Peter Bucheli

Phytochemical profiles of different mulberry (Morus sp.) species from China.

Mulberry is rich in alkaloids, polyphenols, flavonoids, and anthocyanins, which have been suggested to be responsible for health benefits. The concentrations of 1-deoxynojirimycin (DNJ), resveratrol, oxyresveratrol, cyanidin-3-O-beta-glucoside (Cy-3-glu), cyanidin-3-O-beta-rutinoside (Cy-3-rut), and rutin in mulberry juice, fruits, and leaves of 8 species grown in China were examined. It is the first time content determination of DNJ in mulberry juice and oxyresveratrol in mulberry fruits and leaves has been reported. Among the varieties tested, Da 10 (Morus atropurpurea Roxb.) was the most valuable cultivar considering its high content of functional components. Besides, Hetianbaisang (M. alba Linn.), Taiwanguosang (M. atropurpurea Roxb.), Fujian 2 hao (M. alba Linn.), Gaozhoujisang (M. australis Poir.), and Shanxiguosang (M. nigra Linn.) were rich in DNJ, resveratrol, oxyresveratrol, anthocyanins, and flavonoids, respectively. The high contents of functional compounds in mulberry juice, fruits, and leaves implied that they might be potential resources for the development of functional drinks and food.

Inter-Species Transplantation of Gut Microbiota from Human to Pigs

Direct research on gut microbiota for understanding its role as ‘an important organ’ in human individuals is difficult owing to its vast diversity and host specificity as well as ethical concerns. Transplantation of human gut microbiota into surrogate hosts can significantly facilitate the research of human gut ecology, metabolism and immunity but rodents-based model provides results with low relevance to humans. A new human flora-associated (HFA) piglet model was hereby established taking advantage of the high similarity between pigs and humans with respect to the anatomy, physiology and metabolism of the digestive system. Piglets were delivered via cesarean section into a SPF-level barrier system and were inoculated orally with a whole fecal suspension from one healthy 10-year-old boy. The establishment and composition of the intestinal microbiota of the HFA piglets were analyzed and compared with that of the human donor using enterobacterial repetitive intergenic consensus sequence-PCR fingerprinting-based community DNA hybridization, group-specific PCR-temperature gradient gel electrophoresis and real-time PCR. Molecular profiling demonstrated that transplantation of gut microbiota from a human to germfree piglets produced a donor-like microbial community with minimal individual variation. And the microbial succession with aging of those ex-germfree piglets was also similar to that observed in humans. This HFA model provides a significantly improved system for research on gut ecology in human metabolism, nutrition and drug discovery.

Changes to the content of sugars, sugar alcohols, myo-inositol, carboxylic acids and inorganic anions in developing grains from different varieties of Robusta (Coffea canephora) and Arabica (C. arabica) coffees

Changes in concentration of mono- and oligosaccharides, sugar alcohols, myo-inositol, carboxylic acids and inorganic anions in coffee grains were analysed during grain development in three cultivars of Coffea arabica L (Arabica) and two of C. canephora L var. Robusta (Robusta) by high performance anion exchange chromatography coupled to pulsed electrochemical detection (HPAE-PED). The majority of the components analysed either decreased in concentration during the first half of the development period or accumulated steadily during the latter half of the period. The profiles are taken to indicate relationships between the perisperm, the principal tissue in the young grain and the endosperm during maturation. While most of the free sugar in the mature grain is accounted for by sucrose, fructose and glucose are both at higher concentrations in the perisperm. Considerable amounts of myo-inositol (3‐4% dry weight (DW)) are found in young grains, while only the phosphorylated form phytic acid occurs in mature grains (0.3‐0.6% DW). Quinic acid, which is present in very low amounts in mature endosperm, represents between 6 and 16% DW in young grains, this possibly being the major precursor pool for the high amounts of chlorogenic acids (5‐10% DW) which are a characteristic of mature coffee grains. Of the other organic acids analysed, citric and malic acids are dominant in the mature grain, with higher concentrations in Arabica than Robusta. The results are discussed with respect to the potential implications for transport mechanisms in developing coffee grains and also the importance of the compounds analysed for industrial quality and flavour.

Maternal Docosahexaenoic Acid Feeding Protects Against Impairment of Learning and Memory and Oxidative Stress in Prenatally Stressed Rats: Possible Role of Neuronal Mitochondria Metabolism

AIMS Docosahexaenoic acid (22:6n-3; DHA) is known to play a critical role in postnatal brain development. However, no study has been performed to investigate its preventive effect on prenatal stress-induced behavioral and molecular alterations in offspring. In the present study, rats were exposed to restraint stress on days 14-20 of pregnancy, three times a day, 2 hours each time; DHA was given at the doses of 100 and 300 mg/kg/day for two weeks. RESULTS We showed that prenatal restraint stress caused (1) learning and memory impairment, (2) BDNF mRNA level decrease, (3) oxidative damage to proteins, (4) enhanced expression of nitric oxide synthase and apoptosis, and (5) abnormalities in mitochondrial metabolism that included changes in mitochondrial complexes I-V, and enhancement of expression of proteins involved in mitochondrial fusion/fission (Mfn-1, Mfn-2, Drp-1) and autophagy (Atg3, Atg7, Beclin-1, p-Akt, and p-mTOR) in the hippocampus of offspring. INNOVATION Besides the well-known role in child brain development, we reported the novel finding of DHA in protecting prenatal stress-induced cognitive dysfunction involving the modulation of mitochondrial function and dynamics. CONCLUSION Maternal feeding of DHA significantly prevented prenatal stress-induced impairment of learning and memory and normalized the biomarkers of oxidative damage, apoptosis, and mitochondrial metabolism in the hippocampus of both male and female offspring. These results suggest that maternal feeding of DHA exerts preventive effects on prenatal stress-induced brain dysfunction and that modulation of mitochondrial metabolism may play critical role in DHA protection.

Goji berry effects on macular characteristics and plasma antioxidant levels.

Purpose. Goji berry (Lycium barbarum L.) is purported to benefit vision because of its high antioxidant (especially zeaxanthin) content, although this effect has not been demonstrated in high-quality human studies. The purpose of this study was to evaluate the effects of daily supplementation with a proprietary milk-based formulation of goji berry, Lacto-Wolfberry (LWB), on macular characteristics and plasma zeaxanthin and antioxidant capacity levels in elderly subjects. Methods. This was a double-masked, randomized, placebo-controlled trial in healthy elderly subjects (range, 65 to 70 years) receiving 13.7 g/d of LWB (n = 75) or placebo (n = 75) for 90 days. Subjects underwent direct ophthalmic examination to assess pigmentation and soft drusen count in the macula and a blood draw to measure plasma zeaxanthin level and total antioxidant capacity. Results. The placebo group demonstrated hypopigmentation and soft drusen accumulation in the macula, whereas the LWB group remained stable. Both plasma zeaxanthin level and antioxidant capacity increased significantly in the LWB group, by 26% and 57%, respectively, but did not change in the placebo group. No product-related adverse events were reported in either group. Conclusions. Overall, daily dietary supplementation with goji berry for 90 days increases plasma zeaxanthin and antioxidant levels as well as protects from hypopigmentation and soft drusen accumulation in the macula of elderly subjects. However, the mechanism of action is unclear, given the lack of relationship between change in plasma zeaxanthin and change in macular characteristics.

Immunomodulatory Effects of Dietary Supplementation with a Milk-Based Wolfberry Formulation in Healthy Elderly: A Randomized, Double-Blind, Placebo-Controlled Trial

Wolfberry (fruit of Lycium barbarum) has been prized for many years in China for its immunomodulatory property and its high specific antioxidant content. However, clear clinical evidence demonstrating the effect of wolfberry dietary supplementation is still lacking. After our earlier report showing that a proprietary milk-based wolfberry formulation (Lacto-Wolfberry) enhances in vivo antigen-specific adaptive immune responses in aged mice, the present study aimed at demonstrating the effect of dietary Lacto-Wolfberry supplementation on immune functions in the elderly, especially vaccine response known to decline with aging. A 3-month randomized, double-blinded, placebo-controlled study was conducted on 150 healthy community-dwelling Chinese elderly (65-70 years old) supplemented with Lacto-Wolfberry or placebo (13.7 grams/day). Immune response to influenza vaccine was assessed in the study, along with inflammatory and physical status. No serious adverse reactions were reported during the trial, neither symptoms of influenza-like infection. No changes in body weight and blood pressure, blood chemistry or cells composition, as well as autoantibodies levels were observed. The subjects receiving Lacto-Wolfberry had significantly higher postvaccination serum influenza-specific immunoglobulin G levels and seroconversion rate, between days 30 and 90, compared with the placebo group. The postvaccination positive rate was greater in the Lacto-Wolfberry group compared to the placebo group, but did not reach statistical significance. Lacto-Wolfberry supplementation had no significant effect on delayed-type hypersensitivity response and inflammatory markers. In conclusion, long-term dietary supplementation with Lacto-Wolfberry in elderly subjects enhances their capacity to respond to antigenic challenge without overaffecting their immune system, supporting a contribution to reinforcing immune defense in this population.

A Milk-Based Wolfberry Preparation Prevents Prenatal Stress-Induced Cognitive Impairment of Offspring Rats, and Inhibits Oxidative Damage and Mitochondrial Dysfunction In Vitro

Lycium barbarum (Fructus Lycii, Wolfberry, or Gouqi) belongs to the Solanaceae. The red-colored fruits of L. barbarum have been used for a long time as an ingredient in Chinese cuisine and brewing, and also in traditional Chinese herbal medicine for improving health. However, its effects on cognitive function have not been well studied. In the present study, prevention of a milk-based wolfberry preparation (WP) on cognitive dysfunction was tested in a prenatal stress model with rats and the antioxidant mechanism was tested by in vitro experiments. We found that prenatal stress caused a significant decrease in cognitive function (Morris water maze test) in female offspring. Pretreatment of the mother rats with WP significantly prevented the prenatal stress-induced cognitive dysfunction. In vitro studies showed that WP dose-dependently scavenged hydroxyl and superoxide radicals (determined by an electron spin resonance spectrometric assay), and inhibited FeCl2/ascorbic acid-induced dysfunction in brain tissue and tissue mitochondria, including increases in reactive oxygen species and lipid peroxidation and decreases in the activities of complex I, complex II, and glutamate cysteine ligase. These results suggest that dietary supplementation with WP may be an effective strategy for preventing the brain oxidative mitochondrial damage and cognitive dysfunction associated with prenatal stress.

SUGAR ACCUMULATION IN TOMATO AND PARTIAL PURIFICATION OF BUFFER-INSOLUBLE INVERTASE

Abstract Sugar accumulation and invertase activity were studied during fruit development in tomatoes (Lycopersicon esculentum) of two breeding lines. Tomatoes of breeding line A had substantially larger amounts of reducing sugars (38%), glucose (33%) and fructose (23%) at the ripe stage compared to breeding line B. Differences in sugar content between the two breeding lines were significant at all stages of maturation, and higher amounts of sugars were consistently linked to higher invertase activities. Buffer-insoluble acid invertase from pericarp was partially purified (12-fold) and consisted, under denaturing conditions (SDS-PAGE), of four distinct bands with estimated Mr of 56, 32, 21 and 19 k, respectively. Gel filtration chromatography on Sephacryl S-200 and on FPLC Superose 12 indicated a Mr of 50 k for the active invertase enzyme. Western blot analysis with an antibody to carrot cell wall invertase suggested that the two bands with Mr of 56 and 21 k correspond to the invertase enzyme. IEF of purified invertase enzyme gave three major bands with estimated isoelectric points between 5.65 and 5.80. The pH optimum of invertase activity was 5.0, and the activity was stable from pH 3.5 to 7.0.

FT-IR Methodology for Quality Control of Arabinogalactan Protein (AGP) Extracted from Green Tea (Camellia sinensis)

A rapid methodology of quality control was developed for arabinogalactan proteins (AGP) extracted and purified from green tea. Using the vectorial angle method and IR spectrum analysis, the 1200-800 cm(-1) region in second-derivative IR spectra was determined as the key fingerprinting region of green tea AGP, with the 1090-900 cm(-1) region reflecting their conservative and common characteristics. In fact, the key monosaccharides, galactose (Gal) and arabinose (Ara), were shown to have intense peaks at about 1075 and 1045 cm(-1), respectively, and uronic acids at about 1018 cm(-1) in second-derivative IR spectra. The variable region was identified to be at about 1134-1094 and 900-819 cm(-1) and was probably due to compositional and structural differences between AGPs. The constructed methodology was tested on green tea AGP extracted by three treatments and purified to apparent homogeneity as water-extracted Camellia sinensis AGP (CSW-AGP), pectinase-extracted C. sinensis AGP (CSP-AGP), and trypsin-extracted C. sinensis AGP (CST-AGP) with an Ara/Gal ratio of 1.37, 1.57, and 1.82, respectively. Regarding in vitro antioxidant activity, the AGPs (CSW-AGP and CST-AGP) with higher similarity (closer cos theta values calculated for second-derivative IR spectra) exhibited a similar ability of chelating ferrous ions and had a similar capability for scavenging hydroxyl radicals. In conclusion, the combination of second-derivative IR spectrum analysis and the vectorial angle method has allowed a successful characterization of green tea AGPs and was shown to be suitable for their compositional and activity discrimination and rapid quality evaluation.