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Wissenschaftliche Studien deuten auf eine gesundheitsförderndes und gewichtsregulierendes Potential des Kaffees hin, welches vor allem mit dem hohen Gehalt an Antioxidantien in Verbindung gebracht wird. In dieser Arbeit wurden Kaffeeextrakte, -inhaltsstoffe sowie Kaffeegetränke hinsichtlich ihrer antioxidativen Wirksamkeit untersucht. In-vitro wurde die präventive Wirkung von Extrakten aus leicht (AB1), mittel (RI, AC, AB) und stark (AB 2) gerösteten Kaffees mit Markern oxidativer Zellschädigung und Zellantwort in den Kolonkarzinomzelllinien HT-29 und Caco-2 charakterisiert. Vergleichend wurden die ausgewählten originären Kaffeeinhaltstoffe 5-Caffeoylchinasäure (5-CQA) und Trigonellin (TRIG) sowie die Röstprodukte Kaffeesäure (CA), Catechol (Cat), 1,2,4-Trihydroxybenzol (THB), N-Methylpyridinium (NMP) und methylierte NMP-Analoga untersucht. Erfasste Parameter waren zellulärer ROS-Level, (oxidative) DNA-Schädigung und Proteinexpression der ARE-abhängigen Enzyme NQO1, g-GCL und GSR. Zusätzlich wurde die direkte antioxidative Aktivität mittels TEAC- und ORAC-Assay gemessen. Die Ergebnisse zeigten eine radikalabfangende Eigenschaft aller Kaffeeextrakte mit Werten von 0,9-1,5 mM Trolox (TEAC) bzw. 2,5-2,8 mM Trolox (ORAC). Der zelluläre ROS-Level wurde in HT-29 Zellen durch die Extrakte AB, AC, RI und stark gerösteten AB 2 signifikant verringert. Eine konzentrationsabhängige und signifikante Induktion ARE- abhängiger Enzyme (NQO1, g-GCL und GSR) in HT-29 Zellen wurde durch den CQA-reichen AB 1 beobachtet, der NMP-reiche AB 2 war dagegen unwirksam. Von den untersuchten Kaffeeinhaltsstoffen zeigten nur die phenolischen Verbindungen 5-CQA und CA eine ausgeprägte zellfreie antioxidative Aktivität. Der zelluläre ROS-Level konnte durch 5-CQA verringert werden, dagegen waren methylierte NMP-Analoga in der Lage (oxidative) DNA-Schäden in Caco-2 Zellen zu reduzieren. 5-CQA induzierte wie leicht gerösteter AB 1 die Proteinexpression alle untersuchten Enzyme in HT-29 Zellen. Weiterhin wurde in zwei aufeinander folgenden humanen Interventionsstudien das antioxidative Potential unterschiedlicher Kaffeegetränke mit hohem Anteil an Antioxidantien charakterisiert; in der zweiten Studie wurde zusätzlich auf gewichtsregulierende Wirkung des Studienkaffees geprüft. Im Rahmen der ersten Pilotstudie, durchgeführt durch AG. Somoza (DFA Garching) wurde die Modulation von DNA-Schäden im Blut von Probanden nach Konsum zweier Kaffeegetränke erfasst, die reich an Chlorogensäuren oder an N-Methylpyridinium waren (Kaffeekonsum: 0,5 L pro Tag, 4 Wochen). Beide Kaffeegetränke bewirkten im Blut gesunder Probanden eine deutliche Abnahme oxidativer DNA-Schäden. In der zweiten Interventionsstudie nahmen 35 männliche Probanden nach einer vierwöchigen Wash-out Phase über vier Wochen täglich 750 ml eines Antioxidantien reichen Kaffees (580 mg/l CQAs; 71,7 mg/l NMP) auf, anschließend folgte eine vierwöchige Wash-out Phase. Zu Beginn der Studie und am Ende jeder Phase wurden Blutentnahmen zur Bestimmung der Biomarker (oxidative) DNA-Schäden, Glutathion (Gesamtglutathion tGSH, oxidiertes Glutathion GSSG) sowie Bioimpedanzanalysen zur Bestimmung der Körperzusammensetzung durchgeführt. Zusätzlich wurden Energie und Nährstoffaufnahme der Probanden erfasst. Die Ergebnisse zeigten eine signifikante Abnahme (oxidativer) DNA-Schäden (p < 0,001), Anstieg des tGSH-Spiegels (p<0,05) und des GSH-Status (Trend) und eine Erniedrigung des GSSG-Spiegels. Weiterhin wurde eine signifikante Abnahme von Körpergewicht und Körperfett der Probanden während der Kaffeeintervention im Vergleich zur beiden Wash-out Phasen beobachtet, welche vor allem bei Probanden mit BMI < 25 stärker ausgeprägt war. Die Verringerung der Energie- und Nährstoffaufnahme während der Kaffeephase weist auf eine Kaffeeabhängige beeinflussung der Sättigungsregulation hin. Zusammenfassend besitzt der Antioxidantien reiche Studienkaffee ein eindeutiges Potential zur Verringerung oxidativer Zellschädigung sowie gewichtsregulierende Wirkung in gesunden Probanden. Aufgrund der von uns erhaltenen in vitro Daten kann die antioxidative Wirksamkeit zum Teil auf die untersuchten originären Kaffeeinhaltsstoffen (vor allem CQAs) und Röstprodukte zurückgeführt werden. Andere bisher nicht charakterisierte Sustanzen/Stoffgruppen tragen vermutlich ebenfalls zu der beobachteten Wirkung bei.
Hordatines are a characteristic class of secondary metabolites found in barley which have
been reported to be present in barley malt, beer and, recently, brewer ́s spent grain (BSG). However,
little is known about their biological activities such as antioxidative effects in beer or antifungal
activity as their main task within the plants. We conducted an in vitro investigation of the activity
of hordatines isolated from BSG towards enzymes of glucose metabolism. Hordatine-rich fractions
from BSG were prepared by solid-liquid extraction (SLE) with 60% acetone followed by purification
and fractionation. The fractions were characterised and investigated for their in vitro inhibitory
potential on α-glucosidase and glycogen phosphorylase α (GPα). Both enzymes are relevant within
the human glucose metabolism regarding the digestion of carbohydrates as well as the liberation of
glucose from the liver. In total, 10 hordatine-rich fractions varying in the composition of different
hordatines were separated and analysed by mass spectrometry. Hordatine A, B and C, as well as
hydroxylated aglycons and many glycosides, were detected in the fractions. The total hordatine
content was analysed by HPLC-DAD using a semi-quantitative approach and ranged from 60.7 ± 3.1
to 259.6 ± 6.1 μg p-coumaric acid equivalents/mg fraction. Regarding the biological activity of
fractions, no inhibitory effect on GPα was observed, whereas an inhibitory effect on α-glucosidase
was detected (IC50 values: 77.5 ± 6.5–194.1 ± 2.6 μg/mL). Overall, the results confirmed that
hordatines are present in BSG in relatively high amounts and provided evidence that they are potent
inhibitors of α-glucosidase. Further research is needed to confirm these results and identify the active
hordatine structure.
Winery by-products arise in high amounts during winemaking processes. Hence, recovery alternatives are of great interest. In this study, effects of extracts from winery by-products (Vitis vinifera L. cv. Riesling) on mitochondrial functions in human hepatocellular carcinoma (HepG2) cells were examined. Polyphenolic profiles of pomace (PE), stem (SE), vine leaf (VLE), and vine shoot extracts (VSE) were characterized by HPLC-UV/Vis-ESI-MS/MS. The extracts induced dose-dependent cytotoxic effects (PE > SE > VLE > VSE). VSE showed protective effects regarding modulation of tert-butyl hydroperoxide (TBH)-induced intracellular reactive oxygen species (ROS) levels. PE, SE and VLE increased the mitochondrial membrane potential (MMP), whereas VSE decreased it owing to mildly impaired mitochondrial respiration. Cells may try to compensate reduced respiration chain complex activities by increasing the mitochondrial mass, as indicated by enhanced citrate synthase activity and mRNA expression levels after VSE incubation. Thus, winery by-products represent interesting sources of bioactive compounds that exert positive or negative effects on mitochondrial functions.
With a yearly production of about 39 million tons, brewer’s spent grain (BSG) is the
most abundant brewing industry byproduct. Because it is rich in fiber and protein, it is commonly
used as cattle feed but could also be used within the human diet. Additionally, it contains many
bioactive substances such as hydroxycinnamic acids that are known to be antioxidants and potent
inhibitors of enzymes of glucose metabolism. Therefore, our study aim was to prepare different
extracts—A1-A7 (solid-liquid extraction with 60% acetone); HE1-HE6 (alkaline hydrolysis followed
by ethyl acetate extraction) and HA1-HA3 (60% acetone extraction of alkaline residue)—from various
BSGs which were characterized for their total phenolic (TPC) and total flavonoid (TFC) contents,
before conducting in vitro studies on their effects on the glucose metabolism enzymes α-amylase,
α-glucosidase, dipeptidyl peptidase IV (DPP IV), and glycogen phosphorylase α (GPα). Depending
on the extraction procedures, TPCs ranged from 20–350 μg gallic acid equivalents/mg extract
and TFCs were as high as 94 μg catechin equivalents/mg extract. Strong inhibition of glucose
metabolism enzymes was also observed: the IC50 values for α-glucosidase inhibition ranged from
67.4 ± 8.1 μg/mL to 268.1 ± 29.4 μg/mL, for DPP IV inhibition they ranged from 290.6 ± 97.4 to
778.4 ± 95.5 μg/mL and for GPα enzyme inhibition from 12.6 ± 1.1 to 261 ± 6 μg/mL. However, the
extracts did not strongly inhibit α-amylase. In general, the A extracts from solid-liquid extraction
with 60% acetone showed stronger inhibitory potential towards α-glucosidase and GPα than other
extracts whereby no correlation with TPC or TFC were observed. Additionally, DPP IV was mainly
inhibited by HE extracts but the effect was not of biological relevance. Our results show that BSG
is a potent source of α-glucosidase and GPα inhibitors, but further research is needed to identify
these bioactive compounds within BSG extracts focusing on extracts from solid-liquid extraction
with 60% acetone.
Red fruits and their juices are rich sources of polyphenols, especially anthocyanins.
Some studies have shown that such polyphenols can inhibit enzymes of the carbohydrate metabolism,
such as α-amylase and α-glucosidase, that indirectly regulate blood sugar levels. The presented
study examined the in vitro inhibitory activity against α-amylase and α-glucosidase of various
phenolic extracts prepared from direct juices, concentrates, and purees of nine different berries which
differ in their anthocyanin and copigment profile. Generally, the extracts with the highest phenolic
content—aronia (67.7 ± 3.2 g GAE/100 g; cyanidin 3-galactoside; chlorogenic acid), pomegranate
(65.7 ± 7.9 g GAE/100 g; cyanidin 3,5-diglucoside; punicalin), and red grape (59.6 ± 2.5 g GAE/100 g;
malvidin 3-glucoside; quercetin 3-glucuronide)—showed also one of the highest inhibitory activities
against α-amylase (326.9 ± 75.8 µg/mL; 789.7 ± 220.9 µg/mL; 646.1 ± 81.8 µg/mL) and α-glucosidase
(115.6 ± 32.5 µg/mL; 127.8 ± 20.1 µg/mL; 160.6 ± 68.4 µg/mL) and, partially, were even more potent
inhibitors than acarbose (441 ± 30 µg/mL; 1439 ± 85 µg/mL). Additionally, the investigation of single
anthocyanins and glycosylated flavonoids demonstrated a structure- and size-dependent inhibitory
activity. In the future in vivo studies are envisaged.
Phosphodiesterases (PDEs) are essential enzymes for the regulation of pathways mediated
by cyclic adenosine monophosphate (cAMP). Secondary plant compounds like anthocyanins (ACs)
can inhibit PDE activity and, consequently, may be beneficial for lipid metabolism. This study
investigated 18 AC-rich juice extracts and pure reference compounds from red fruits for potential
inhibitory effects on PDE 3B activity. Extracts were obtained through adsorption on Amberlite® XAD
7 resin. Based on this screening, the chokeberry, blueberry, pomegranate, and cranberry extracts
were active, with half maximal inhibitory concentrations (IC50) ranging from 163 ± 3 µg/mL to
180 ± 3 µg/mL. The ACs in these extracts, peonidin-3-glucoside and cyanidin-3-arabinoside, were the
most active single compounds (IC50 = 56 ± 20 µg/mL, 108 ± 6 µg/mL). All extracts comprised high
amounts of phenolic compounds, as determined by the Folin–Ciocalteu assay, ranging from 39.8 ± 1.5
to 73.5 ± 4.8 g gallic acid equivalents (GAE)/100 g extract. Pomegranate and chokeberry extracts
exhibited the largest amounts of polyphenols (72.3 ± 0.7 g GAE/100 g, 70.6 ± 4.1 g GAE/100 g,
respectively). Overall, our results showed that fruit juice extracts and their ACs can inhibit PDE
activity. Any potential health benefits in vivo will be investigated in the future.
Sustained Human Background Exposure to Acrolein Evidenced by Monitoring Urinary Exposure Biomarkers
(2019)
Scope
This study investigates a potential correlation between the intake of heat-processed food and the excretion of the acrolein (AC) biomarkers N-acetyl-S-(3-hydroxypropyl)-l-cysteine (HPMA) and N-acetyl-S-(carboxyethyl)-l-cysteine (CEMA) based on two human studies.
Methods and Results
Human exposure to AC is monitored using the AC-related mercapturic acids HPMA and CEMA in the urine of a) non-smoking volunteers under defined living conditions and b) of non-smoking volunteers on unrestricted or vegan diet under free living conditions. Free living volunteers in part show markedly enhanced urinary excretions of HPMA and CEMA. The intake of heat-processed food does not influence AC-related biomarker excretion. Incidentally enhanced urinary exposure biomarker levels appear to suggest AC exposure possibly from open fire, barbecuing, or tobacco smoke. However, kinetics of urinary biomarkers related to tobacco and other potential smoke exposure, do not correlate with those observed for HPMA and CEMA.
Conclusion
This study is the first to convincingly show a sustained and substantial background exposure to AC in non-smoking humans, clearly independent from uptake of heat-processed foods. The data strongly point to endogenous AC generation by pathways of mammalian and/or microbial metabolism as yet not taken into consideration.
Purpose
We investigated the cytosolic and membrane-associated contents of polyphenols after 4 hours of incubation (50 μM of each polyphenol) in the colon carcinoma cell line T84 using a novel, rapid, and convenient method based on permeabilization of the cell membrane using digitonin. The colon carcinoma cell line was used to investigate the intestinal uptake of polyphenols present in apple products.
Recent Findings
The results showed that hydroxycinnamic acids (caffeic and 5-caffeoylquinic acid) were only detected in the cytosolic fractions. In contrast, 0.3 to 8.2% of the initial concentrations (50 μM) of the flavonoids phloretin, quercetin, phloretin 2′-O-glucoside, and quercetin 3-O-rhamnoside were found in the membrane-associated fractions. In the cytosolic fractions, 0.2–2.9% of these compounds were detected, corresponding to 25 to 40% of the total cell-associated (cytosolic plus membrane-associated fractions) polyphenol content.
Summary
Our results showed that after uptake, polyphenols were present in the cytosolic fraction of the cells as well as associated with the cell membrane. The presented method provides a useful in vitro tool for determining biologically active compounds in cellular fractions.