Imprinted gene alterations in the kidneys of growth restricted offspring may be mediated by a long non-coding RNA.

Altered epigenetic mechanisms have been previously reported in growth restricted offspring whose mothers experienced environmental insults during pregnancy in both human and rodent studies. We previously reported changes in the expression of the DNA methyltransferase Dnmt3a and the imprinted genes Cdkn1c (Cyclin-dependent kinase inhibitor 1C) and Kcnq1 (Potassium voltage-gated channel subfamily Q member 1) in the kidney tissue of growth restricted rats whose mothers had uteroplacental insufficiency induced on day 18 of gestation, at both embryonic day 20 (E20) and postnatal day 1 (PN1). To determine the mechanisms responsible for changes in the expression of these imprinted genes, we investigated DNA methylation of KvDMR1, an imprinting control region (ICR) that includes the promoter of the antisense long non-coding RNA Kcnq1ot1 (Kcnq1 opposite strand/antisense transcript 1). Kcnq1ot1 expression decreased by 51% in growth restricted offspring compared to sham at PN1. Interestingly, there was a negative correlation between Kcnq1ot1 and Kcnq1 in the E20 growth restricted group (Spearman's ρ = 0.014). No correlation was observed between Kcnq1ot1 and Cdkn1c expression in either group at any time point. Additionally, there was a 11.25% decrease in the methylation level at one CpG site within KvDMR1 ICR. This study, together with others in the literature, supports that long non-coding RNAs may mediate changes seen in tissues of growth restricted offspring.

Novel constituents of Salvia hispanica L. (chia) nutlet mucilage and the improved in vitro fermentation of nutlets when ground.

Upon wetting, chia (Salvia hispanica L.) nutlets produce a gel-like capsule of polysaccharides called mucilage that comprises a significant part of their dietary fibre content. Seed/nutlet mucilage is often used as a texture modifying hydrocolloid and bulking dietary fibre due to its water-binding ability, though the utility of mucilage from different sources is highly structure-function dependent. The composition and structure of chia nutlet mucilage is poorly defined, and a better understanding will aid in exploiting its dietary fibre functionality, particularly if, and how, it is utilised by gut microbiota. In this study, microscopy, chromatography, mass spectrometry and glycome profiling techniques showed that chia nutlet mucilage is highly complex, layered, and contains several polymer types. The mucilage comprises a novel xyloamylose containing both ß-linked-xylose and α-linked-glucose, a near-linear xylan that may be sparsely substituted, a modified cellulose domain, and abundant alcohol-soluble oligosaccharides. To assess the dietary fibre functionality of chia nutlet mucilage, an in vitro cumulative gas production technique was used to determine the fermentability of different chia nutlet preparations. The complex nature of chia nutlet mucilage led to poor fermentation where the oligosaccharides appeared to be the only fermentable substrate present in the mucilage. Of note, ground chia nutlets were better fermented than intact whole nutlets, as judged by short chain fatty acid production. Therefore, it is suggested that the benefits of eating chia as a "superfood", could be notably enhanced if the nutlets are ground rather than being consumed whole, improving the bioaccessibility of key nutrients including dietary fibre.

A chromosome-level genome assembly of Plantago ovata.

Plantago ovata is cultivated for production of its seed husk (psyllium). When wet, the husk transforms into a mucilage with properties suitable for pharmaceutical industries, utilised in supplements for controlling blood cholesterol levels, and food industries for making gluten-free products. There has been limited success in improving husk quantity and quality through breeding approaches, partly due to the lack of a reference genome. Here we constructed the first chromosome-scale reference assembly of P. ovata using a combination of 5.98 million PacBio and 636.5 million Hi-C reads. We also used corrected PacBio reads to estimate genome size and transcripts to generate gene models. The final assembly covers ~ 500 Mb with 99.3% gene set completeness. A total of 97% of the sequences are anchored to four chromosomes with an N50 of ~ 128.87 Mb. The P. ovata genome contains 61.90% repeats, where 40.04% are long terminal repeats. We identified 41,820 protein-coding genes, 411 non-coding RNAs, 108 ribosomal RNAs, and 1295 transfer RNAs. This genome will provide a resource for plant breeding programs to, for example, reduce agronomic constraints such as seed shattering, increase psyllium yield and quality, and overcome crop disease susceptibility.

Industrial hemp seed: from the field to value-added food ingredients.

Industrial hemp, with low levels of the intoxicating cannabinoid tetrahydrocannabinol (THC), is grown for fibre and seeds. The industrial hemp industry is poised for expansion. The legalisation of industrial hemp as an agricultural commodity and the inclusion of hemp seed in foods is helping to drive the expansion of the hemp food ingredients industry. This paper discusses the opportunity to build an industrial hemp industry, with a focus on the prospects of hemp seed and its components in food applications. The market opportunities for industrial hemp products are examined. Various aspects of the science that underpins the development of an industrial hemp industry through the food supply chain are presented. This includes a discussion on the agronomy, on-farm and post-harvest considerations and the various types of food ingredients that can be made from hemp seed. The characteristics of hemp seed meal, hemp seed protein and hemp seed oil are reviewed. Different processes for production of value-added ingredients from hemp seed, hemp seed oil and hemp seed protein, are examined. The applicability of hemp seed ingredients in food applications is reviewed. The design of hemp seed ingredients that are fit-for-purpose for target food applications, through the selection of varieties and processing methods for production of various hemp seed ingredients, needs to consider market-led opportunities. This will require an integrated through chain approach, combined with the development of on-farm and post-farm strategies, to ensure that the hemp seed ingredients and foods containing hemp seed are acceptable to the consumer.

Convalescent plasma in the treatment of moderate to severe COVID-19 pneumonia: a randomized controlled trial (PROTECT-Patient Trial).

There is a need for effective therapy for COVID-19 pneumonia. Convalescent plasma has antiviral activity and early observational studies suggested benefit in reducing COVID-19 severity. We investigated the safety and efficacy of convalescent plasma in hospitalized patients with COVID-19 in a population with a high HIV prevalence and where few therapeutic options were available. We performed a double-blinded, multicenter, randomized controlled trial in one private and three public sector hospitals in South Africa. Adult participants with COVID-19 pneumonia requiring non-invasive oxygen were randomized 1:1 to receive a single transfusion of 200 mL of either convalescent plasma or 0.9% saline solution. The primary outcome measure was hospital discharge and/or improvement of ≥ 2 points on the World Health Organisation Blueprint Ordinal Scale for Clinical Improvement by day 28 of enrolment. The trial was stopped early for futility by the Data and Safety Monitoring Board. 103 participants, including 21 HIV positive individuals, were randomized at the time of premature trial termination: 52 in the convalescent plasma and 51 in the placebo group. The primary outcome occurred in 31 participants in the convalescent plasma group and and 32 participants in the placebo group (relative risk 1.03 (95% CI 0.77 to 1.38). Two grade 1 transfusion-related adverse events occurred. Participants who improved clinically received convalescent plasma with a higher median anti-SARS-CoV-2 neutralizing antibody titre compared with those who did not (298 versus 205 AU/mL). Our study contributes additional evidence for recommendations against the use of convalescent plasma for COVID-19 pneumonia. Safety and feasibility in this population supports future investigation for other indications.

Nutritional properties of selected superfood extracts and their potential health benefits.

BACKGROUND: The term 'superfoods' is used to market foods considered to have significant health benefits. 'Superfoods' are claimed to prevent diseases as well as improving overall health, though the lack of explicit criteria means that any food can be labelled 'super' without support from scientific research. Typically, these 'superfoods' are rich in a particular nutrient for example antioxidants or omega-3 fatty acids. The objective of this study was to investigate the nutritional properties of a selection of superfood seeds: flax, chia, hulled sunflower and two types of processed hemp seeds and determine whether they may have potential health benefits. METHODS: We developed a simple aqueous extraction method for ground seeds and analysed their composition by mineral, protein and monosaccharide analyses. Cell viability assays were performed on Caco-2 and IEC-6 intestinal epithelial cells using increasing doses of the prepared extracts. RESULTS: Increased cell viability was observed in both cell lines with increasing concentrations of the flax seed, chia seed or hulled sunflower extracts (P < 0.05). Compositional analyses revealed the presence of polysaccharides, proteins and essential minerals in the aqueous extracts and in vitro assays showed sunflower had the highest antioxidant activity. However, differences in extract composition and antioxidant properties could not be directly related to the observed increase in cell viability suggesting that other components in the extracts may be responsible. Future studies will further characterize these extracts and investigate whether they are beneficial for gastrointestinal health.

Rapid and Successful Implementation of a COVID-19 Convalescent Plasma Programme-The South African Experience.

BACKGROUND: COVID-19 convalescent plasma (CCP) has been considered internationally as a treatment option for COVID-19. CCP refers to plasma collected from donors who have recovered from and made antibodies to SARS-CoV-2. To date, convalescent plasma has not been collected in South Africa. As other investigational therapies and vaccination were not widely accessible, there was an urgent need to implement a CCP manufacture programme to service South Africans. METHODS: The South African National Blood Service and the Western Cape Blood Service implemented a CCP programme that included CCP collection, processing, testing and storage. CCP units were tested for SARS-CoV-2 Spike ELISA and neutralising antibodies and routine blood transfusion parameters. CCP units from previously pregnant females were tested for anti-HLA and anti-HNA antibodies. RESULTS: A total of 987 CCP units were collected from 243 donors, with a median of three donations per donor. Half of the CCP units had neutralising antibody titres of >1:160. One CCP unit was positive on the TPHA serology. All CCP units tested for anti-HLA antibodies were positive. CONCLUSION: Within three months of the first COVID-19 diagnosis in South Africa, a fully operational CCP programme was set up across South Africa. The infrastructure and skills implemented will likely benefit South Africans in this and future pandemics.

The composition of Australian Plantago seeds highlights their potential as nutritionally-rich functional food ingredients.

When wetted, Plantago seeds become covered with a polysaccharide-rich gel called mucilage that has value as a food additive and bulking dietary fibre. Industrially, the dry husk layer that becomes mucilage, called psyllium, is milled off Plantago ovata seeds, the only commercial-relevant Plantago species, while the residual inner seed tissues are either used for low value animal feed or discarded. We suggest that this practice is potentially wasting a highly nutritious resource and here describe the use of histological, physicochemical, and chromatographic analyses to compare whole seed composition/characteristics of P. ovata with 11 relatives already adapted to harsh Australian conditions that may represent novel commercial crop options. We show that substantial interspecific differences in mucilage yield and macromolecular properties are mainly a consequence of differences in heteroxylan and pectin composition and probably represent wide differences in hydrocolloid functionality that can be exploited in industry. We also show that non-mucilage producing inner seed tissues contain a substantial mannan-rich endosperm, high in fermentable sugars, protein, and fats. Whole seed Plantago flour, particularly from some species obtained from harsh Australian environments, may provide improved economic and health benefits compared to purified P. ovata psyllium husk, by retaining the functionality of the seed mucilage and providing additional essential nutrients.

The goo-d stuff: Plantago as a myxospermous model with modern utility.

Mucilage, a gel-like layer formed around wetted seeds in a process called myxospermy, has importance as a proxy for studying cell wall polysaccharide biosynthesis and interactions and as a source of valuable health supplements and hydrocolloids. Arabidopsis thaliana has provided unrivalled insight into mucilage/cell wall synthesis, but its lack of commercial utility presents an opportunity to develop an alternative myxospermous model linking genetics, chemistry and functionality. Here, we discuss recent advances in the understanding of mucilage production, composition and properties of Plantago, a promising candidate as an alternative model with economic relevance. We outline how genomic/transcriptomic and chemical analysis advances could be made to strengthen Plantago's use as a model system, through challenging but achievable approaches.

The novel features of Plantago ovata seed mucilage accumulation, storage and release.

Seed mucilage polysaccharide production, storage and release in Plantago ovata is strikingly different to that of the model plant Arabidopsis. We have used microscopy techniques to track the development of mucilage secretory cells and demonstrate that mature P. ovata seeds do not have an outer intact cell layer within which the polysaccharides surround internal columellae. Instead, dehydrated mucilage is spread in a thin homogenous layer over the entire seed surface and upon wetting expands directly outwards, away from the seed. Observing mucilage expansion in real time combined with compositional analysis allowed mucilage layer definition and the roles they play in mucilage release and architecture upon hydration to be explored. The first emergent layer of hydrated mucilage is rich in pectin, extremely hydrophilic, and forms an expansion front that functions to 'jumpstart' hydration and swelling of the second layer. This next layer, comprising the bulk of the expanded seed mucilage, is predominantly composed of heteroxylan and appears to provide much of the structural integrity. Our results indicate that the synthesis, deposition, desiccation, and final storage position of mucilage polysaccharides must be carefully orchestrated, although many of these processes are not yet fully defined and vary widely between myxospermous plant species.

A small-scale fractionation pipeline for rapid analysis of seed mucilage characteristics.

BACKGROUND: Myxospermy is a process by which the external surfaces of seeds of many plant species produce mucilage-a polysaccharide-rich gel with numerous fundamental research and industrial applications. Due to its functional properties the mucilage can be difficult to remove from the seed and established methods for mucilage extraction are often incomplete, time-consuming and unnecessarily wasteful of precious seed stocks. RESULTS: Here we tested the efficacy of several established protocols for seed mucilage extraction and then downsized and adapted the most effective elements into a rapid, small-scale extraction and analysis pipeline. Within 4 h, three chemically- and functionally-distinct mucilage fractions were obtained from myxospermous seeds. These fractions were used to study natural variation and demonstrate structure-function links, to screen for known mucilage quality markers in a field trial, and to identify research and industry-relevant lines from a large mutant population. CONCLUSION: The use of this pipeline allows rapid analysis of mucilage characteristics from diverse myxospermous germplasm which can contribute to fundamental research into mucilage production and properties, quality testing for industrial manufacturing, and progressing breeding efforts in myxospermous crops.

A mathematical model of sonoporation using a liquid-crystalline shelled microbubble.

In recent years there has been a great deal of interest in using thin shelled microbubbles as a transportation mechanism for localised drug delivery, particularly for the treatment of various types of cancer. The technique used for such site-specific drug delivery is sonoporation. Despite there being numerous experimental studies on sonoporation, the mathematical modelling of this technique has still not been extensively researched. Presently there exists a very small body of work that models both hemispherical and spherical shelled microbubbles sonoporating due to acoustic microstreaming. Acoustic microstreaming is believed to be the dominant mechanism for sonoporation via shelled microbubbles. Rather than considering the shell of the microbubble to be composed of a thin protein, which is typical in the literature, in this paper we consider the shell to be a liquid-crystalline material. Up until now there have been no studies reported in the literature pertaining to sonoporation of a liquid-crystalline shelled microbubble. A mathematical expression is derived for the maximum wall shear stress, illustrating its dependency on the shell's various material parameters. A sensitivity analysis is performed for the wall shear stress considering the shell's thickness; its local density; the elastic constant of the liquid-crystalline material; the interfacial surface tension and; the shell's viscoelastic properties. In some cases, our results indicate that a liquid-crystalline shelled microbubble may yield a maximum wall shear stress that is two orders of magnitude greater than the stress generated by commercial shelled microbubbles that are currently in use within the scientific community. In conclusion, our preliminary analysis suggests that using liquid-crystalline shelled microbubbles may significantly enhance the efficiency of site-specific drug delivery.