Impact of the COVID-19 pandemic on the real-world diagnostic infrastructure for tuberculosis-An ESGMYC collaborative study

We determined the impact of the COVID-19 pandemic on mycobacterial diagnostic services. 40 laboratories from 22 countries completed an online questionnaire covering the redeployment of the laboratory infrastructure and/or staff for SARS-CoV-2 testing, staff shortages and supply chain disruptions. 28 laboratories reported monthly numbers of samples processed for mycobacterial investigations and monthly numbers of M. tuberculosis complex (MTBC) PCRs performed between October 1st 2018 and October 31st 2020. More than half (23/40) of the participating TB laboratories reported having performed COVID-19 diagnostics in the early phase of the pandemic, in part with negative impact on the mycobacterial service activities. All participating laboratories reported shortages of consumables and laboratory equipment due to supply chain issues. Average monthly sample numbers decreased by 24% between January 2020 and October 2020 compared to pre-pandemic averages. At the end of the study period, most participating laboratories had not returned to pre-pandemic average MTBC PCR throughput

Robust estimation of survival and contribution of captive-bred Mallards Anas platyrhynchos to a wild population in a large-scale release programme

International audienceThe survival of captive-bred individuals from release into the wild to their first breeding season is crucial to assess the success of reintroduction or translocation programmes, and to assess their potential impact of wild populations. However, assessing the survival of captive-bred individuals following their release is often complicated by immediate dispersal once in the wild. Here, we apply Lindberg's robust design model, a method that incorporates emigration from the study site, to obtain true estimates of survival of captive-bred Mallards Anas platyrhynchos, a common duck species released on a large scale in Europe since the 1970s. Overall survival rate from release in July until the onset of the next breeding season in April was low (0.18 ± 0.07 se) and equivalent to half the first-year survival of local wild Mallards. Higher overall detectability and temporary emigration during the hunting period revealed movements in response to hunting pressure. Such low survival of released Mallards during their first year may help prevent large-scale genetic mixing with the wild population. Nevertheless, by combining our results with regional waterfowl counts, we estimated that a minimum of 34% of the Mallards in the region were of captive origin at the onset of the breeding season. Although most released birds quickly die, restocking for hunting may be of sufficient magnitude to affect the wild population through genetic homogenization or loss of local adaptation. Robust design protocols allow for the estimation of true survival estimates by controlling for permanent and temporary emigration and may require only a moderate increase in fieldwork effort

Contribution of released captive-bred Mallards to the dynamics of the natural population

The consequences of releasing captive-bred game animals into the wild have received little attention, despite their potential demographic impact, as well as costs and/or benefits for recipient populations. If restocking aims at increasing harvest opportunities, increased hunting pressure is expected, which would then be supported by either wild or released individuals. On the other hand, the wild recipient population may benefit from the release of captive-bred conspecifics if this reduces hunting pressure on the former through dilution of risk or selective harvesting of captive-bred individuals. Here, we modelled a Mallard (Anas platyrhynchos) population consisting of wild individuals supplemented by captive-bred conspecifics, a very common practice in Europe over the last 40 years. The objective was to test the effect of an increase of harvest rate on released and wild individuals, respectively. Our results show that, due to the low reproductive value of the released Mal-lards, the population was hardly affected by a change in harvest of these low performance individuals. Conversely, a 15 percent increase in harvest rate of the wild individuals would lead to a quick decline of the population. We discuss these results in the context of the Camargue population, located in the South of France, which has experienced an increase in Mallard harvest without apparent reduction of population size. We suggest that this has only been possible due to the release of captive-bred Mallards

MEG8 regulates Tissue Factor Pathway Inhibitor 2 (TFPI2) expression in the endothelium

A large portion of the genome is transcribed into non-coding RNA, which does not encode protein. Many long non-coding RNAs (lncRNAs) have been shown to be involved in important regulatory processes such as genomic imprinting and chromatin modification. The 14q32 locus contains many non-coding RNAs such as Maternally Expressed Gene 8 (MEG8). We observed an induction of this gene in ischemic heart disease. We investigated the role of MEG8 specifically in endothelial function as well as the underlying mechanism. We hypothesized that MEG8 plays an important role in cardiovascular disease via epigenetic regulation of gene expression. Experiments were performed in human umbilical vein endothelial cells (HUVECs). In vitro silencing of MEG8 resulted in impaired angiogenic sprouting. More specifically, total sprout length was reduced as was proliferation, while migration was unaffected. We performed RNA sequencing to assess changes in gene expression after loss of MEG8. The most profoundly regulated gene, Tissue Factor Pathway Inhibitor 2 (TFPI2), was fivefold increased following MEG8 silencing. TFPI2 has previously been described as an inhibitor of angiogenesis. Mechanistically, MEG8 silencing resulted in a reduction of the inhibitory histone modification H3K27me3 at the TFPI2 promoter. Interestingly, additional silencing of TFPI2 partially restored angiogenic sprouting capacity but did not affect proliferation of MEG8 silenced cells. In conclusion, silencing of MEG8 impairs endothelial function, suggesting a potential beneficial role in maintaining cell viability. Our study highlights the MEG8/TFPI2 axis as potential therapeutic approach to improve angiogenesis following ischemia

MEG8 regulates Tissue Factor Pathway Inhibitor 2 (TFPI2) expression in the endothelium

A large portion of the genome is transcribed into non-coding RNA, which does not encode protein. Many long non-coding RNAs (lncRNAs) have been shown to be involved in important regulatory processes such as genomic imprinting and chromatin modification. The 14q32 locus contains many non-coding RNAs such as Maternally Expressed Gene 8 (MEG8). We observed an induction of this gene in ischemic heart disease. We investigated the role of MEG8 specifically in endothelial function as well as the underlying mechanism. We hypothesized that MEG8 plays an important role in cardiovascular disease via epigenetic regulation of gene expression. Experiments were performed in human umbilical vein endothelial cells (HUVECs). In vitro silencing of MEG8 resulted in impaired angiogenic sprouting. More specifically, total sprout length was reduced as was proliferation, while migration was unaffected. We performed RNA sequencing to assess changes in gene expression after loss of MEG8. The most profoundly regulated gene, Tissue Factor Pathway Inhibitor 2 (TFPI2), was fivefold increased following MEG8 silencing. TFPI2 has previously been described as an inhibitor of angiogenesis. Mechanistically, MEG8 silencing resulted in a reduction of the inhibitory histone modification H3K27me3 at the TFPI2 promoter. Interestingly, additional silencing of TFPI2 partially restored angiogenic sprouting capacity but did not affect proliferation of MEG8 silenced cells. In conclusion, silencing of MEG8 impairs endothelial function, suggesting a potential beneficial role in maintaining cell viability. Our study highlights the MEG8/TFPI2 axis as potential therapeutic approach to improve angiogenesis following ischemia

Contribution of released captive-bred Mallards to the dynamics of natural populations

International audienceThe consequences of releasing captive-bred game animals into the wild have received little attention, despite their potential demographic impact, as well as costs and/or benefits for recipient populations. If restocking aims at increasing harvest opportunities, increased hunting pressure is expected, which would then be supported by either wild or released individuals. On the other hand, the wild recipient population may benefit from the release of captive-bred conspecifics if this reduces hunting pressure on the former through dilution of risk or selective harvesting of captive-bred individuals. Here, we modelled a Mallard (Anas platyrhynchos) population consisting of wild individuals supplemented by captive-bred conspecifics, a very common practice in Europe over the last 40 years. The objective was to test the effect of an increase of harvest rate on released and wild individuals, respectively. Our results show that, due to the low reproductive value of the released Mal-lards, the population was hardly affected by a change in harvest of these low performance individuals. Conversely, a 15 percent increase in harvest rate of the wild individuals would lead to a quick decline of the population. We discuss these results in the context of the Camargue population, located in the South of France, which has experienced an increase in Mallard harvest without apparent reduction of population size. We suggest that this has only been possible due to the release of captive-bred Mallards

Long non-coding RNA MEG8 induces endothelial barrier through regulation of microRNA-370 and -494 processing

The 14q32 locus is an imprinted region in the human genome which contains multiple noncoding RNAs. We investigated the role of Maternally Expressed Gene 8 (MEG8) in endothelial function and the underlying mechanism. A 5-fold increase in MEG8 was observed with increased passage number in Human Umbilical Vein Endothelial Cells, suggesting MEG8 is induced during aging. MEG8 knockdown resulted in a 1.8-fold increase in senescence, suggesting MEG8 might be protective during aging. Endothelial barrier was impaired after MEG8 silencing. MEG8 knockdown resulted in reduced expression of miRNA-370 and -494 but not -127, -487b and -410. Overexpression of miRNA-370/-494 partially rescued MEG8-silencing-induced barrier loss. Mechanistically, MEG8 regulates expression of miRNA-370 and -494 at the mature miRNA level through interaction with RNA binding proteins Cold Inducible RNA Binding Protein (CIRBP) and Hydroxyacyl-CoA Dehydrogenase Trifunctional Multi-enzyme Complex Subunit Beta (HADHB). Precursor and mature miRNA-370/-494 were shown to interact with HADHB and CIRBP respectively. CIRBP/HADHB silencing resulted in downregulation of miRNA-370 and induction of miRNA-494. These results suggest MEG8 interacts with CIRBP and HADHB and contributes to miRNA processing at the post-transcriptional level

The potential role of drug transporters and amikacin modifying enzymes in M. avium

ABSTRACT: Objectives: Mycobacterium avium (M. avium) complex bacteria cause opportunistic infections in humans. Treatment yields cure rates of 60% and consists of a macrolide, a rifamycin, and ethambutol, and in severe cases, amikacin. Mechanisms of antibiotic tolerance remain mostly unknown. Therefore, we studied the contribution of efflux and amikacin modification to antibiotic susceptibility. Methods: We characterised M. avium ABC transporters and studied their expression together with other transporters following exposure to clarithromycin, amikacin, ethambutol, and rifampicin. We determined the effect of combining the efflux pump inhibitors berberine, verapamil and CCCP (carbonyl cyanide m-chlorophenyl hydrazone), to study the role of efflux on susceptibility. Finally, we studied the modification of amikacin by M. avium using metabolomic analysis. Results: Clustering shows conservation between M. avium and M. tuberculosis and transporters from most bacterial subfamilies (2–6, 7a/b, 10–12) were found. The largest number of transporter encoding genes was up-regulated after clarithromycin exposure, and the least following amikacin exposure. Only berberine increased the susceptibility to clarithromycin. Finally, because of the limited effect of amikacin on transporter expression, we studied amikacin modification and showed that M. avium, in contrast to M. abscessus, is not able to modify amikacin. Conclusion: We show that M. avium carries ABC transporters from all major families important for antibiotic efflux, including homologues shown to have affinity for drugs included in treatment. Efflux inhibition in M. avium can increase susceptibility, but this effect is efflux pump inhibitor– and antibiotic-specific. Finally, the lack of amikacin modifying activity in M. avium is important for its activity