33 research outputs found
Starch based systems for the colonic delivery of bioactive peptides
Bioactive peptides have numerous health benefits, although if taken orally they may be digested during gastrointestinal (GI) transit. Encapsulation is an established method for oral delivery of bioactives. However, many current approaches arise from pharmaceutical applications and may be unsuitable for food due to the materials used, cost and scale of production. Therefore, in this project we set out to create a simple and clean-label encapsulation system, suitable for use in the food industry, which could deliver bioactive peptides to the colon. One potential clean-label entrapment material is resistant starch, which is the portion of starch that resists digestion in the upper GI tract but can be digested by bacteria in the colon. As a model bioactive peptide, the well characterised antimicrobial peptide nisin was used; this peptide is normally digested during GI transit. To prepare the nisin a simple purification process was developed, which produced a powder containing ∼33% nisin from a nisin producing culture and also enriched a commercial nisin preparation over 30-fold to a purity of ∼58%. The digestion of nisin was characterised (in vitro) and 6 nisin fragments (4 of which are bioactive) were identified in the digestion products; it was also observed that nisin formed a complex with bile salts that effected its digestion. Nisin was entrapped in starch through multiple approaches based on spray coating, co-spray drying and gel entrapment. A simple approach based on gel entrapment was the most successful and it was shown in vitro to be capable of protecting a portion of the entrapped nisin during transit in the upper GI tract. Using a murine model, it was determined in vivo that a nisin entrapped in starch gel diet significantly (p < 0.001, n = 10) affected the relative abundance of 3 times as many genera from the lower GI tract than a control nisin in starch diet, despite the mice consuming 3-fold less nisin than the control diet
Simulated gastrointestinal digestion of nisin and interaction between nisin and bile
peer-reviewedNisin, an antimicrobial peptide showing activity against many Gram positive bacteria, is widely used as a food preservative. The simulated gastrointestinal digestion of nisin (variant A) was studied using the in vitro INFOGEST digestion method. Following oral, gastric and small intestinal digestion, there was no intact nisin in the system and the nisin was primarily digested by pancreatin. After digestion, six nisin fragments (1–11, 1–12, 1–20, 1–21, 1–29 and 1–32) were identified by reversed phase high performance liquid chromatography and mass spectroscopy and four of these nisin fragments (1–20, 1–21, 1–29 and 1–32) demonstrated low antibacterial activity against Lactococcus lactis HP in agar diffusion activity assays. Additionally, it was observed that bile salts form a complex with nisin. This was examined by atomic force microscopy, turbidity and dynamic light scattering, which showed that this interaction resulted in significantly larger bile salt micelles. The presence of bile salts at physiological levels significantly altered the relative amounts of the nisin fragments 1–12, 1–20 and 1–29 produced during an in vitro digestion. This study highlights the importance of including bile in simulated digestions of antimicrobial peptides in order to obtain a more accurate simulation of the in vivo digestion products and their activity
Oral Delivery of Nisin in Resistant Starch Based Matrices Alters the Gut Microbiota in Mice
peer-reviewedThere is a growing recognition of the role the gastrointestinal microbiota plays in health and disease. Ingested antimicrobial proteins and peptides have the potential to alter the gastrointestinal microbiota; particularly if protected from digestion. Nisin is an antimicrobial peptide that is used as a food preservative. This study examined the ability of nisin to affect the murine microbiota when fed to mice in two different starch based matrices; a starch dough comprising raw starch granules and a starch gel comprising starch that was gelatinized and retrograded. The effects of the two starch matrices by themselves on the microbiota were also examined. Following 16S rRNA compositional sequencing, beta diversity analysis highlighted a significant difference (p = 0.001, n = 10) in the murine microbiota between the four diet groups. The differences between the two nisin containing diets were mainly attributable to differences in the nisin release from the starch matrices while the differences between the carriers were mainly attributable to the type of resistant starch they possessed. Indeed, the differences in the relative abundance of several genera in the mice consuming the starch dough and starch gel diets, in particular Akkermansia, the relative abundance of which was 0.5 and 11.9%, respectively (p = 0.0002, n = 10), points to the potential value of resistance starch as a modulator of beneficial gut microbes. Intact nisin and nisin digestion products (in particular nisin fragment 22–31) were detected in the feces and the nisin was biologically active. However, despite a three-fold greater consumption of nisin in the group fed the nisin in starch dough diet, twice as much nisin was detected in the feces of the group which consumed the nisin in starch gel diet. In addition, the relative abundance of three times as many genera from the lower gastrointestinal tract (GIT) were significantly different (p < 0.001, n = 10) to the control for the group fed the nisin in starch gel diet, implying that the starch gel afforded a degree of protection from digestion to the nisin entrapped within it
About the p-mode frequency shifts in HD 49933
We study the frequency dependence of the frequency shifts of the low-degree p
modes measured in the F5V star HD 49933, by analyzing the second run of
observations collected by the CoRoT satellite. The 137-day light curve is
divided into two subseries corresponding to periods of low and high stellar
activity. The activity-frequency relationship is obtained independently from
the analysis of the mode frequencies extracted by both a local and a global
peak-fitting analyses, and from a cross-correlation technique in the frequency
range between 1450 microHz and 2500 microHz. The three methods return
consistent results. We show that the frequency shifts measured in HD 49933
present a frequency dependence with a clear increase with frequency, reaching a
maximal shift of about 2 microHz around 2100 microHz. Similar variations are
obtained between the l=0 and l=1 modes. At higher frequencies, the frequency
shifts show indications of a downturn followed by an upturn, consistent between
the l=0 and 1 modes. We show that the frequency variation of the p-mode
frequency shifts of the solar-like oscillating star HD 49933 has a comparable
shape to the one observed in the Sun, which is understood to arise from changes
in the outer layers due to its magnetic activity.Comment: 5 pages, 3 figures, 1 table, Accepted for publication in A\&
Large expert-curated database for benchmarking document similarity detection in biomedical literature search
Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe
Point-of-care testing in paediatric settings in the UK and Ireland: A cross-sectional study
Background: Point-of-care testing (POCT) is diagnostic testing performed at or near to the site of the patient. Understanding the current capacity, and scope, of POCT in this setting is essential in order to respond to new research evidence which may lead to wide implementation. Methods: A cross-sectional online survey study of POCT use was conducted between 6th January and 2nd February 2020 on behalf of two United Kingdom (UK) and Ireland-based paediatric research networks (Paediatric Emergency Research UK and Ireland, and General and Adolescent Paediatric Research UK and Ireland). Results: In total 91/109 (83.5%) sites responded, with some respondents providing details for multiple units on their site based on network membership (139 units in total). The most commonly performed POCT were blood sugar (137/139; 98.6%), urinalysis (134/139; 96.4%) and blood gas analysis (132/139; 95%). The use of POCT for Influenza/Respiratory Syncytial Virus (RSV) (45/139; 32.4%, 41/139; 29.5%), C-Reactive Protein (CRP) (13/139; 9.4%), Procalcitonin (PCT) (2/139; 1.4%) and Group A Streptococcus (5/139; 3.6%) and was relatively low. Obstacles to the introduction of new POCT included resources and infrastructure to support test performance and quality assurance. Conclusion: This survey demonstrates significant consensus in POCT practice in the UK and Ireland but highlights specific inequity in newer biomarkers, some which do not have support from national guidance. A clear strategy to overcome the key obstacles of funding, evidence base, and standardising variation will be essential if there is a drive toward increasing implementation of POCT
A Simple Method for the Purification of Nisin
peer-reviewedNisin, an antimicrobial peptide showing activity against a broad range of Gram-positive bacteria, is widely used as a food preservative and has potential as a therapeutic for a range of infectious diseases. Here, we present a simple purification method, based on a salting-out approach, which can produce a powder containing ∼33% nisin, from a nisin-producing culture in a whey permeate-based medium. This process removes over 99% of the lactic acid, NaCl, lactose and non-nisin proteins from the cell-free culture supernatant. The approach can also enrich a commonly used commercial nisin preparation over 30-fold to a purity of ∼58%. These are higher purities than comparable published methods. The simplicity of this approach facilitates its use in research and also its scale-up
Simulated gastrointestinal digestion of nisin and interaction between nisin and bile
Nisin, an antimicrobial peptide showing activity against many Gram positive bacteria, is widely used as a food preservative. The simulated gastrointestinal digestion of nisin (variant A) was studied using the in vitro INFOGEST digestion method. Following oral, gastric and small intestinal digestion, there was no intact nisin in the system and the nisin was primarily digested by pancreatin. After digestion, six nisin fragments (1–11, 1–12, 1–20, 1–21, 1–29 and 1–32) were identified by reversed phase high performance liquid chromatography and mass spectroscopy and four of these nisin fragments (1–20, 1–21, 1–29 and 1–32) demonstrated low antibacterial activity against Lactococcus lactis HP in agar diffusion activity assays. Additionally, it was observed that bile salts form a complex with nisin. This was examined by atomic force microscopy, turbidity and dynamic light scattering, which showed that this interaction resulted in significantly larger bile salt micelles. The presence of bile salts at physiological levels significantly altered the relative amounts of the nisin fragments 1–12, 1–20 and 1–29 produced during an in vitro digestion. This study highlights the importance of including bile in simulated digestions of antimicrobial peptides in order to obtain a more accurate simulation of the in vivo digestion products and their activity
Oral Delivery of Nisin in Resistant Starch Based Matrices Alters the Gut Microbiota in Mice
There is a growing recognition of the role the gastrointestinal microbiota plays in health and disease. Ingested antimicrobial proteins and peptides have the potential to alter the gastrointestinal microbiota; particularly if protected from digestion. Nisin is an antimicrobial peptide that is used as a food preservative. This study examined the ability of nisin to affect the murine microbiota when fed to mice in two different starch based matrices; a starch dough comprising raw starch granules and a starch gel comprising starch that was gelatinized and retrograded. The effects of the two starch matrices by themselves on the microbiota were also examined. Following 16S rRNA compositional sequencing, beta diversity analysis highlighted a significant difference (p = 0.001, n = 10) in the murine microbiota between the four diet groups. The differences between the two nisin containing diets were mainly attributable to differences in the nisin release from the starch matrices while the differences between the carriers were mainly attributable to the type of resistant starch they possessed. Indeed, the differences in the relative abundance of several genera in the mice consuming the starch dough and starch gel diets, in particular Akkermansia, the relative abundance of which was 0.5 and 11.9%, respectively (p = 0.0002, n = 10), points to the potential value of resistance starch as a modulator of beneficial gut microbes. Intact nisin and nisin digestion products (in particular nisin fragment 22–31) were detected in the feces and the nisin was biologically active. However, despite a three-fold greater consumption of nisin in the group fed the nisin in starch dough diet, twice as much nisin was detected in the feces of the group which consumed the nisin in starch gel diet. In addition, the relative abundance of three times as many genera from the lower gastrointestinal tract (GIT) were significantly different (p < 0.001, n = 10) to the control for the group fed the nisin in starch gel diet, implying that the starch gel afforded a degree of protection from digestion to the nisin entrapped within it
Construction of a Natural Mucin Microarray and Interrogation for Biologically Relevant Glyco-Epitopes
Mucins are the principal components of mucus, and mucin
glycosylation
has important roles in defense, microbial adhesion, immunomodulation,
inflammation, and cancer. Mucin expression and glycosylation are dynamic,
responding to changes in local environment and disease. Potentially
hundreds of heterogeneous glycans can substitute one mucin molecule,
and it is difficult to identify biologically accessible glyco-epitopes.
Thirty-seven mucins, from the reproductive and gastrointestinal (GI)
tracts of six species (bovine, ovine, equine, porcine, chicken, and
deer) and from two human-derived cell lines, were purified. Following
optimization of mucin printing and construction of a novel mucin microarray,
the glycoprofiles of the whole mucins on the microarray were compared
using a panel of lectins and one antibody. Accessible glyco-motifs
of GI mucins varied according to species and localization of mucin
origin, with terminal fucose, the sialyl T-antigen, and <i>N</i>-linked oligosaccharides identified as potentially important. The
occurrence of T- and sialyl T-antigen varied in bovine and ovine reproductive
tract mucins, and terminal <i>N</i>-acetylgalactosamine
(GalNAc) and sulfated carbohydrates were detected. This study introduces
natural mucin microarrays as an effective tool for profiling mucin
glyco-epitopes and highlights their potential for discovery of biologically
important motifs in bacterial–host interactions and fertility