8. Health, Nutrition and Food Science
Transcript of 8. Health, Nutrition and Food Science
123
8. Health, Nutrition and Food Science
124
Health, Nutrition and Food Science
125
Influence of molecular weight of ethylcellulose in aqueous based coatings
Jurgita Kazlauskeab, Sara Almera and Anette Larssonab
aPharmaceutical Technology, Applied Chemistry, Department of Chemistry and Chemical Engineering, Chalmers University of Technology, 412 96 Goteborg,
Sweden bSuMo Biomaterials, a VINN Excellent Center at Chalmers University of Technology,
412 96 Goteborg, Sweden
[email protected] and [email protected]
One of the most commonly used polymers for controlled release from aqueous-
based coatings is ethylcellulose, a water-insoluble cellulose derivative. Previous
investigations on aqueous-based ethylcellulose dispersions are done by using an
ethylcellulose viscosity grade of 20 cps (1-3). Since different molecular weights
of ethylcellulose correlates with different properties for the pure material, it is
natural to think that the properties of aqueous-based coatings produced with
different molecular weights of ethylcellulose will also differ from each other.
Previous work in our group revealed that the molecular weight of ethylcellulose
was important for organic solvent based coatings, there for example mixtures of
hydroxypropyl cellulose and ethylcellulose of different molecular weight gave
different microstructural features and permeabilities (4).
The study shows that the coalescence is decreasing with increasing molecular
weight and the permeability of water is increasing. These findings are important
features in the development of new pharmaceutics with drug release rates
controlled by aqueous-based latex coatings.
References.
1. Frohoff-Hülsmann MA et al., International journal of pharmaceutics. 1999;177(1):69–82. 2. Parikh NH et al., Pharmaceutical research. 1993;10(4):525–34. 3. Muschert S et al., International journal of pharmaceutics. 2009;368(1):138–45. 4. Andersson H et al. European journal of pharmaceutical sciences 2013;48(1–2):240–8.
Health, Nutrition and Food Science
126
Temporal transcriptional response to insulin stimulation of myocytes: Impact of type 2 diabetes and obesity
Leif Väremoa, Camilla Scheeleb, Christa Broholmb, Adil Mardinoglua, Intawat Nookaewa, Mathias Uhlénc, Bente Klarlund Pedersenb, Jens Nielsena,c
aBiology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
bThe Centre of Inflammation and Metabolism and the Centre for Physical Activity Research, Department of Infectious Diseases, Rigshospitalet, University of
Copenhagen, Denmark cScience for Life Laboratory, Royal Institute of Technology (KTH), Stockholm,
Sweden
Skeletal muscle is the major target tissue for insulin resistance preceding the
development of type 2 diabetes (T2D). To unravel how this development can be
prevented it is important to map the interaction between insulin and insulin
stimulated responses in human skeletal muscle cells. Therefore, the aim of this
study is to elucidate time-dependent transcriptional responses invoked by insulin
stimulation and in particular to investigate how these temporal profiles manifest
in molecular mechanisms at the level of cellular metabolism and hereby confer
insulin resistance and possible T2D. This is achieved by deep RNA-sequencing
of 96 samples from primary human muscle cells from a clinically well-defined
cohort (covering healthy and diabetic, obese and non-obese, males and females)
and performing thorough data analysis and integration with genome-scale
metabolic modeling. Preliminary results point at a major activity 2 hours after
insulin stimulation and a distinction of implicated processes in the different sub-
groups.
Figure 1. An outline of the experimental design. From a larger cohort, 24 subjects were selected and
divided into four groups (NGT/OB, NGT/non-OB, T2D/OB, T2D/non-OB; NGT=normal glucose tolerant,
OB=obese, T2D=type 2 diabetic) with three females and three males in each group. A vastus lateralis
biopsy was taken from each subject (1), and myoblasts were isolated and differentiated in vitro (2). Fully
differentiated myocytes were serum starved for 2h and stimulated with insulin (3). RNA samples for
sequencing were taken at four time points (4).
Health, Nutrition and Food Science
127
Taurine bioaccessibility but not bioavailability in Caco-2 cells is increased by the omega-3 fatty acids eicosapentaenoic acid and
docosahexaenoic acid
Andrew Vincent, Nathalie Scheers and Ann-Sofie Sandberg
Food Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
Email: [email protected]
BACKGROUND: A diet rich in fish reduces risk factors for cardiovascular
disease. This has been mainly attributed to ω-3 fatty acids but fish contain several
other functional compounds that have proved to be important for cardiovascular
health. Fish also contains high amounts of taurine, which has been shown to
improve vascular health by regulating calcium transport and acting to maintain
membrane stability and redox homeostasis. As these nutrients are seldom
consumed in isolation, it is important to study combinations of nutrients to
understand how they may work synergistically with each other. Omega-3 fatty
acids may influence membrane fluidity and transporters, with a potential impact
on nutrient bioavailability, which could affect the uptake of other nutrients from
fish, including taurine.
OBJECTIVES: To determine if taurine bioaccessibility (uptake) and
bioavailability (basal efflux) is influenced by the ω-3 fatty acids, eicosapentaenoic
acid (EPA) and docosahexaenoic acid (DHA).
METHODS: The human intestinal Caco-2 cell model was used to estimate the
bioaccessibility and bioavailability of taurine. Transport of taurine (20 mM), as
[3H]-taurine (18.5 kBq mL-1), in the presence of DHA (100 μM) and EPA (100
μM) was measured by liquid scintillation counting.
RESULTS: The results indicate that taurine uptake was significantly increased in
the presence of DHA (28 %) and EPA (22 %). Taurine cellular efflux was reduced
by EPA (14 %) and DHA (8 %).
CONCLUSIONS: EPA and DHA increase the bioaccesibility of taurine but do
not affect the efflux, therefore even though local taurine concentrations may be
increased, there does not appear to be an effect on taurine bioavailability.
Health, Nutrition and Food Science
128
Effect of solvent system and biomass pretreatment method on the yield and quality of fatty acids from Saccharina latissima
Giorgia Tibalderoa,b,c, Joshua Mayersb, Eva Albersb, Ingrid Undelanda
Dept. Biology and Biological Engineering, aDiv. Food and Nutrition Science & bIndustrial Biotechnology, Chalmers University of Technology, Göteborg, Sweden
cUniversità degli studi di Torino, Dept. Scienze della Vita e Biologia dei sistemi, Unit Biochemistry, Turin Italy
Email: [email protected], [email protected]
Although levels of fatty acids in seaweeds are generally low (<10 mg g DW-1),
this fraction is enriched in long chain n-3 polyunsaturated fatty acids. In addition,
this fraction can contain considerable quantities of hydrophobic compounds such
as vitamins, sterols and polyphenols, which still mark seaweed lipids as an
interesting nutritional resource. In the SEAFARM project, Saccharina latissima
harvested from Sweden’s West coast will be subjected to biorefining, with the
aim of maximizing its value as a raw material for food/feed ingredients, chemicals
and energy.
A possible limiting step in this field is the compositional characterization of the
structurally complex seaweed biomass, not least its lipid fraction. The present
study aims to develop extraction methods, which maximize accurate measurement
of this pool. Furthermore, the applicability of different extraction methods to
large-scale biorefinery scenarios is assessed, taking into account that ‘wet-
biomass’ will mostly likely be utilised in such cases.
Firstly, chloroform & methanol systems were tested for fatty acid extraction
(Folch vs. Bligh & Dyer methods) on wet and freeze-dried material, in
conjunction with several pre-treatment methods (mechanical disruption, ultra-
sonication and microwave-assisted extraction). The best pretreatment was then
tested using ethanol and 1-butanol. Direct transesterification was conducted as a
reference method and other techniques appraised with respect to the yield and
profile of extracted fatty acids.
Overall, the modified Bligh & Dyer chloroform:methanol method was the most
effective solvent system in unassisted extractions (5.3 mg g fatty acids DW-1);
microwave and sonication pre-treatments significantly improved fatty acid yields
(7.3 mg g DW-1). Ultra-sonication assisted ethanol extraction performed equally
as well as Bligh & Dyer with pre-treatment (7.6 mg g DW-1). Yields from wet
biomass were generally comparable to those from freeze-dried material. Most
extracts were rich in polyunsaturated fatty acids (40-50%), with up to 0.82 and
0.72 mg g DW-1 being stearidonic and eicosapentaenoic acid, respectively.
Further results and insights into lipid extraction will be presented.
Health, Nutrition and Food Science
129
Genome Scale Metabolic Modeling of Child Growth, 0-6 months
Avlant Nilssona, Adil Mardinoglua and Jens Nielsen 3a
a Department for Biology and Biological Engineering
Malnutrition of children is a global challenge. Improving our understanding of
child growth may translate in to faster detection and to cost efficient diet
supplements. Genome Scale Metabolic Models (GEMs) have been successfully
applied to many biological systems. Here we make use of a GEM, HMR 2.0 [1],
to model the effect of co-factor deficiency in infants. The concentrations of
metabolites in breast milk are given as input, and the growth of biomass is the
output. The model also includes, weight dependent, maintenance energy
expenditure. Co-factor deficiency is modelled by reducing the metabolic flux
through reactions that require a given cofactor (Figure 1).
Figure 1. Growth trajectories for deficiency of 3 different co factors. The flux through their associated
reactions was limited to 90% of the optimal flux. The 5th, 50th and 95th percentile of WHO growth data
for boys as reference.
References.
[1] Adil Mardinoglu, Rasmus Agren, Caroline Kampf, Anna Asplund, Mathias Uhlen, and Jens
Nielsen. Genome-scale metabolic modelling of hepatocytes reveals serine deficiency in
patients
with non-alcoholic fatty liver disease. Nature communications, 5(May 2013):3083, 2014.
Health, Nutrition and Food Science
130
In vitro study of gastrointestinal oxidation of marine long chain n-3 polyunsaturated fatty acids – comparison between models based
on human fluids or porcine extracts
Tullberg, C.a, Vegarud, G.b and Undeland, Ia
aDivision of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Sweden
bDivision of Food proteins; Structure and biological function, Department of Chemistry, Biotechnology and Food Science, Norwegian University of
Life Sciences, Norway
Marine long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) have attracted
public awareness due to the positive health effects they have been associated with.
LC n-3 PUFA intake has been connected with reduced risk for many diseases, e.g.
cardiovascular and inflammatory diseases. In vitro studies have shown that fish
lipids can oxidize under gastrointestinal tract (GIT) conditions when using
digestive enzymes of porcine origin (1). The lipid oxidation reaction is known to
generate highly reactive oxidation products that may interact with DNA and
proteins, leading e.g. to various effects on cell function. Some of these oxidation
products are malondialdehyde (MDA) and the α,β-unsaturated aldehydes 4-
hydroxy-trans-2-nonenal (HNE) and 4-hydroxy-trans-2-hexenal (HHE), but little
is known about formation of these compounds in the GIT.
In this study, two GIT in vitro models were set up to study the effect of digestion
on lipid oxidation in cod liver oil. In the first model, digestive enzymes and bile
of porcine origin were used, and in the second model, human digestive juices were
used. The standardised Infogest protocol (2) was applied to the models and
enzymatic activities were matched between them. The digesta was analysed for
reactive oxidation products (MDA, HNE and HHE) by Liquid Chromatography–
Mass Spectrometry (LC-MS) and for free fatty acids by Gas Chromatography-
Mass Spectrometry (GC-MS). Results show that lipid oxidation occurs in both
models under gastric as well as intestinal conditions. However, both time
dependence and maximum levels of oxidation products differed, something which
will be discussed.
References.
1. Larsson, K. et al., J. Agric. Food Chem. (2012) 60:7556-7564
2. Minekus, M. et al., Food & Function (2014) 5:1113-24
Health, Nutrition and Food Science
131
Eicosapentaenoic and docosahexaenoic acid-enriched high fat diet delays the development of fatty liver in mice
Nikul Soni1, Intawat Nookaew2, 3, Ann-Sofie Sandberg1*, Britt G Gabrielsson1
1Divisions of Food Science and Nutrition and 2Systems Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296
Gothenburg, Sweden. Present address: 3Comparative Genomics Group, Biosciences division, Oak Ridge National Library, Oak Ridge, TN 37831, USA.
Scope: To investigate the effects of quality of fat in a high fat diet HFD)
over time on hepatic lipid storage and transcriptome in mice.
Methods and results: Male C57BL/6J mice were fed control, HFD-
EPA/DHA or HFD-corn oil diet for 8 or 12 weeks. Body weights, body
composition, plasma and hepatic triglyceride content were measured. Hepatic
transcriptomes were analysed by microarray followed by gene-set enrichment
analyses. At 8 weeks, the HFD-corn oil mice had higher body weight and
adipose depot mass than the HFD-EPA/DHA but there were no differences
at 12 weeks. Hepatic triglyceride content was low in HFD-EPA/DHA fed
mice at both time- points. Enrichment analyses showed that lipid/fatty acid
biosynthesis; transport and homeostasis were lower in the HFD-corn oil fed
compared with the HFD-EPA/DHA fed mice. Genes encoding proteins
associated to cytoplasmic lipid droplets were expressed at higher levels in
livers from the HFD-corn oil compared to HFD-EPA/DHA mice.
Conclusions: EPA and DHA maintained lower hepatic triglyceride content
despite being fed HFD. The liver transcriptome data implicate that the quality
of dietary fat could modulate Ppar-related gene expression that in turn affects
hepatic lipid storage and maintenance of metabolic health.
Health, Nutrition and Food Science
132
Serum proportions of eicosapentaenoic acid (EPA) in infants may decrease allergy risk: results from the FARMFLORA birth-cohort
Karin Jonsson1, Malin Barman1, Sara Moberg1, Hilde K Brekke2, Agnes Wold3, and Ann-Sofie Sandberg4
1Food Science, Dept. of Biology and Biological Engineering, Chalmers 2Dept. of Internal Medicine and Clinical Nutrition, University of Gothenburg
3Dept. of Infectious medicine, Clinical Bacteriology section, University of Gothenburg
Email: [email protected]
Background and aim: The allergy prevalence has increased drastically in
countries with a Western lifestyle, and the cause is largely unknown. Some
protective factors have been identified, such as growing up on farms. Data also
suggest that serum composition of polyunsaturated fatty acids (PUFAs) may
affect allergy development. The aims of this study were to: 1) relate fatty acid
composition in cord and infant serum to allergy development; 2) correlate infant
serum fatty acids to maternal diet during pregnancy as well as to diet and breast
milk fatty acids during lactation; 3) relate differences in serum fatty acid
composition to farm residence.
Methods: Twenty-eight farm and 37 non-farm mother-infant pairs were followed
in the FARMFLORA birth-cohort. Fatty acid proportions were analyzed in cord
serum and in child serum and breast milk four months postpartum. Maternal diet
during pregnancy was assessed by food-frequency questionnaires; diet during the
fourth month of lactation was recorded by 24-hour dietary recalls followed by 24-
hour food diaries. Serum fatty acid composition was related to doctors’ diagnosis
of allergy at three years of age as well as to farm residence, and correlated to diet
and breast milk fatty acids.
Results: Proportions in serum at four months of eicosapentaenoic acid (EPA, a
long-chain omega-3 PUFA) were higher in healthy compared to allergic children
(0.48% and 0.16%, respectively, P = .001). The Odds Ratio was 0.47 (P = .01,
95% CI: 0.27-0.83) for every 0.1% increase in EPA, and did not change
substantially after being adjusted for potential confounders. Serum EPA
proportions at four months correlated to maternal fatty fish intake during
pregnancy (rho = 0.4, P = .01) as well as to breast milk proportions and maternal
intake of long-chain omega-3 PUFAs during lactation (rho = 0.4 and 0.5, P = .05
and .01, resp.). When farm children were compared to non-farm children, higher
proportions of arachidonic acid were found in cord serum (14% and 12%,
respectively, P = .001).
Conclusion: Our preliminary data suggest that low proportions of EPA in infants’
serum may increase the risk of developing allergy. The protective effects could
be mediated by EPA per se or by a higher fish intake, which might offer protection
by other means than solely its EPA content.
Health, Nutrition and Food Science
133
Supercritical CO2 extraction of bilberry (Vaccinium myrtillus L.) seeds: composition and antioxidant activity
Graziele Gustinelli Arantes de Carvalhoab, Marie Almingerb and Lilia Ahrnéab
aSP Technical Research Institute of Sweden bDept. of Biology and Biologial Engineering
Chalmers University of Technology
Bilberries (Vaccinium myrtillus L.) are commonly used as raw material for food
and drinks in Europe, generating seeds as by-products, which contain valuable bio
compounds. The objective of this study was to evaluate the properties of bilberry
seed oil extracted by supercritical fluid extraction.
Extractions were carried out using 50 ±0.1 g of bilberry seeds (milled for 30 s and
moisture content of 4.49%), using a flow rate of 40g CO2/min, for 120 min (to
determine yield) and 80 min (to use for analysis). Pressure at 200, 350 and 500
bars and temperature at 40, 50 and 60°C were selected. Analysis of fatty acid
profile, vitamin E, free radical scavenging activity (DPPH) and peroxide content
were carried out. The mean values of duplicates were compared by one-way
ANOVA and when different, T-test was applied (p<0.05 for both tests).
The yields obtained at 80 min were 8.0-21.4%. Bilberry seed oil showed high
amount of fatty acids (75.1-76.2 g/100 g of oil) and had similar composition
between the different conditions. All extracts had high amount values of 18:2 n-6
(ω6) and 18:3 n-3 (ω3) and the ratio ω6/ω3 < 1. The samples contained 47.1-105.3
mg vitamin E, gamma-tocotrienol/100g of oil. The values for Antioxidant
Activity Index (AAI) obtained were 2.1-3.57. Peroxide values were 1.1-2.0
mekv/kg of oil. The different extraction conditions resulted in significant different
values of vitamin E, DPPH and peroxides. Oils extracted from different conditions
obtained statistical differences in all analysis, except in total fatty acids.
The high amount of ω6 and ω3 and low ω6/ω3 ratio present in all extracts, indicate
high quality. The results for antioxidant capacity, amount of vitamin E show
antioxidative properties. The obtained peroxide values were lower than those
recommended for commercial vegetable oils (≤10) suggesting the extracts are
stable.
Health, Nutrition and Food Science
134
Sourdough fermentation of wheat flour does not prevent the interaction of transglutaminase 2 with α2-gliadin or gluten
Niklas Engströma, Ann-Sofie Sandberga, and Nathalie Scheersa
aDepartment of Biology and Biological Engineering, Food and Nutrition Science, Chalmers University of Technology
Email: [email protected]
Tissue transglutaminase 2 (TG2) plays a critical role in the initiation of celiac
disease by catalyzing the deamidation of gluten peptides. These deamidated
peptides can initiate an immune response, which leads to intestinal inflammation
and symptoms characteristic of celiac disease. .
Several studies have focused on lactic acid fermentation, often in combination
with enzymes, as a way to degrade gluten in order to alleviate its toxicity to
celiacs. However, incomplete degradation of gluten may lead to an increase in
TG2 binding sites. Therefore, we have investigated how lactic acid fermentation,
similar to that of a normal sourdough, affects the potential binding of TG2 to
gluten protein in wheat flour.
We observed that lactic fermentation of wheat flour, as slurry or as part of
sourdough bread, did not decrease the TG2-mediated transamidation or the
available TG2 binding motif QLP in α2-gliadin to a sufficient extent to be useful
for celiac safe food.
Health, Nutrition and Food Science
135
Did you eat sourdough bread for supper? A pilot validation study on plasma
Katharina Dihma,b, Henrik Sundéna, Alastair Rossb and Otto Savolainenb
aDivision of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology
bDivision of Organic Chemistry, Department of Chemistry and Chemical Engineering, Chalmers University of Technology
Food intake biomarkers are a valuable tool in clinical trials and observational
studies aiming to assess the effects of different diets on human health. Recent
work employing metabolomics has identified hydroxy-N-(2-hydroxyphenyl)
acetamide (HHPAA) and N-(2-hydroxyphenyl) acetamide (HPAA) as potential
specific biomarkers for sourdough rye bread intake. However, before being
accepted as validated biomarkers these markers have to be tested across many
studies to assess their performance in different study designs and intake levels.
Our present study aims to validate HPAA and HHPAA as biomarkers and
includes development of a bioanalytical UPLC-QqQ-MS method for measuring
concentration of the proposed biomarkers from plasma and food matrices. The
clinical trial will measure both postprandial kinetics and fasting plasma response
to daily sourdough bread intake over one week. Validation of the proposed
biomarkers can add to the tools available for unravelling the interactions of health
and diet and form the basis for understanding whether they are sufficiently
bioavailable to be of interest as bioactive compounds for human health.
Health, Nutrition and Food Science
136
Synthesis of novel bioactive compounds produced during sourdough fermentation
Katharina Dihma, b, Henrik Sundéna, Otto Savolainenb and Alastair Rossb
aDivision of Organic Chemistry, Department of Chemistry and Chemical Engineering, Chalmers University of Technology
bDivision of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology
Benzoxazinoids, a group of bioactive compounds found in wholegrain cereals, are
thought to be partially responsible for a variety of protective health effects that a
wholegrain diet is known for.1,2,3 In order to understand more about these
compounds some relevant Benzoxazinoids and their glycosides were synthesised
(see Fig. 1).
A synthesis using HBOA as a building block in order to form HBOA-glucoside
and HHPAA was found. HBOA was made in a two-step synthesis from 2-
aminophenol and dichloroacetyl chloride. Reduction of HBOA with NaBH4 in
methanol gave HHPAA which was purified by HPLC. Selectively conjugating
HBOA with glucose proved to be difficult but could be accomplished with a
Hg(CN)2 mediated coupling between 2-bromo-2H-1,4-benzoxazin-3(4H)-one
and 2,3,4,6-tetra-O-acetyl-beta-D-glucose. The synthesis of 1, 2 and 3 was
successful and incorporation of deuterium will be attempted using deuterated
nitrophenol as a starting material. This will enable the quantification of 1, 2 and 3
in biological samples.
Fig. 1.
References.
1. A. Esmaillzadeh, P. Mirmiran and F. Azizi, European J. of Clinical Nutrition, 2005, 59, 353-362.
2. K. B. Adhikari et. Al., Mol. Nutr. Food Res. 2013, 57, 1847-1858. 3. K. B. Adhikari et. Al., J. Agric. Food Chem., 2012, 60, 2497-2906.
Health, Nutrition and Food Science
137
In vitro digestion of rye products: impact of microstructure on glucose and maltose release and digesta viscosity
Daniel Johanssona, José Vázquez Gutiérreza, Marie Almingerb, Rikard Landberga,c
and Maud Langtona
a Dept. of Food Science, Swedish University of Agricultural Sciences, Uppsala b Dept. of Biology and Biological engineering, Chalmers University of Technology
cKarolinska Institutet, Stockholm
Development of tailored foods, which increase fullness and reduce hunger, could
be one way to address the issue of overconsumption of foods which is a strong
contributor to obesity and related disorders. Processing of whole grain rye crisp
breads have been shown to be of importance for postprandial responses in
humans, possibly due to changes in microstructure and physiochemical
composition influencing absorption kinetics of nutrients. The aim of the current
work was to investigate to what extent microstructure and digesta viscosity affect
in vitro starch digestibility.
The digestion of a range of rye products, including porridge, fermented and
unfermented crisp bread, sourdough bread and an extruded product, based on the
same raw material, and a refined wheat bread was studied using two in vitro
methods. A dynamic in vitro model, TIM-1, was used to simulate digestion in the
gastric compartment and the small intestine. Samples were collected during 180
min for characterization of microstructure and to measure release of glucose and
maltose, representing total glucose available for absorption. A static in vitro
method was used for characterization of digesta viscosity after gastric digestion.
There were clear differences in microstructure between the products. The
extruded product had a continuous starch matrix while the other products had a
higher degree of swollen but intact starch granules. In the sourdough bread and to
some extent in the crisp breads, amylose formed a film around the starch granules.
The wheat bread had a continuous protein network and less swollen starch
granules compared to all rye products. In vitro starch digestibility differed
significantly between products (P<0.05) and may be influenced by the presence
of an amylose film as well as starch retrogradation. Further, digesta viscosity was
found to be lower for fermented products, likely due to the degradation of viscous
fibers during the fermentation process.
Health, Nutrition and Food Science
138
3D Bioprinting of Human Skin
Daphne Hingerta, Sandra Ferreyra Vegaa, Daniel Hägga, Vladimir Kirejevb, Marica Ericsonb,c, and Paul Gatenholma
a Biopolymer Technology, Dept. of Chemistry and Chemical Engineering, Chalmers University of Technology, Göteborg, Sweden
bBiomedical photonics group, Dept. of Chemistry and Molecular Biology, University of Gothenburg, Sweden
cSkinResQU, Centre for Skin Research, University of Gothenburg & Chalmers University of Technology, Göteborg, Sweden
Annually, 11 million people worldwide suffer from burns so severe that they required medical attention1. Out of these 265,000 cases are fatal, this leaves millions of surviving burn victims each year suffering from the disfigurement, disabilities and associated stigma and trauma that follows with the condition. In many countries, burns are among the top causes of disability-adjusted life-years, possibly because young children are at an increased risk of burns. In the year 2000, $211 million were spent on the care of children who had suffered from burns1. Ultimately, by the development of 3D bioprinting technology, patient specific living tissue and organs can be biofabricated with high accuracy using the patients’ own cells. In this project, we explore the feasibility of obtaining 3D bioprinted human skin. Proof-of-principle skin constructs are being developed by alternating one-by-one layer of supporting material and human cells. Firstly, bioink based on nanocellulose and alginate is combined with well-defined density of fibroblasts. Then, the several layers of keratinocytes premixed with melanocytes are deposited by 3D bioprinter. Image data acquired from two photon fluorescence microscopy images of human skin will in the future be used as input for creating a computer-aided design (CAD) file for 3D bioprinting. The biofabricated skin will be stratified in the tailor-made bioreactor. We believe that this emerging technology offers unique opportunity for biofabrication of constructs with the ability to exactly mimic the microarchitecture and thus properties of human skin. The approach can potentially be used in clinics to increase quality of life of burn patients or applied for pharmaceutical and toxicology testing.
Figure 1. Schematic illustration of human skin and 3D bioprinting process.
Acknowledgment Per Börjesson, Jessica Ryler, Jakob Obermüller
References.1. World Health Organisation,
Burns, accessed: March 3, 2015
http://www.who.int/mediacentre/factsheets/
fs365/en/.
Health, Nutrition and Food Science
139
Postprandial plasma differences after beef and herring based meals in 2-aminoaidipic acid, β-alanine, 4-hydroxyproline, cetoleic acid
and docosahexaenoic acid: a metabolomics study
Ross, A.B.a,b Svelander, C.a,b, Pinto, R.c, Sandberg, A.S.a
a Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
bChalmers Metabolomics Centre, Chalmers University of Technology, Gothenburg, Sweden
c Bioinformatics Infrastructure for Life Sciences (BILS), Linköpings Universitet, Linköping, Sweden
Dietary guidelines generally recommend increased intake of fish, and reduced
intake of red meat for better long-term health. However, few studies have
compared the metabolic differences between eating meat and fish. We studied the
metabolic response of humans to fish or red meat in 17 overweight males in a
randomised cross-over intervention study. Subjects ate baked herring, pickled
herring or baked beef based meals and post-prandial blood plasma samples were
taken over 7 h. Metabolic profile was measured using gas chromatography-mass
spectrometry. A total of 50 metabolite differences were found for the
comparisons between baked herring and baked beef, or baked herring with pickled
herring. 2-aminoadipic acid, a suggested marker of diabetes risk, was elevated
after the beef meal compared to the herring meals, preceded by a similar rise in
the branch chain amino acid leucine, also suggested to be an early marker of
diabetes risk. Furthermore we found marked rises in β-alanine and 4-
hydroxyproline after beef intake. Herring intake led to greater plasma
concentrations of docosahexaenoic acid (DHA) and cetoleic acid (fatty acid C22:1
n-11), while hippuric acid and benzoic acid differentiated between baked and
pickled herring intake. These results confirm that DHA and cetoleic acid are
biomarkers of herring intake, while β–alanine and 4-hydroxyproline are potential
biomarkers for beef intake. The greater postprandial rise in 2-aminoadipic acid
and leucine suggests a potential role for beef in stimulating insulin secretion,
which may have importance in the context of red meat intake and increased
diabetes risk.
Health, Nutrition and Food Science
140
A simultaneous metabolic profiling and quantitative multimetabolite metabolomic method for human plasma using gas-chromatography
tandem mass spectrometry
Savolainen, O.-I. Sandberg, A.-S. Ross, A B.
Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology
Metabolomics is an increasingly common tool in the biosciences, yet balancing
the need for broad metabolome coverage and quantification remains a challenge.
To address this challenge, we developed a gas chromatography tandem mass
spectrometry (GC-MS/MS) metabolomics method using a high scanning speed
(20 000 Da/second) GC-MS/MS system that enables simultaneous data
acquisition of both non-targeted full scan and targeted quantitative tandem mass
spectrometry data. Although metabolomics analytics has previously been divided
into either targeted or non-targeted, the method presented herein suggests that
both of these approaches can be brought together into one effective method
allowing reproducible quantification of at least 27 metabolites using multiple
reaction monitoring (MRM) and full mass spectral scan-based detection of 601
metabolic features from human plasma. The method showed good linearity over
normal concentrations in plasma (0.18 - 720 to 4 - 14 400 μM depending on the
metabolite) and good intra- and inter-batch precision (1,8 – 16,7 and 2,6 – 25,1
RSD%). Based on the parameters determined for this method, targeted
quantification using MRM can be expanded to cover at least 508 metabolites. The
new simultaneous targeted and non-targeted metabolomics method will enable
more sensitive and accurate detection of metabolites and biomarkers of interest,
while still allowing detection and identification of unknown metabolites. This
GC-MS/MS method can result in time and cost savings for metabolomics analyses
and demonstrates the utility of GC-MS/MS with high scanning rates for complex
analyses.
Health, Nutrition and Food Science
141
The origin of symmetry in the metabolism of cancer
Francesco Gattoa, Almut Schulzeb, and Jens Nielsena
aDepartment of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden
bTheodor-Boveri-Institute, Biocenter, Würzburg, Germany
With the advent of next-generation sequencing technologies, it has been
increasingly appreciated the extent of the heterogeneity of cancer genome. The
complexity of cancer genome is daunting for a rational treatment of the disease.
Nevertheless, all cancers converge to some emerging properties, e.g. invasion (1).
Mutations are central in the evolution of most cancers and they are responsible to
select clones that reprogrammed the cellular gene expression to trigger an
outstanding regulation of these emerging properties (2). In this study, we derived
genome-wide associations between cancer-associated mutations and gene
expression in 1082 human primary tumors with the aim to verifying whether
mutations converge in the transcriptional regulation of known or novel molecular
processes, which are hence likely to constitute a universal criterion for selection
in cancer.
We show that 12 cancer mutations converge independently and primarily on the
regulation of metabolic processes, regardless of the cancer type. In particular, we
derived a network of reactions, termed AraX, that involve the glutathione- and
oxygen-mediated metabolism of arachidonic acid and xenobiotics, whose
deregulation is overrepresented by all 12 mutations, an overrepresentation unseen
in any other biological pathway. Finally, we observed that, among all metabolic
pathways, AraX deregulation represents the strongest predictor for survival in
cancer. These findings suggest that mutations independently select for clones with
highly deregulated AraX to gain a selective advantage in cancer evolution.
References.
1. D. Hanahan, R. A. Weinberg, Hallmarks of cancer: the next generation. Cell 144, 646-674 (2011); published online EpubMar 4 (10.1016/j.cell.2011.02.013).
2. B. Vogelstein, K. W. Kinzler, Cancer genes and the pathways they control. Nature medicine 10,
789-799 (2004); published online EpubAug (Doi 10.1038/Nm1087).
Health, Nutrition and Food Science
142
Effects of pH-shift processing on the bioaccessibility of lipids in the microalgae Nannochloropsis – an in vitro model approach
Lillie Cavoniusa,b, Eva Albersb and Ingrid Undelanda
aChalmers University of Technology, Department of Biology and Biological Engineering, Division of Food and Nutrition Science
bChalmers University of Technology, Department of Biology and Biological Engineering, Division of Industrial Biotechnology
Email: [email protected]
Microalgae are a potential source of human nutrition and animal feed due to the
high content of long-chained n-3 fatty acids (LC n-3 PUFA) and favorable amino
acid profile of some species. Furthermore, microalgae can be cultivated with
minimal nutrient requirements on otherwise non-arable land. We have previously
shown that application of the pH-shift process on Nannochloropsis yields a
product enriched in essential amino acids and with the original amount of LC n-3
PUFA retained (120 mg/g dry weight). Therefore, the product has potential as a
functional food and/or feed ingredient. However, it is not known to which extent
the nutrients are bio-accessible; earlier work has suggests that algal cell walls limit
digestibility in the gastrointestinal tract. Based on the physical changes that take
place during pH-shift processing, e.g. cell lysis and protein
solubilization/precipitation, we hypothesize that the pH-shift process renders the
algal nutrients more available for hydrolysis. The aim of the present study is to
test this hypothesis using a static in vitro digestion model.
Nannochloropsis is subjected to the pH-shift process and nutrient content (fatty
acids, protein, amino acids) is determined before/after processing. To determine
the bio-accessibility of Nannochloropsis processed in various ways, the
microalgae is digested using a static in vitro model of human digestion, based on
the recent international consensus method emerging from the COST FA
1005 Infogest. Intestinal digesta will then be analyzed in respect to degree of
lipolysis using solid phase extraction (SPE) combined with gas chromatography-
mass spectrometry (GC-MS).
Health, Nutrition and Food Science
143
Antioxidant potential of chia seeds (Salvia hispanica L.)
Renata Adriana LABANCAa,b; Cecilia SVELANDERb; Marie ALMINGERb
aDepartment of Food Science, Federal University of Minas Gerais, Brazil bDepartment of Biology and Biological Engineering, Food and Nutrition Science,
Chalmers University of Technology, Gothenburg, Sweden
E-mail: [email protected]
Human intervention studies have provided evidence for protective effects of
various polyphenol rich foods against chronic diseases. Phenolic compounds are
found in a wide variety of cereals and also in chia seeds, an oilseed native of
Mesoamerica, that are considered pseudocereals. The objective of this study was
to characterize chia (Salvia Hispanica L.) samples (seeds and stabilized flour) by
evaluation of their antioxidant properties using DPPH and ABTS radical
scavenging capacity assay, oxygen radical absorbance capacity (ORAC) assay
and ferric reducing antioxidant potential (FRAP) assays. Total phenolic content
was determined by the Folin-Ciocalteu method. Fatty acid composition and
vitamin E were also analyzed by gas chromatography and mass spectrometry
(GC-MS) and reverse phase high-performance liquid chromatography (HPLC)
respectively. The experiments were carried out using chia samples obtained in
Brazil, Sweden and the Netherlands. Antioxidant properties and total phenolic
content was found to differ significantly between samples. Chia seeds from Brazil
showed higher antioxidant capacity and higher phenolic content (3.56±0.20 mg
GAE/g) compared with the chia seed sample from Sweden (2.00±0.09 mg GAE/g)
and the Netherlands (1.65±0.09 mg GAE/g). The flour from Brazil had the lowest
total content of phenolic compounds (1.20±0.16 mg GAE/g), as well as the lowest
antioxidant capacities in most of the applied assays. It could thus be inferred that
the processing for production of stabilized flour affects these parameters. Within
the samples, the order of antioxidant capacity depended on the assay used. A
significant relationship between antioxidant capacity and total phenolic content
was found, indicating that phenolic compounds are the major contributors to the
antioxidant properties of these cereals. To assess the antioxidant activities of
single compounds or the antioxidant capacity of food extracts, a variety of
methods based on different mechanistic principles must be used in parallel,
because different methods often give different results.
Health, Nutrition and Food Science
144
Genome-wide identification of mononuclear cell DNA methylation sites potentially affected by infant fish oil supplementation from 9
to 18 months
Mads V. Linda,b, David Martinoc, Laurine BS. Harsløfb, Zdenka O. Kyjovskad, Mette Kristensenb, Lotte Lauritzenb
a Food and Nutritional Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
bDepartment of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark
cMurdoch Childrens Research Institute, Royal Children's Hospital, University of Melbourne, Australia
dNational Research Centre for the Working Environment, Copenhagen, Denmark
Long-chain n-3 fatty acid (n-3LCPUFA) intake affects markers of metabolic
syndrome, immune function and brain development. Recent evidence suggests
that some of the effects of n-3LCPUFA might be mediated through epigenetic
mechanisms, especially DNA-methylation. Epigenetic alterations are of particular
interest during pregnancy and early life, since these have been suggested as a
mechanism for programming of later diseases. A parallel randomized control trial
was conducted in 133 9-mo-old, Danish infants who received a teaspoon of fish
oil (FO) or sunflower oil (SO) for 9 mo (median intake=3.8 g/day). DNA was
extracted from buffy coat before and after intervention. Illumina Human
Methylation 450K-arrays were used to explore possible differentiation between
the FO and SO supplemented groups on genome-wide DNA-methylation in a
subset of 12 children. The FO intervention gave rise to a 3-fold increase in
erythrocyte n-3LCPUFA, and reduced plasma triacylglycerol, blood pressure and
ex vivo stimulated interleukin-6 production. Analysis of genome-wide
methylation levels by regression did not find significant differences between
groups after adjustment for multiple testing. Analysis of the top-ranked CpG sites
found 43 CpG’s that appeared to be modified by the intervention with an absolute
difference in methylation of at least 10% (unadjusted P-value <0.01). Methylation
levels at these sites were associated with phenotypic changes mainly in blood
pressure. Our analyses suggest potential epigenome effects associated with
functional outcomes, yet the effect sizes were small and did not reach genome-
wide significance. Thus, while the hypothesis that supplementing infants with n-
3LCPUFA leads to changes in DNA-methylation appears to be well founded,
additional investigation is required as to the overall importance of these changes.