Extremophiles in an Antarctic Marine Ecosystem

Recent attempts to explore marine microbial diversity and the global marine microbiome have indicated a large proportion of previously unknown diversity. However, sequencing alone does not tell the whole story, as it relies heavily upon information that is already contained within sequence databases. In addition, microorganisms have been shown to present small-to-large scale biogeographical patterns worldwide, potentially making regional combinations of selection pressures unique. Here, we focus on the extremophile community in the boundary region located between the Polar Front and the Southern Antarctic Circumpolar Current in the Southern Ocean, to explore the potential of metagenomic approaches as a tool for bioprospecting in the search for novel functional activity based on targeted sampling efforts. We assessed the microbial composition and diversity from a region north of the current limit for winter sea ice, north of the Southern Antarctic Circumpolar Front (SACCF) but south of the Polar Front. Although, most of the more frequently encountered sequences were derived from common marine microorganisms, within these dominant groups, we found a proportion of genes related to secondary metabolism of potential interest in bioprospecting. Extremophiles were rare by comparison but belonged to a range of genera. Hence, they represented interesting targets from which to identify rare or novel functions. Ultimately, future shifts in environmental conditions favoring more cosmopolitan groups could have an unpredictable effect on microbial diversity and function in the Southern Ocean, perhaps excluding the rarer extremophiles

Volatile organic compounds associated with neonectria ditissima infection in apples (Malus pumila cv Gala)

Postharvest diseases in apples during long term storage result in loss and waste. This is mainly caused by fungal pathogens. Fungal contamination and rot can change some of the volatile organic compounds (VOC) emitted by apple fruits. In this study, disease free Gala apples were inoculated with Neonectria ditissima. The aim was to identify VOCs associated with N. ditissima infection in gala apples. The inoculated apples were placed in 5L glass flask, sealed, and incubated at 20oC for one hour after which a charcoal filtered airflow of 1 L/min was maintained for one hour through the Volatile Capture Trap (VCT) with volatile emissions captured on a porapak-Q absorbent filter. Captured volatiles were eluted using 1 mL of dichloromethane (DCM) into a standard Agilent 1.5 mL HPLC vial. Eluted volatiles were analysed using Gas Chromatography coupled with Mass Spectrometry (GC/MS). Volatiles were capture in three replicates for both inoculated and healthy control groups at 2 days, 8 days, 14 days, 21 days, 28 days, 35 days, and 42 days post-inoculation. The N. ditissima discriminatory volatile were identified/discriminated qualitatively based on the unique volatile compounds detected and quantitatively based on variation in peak area of certain combinations of volatile compounds. Some of the discriminatory volatiles such as dodecyl hexanoate, 9-decen-1-yl hexanoate, hexyl butanoate and pentyl acetate were detected in the early stages of the infection. Styrene, terpinene-4-ol, ethyl hexanoate, ethyl butanoate, ethyl pentanoate and 2-methylpentyl formate constituted the main VOCs emitted during apple fruit decay. Other compounds such as alpha Farnesene and hexyl acetate were common to both healthy and inoculated apples but the peak areas in the healthy apples were well above the peak areas in the inoculated apples. However, these compounds expressed a decline in peak area over time. Apples are stored commercially in sealed stores for months making visual observation for early detection of disease almost impossible. Disease of stored apples are most times only detected at advanced stages when it has become nearly impossible to prevent losses. These discriminatory volatile metabolites detected at early stages of infection are important for early non-visual detection of N. ditissima in stored apples. Further research is recommended to the use of these compounds in early detection of the disease caused by N. ditissima

eFluorination for the rapid synthesis of Carbamoyl fluorides from oxamic acids

In this letter, we disclose the anodic oxidation of oxamic acids in the presence of Et3N·3HF as a practical, scalable, and robust method to rapidly access carbamoyl fluorides from readily available and stable precursors. The simplicity of this method also led us to develop the first flow electrochemical preparation of carbamoyl fluorides, demonstrating scale-up feasibility as a proof of concept

Machine learning to detect fungal infections in stored pome fruits via mass spectrometry data: industry, economic, and social implications

Pome fruits, notably apples and pears, experience decay during storage due to fungal infections. The timely discernment of these infections is imperative to avert the deterioration of these fruits within warehouse confines. In an experimental setup, two distinct apple cultivars, Braeburn and Gala, were inoculated with fungi Monilinia laxa, Neonectria ditissima, and Botrytis cinerea. As the infection progresses, the apples release chemical volatile components, which are measured using mass spectrometry in both positive and negative ion modes, recording mass-charge ratios ranging from m/z 30 to m/z 900 with a 0.3 Dalton difference between each measurement. The dataset is then partitioned into 24 sets of three-dimensional data, encompassing attributes related to two types of apples, three types of fungi, and two types of ions. They are analyzed using various machine learning algorithms, including Logistic Regression, Support Vector Machines, XGBoost, Random Forest, and four distinct customised Neural Networks, to classify infected and uninfected apples. The outcomes from the different machine learning algorithms across the 12 combinations of Apple-FungiIon are recorded, revealing that certain algorithms excel in different combinations. The performance metrics namely True Positive, True Negative, False Positive, False Negative, Accuracy are closely analysed and the algorithms that produces the highest and second-highest accuracy are highlighted. Upon thorough analysis of the 12 combinations, it is observed that Logistic Regression and SVM with a linear kernel achieve the highest accuracy in approximately 11 combinations. Specifically, Logistic Regression achieves a precision of 98% for Braeburn apples, while SVM attains a 99% accuracy for Gala apples. This research project has a triple impact on industry, economy, and society. On an industrial level, the precision and early predictions of the proposed work can effectively safeguard large quantities of apples in storage bins. Economically, it has the potential to avert substantial monetary losses. Societally, it plays a crucial role in determining the ideal timing to release fruits to the market for consumption without jeopardizing human health

Investigating the utility of saliva immunoglobulins for the detection of myeloma and using myeloma proteins to clarify partition between oral and systemic immunity

OBJECTIVES Myeloma is characterised by the presence of monoclonal immunoglobulin (M-protein) and the free light chain (FLC) in blood. We investigated whether these M-proteins and FLC are detectable in myeloma patients' saliva to evaluate its utility for non-invasive screening and monitoring of haematological malignancies. METHODS A total of 57 patients with monoclonal gammopathy and 26 age-matched healthy participants provided paired serum and saliva samples for immunoglobulin characterisation and quantification. RESULTS Myeloma patients had IgG or IgA M-protein levels ranging up to five times and FLC levels up to a thousand times normal levels of polyclonal immunoglobulins. Despite these highly elevated levels, only two IgG and no IgA M-proteins or FLC could be detected in paired saliva samples. Most patients had reduced levels of serum polyclonal immunoglobulins, but all had normal levels of salivary IgA. CONCLUSIONS Immunoglobulin transfer from blood is not determined by levels in the systemic circulation and more likely dictated by periodontal inflammation and the integrity of the oral epithelium. Immunoglobulins secreted by bone marrow plasma cells do not substantially enter saliva, which represents a poor medium for myeloma diagnosis. These findings, along with normal salivary IgA levels despite systemic immunoparesis, support a strong partitioning of oral from systemic humoral immunity

Anodic oxidation of dithiane carboxylic acids: a rapid and mild 2 way to access functionalized orthoesters

A new electrochemical methodology has been developed for the preparation of a wide variety of functionalized orthoesters under mild and green conditions from easily accessible dithiane derivatives. The new methodology also offers an unprecedented way to access tri(fluorinated) orthoesters, a class of compound that has never been studied before. This provides the community with a rapid and general method to prepare libraries of functionalized orthoesters from simple and readily available starting materials

Hardness variation in inconel 718 produced by laser directed energy deposition

Directed energy deposition (DED) of Inconel 718 is of critical importance for the repair of aerospace components, which have tight tolerances for certification, particularly on mechanical properties. Significant hardness variation has been seen throughout DED manufactured Inconel 718 components, suggestive of variation in mechanical properties, which must be understood such that the variation can either be removed, or implemented within the design in line with regulatory guidance. In this work, γʹ precipitation was theorised to be the cause of hardness variation throughout the component, despite Inconel 718 conventionally being regarded as a γʺ strengthened alloy. A simple precipitation potential model based on a moving heat source was found to correlate with the measured hardness and explain the hardness distribution observed. In addition, it has been shown that sections under a critical thickness of 2 mm never reach the peak hardness in the as-built condition. This understanding allows for the development of in-situ heat treatment strategies to be developed for microstructural, and hence, mechanical property optimisation, necessary for repair technologies where post processing steps are limited

Hardness variation in inconel 718 produced by laser directed energy deposition

Directed energy deposition (DED) of Inconel 718 is of critical importance for the repair of aerospace components, which have tight tolerances for certification, particularly on mechanical properties. Significant hardness variation has been seen throughout DED manufactured Inconel 718 components, suggestive of variation in mechanical properties, which must be understood such that the variation can either be removed, or implemented within the design in line with regulatory guidance. In this work, γʹ precipitation was theorised to be the cause of hardness variation throughout the component, despite Inconel 718 conventionally being regarded as a γʺ strengthened alloy. A simple precipitation potential model based on a moving heat source was found to correlate with the measured hardness and explain the hardness distribution observed. In addition, it has been shown that sections under a critical thickness of 2 mm never reach the peak hardness in the as-built condition. This understanding allows for the development of in-situ heat treatment strategies to be developed for microstructural, and hence, mechanical property optimisation, necessary for repair technologies where post processing steps are limited

Exosomes in the diagnosis and treatment of renal cell cancer

Renal cell carcinoma (RCC) is the most prevalent type of kidney cancer originating from renal tubular epithelial cells, with clear cell RCC comprising approximately 80% of cases. The primary treatment modalities for RCC are surgery and targeted therapy, albeit with suboptimal efficacies. Despite progress in RCC research, significant challenges persist, including advanced distant metastasis, delayed diagnosis, and drug resistance. Growing evidence suggests that extracellular vesicles (EVs) play a pivotal role in multiple aspects of RCC, including tumorigenesis, metastasis, immune evasion, and drug response. These membrane-bound vesicles are released into the extracellular environment by nearly all cell types and are capable of transferring various bioactive molecules, including RNA, DNA, proteins, and lipids, aiding intercellular communication. The molecular cargo carried by EVs renders them an attractive resource for biomarker identification, while their multifarious role in the RCC offers opportunities for diagnosis and targeted interventions, including EV-based therapies. As the most versatile type of EVs, exosomes have attracted much attention as nanocarriers of biologicals, with multi-range signaling effects. Despite the growing interest in exosomes, there is currently no widely accepted consensus on their subtypes and properties. The emerging heterogeneity of exosomes presents both methodological challenges and exciting opportunities for diagnostic and clinical interventions. This article reviews the characteristics and functions of exosomes, with a particular reference to the recent advances in their application to the diagnosis and treatment of RCC

Energy absorption in lattice structures in dynamics: Experiments

Lattice structures offer the potential to relatively easily engineer specific (meso-scale properties (cell level)), to produce desirable macro-scale material properties for a wide variety of engineering applications including wave filters, blast and impact protection systems, thermal insulation, structural aircraft and vehicle components, and body implants. The work presented here focuses on characterising the quasi-static and, in particular, the dynamic load-deformation behaviour of lattice samples. First, cubic, diamond and re-entrant cube lattice structures were tested under quasi-static conditions to investigate failure process and stress–strain response of such materials. Following the quasi-static tests, Hopkinson pressure bar (HPB) tests were carried out to evaluate the impact response of these materials under high deformation rates. The HPB tests show that the lattice structures are able to spread impact loading in time and to reduce the peak impact stress. A significant rate dependency of load-deformation characteristics was identified. This is believed to be the first published results of experimental load-deformation studies of additively manufactured lattice structures. The cubic and diamond lattices are, by a small margin, the most effective of those lattices investigated to achieve this