Development of a fermented quinoa-based beverage.

Quinoa is a crop that originated from the Andes. It has high nutritional value, outstanding agro-ecological adaptability, and low water requirements. Quinoa is an excellent crop alternative to help overcome food shortages, and it can also have a role in the prevention of developed world lifestyle diseases, such as type-2 diabetes, cardiovascular diseases, osteoporosis, inflammatory and autoimmune diseases, etc. In order to expand the traditional uses of quinoa and to provide new, healthier and more nutritious food products, a fermented quinoa-based beverage was developed. Two quinoa varieties (Rosada de Huancayo and Pasankalla) were studied. The fermentation process, viscosity, acidity, and metabolic activity during the preparation and storage of the drink were monitored, as well as the preliminary organoleptic acceptability of the product. The drink had viable and stable microbiota during the storage time and the fermentation proved to be mostly homolactic. Both quinoa varieties were suitable as base for fermented products; Pasankalla, however, has the advantage due to higher protein content, lower saponin concentration, and lower loss of viscosity during the fermentation process. These results suggest that the differences between quinoa varieties may have substantial effects on food processes and on the properties of final products. This is a factor that should be taken into account when planning novel products based on this grain.

Aspergillus flavus growth inhibition by Lactobacillus strains isolated from traditional fermented Kenyan milk and maize products.

Certain strains of lactic acid bacteria have been reported to inhibit fungal growth and may so be potential as biocontrol agents. In this study, 171 LAB strains were isolated from traditional fermented Kenyan milk and maize products and tested against aflatoxin-producing A. flavus fungi. The three LAB strains showing highest antifungal activity were identified as Lactobacillus plantarum. None of the strains were able to completely inhibit fungal growth under conditions favorable for fungi and suboptimal for LAB. These conditions probably reduced the growth and metabolic activity of some LAB isolates, as several growth-related aspects like production of antifungal biomolecules and other metabolites contribute to the inhibiting activity. The results suggest that certain LAB strains could be employed in food to control the growth of aflatoxigenic fungi. Further studies to establish the efficacy of the potential LAB strains in fermented products are in progress.

Potential of lactic acid bacteria in aflatoxin risk mitigation.

Aflatoxins (AF) are ubiquitous mycotoxins contaminating food and feed. Consumption of contaminated food and feed can cause a severe health risk to humans and animals. A novel biological method could reduce the health risks of aflatoxins through inhibiting mold growth and binding aflatoxins. Lactic acid bacteria (LAB) are commonly used in fermented food production. LAB are known to inhibit mold growth and, to some extent, to bind aflatoxins in different matrices. Reduced mold growth and aflatoxin production may be caused by competition for nutrients between bacterial cells and fungi. Most likely, binding of aflatoxins depends on environmental conditions and is strain-specific. Killed bacteria cells possess consistently better binding abilities for aflatoxin B1 (AFB1) than viable cells. Lactobacilli especially are relatively well studied and provide noticeable possibilities in binding of aflatoxin B1 and M1 in food. It seems that binding is reversible and that bound aflatoxins are released later on (Haskard et al., 2001; Peltonen et al., 2001). This literature review suggests that novel biological methods, such as lactic acid bacteria, show potential in mitigating toxic effects of aflatoxins in food and feed.

Interfacial sliding properties of bone screw materials and their effect on screw fixation strength.

BACKGROUND: This study examined the effect of interfacial sliding and test material properties on the fixation strength and insertional properties of self-tapping bone screws. Various substitute materials (polyacetal [POM], poly(methyl methacrylate) [PMMA] and E-glass-filled Epoxy [Sawbones®]) for human bone were evaluated, and the results were compared with the findings for cadaver bone.
 METHODS: Initial coefficient of friction (CoF) of the screw material stainless steel AISI316 was tested using a pin-on-disk apparatus, and the screws were exposed to pullout tests after insertion torque tests. The effect of a smooth diamond-like carbon (DLC) coating was studied by applying the coating on both CoF test balls and bone screws.
 RESULTS: Mechanical properties of test blocks strongly correlated to both pullout strength and insertion torque of the screws: for noncoated 2.7-mm screws, tensile strength correlated to pullout strength and insertion torque, with Pearson correlation coefficients r=0.977 and r=0.738, respectively. In contrast, CoF correlated strongly to screw insertion torque but not to pullout strength in bone substitute materials (for noncoated 2.7-mm screws, r=0.652 and r=0.248, respectively). There were no significant differences in CoF using noncoated and DLC-coated screw materials against bone substitutes.
 CONCLUSIONS: Proper materials for in vitro testing help in evaluating the biomechanics of the implants in advance. However, choosing the material needs attention, as their ability to model human bone depends on test type.

Osteogenesis of human mesenchymal stem cells on micro-patterned surfaces.

Osteogenic responses of human mesenchymal stromal cells (hMSCs) were compared on square-patterned, inverse square-patterned, and planar titanium, chromium, diamond-like carbon (DLC), and tantalum; hypothesis was that both the materials and patterns affect osteogenesis. Samples were produced using photolithography and physical vapor deposition. Early-marker alkaline phosphatase (ALP) and mid-markers, small body size and mothers against decapentaplegic-related protein-1 (SMAD1), runt-related transcription factor-2 (RUNX2), and osteopontin were studied using quantitative real-time polymerase chain reaction. ALP and hydroxyapatite, were colorimetrically studied. ALP reached highest values on both patterned titanium samples, but mid-markers disclosed that it was already lagging behind planar and inverse patterned tantalum. Hydroxyapatite formation disclosed that osteo-induced hMSCs passed all the differentiation stages (except on planar chromium). Presence of hydroxyapatite disclosed that both types of patterning promoted (p < 0.001) osteogenesis compared to planar samples. Results suggest that the osseocompatibility/integration of implants could be improved by changing the monotonous and featureless implant-host interface into micro-patterned interface to provide physical differentiation cues.

Formation and retention of staphylococcal biofilms on DLC and its hybrids compared to metals used as biomaterials.

Staphylococcus epidermidis and Staphylococcus aureus cause most of the implant-related infections. Antibiotic treatment often fails and cure requires surgical intervention. It was hypothesized that biomaterial coatings resistant to biofilms offer a preventive option. Physical vapour deposited diamond-like carbon (DLC) and its polytetrafluoroethylene (DLC-PTFE-h) and polydimethylsiloxane (DLC-PDMS-h) hybrids were compared to titanium (Ti), tantalum (Ta) and chromium (Cr) thin films on silicon wafers for their resistance against formation and/or retention of biofilms produced by S. epidermidis and S. aureus in vitro. Sample surfaces were characterized for surface topography, contact angle and zeta-potential, because such properties might affect the biofilm. Biofilm was stained using calcofluor white and analysed in fluorescence microscopy using morphometry. Sixteen hour incubation was selected in pilot tests; at this checkpoint Ti, Ta, Cr and DLC-PDMS-h were almost fully covered by biofilm, but DLC and DLC-PTFE-h were only partially biofilm coated by S. epidermidis (88±26%, p<0.001 and 56±39%, p<0.001, respectively) or S. aureus (81±24%, p<0.001 and 51±26%, p<0.001, respectively). DLC and its PTFE hybrid offer a potential biofilm hostile surface coating for implants and medical devices. This ability to resist biofilm formation and attachment could not be explained by only one factor, but it seems to be related to a combination of various properties, with electrokinetic streaming potential and protein coating being particularly important for its outcome.

Toxigenic potential of Aspergillus species occurring on maize kernels from two agro-ecological zones in Kenya.

Two agro-ecological zones in Kenya were selected to compare the distribution in maize of Aspergillus spp. and their toxigenicity. These were Nandi County, which is the main maize growing region in the country but where no human aflatoxicoses have been reported, and Makueni County where most of the aflatoxicosis cases have occurred. Two hundred and fifty-five households were sampled in Nandi and 258 in Makueni, and Aspergillus was isolated from maize. Aspergillus flavus and A. parasiticus isolates were tested for the presence of aflD and aflQ genes. Positive strains were induced to produce aflatoxins on yeast extract sucrose and quantified using liquid chromatography-tandem mass spectrometry (LCMSMS). Aspergillus flavus was the most common contaminant, and the incidence of occurrence in Nandi and Makueni was not significantly different (82.33% and 73.26%, respectively). Toxigenic strains were more prevalent than non-toxigenic strains. All the toxigenic strains from Makueni were of the S-type while those from Nandi belonged to the l-type. Quantitative differences in aflatoxin production in vitro between isolates and between strains were detected with S strains producing relatively larger amounts of total aflatoxins, B toxins and lower values for G toxins. This was in accord with the frequent aflatoxicosis outbreaks in Makueni. However some L strains produced considerable amounts of B toxins. Given the widespread distribution of toxigenic strains in both regions, the risk of aflatoxin poisoning is high when favorable conditions for toxin production occur.

Early bone growth on the surface of titanium implants in rat femur is enhanced by an amorphous diamond coating.

BACKGROUND AND PURPOSE: Amorphous diamond (AD) is a durable and compatible biomaterial for joint prostheses. Knowledge regarding bone growth on AD-coated implants and their early-stage osseointegration is poor. We investigated bone growth on AD-coated cementless intramedullary implants implanted in rats. Titanium was chosen as a reference due to its well-known performance. MATERIALS AND METHODS: We placed AD-coated and non-coated titanium implants (R(a) ≈ 0.2 µm) into the femoral bone marrow of 25 rats. The animals were divided in 2 groups according to implant coating and they were killed after 4 or 12 weeks. The osseointegration of the implants was examined from hard tissue specimens by measuring the new bone formation on their surface. RESULTS: 4 weeks after the operation, the thickness of new bone in the AD-coated group was greater than that in the non-coated group (15.3 (SD 7.1) µm vs. 7.6 (SD 6.0) µm). 12 weeks after the operation, the thickness of new bone was similar in the non-coated group and in the AD-coated group. INTERPRETATION: We conclude that AD coating of femoral implants can enhance bone ongrowth in rats in the acute, early stage after the operation and might be an improvement over earlier coatings.

Chemiluminescence of non-differentiated THP-1 promonocytes: developing an assay for screening anti-inflammatory milk proteins and peptides.

Human leukemic THP-1 promonocytes are widely used as a model for peripheral blood monocytes. However, superoxide production during respiratory burst (RB) of non-differentiated THP-1 (nd-THP-1) cells is very low. Here we present a rapid and low-cost method for measuring the chemiluminescence (CL) of opsonized zymosan (OZ) induced RB which allows detection of Escherichia coli lipopolysaccharide (LPS) induced priming of nd-THP-1 cells on the basis of CL reaction kinetics. Maximum CL intensity obtained was 2.20 ± 0.25 and 1.30 ± 0.11 relative light units, while CL peak time was achieved at 18.1 ± 2.6 and 28.7 ± 1.3 min in primed and non-primed cells, respectively. The priming of nd-THP-1 cells with LPS evoked typical TNF-α and IL-6 production. We tested the effects of bovine lactoferrin and protein fractions from Lactobacillus helveticus BGRA43 fermented milk for potential anti-inflammatory effects on LPS primed nd-THP-1 cells. Four fractions were found to inhibit the OZ-induced CL in a dose-dependent manner (IC(50) 3-30 µg/mL), while lactoferrin inhibited CL to a lesser extent (IC(50) 270 µg/mL). These results suggest that measuring CL response of nd-THP-1 cells can serve as a method for screening anti-inflammatory compounds which could be used in reducing the risk of phagocyte-mediated inflammatory diseases.

Interactions of human bone cells with diamond-like carbon polymer hybrid coatings.

Diamond-like carbon (DLC) coatings produced using the plasma-accelerating filtered pulsed arc discharge (FPAD) method display excellent adherence to the substrate and improve its corrosion resistance. This article reports the interactions of human osteoblastic cells with DLC and two DLC polymer hybrid (DLC-p-h) coatings deposited on smooth, matt and rough silicon wafers by the FPAD method. The DLC-p-h materials were DLC-polytetrafluoroethylene hybrid (DLC-PTFE-h) and DLC-polydimethylsiloxane hybrid (DLC-PDMS-h) coatings. The biocompatibility of the coatings was assayed by using mesenchymal stem cells, primary osteoblasts and Saos-2 cells. Human mesenchymal stem cells proliferated when cultured on DLC and DLC-PTFE-h, but their numbers diminished on DLC-PDMS-h. In all three cell types studied, phalloidin-TRITC staining disclosed cell-type organization typical of an actin cytoskeleton on DLC and DLC-PTFE-h, but minimal and disorganized stress fibers on cells cultured on DLC-PDMS-h. The microtubular cytoskeleton was similarly disorganized on DLC-PDMS-h. Cells on DLC-PDMS-h developed a peculiar form of membrane damage, with nuclear staining by propidium iodide associated with granular calcein staining of the cytoplasm. Active caspase-3 labeling was only seen in cells cultured on DLC-PDMS-h, indicating that these cells undergo apoptosis induced by defective cell adhesion. Results suggest that DLC-PDMS-h coatings might be useful in orthopedic applications where an implant or implant-facet should be protected against bone overgrowth while DLC and DLC-PTFE-h coatings might improve osseointegration.

Surface characterization and in vitro biocompatibility assessment of photosensitive polyimide films.

Polyimide (PI) is a commonly used polymer in microelectronics. Recently, numerous PI-based flexible neural interfaces have been developed for reducing mechanical mismatch between rigid implant and soft neural tissue. Most approaches employ non-photosensitive PI, which has been proven earlier to be biocompatible. However, photosensitive polyimide (PSPI) would simplify device fabrication remarkably, but its biocompatibility has been only sparsely reported. In this study, cytotoxicity of spin-coated PSPI (HD Microsystems PI-2771) and conventional PI (HD Microsystems PI-2525) films were evaluated in vitro using BHK-21 fibroblasts according to the ISO-10993-5 standard. PSPIs were tested as cured at a temperature of 200 degrees C (PI-2771-200) and 350 degrees C (PI-2771-350). The PI film surfaces were characterized in terms of their roughness, energy and zeta potential which are hypothesized to affect cell-material interactions. The values of the total surface free energy (SFE), and its polar and dispersive component, were significantly (p<0.001) greater for the PI-2525 film (SFE: 47.3 mJ/m2) than for the PI-2771-200 (25.6 mJ/m2) or PI-2771-350 films (26.2 mJ/m2). The curing temperature of the PI-2771 had a significant effect on the zeta potential values (p<0.001), but not on surface energy (p=0.091) or roughness (p=0.717). The results from the MTS proliferation assays and live/dead staining revealed that PSPI is almost as non-cytotoxic as conventional PI and polyethylene (negative control). The morphology and spreading of BHK-21 cells were similar on all the PI materials tested. In conclusion, PSPI seems to be a promising biocompatible material, while further studies in vitro and in vivo are needed to clarify the long-term effects.

Adhesion, spreading and osteogenic differentiation of mesenchymal stem cells cultured on micropatterned amorphous diamond, titanium, tantalum and chromium coatings on silicon.

It was hypothesized that human mesenchymal stromal cell (hMSC) can be guided by patterned and plain amorphous diamond (AD), titanium (Ti), tantalum (Ta) and chromium (Cr) coatings, produced on silicon wafer using physical vapour deposition and photolithography. At 7.5 h hMSCs density was 3.0-3.5 x higher (P < 0.0003, except Ti) and cells were smaller (68 vs. 102 microm, P 0.000006-0.02) on patterns than on silicon background. HMSC-covered surface of the background silicon was lower on Ti than AD patterns (P = 0.015), but at 5 days this had reversed (P = 0.006). At 7.5 h focal vinculin adhesions and actin cytoskeleton were outgoing from pattern edges so cells assumed geometric square shapes. Patterns allowed induced osteogenesis, but less effectively than plain surfaces, except for AD, which could be used to avoid osseointegration. All these biomaterial patterns exert direct early, intermediate and late guidance on hMSCs and osteogenic differentiation, but indirect interactions exist with cells on silicon background.

Improved adherence and spreading of Saos-2 cells on polypropylene surfaces achieved by surface texturing and carbon nitride coating.

The adhesion and contact guidance of human primary osteogenic sarcoma cells (Saos-2) were characterized on smooth, microstructured (MST) and micro- and nano-structured (MNST) polypropylene (PP) and on the same samples with a silicon-doped carbon nitride (C(3)N(4)-Si) coating. Injection molding was used to pattern the PP surfaces and the coating was obtained by using ultra-short pulsed laser deposition (USPLD). Surfaces were characterized using atomic force microscopy and surface energy components were calculated according to the Owens-Wendt model. The results showed C(3)N(4)-Si coated surfaces to be significantly more hydrophilic than uncoated ones. In addition, there were 86% more cells in the smooth C(3)N(4)-Si coated PP compared to smooth uncoated PP and 551%/476% more cells with MST/MNST C(3)N(4)-Si coated PP than could be obtained with MST/MNST uncoated PP. Thus the adhesion, spreading and contact guidance of osteoblast-like cells was effectively improved by combining surface texturing and deposition of osteocompatible C(3)N(4)-Si coating.

Fabrication and testing of polyimide-based microelectrode arrays for cortical mapping of evoked potentials.

Modern microfabrication techniques make it possible to develop microelectrode arrays that may be utilized not only in neurophysiological research but also in the clinic, e.g. in neurosurgery and as elements of neural prostheses. The aim of this study was to test whether a flexible microelectrode array is suitable for recording cortical surface field potentials in rats. Polyimide-based microelectrode arrays were fabricated by utilizing microfabrication techniques e.g. photolithography and magnetron sputter deposition. The present microelectrode array consists of eight platinum microelectrodes (round-shaped, Ø: 200 microm), transmission lines and connector pads sandwiched between two thin layers of biocompatible polyimide. The microelectrode arrays were electrochemically characterized by impedance spectroscopy in physiological saline solution and successfully tested in vivo by conducting acute and chronic measurements of evoked potentials on the surface of rat cortex. The arrays proved excellent flexibility and mechanical strength during handling and implantation onto the surface of cortex. The excellent electrochemical characteristics and stable in vivo recordings with high spatiotemporal resolution highlight the potential of these arrays. The fabrication protocol described here allows implementation of several other neural interfaces with different layouts, material selections or target areas either for recording or stimulation purposes.

Inhibition of Angiotensin converting enzyme I caused by autolysis of potato proteins by enzymatic activities confined to different parts of the potato tuber.

Autolysis of protein isolates from vascular bundle and inner tuber tissues of potato (Solanum tuberosum) enhanced the inhibition of the angiotensin converting enzyme I (ACE), a biochemical factor affecting blood pressure (hypertension). The physiological age of the tuber affected the strength of ACE inhibition, the rate of its increase during autolysis, and the tuber tissue where ACE inhibition was most pronounced. The highest inhibitory activities (50% reduction in ACE activity achieved following autolysis at a protein concentration of 0.36 mg mL (-1)) were measured in tubers after 5-6 months of storage prior to sprouting. The rate of ACE inhibition was positively correlated with protease activity in tuber tissues. Amendment of the autolysis reaction with protein substrates from which bioactive ACE-inhibitory peptides may be released, for example, a purified recombinant protein or a concentrate of total tuber proteins, also enhanced ACE inhibition. Many tuber proteins including aspartic protease inhibitors were degraded during autolysis. The data provide indications of differences in the enzymatic activities confined to different parts of the potato tuber at different physiological stages. Results suggest that native enzymes and substrate proteins of potato tubers can be utilized in search of dietary tools to manage elevated blood pressure.

ACE-inhibitory and antioxidant properties of potato (Solanum tuberosum).

Proteins were isolated from potato tubers (Solanum tuberosum) at different physiological states, and by-products from the potato industry were used to evaluate their ACE-inhibitory and radical-scavenging potencies. Protein isolates and by-products were autolysed or hydrolysed by alcalase, neutrase and esperase. Hydrolysis increased the inhibition of the angiotensin-converting enzyme (ACE) and the radical-scavenging activity. The ACE-inhibitory potencies of the hydrolysates were high (IC50=0.018-0.086) and the by-product fractions showed ACE-inhibition also before hydrolysis. All samples exhibited low radical-scavenging activity, and hydrolysis for 2h with proteases was needed to produce an increase in the activity. Ultrafiltration through 10-3kDa membranes efficiently separated the ACE-inhibitory compounds into permeate fractions. The results of this study suggest that potato is a promising source for the production of bioactive compounds as ingredients for developing functional foods with a beneficial impact on cardiovascular health.

Development of flexible microelectrode arrays for recording cortical surface field potentials.

The fabrication, characterization and application of an 8-channel flexible microelectrode array for recording cortical surface field potentials is described. Polyimide-based microelectrode arrays were fabricated by using photolithography and physical vapour deposition (PVD) methods. Polyimide was chosen as the substrate and insulation material due to its suitable mechanical properties and biocompatibility. Electrodes and transmission lines were formed by sputter-coating of platinum thin films. Microelectrode arrays were characterized and tested successfully in vitro by impedance spectroscopy and in vivo by somatosensory evoked potential (SEP) recordings in rats. These tests indicated good performance and the potential of microelectrode arrays.

Effect of processing and storage on the stability of flaxseed lignan added to dairy products.

This study investigated the effects of processing and storage on the stability of purified, flaxseed-derived secoisolariciresinol diglucoside (SDG) added to milk prior to the manufacture of different dairy products. We analyzed the effect of high-temperature pasteurization, fermentation, and milk renneting as well as storage on the stability of SDG added to milk, yogurt, and cheese. Also, the stability of SDG in whey-based drinks was studied. Added SDG was found to withstand the studied processes well. In edam cheese manufacture, most of the added SDG was retained in the whey fraction and 6% was found in the cheese curd. SDG was also relatively stable in edam cheese during ripening of 6 weeks at 9 degrees C and in yogurt during storage of 21 days at 4 degrees C. Up to 25% of added SDG was lost in whey-based drinks during storage of 6 months at 8 degrees C. We conclude that SDG can be successfully supplemented in dairy-based products.

Effect of processing and storage on the stability of flaxseed lignan added to bakery products.

The study focused on the effects of processing and storage on the stability of flaxseed-derived secoisolariciresinol diglucoside (SDG) added to various bakery products. The SDG concentration of doughs, baked rye breads, graham buns, and muffins was analyzed by high-performance liquid chromatography-diode array detection; the baked products were analyzed immediately after baking and upon storage at room temperature for 1 week and at -25 degrees C for 1 and 2 months, respectively. Added SDG was found to withstand normal baking temperatures in all bakery products. SDG also was a relatively stable compound during storage. Similarly, the content of SDG in flax buns containing fat-free flaxseed meal was unaffected by storage. We conclude that cereal-based bakery products can be supplemented with flaxseed-derived SDG.

Milk immunoglobulins for health promotion.

The biological function of bovine colostral immunoglobulins is to provide the newborn calf with adequate passive immune protection against microbial infections. Immunoglobulin preparations designed for farm animals are commercially available, and some colostrum-based products are marketed also for humans as dietary supplements. The concentration of specific antibodies against a certain pathogenic microorganism can be raised in colostrum and milk by immunizing cows with this pathogen or its antigen. Advances in bioseparation and chromatographic techniques have made it possible to fractionate and enrich these antibodies and formulate so-called hyperimmune colostral or milk preparations. Their efficacy in prevention and treatment of various microbial infections has been evaluated in numerous studies. Immune milk preparations have proven effective in prophylaxis against infections caused by a variety of gastrointestinal pathogens. Their therapeutic efficacy, however, seems more limited. A few commercial immune milk products are already on market and more applications can be expected in the coming years. This article reviews the recent progress made in isolation techniques of bovine immunoglobulins and the application of colostral and immune milk preparations in fighting various microbial infectious diseases in humans.