Comparative genomics points to tandem duplications of SAD gene clusters as drivers of increased α-linolenic (ω-3) content in S. hispanica seeds.

Salvia hispanica L. (chia) is a source of abundant ω-3 polyunsaturated fatty acids (ω-3-PUFAs) that are highly beneficial to human health. The genomic basis for this accrued ω-3-PUFA content in this emerging crop was investigated through the assembly and comparative analysis of a chromosome-level reference genome for S. hispanica. The highly contiguous 321.5-Mbp genome assembly covering all six chromosomes enabled the identification of 32,922 protein-coding genes. Two whole-genome duplications (WGD) events were identified in the S. hispanica lineage. However, these WGD events could not be linked to the high α-linolenic acid (ALA, ω-3) accumulation in S. hispanica seeds based on phylogenomics. Instead, our analysis supports the hypothesis that evolutionary expansion through tandem duplications of specific lipid gene families, particularly the stearoyl-acyl carrier protein desaturase (ShSAD) gene family, is the main driver of the abundance of ω-3-PUFAs in S. hispanica seeds. The insights gained from the genomic analysis of S. hispanica will help establish a molecular breeding target that can be leveraged through genome editing techniques to increase ω-3 content in oil crops.

Rounding up the annual ryegrass genome: High-quality reference genome of Lolium rigidum.

The genome of the major agricultural weed species, annual ryegrass (Lolium rigidum) was assembled, annotated and analysed. Annual ryegrass is a major weed in grain cropping, and has the remarkable capacity to evolve resistance to herbicides with various modes of action. The chromosome-level assembly was achieved using short- and long-read sequencing in combination with Hi-C mapping. The assembly size is 2.44 Gb with N50 = 361.79 Mb across 1,764 scaffolds where the seven longest sequences correspond to the seven chromosomes. Genome completeness assessed through BUSCO returned a 99.8% score for complete (unique and duplicated) and fragmented genes using the Viridiplantae set. We found evidence for the expansion of herbicide resistance-related gene families including detoxification genes. The reference genome of L. rigidum is a critical asset for leveraging genetic information for the management of this highly problematic weed species.

Addressing smoking in sheltered homelessness with intensive smoking treatment (ASSIST project): A pilot feasibility study of varenicline, combination nicotine replacement therapy and motivational interviewing.

BACKGROUND: This pilot study aimed to test the feasibility of providing varenicline in combination with nicotine replacement therapy (NRT) and motivational interviewing (MI) to adult male smokers attending a clinic in a hostel for homeless people. METHODS: A single group pre- and post-treatment (12 weeks following intervention commencement) design with embedded process evaluation (at weekly counselling and fortnightly safety check-ins). Participants were 20 male smokers attending a health clinic within a homelessness service in Sydney, Australia, between December 2019 and March 2020. Participants set a target quit date 7-days post intervention commencement. Adverse events, self-reported abstinence, cigarettes per day, treatment adherence and acceptability of the study interventions were assessed 12 weeks post intervention commencement. Abstinence was biochemically verified. Results are complete cases. RESULTS: Retention was 65% at 12-weeks post-intervention commencement (n = 13). No related adverse events were reported. Three participants (15%) reported continuous abstinence. Two participants self-reported 30-day point prevalence abstinence (10%), confirmed by CO level. Participants who did not quit smoking (n = 10), reported a significant reduction in the number of cigarettes smoked per day (19.4 vs 4.7, p < .01). Cravings, withdrawal symptoms, and psychological distress significantly decreased from baseline to 12-week follow-up (all < 0.01). Adherence to the pharmacological interventions was good, most used combination NRT and varenicline. Adherence to the counselling sessions was low, attending three of 12 sessions. Both NRT and MI were rated as highly acceptable. Some participants expressed concerns about the safety of varenicline. CONCLUSIONS: The intervention was feasible and acceptable and associated with short-term smoking cessation and significant reductions in the number of cigarettes smoked-per-day.

Effect of recording site on pulse oximetry readings.

BACKGROUND: Ambulatory oxygen requirements are routinely assessed and titrated using portable finger pulse oximetry. However, movement artefacts from hands moving or reduced circulation may adversely affect the accuracy of measurement. At Wythenshawe Hospital, we decided to use an ear oximeter routinely in addition to the finger oximeter. AIM: To evaluate and compare the use of portable oximetry measurements taken from the ear and finger during ambulatory assessments. METHOD: Before, during and after a six-minute walking test, 304 patients had ear and finger pulse oximetry measurements recorded. RESULTS: There was a significant difference between ear and finger measurements before and after exercise (p=<0.001 for both). The differences would have altered the clinical outcome in 27% of those assessed.

Stretch and CO2 modulate the inflammatory response of alveolar macrophages through independent changes in metabolic activity.

Ventilatory-induced strain can exacerbate acute lung injury (ALI). Current ventilation strategies favour low tidal volumes and high end-expiratory volumes to 'rest' the lung, but can lead to an increase in CO2. Alveolar macrophages (AM) play a pivotal role in ALI through the release of inflammatory mediators. The effect of physical strain and CO2 on the release of pro-inflammatory mediators was examined in isolated rat AM. AM were cultured on IgG-coated silastic membranes with or without lipopolysaccharide (LPS) and 5% or 20% CO2 and subjected to a repetitive sinusoidal mechanical strain (30%, 60 cycles/min) for 4 h. Cell viability and metabolic activity were assessed. In both the presence and absence of LPS, physical strain increased metabolic activity by approximately 5%, while 20% CO2 decreased metabolic activity by approximately 40%. Twenty per cent CO2 decreased TNF-alpha secretion by approximately 45%, without affecting cell viability. Physical strain enhanced LPS-induced secretion of TNF-alpha by 1.5%, but not IL-6 or CINC-1. Hence, the effects of both CO2 and physical strain are mediated independently through changes in AM metabolic activity. Physical strain is not a major determinant of TNF-alpha, IL-6 or CINC-1 in AM. Our results confirm that high CO2 can lessen the TNF-alpha inflammatory response of AM.

GPCR screening via ERK 1/2: a novel platform for screening G protein-coupled receptors.

Discovery of novel agonists and antagonists for G protein-coupled receptors (GPCRs) relies heavily on cell-based assays because determination of functional consequences of receptor engagement is often desirable. Currently, there are several key parameters measured to achieve this, including mobilization of intracellular Ca2+ and formation of cyclic adenosine monophosphate or inositol triphosphate. However, no single assay platform is suitable for all situations, and all of the assays have limitations. The authors have developed a new high-throughput homogeneous assay platform for GPCR discovery as an alternative to current assays, which employs detection of phosphorylation of the key signaling molecule p42/44 MAP kinase (ERK 1/2). The authors show that ERK 1/2 is consistently activated in cells stimulated by Gq-coupled GPCRs and provides a new high-throughput platform for screening GPCR drug candidates. The activation of ERK 1/2 in Gq-coupled GPCR systems generates comparable pharmacological data for receptor agonist and antagonist data obtained by other GPCR activation measurement techniques.