Effect of Freezing Process on the Microstructure of Gelatin Methacryloyl Hydrogels

Gelatin methacryloyl (GelMA) hydrogels have aroused considerable interests in the field of tissue engineering due to tunable physical properties and cell response parameters. A number of works have studied the impact of GelMA concentration, photo-initiator concentration, methacrylic anhydride (MA) concentration, cooling rate and temperature gradient on GelMA hydrogel generation, but little attention has been paid to the effect of the freezing temperatures and freezing time of GelMA prepolymer solution during preparation. In this study, GelMA hydrogels were synthesized with different freezing temperatures and time. It was found that the lower freezing temperatures and longer freezing time caused smaller pore sizes that realized higher cell viability and proliferation of MC3T3-E1 cells. The results showed that tunable microstructure of GelMA could be achieved by regulating the freezing conditions of GelMA, which provided a broad prospect for the applications of GelMA hydrogels in tissue engineering.

Active virus-host interactions at sub-freezing temperatures in Arctic peat soil

Background Winter carbon loss in northern ecosystems is estimated to be greater than the average growing season carbon uptake and is primarily driven by microbial decomposers. Viruses modulate microbial carbon cycling via induced mortality and metabolic controls, but it is unknown whether viruses are active under winter conditions (anoxic and sub-freezing temperatures). Results We used stable isotope probing (SIP) targeted metagenomics to reveal the genomic potential of active soil microbial populations under simulated winter conditions, with an emphasis on viruses and virus-host dynamics. Arctic peat soils from the Bonanza Creek Long-Term Ecological Research site in Alaska were incubated under sub-freezing anoxic conditions with H218O or natural abundance water for 184 and 370 days. We sequenced 23 SIP-metagenomes and measured carbon dioxide (CO2) efflux throughout the experiment. We identified 46 bacterial populations (spanning 9 phyla) and 243 viral populations that actively took up 18O in soil and respired CO2 throughout the incubation. Active bacterial populations represented only a small portion of the detected microbial community and were capable of fermentation and organic matter degradation. In contrast, active viral populations represented a large portion of the detected viral community and one third were linked to active bacterial populations. We identified 86 auxiliary metabolic genes and other environmentally relevant genes. The majority of these genes were carried by active viral populations and had diverse functions such as carbon utilization and scavenging that could provide their host with a fitness advantage for utilizing much-needed carbon sources or acquiring essential nutrients. Conclusions Overall, there was a stark difference in the identity and function of the active bacterial and viral community compared to the unlabeled community that would have been overlooked with a non-targeted standard metagenomic analysis. Our results illustrate that substantial active virus-host interactions occur in sub-freezing anoxic conditions and highlight viruses as a major community-structuring agent that likely modulates carbon loss in peat soils during winter, which may be pivotal for understanding the future fate of arctic soils' vast carbon stocks.

Cold-Triggered Induction of ROS- and Raffinose Metabolism in Freezing-Sensitive Taproot Tissue of Sugar Beet

Sugar beet (Beta vulgaris subsp. vulgaris) is the exclusive source of sugar in the form of sucrose in temperate climate zones. Sugar beet is grown there as an annual crop from spring to autumn because of the damaging effect of freezing temperatures to taproot tissue. A collection of hybrid and non-hybrid sugar beet cultivars was tested for winter survival rates and freezing tolerance. Three genotypes with either low or high winter survival rates were selected for detailed study of their response to frost. These genotypes differed in the severity of frost injury in a defined inner region in the upper part of the taproot, the so-called pith. We aimed to elucidate genotype- and tissue-dependent molecular processes during freezing and combined analyses of sugar beet anatomy and physiology with transcriptomic and metabolite profiles of leaf and taproot tissues at low temperatures. Freezing temperatures induced strong downregulation of photosynthesis in leaves, generation of reactive oxygen species (ROS), and ROS-related gene expression in taproots. Simultaneously, expression of genes involved in raffinose metabolism, as well as concentrations of raffinose and its intermediates, increased markedly in both leaf and taproot tissue at low temperatures. The accumulation of raffinose in the pith tissue correlated with freezing tolerance of the three genotypes. We discuss a protective role for raffinose and its precursors against freezing damage of sugar beet taproot tissue.

1479 Trench Foot: A Rare and Unusual Presentation of a World War I Disease

Introduction 'Trench foot' is a serious disorder of the lower limb, involving damage to the skin, nerves, and muscle, previously described in World War I soldiers during trench warfare. We present a rare and unusual case of ‘trench foot' sustained at home in a 20-year-old female. Case Report A 20-year-old woman presented to the Plastic Surgery department with pain and numbness affecting both feet, following failed trials of antibiotics. She managed her symptoms at home by immersing her feet in ice baths for 18-23 hours per day. She was tachycardic with raised inflammatory markers. Examination revealed breakdown of the skin bilaterally, with full thickness eschar. MRI showed extensive subcutaneous oedema and myositis. Initial surgical plan consisted of antibiotics and debridement of necrotic tissue with a view to grafting the skin later. Intra-operative findings included necrosis of the subcutaneous tissues and muscles. The patient deteriorated post-operatively with sepsis and underwent urgent Guillotine-style bilateral amputations. She was discharged home 18 days later. Discussion 'Trench foot' can be mistaken for soft tissue infections or frostbite. It is attributed to vasoconstriction followed by neurovascular changes and repeated cycles of thawing and freezing, acquired above freezing temperatures, unlike frostbite which occurs below freezing temperatures. The patient was initially reviewed by non-specialist teams without a clear diagnosis and sustained more pervasive tissue destruction than was originally apparent on examination. Prevention remains the best cure for ‘trench foot'. It is therefore important to familiarise ourselves with this rare disease. Where prevention or early detection is not possible, amputation can reduce the morbidity and mortality of the ensuing sepsis.