Favorable Antiviral Effect of Metformin on Severe Acute Respiratory Syndrome Coronavirus 2 Viral Load in a Randomized, Placebo-Controlled Clinical Trial of Coronavirus Disease 2019.

BACKGROUND: Metformin has antiviral activity against RNA viruses including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The mechanism appears to be suppression of protein translation via targeting the host mechanistic target of rapamycin pathway. In the COVID-OUT randomized trial for outpatient coronavirus disease 2019 (COVID-19), metformin reduced the odds of hospitalizations/death through 28 days by 58%, of emergency department visits/hospitalizations/death through 14 days by 42%, and of long COVID through 10 months by 42%. METHODS: COVID-OUT was a 2 × 3 randomized, placebo-controlled, double-blind trial that assessed metformin, fluvoxamine, and ivermectin; 999 participants self-collected anterior nasal swabs on day 1 (n = 945), day 5 (n = 871), and day 10 (n = 775). Viral load was quantified using reverse-transcription quantitative polymerase chain reaction. RESULTS: The mean SARS-CoV-2 viral load was reduced 3.6-fold with metformin relative to placebo (-0.56 log10 copies/mL; 95% confidence interval [CI], -1.05 to -.06; P = .027). Those who received metformin were less likely to have a detectable viral load than placebo at day 5 or day 10 (odds ratio [OR], 0.72; 95% CI, .55 to .94). Viral rebound, defined as a higher viral load at day 10 than day 5, was less frequent with metformin (3.28%) than placebo (5.95%; OR, 0.68; 95% CI, .36 to 1.29). The metformin effect was consistent across subgroups and increased over time. Neither ivermectin nor fluvoxamine showed effect over placebo. CONCLUSIONS: In this randomized, placebo-controlled trial of outpatient treatment of SARS-CoV-2, metformin significantly reduced SARS-CoV-2 viral load, which may explain the clinical benefits in this trial. Metformin is pleiotropic with other actions that are relevant to COVID-19 pathophysiology. CLINICAL TRIALS REGISTRATION: NCT04510194.

Metformin reduces SARS-CoV-2 in a Phase 3 Randomized Placebo Controlled Clinical Trial.

Current antiviral treatment options for SARS-CoV-2 infections are not available globally, cannot be used with many medications, and are limited to virus-specific targets.1-3 Biophysical modeling of SARS-CoV-2 replication predicted that protein translation is an especially attractive target for antiviral therapy.4 Literature review identified metformin, widely known as a treatment for diabetes, as a potential suppressor of protein translation via targeting of the host mTor pathway.5 In vitro, metformin has antiviral activity against RNA viruses including SARS-CoV-2.6,7 In the COVID-OUT phase 3, randomized, placebo-controlled trial of outpatient treatment of COVID-19, metformin had a 42% reduction in ER visits/hospitalizations/death through 14 days; a 58% reduction in hospitalizations/death through 28 days, and a 42% reduction in Long COVID through 10 months.8,9 Here we show viral load analysis of specimens collected in the COVID-OUT trial that the mean SARS-CoV-2 viral load was reduced 3.6-fold with metformin relative to placebo (-0.56 log10 copies/mL; 95%CI, -1.05 to -0.06, p=0.027) while there was no virologic effect for ivermectin or fluvoxamine vs placebo. The metformin effect was consistent across subgroups and with emerging data.10,11 Our results demonstrate, consistent with model predictions, that a safe, widely available,12 well-tolerated, and inexpensive oral medication, metformin, can be repurposed to significantly reduce SARS-CoV-2 viral load.

A rapid, cost-effective tailed amplicon method for sequencing SARS-CoV-2.

BACKGROUND: The global COVID-19 pandemic has led to an urgent need for scalable methods for clinical diagnostics and viral tracking. Next generation sequencing technologies have enabled large-scale genomic surveillance of SARS-CoV-2 as thousands of isolates are being sequenced around the world and deposited in public data repositories. A number of methods using both short- and long-read technologies are currently being applied for SARS-CoV-2 sequencing, including amplicon approaches, metagenomic methods, and sequence capture or enrichment methods. Given the small genome size, the ability to sequence SARS-CoV-2 at scale is limited by the cost and labor associated with making sequencing libraries. RESULTS: Here we describe a low-cost, streamlined, all amplicon-based method for sequencing SARS-CoV-2, which bypasses costly and time-consuming library preparation steps. We benchmark this tailed amplicon method against both the ARTIC amplicon protocol and sequence capture approaches and show that an optimized tailed amplicon approach achieves comparable amplicon balance, coverage metrics, and variant calls to the ARTIC v3 approach. CONCLUSIONS: The tailed amplicon method we describe represents a cost-effective and highly scalable method for SARS-CoV-2 sequencing.

Ancient DNA reveals the timing and persistence of organellar genetic bottlenecks over 3,000 years of sunflower domestication and improvement.

Here, we report a comprehensive paleogenomic study of archaeological and ethnographic sunflower remains that provides significant new insights into the process of domestication of this important crop. DNA from both ancient and historic contexts yielded high proportions of endogenous DNA, and although archaeological DNA was found to be highly degraded, it still provided sufficient coverage to analyze genetic changes over time. Shotgun sequencing data from specimens from the Eden's Bluff archaeological site in Arkansas yielded organellar DNA sequence from specimens up to 3,100 years old. Their sequences match those of modern cultivated sunflowers and are consistent with an early domestication bottleneck in this species. Our findings also suggest that recent breeding of sunflowers has led to a loss of genetic diversity that was present only a century ago in Native American landraces. These breeding episodes also left a profound signature on the mitochondrial and plastid haplotypes in cultivars, as two types were intentionally introduced from other Helianthus species for crop improvement. These findings gained from ancient and historic sunflower specimens underscore how future in-depth gene-based analyses can advance our understanding of the pace and targets of selection during the domestication of sunflower and other crop species.

Response to joint selection on germination and flowering phenology depends on the direction of selection.

Flowering and germination time are components of phenology, a complex phenotype that incorporates a number of traits. In natural populations, selection is likely to occur on multiple components of phenology at once. However, we have little knowledge of how joint selection on several phenological traits influences evolutionary response. We conducted one generation of artificial selection for all combinations of early and late germination and flowering on replicated lines within two independent base populations in the herb Campanula americana. We then measured response to selection and realized heritability for each trait. Response to selection and heritability were greater for flowering time than germination time, indicating greater evolutionary potential of this trait. Selection for earlier phenology, both flowering and germination, did not depend on the direction of selection on the other trait, whereas response to selection to delay germination and flowering was greater when selection on the other trait was in the opposite direction (e.g., early germination and late flowering), indicating a negative genetic correlation between the traits. Therefore, the extent to which correlations shaped response to selection depended on the direction of selection. Furthermore, the genetic correlation between timing of germination and flowering varies across the trait distributions. The negative correlation between germination and flowering time found when selecting for delayed phenology follows theoretical predictions of constraint for traits that jointly determine life history schedule. In contrast, the lack of constraint found when selecting for an accelerated phenology suggests a reduction of the covariance due to strong selection favoring earlier flowering and a shorter life cycle. This genetic architecture, in turn, will facilitate further evolution of the early phenology often favored in warm climates.

Transitions in photoperiodic flowering are common and involve few loci in wild sunflowers (Helianthus; Asteraceae).

UNLABELLED: • PREMISE OF THE STUDY: Evolutionary changes in how flowering time responds to photoperiod cues have been instrumental in expanding the geographic range of agricultural production for many crop species. Locally adaptive natural variation in photoperiod response present in wild relatives of crop plants could be leveraged to further improve the present and future climatic ranges of cultivation or to increase region-specific yields. Previous work has demonstrated ample variability in photoperiod response among wild populations of the common sunflower, Helianthus annuus. Here, we characterize patterns of photoperiod response variation throughout the genus and examine the genetic architecture of intraspecific divergence.• METHODS: The requirement of short day lengths for floral induction was characterized for a phylogenetically dispersed sample of Helianthus species. In addition, flowering time was assessed under short days and long days for a population of F3 individuals derived from crosses between day-neutral and short-day, wild H. annuus parents.• KEY RESULTS: An obligate requirement for short-day induced flowering has evolved repeatedly in Helianthus, and this character was correlated with geographic ranges restricted to the southern United States. Parental flowering times under long days were recovered in high proportion in the F3 generation.• CONCLUSIONS: Together, these findings (1) reveal that substantial variation in the nature of flowering time responses to photoperiod cues has arisen during the evolution of wild sunflowers and (2) suggest these transitions may be largely characterized by simple genetic architectures. Thus, introgression of wild alleles may be a tractable means of genetically tailoring sunflower cultivars for climate-specific production.

A comparison of Japanese and Australian consumers' sensory perceptions of beef.

This paper presents a comparison of Japanese and Australian consumer sensory responses to beef, based on Meat Standards Australia methodology. Japanese and Australian consumers evaluated paired beef samples according to four sensory traits, and the weighted results were combined to produce a Meat Quality score (MQ4). The consumers also categorized the beef samples to one of four grades (unsatisfactory, good everyday, better than everyday and premium). The proportion of samples assigned to each grade was similar for Japanese and Australian consumers for yakiniku and shabu shabu cooking methods; however, Japanese consumers assigned lower scores to the grill samples. In terms of the MQ4 boundary scores between grades, these were very similar for both Japanese and Australian consumers across all cooking methods. In terms of the weightings for the four sensory traits, juiciness was more important for Japanese consumers than Australian for grill and shabu shabu cooking methods. Flavor had the highest weighting for both consumer groups. This study showed that a beef description system based on the MQ4 score, with some adjustments to the weightings and cut-off values, could be useful in describing the eating quality of beef for the Japanese consumer.

The Embryonic Transcriptome of the Red-Eared Slider Turtle (Trachemys scripta).

The bony shell of the turtle is an evolutionary novelty not found in any other group of animals, however, research into its formation has suggested that it has evolved through modification of conserved developmental mechanisms. Although these mechanisms have been extensively characterized in model organisms, the tools for characterizing them in non-model organisms such as turtles have been limited by a lack of genomic resources. We have used a next generation sequencing approach to generate and assemble a transcriptome from stage 14 and 17 Trachemys scripta embryos, stages during which important events in shell development are known to take place. The transcriptome consists of 231,876 sequences with an N50 of 1,166 bp. GO terms and EC codes were assigned to the 61,643 unique predicted proteins identified in the transcriptome sequences. All major GO categories and metabolic pathways are represented in the transcriptome. Transcriptome sequences were used to amplify several cDNA fragments designed for use as RNA in situ probes. One of these, BMP5, was hybridized to a T. scripta embryo and exhibits both conserved and novel expression patterns. The transcriptome sequences should be of broad use for understanding the evolution and development of the turtle shell and for annotating any future T. scripta genome sequences.

Japanese consumer categorisation of beef into quality grades, based on Meat Standards Australia methodology.

The objective was to evaluate the sensory categorisation of beef by Japanese consumers, based on Meat Standards Australia methodology. Various cuts of beef, with a wide range of quality (from Australian and Japanese cattle) and three cooking methods (grill, yakiniku, shabu shabu), were evaluated by 1620 Japanese consumers in Tokyo and Osaka. Consumers rated each sample for four sensory attributes (tenderness, juiciness, flavor and overall satisfaction), then selected one of four grades (unsatisfactory/2-star, good everyday/3-star, better than everyday/4-star, and premium quality/5-star), based on the quality of the beef within each cooking method. Meat quality scores, denoted as MQ4 scores (weighted combination of the four sensory attributes) were calculated from the Japanese consumer test results, to describe the Japanese consumer rating of beef. The distribution of the Japanese consumer MQ4 scores showed a clear distinction between grades, with the majority of scores being included within the boundaries of each grade. The MQ4 score allocated approximately 64% of the samples to their correct consumer grades. The MQ4 score showed potential to be used as a tool in developing and monitoring a consumer-focused grading system that is able to predict Japanese consumer satisfaction of individual beef cuts prepared by different cooking methods.

Assessing the impact of suicide exclusion periods on life insurance.

BACKGROUND: We study the impact of suicide-exclusion periods, common in life insurance policies in Australia, on suicide and accidental death rates for life-insured individuals. If a life-insured individual dies by suicide during the period of suicide exclusion, commonly 13 months, the sum insured is not paid. AIMS: We examine whether a suicide-exclusion period affects the timing of suicides. We also analyze whether accidental deaths are more prevalent during the suicide-exclusion period as life-insured individuals disguise their death by suicide. We assess the relationship between the insured sum and suicidal death rates. METHODS: Crude and age-standardized rates of suicide, accidental death, and overall death, split by duration since the insured first bought their insurance policy, were computed. RESULTS: There were significantly fewer suicides and no significant spike in the number of accidental deaths in the exclusion period for Australian life insurance data. More suicides, however, were detected for the first 2 years after the exclusion period. Higher insured sums are associated with higher rates of suicide. CONCLUSIONS: Adverse selection in Australian life insurance is exacerbated by including a suicide-exclusion period. Extension of the suicide-exclusion period to 3 years may prevent some "insurance-induced" suicides - a rationale for this conclusion is given.

Delirium in the elderly. A survey of environmental policies and procedures in Melbourne hospitals.

This study aimed to review the presence of policies for management of behavioural symptoms and physical restraints, the availability of delirium management protocols and educational programs, and accessibility of a physical environment appropriate for the management of delirium in Melbourne hospitals. A structured survey tool was developed, and 70 Melbourne hospitals were surveyed seeking responses from a senior member of the nursing staff. Overall, 90% of Melbourne hospitals responded to the survey. It was found that smaller hospitals have fewer policies relating to the management of behavioural symptoms, and fewer delirium management protocols. Some education is available for nursing staff; however, less for the night staff, who often manage behavioural symptoms associated with delirium. Physical restraint policies exist at most hospitals. Single rooms and night lights are generally available, but low-low beds and orientation devices are relatively uncommon.

The minimum capture proportion for reliable estimation in capture-recapture models.

We derive estimates of the minimum capture proportion required to obtain a reliable estimate of the population size for several continuous and discrete-time capture-recapture models. The models considered are M(0), M(t), M(b), M(h), M(ht), and M(tb) in the notation of Otis et al., (1978, Wildlife Monograph62, 1-135). Numerical results with simulation studies are given, and two real examples for the model M(h) are also considered. Potential applications of these results are suggested.

A unified likelihood-based approach for estimating population size in continuous-time capture-recapture experiments with frailty.

A unified likelihood-based approach is proposed to estimate population size for a continuous-time closed capture-recapture experiment with frailty. The frailty model allows the capture intensity to vary with individual heterogeneity, time, and behavioral response. The individual heterogeneity effect is modeled as being gamma distributed. The first-capture and recapture intensities are assumed to be in constant proportion but may otherwise vary arbitrarily through time. The approach is also extended to capture-recapture experiments with possible random removals. Simulation studies are conducted to examine the performance of the proposed estimators. By asymptotic efficiency comparison and simulation studies, the proposed estimators have been shown to be superior to their discrete-time model counterparts in genuine continuous-time capture-recapture experiments.

Estimation of vaccine efficacy and the vaccination threshold.

This paper considers the effect of imperfect vaccination in a susceptible-infected-removal (SIR) epidemic model. The minimum proportion of the population that needs to be vaccinated to prevent a major epidemic depends on the vaccine efficacy and the basic reproductive rate for the SIR model, allowing for imperfect and variable vaccination. Martingale theory is used to derive estimates and associated standard errors for these parameters. Asymptotic properties of the resulting estimators are investigated. Data for a mumps outbreak are used as an illustrative example.