Castillo
Proceedings 2020, 61(1), 31; https://doi.org/10.3390/IECN2020-06999 - 30 Oct 2020
Cited by 10 | Viewed by 1189
Abstract
Citrus (mandarin and orange) pomace is an agro-food industrial waste rich in polyphenols and dietary fiber with great potential as a functional ingredient. In this work, the chemical composition and in vitro bioaccessibility of health-promoting compounds present in raw citrus pomaces (Clemenule and [...] Read more.
Citrus (mandarin and orange) pomace is an agro-food industrial waste rich in polyphenols and dietary fiber with great potential as a functional ingredient. In this work, the chemical composition and in vitro bioaccessibility of health-promoting compounds present in raw citrus pomaces (Clemenule and Ortanique mandarins and Navel and Valencia oranges) were studied. In addition, the by-products were employed as food ingredients in cookies and the effect of the food matrix on the bioaccessibility of their bioactive compounds was evaluated. Nobiletin, hesperidin/neohesperidin, tangeretin, heptamethoxyflavone, tetramethylscutellarein, and naringin/narirutin were detected in the citrus samples by UHPLC-MS. Citrus pomaces were in vitro digested mimicking the human oral gastrointestinal conditions and the bioactivity of the digests (antioxidant, carbohydrases modulation, and anti-inflammatory effects) was assessed. The bioaccessibility of the antioxidants in the by-products was confirmed by Total Polyphenol Content (TPC) (6.6–11.0 mg GAE/g digest), ABTS (65.5–97.1 µmol TE/g digest), ORAC-FL assays (135.5–214.8 µmol TE/g digest), and inhibition of Reactive Oxygen Species (ROS) formation induced by treatment with tert-butyl hydroperoxide 1 mM in intestinal IEC-6 (19–45%) and CCD-18Co (28–45%) cells pretreated with the digests at concentrations ranging between 5 and 25 µg/mL. Inhibitors of the enzymatic activity of α-glucosidase (IC50 3.97–11.42 mg/mL) and α-amylase (IC50 58.04–105.68 mg/mL) also remained bioaccessible after in vitro digestion of citrus pomaces. In addition, the bioaccessible compounds in orange pomace samples significantly reduced (p < 0.05) the lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 macrophages. The digests of orange pomace cookies with the nutrition claims “no-added sugars” and “source of fiber” presented antioxidant and anti-diabetic properties, and good sensory quality (6.9–6.7 on a scale of 1 to 9). The results obtained support the feasibility of unfractionated orange pomace as a functional ingredient for reducing the risk of diabetes. The health-promoting benefits observed in the present research might be, at least partially, associated with flavonoids. Full article
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<p>Induced ROS formation in normal human intestinal cells (CCD-18Co) treated with digests of Clemenule and Ortanique mandarin and Navel and Valencia orange pomaces prior to the oxidative damage. Normal human colon cells were treated with samples for 24 h followed by co-administration of the oxidative agent (tert-butyl hydroperoxide) and sample for 30 min. Bars represent mean values while error bars denote standard error of the mean (SEM). * indicate statistical differences between mean values with <span class="html-italic">p</span> &lt; 0.05 by Tukey test.</p>
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<p>Effect of the orange pomace bioaccessible compounds (250 µg/mL) in the formation of NO in RAW264.7 macrophages. Cells were treated with samples during 24 h followed by co-administration of LPS and sample during 24 h. Bars represent mean values while error bars denote standard error of the mean (SEM). * indicate statistical differences between mean values with <span class="html-italic">p</span> &lt; 0.05 by Tukey test.</p>
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