Mitochondrial dysfunction in sepsis:identifying mechanisms and novel therapies

Sepsis is a life-threatening condition with an in-hospital mortality rate of up to 30%. The global incidence of sepsis is estimated to be approximately 50 million cases. Sepsis is on the rise due to increased antibiotic resistance, lifespans, chronic diseases, the use of immunosuppressive therapy, chemotherapy, and invasive procedures (e.g., mechanical ventilation and dialyses). Currently, the treatment options for sepsis are limited to antibiotics and supportive care, with no targeted treatment available. Furthermore, survivors of sepsis face a significant risk of cardiovascular events, with a 3-year mortality rate as high as 47% after hospital discharge. This places sepsis as a major burden on healthcare and a significant global cause of death.This thesis is shedding light on new mechanisms behind sepsis and potential therapeutics, with a specific focus on mitochondrial dysfunction in sepsis. Mitochondria are the powerhouses of cells responsible for the body's energy production and organ function. This research revealed increased mitochondrial damage and damage markers (oxidative stress) in patients with sepsis and acute kidney injury, which is linked to sepsis severity and long-term mortality. Further, this research showed endothelial cells, lining the blood vessels, involved in the development of acute kidney injury in sepsis. Furthermore, a direct interaction between bacterial endotoxin and mitochondria in endothelial cells and kidney was demonstrated, inducing mitochondrial dysfunction. Therapeutic interventions showed promising results to preserve mitochondria using hydrogen sulfide (H2S) donors and SUL138 in pre-clinical sepsis models. This research revealed mitochondrial therapeutic targets to develop new therapies in patients with sepsis to improve outcome

Ancient DNA reveals the chronology of walrus ivory trade from Norse Greenland.

The importance of the Atlantic walrus ivory trade for the colonization, peak, and collapse of the medieval Norse colonies on Greenland has been extensively debated. Nevertheless, no studies have directly traced medieval European ivory back to distinct Arctic populations of walrus. Analysing the entire mitogenomes of 37 archaeological specimens from Europe, Svalbard, and Greenland, we here discover that Atlantic walrus comprises two monophyletic mitochondrial (MT) clades, which diverged between 23 400 and 251 120 years ago. Our improved genomic resolution allows us to reinterpret the geographical distribution of partial MT data from 306 modern and nineteenth-century specimens, finding that one of these clades was exclusively accessible to Greenlanders. With this discovery, we ascertain the biological origin of 23 archaeological specimens from Europe (most dated between 900 and 1400 CE). These results reveal a significant shift in trade from an early, predominantly eastern source towards a near exclusive representation of Greenland ivory. Our study provides empirical evidence for how this remote Arctic resource was progressively integrated into a medieval pan-European trade network, contributing to both the resilience and vulnerability of Norse Greenland society.This work was supported by the Nansenfondet, the Research Council of Norway projects 262777 and 230821, and the Leverhulme Trust project ‘Northern Journeys’ MRF-2013-065

Forest edges and other semi-natural habitat edges increase wild bee species richness and habitat connectivity in intensively managed temperate landscapes

Pollinator conservation schemes are typically focused on conserving existing-, restoring degraded- or establishing new wild bee habitats. The effectiveness of such conservation schemes depends on the presence of dispersal corridors that allow habitat colonization by bees. Nonetheless, we lack an understanding of the role of semi-natural habitats edges on the connectivity of pollinator communities across intensively managed landscapes. Here, we use data from wild bee communities comprising 953 occurrences from 79 species of non-parasitic bees, sampled at 68 locations distributed across a Norwegian and a Danish landscape to show that the proportion of semi-natural habitat edges is positively correlated to bee species richness and habitat connectivity. Specifically, we found that wild bee species richness sampled along roadsides increased with the proportion of semi-natural habitat edges within1.5 km of the study sites and with local plant species richness. We combined maps showing the proportion of seminatural habitat edges with least cost path analysis to find the most likely dispersal route between our bee communities. We find that these least cost path lengths provide better models of bee species compositional similarity than geographic distance (|ΔAICc| > 2), suggesting that seminatural habitat edges act as dispersal corridors in intensively managed landscapes. However, we also find that compositional similarity between communities depend on site-specific plant species richness stressing the importance of improving the habitat quality of edge habitats if they are to function as dispersal corridors. We discuss potential management options for improving wild bee habitat conditions along seminatural habitat edges and illustrate how maps of least cost paths can be used to identify dispersal corridors between pollinator habitats of conservation priority. Maps of dispersal corridors can be used to direct wild bee habitat management actions along seminatural habitat edges to facilitate the dispersal of bees between larger grassland habitats. Bees, Connectivity, Conservation planning, Pollinators, Restoration, GrasslandacceptedVersio

Sepsis is associated with mitochondrial DNA damage and a reduced mitochondrial mass in the kidney of patients with sepsis-AKI

BACKGROUND: Sepsis is a life-threatening condition accompanied by organ dysfunction subsequent to a dysregulated host response to infection. Up to 60% of patients with sepsis develop acute kidney injury (AKI), which is associated with a poor clinical outcome. The pathophysiology of sepsis-associated AKI (sepsis-AKI) remains incompletely understood, but mitochondria have emerged as key players in the pathogenesis. Therefore, our aim was to identify mitochondrial damage in patients with sepsis-AKI. METHODS: We conducted a clinical laboratory study using "warm" postmortem biopsies from sepsis-associated AKI patients from a university teaching hospital. Biopsies were taken from adult patients (n = 14) who died of sepsis with AKI at the intensive care unit (ICU) and control patients (n = 12) undergoing tumor nephrectomy. To define the mechanisms of the mitochondrial contribution to the pathogenesis of sepsis-AKI, we explored mRNA and DNA expression of mitochondrial quality mechanism pathways, DNA oxidation and mitochondrial DNA (mtDNA) integrity in renal biopsies from sepsis-AKI patients and control subjects. Next, we induced human umbilical vein endothelial cells (HUVECs) with lipopolysaccharide (LPS) for 48 h to mimic sepsis and validate our results in vitro. RESULTS: Compared to control subjects, sepsis-AKI patients had upregulated mRNA expression of oxidative damage markers, excess mitochondrial DNA damage and lower mitochondrial mass. Sepsis-AKI patients had lower mRNA expression of mitochondrial quality markers TFAM, PINK1 and PARKIN, but not of MFN2 and DRP1. Oxidative DNA damage was present in the cytosol of tubular epithelial cells in the kidney of sepsis-AKI patients, whereas it was almost absent in biopsies from control subjects. Oxidative DNA damage co-localized with both the nuclei and mitochondria. Accordingly, HUVECs induced with LPS for 48 h showed an increased mnSOD expression, a decreased TFAM expression and higher mtDNA damage levels. CONCLUSION: Sepsis-AKI induces mitochondrial DNA damage in the human kidney, without upregulation of mitochondrial quality control mechanisms, which likely resulted in a reduction in mitochondrial mass

GYY4137-Derived Hydrogen Sulfide Donates Electrons to the Mitochondrial Electron Transport Chain via Sulfide:Quinone Oxidoreductase in Endothelial Cells

The protective effects of hydrogen sulphide (H2S) to limit oxidative injury and preserve mitochondrial function during sepsis, ischemia/reperfusion, and neurodegenerative diseases have prompted the development of soluble H2S-releasing compounds such as GYY4137. Yet, the effects of GYY4137 on the mitochondrial function of endothelial cells remain unclear, while this cell type comprises the first target cell after parenteral administration. Here, we specifically assessed whether human endothelial cells possess a functional sulfide:quinone oxidoreductase (SQOR), to oxidise GYY4137-released H2S within the mitochondria for electron donation to the electron transport chain. We demonstrate that H2S administration increases oxygen consumption by human umbilical vein endothelial cells (HUVECs), which does not occur in the SQOR-deficient cell line SH-SY5Y. GYY4137 releases H2S in HUVECs in a dose- and time-dependent fashion as quantified by oxygen consumption and confirmed by lead acetate assay, as well as AzMC fluorescence. Scavenging of intracellular H2S using zinc confirmed intracellular and intramitochondrial sulfur, which resulted in mitotoxic zinc sulfide (ZnS) precipitates. Together, GYY4137 increases intramitochondrial H2S and boosts oxygen consumption of endothelial cells, which is likely governed via the oxidation of H2S by SQOR. This mechanism in endothelial cells may be instrumental in regulating H2S levels in blood and organs but can also be exploited to quantify H2S release by soluble donors such as GYY4137 in living systems.</p

An accurate assignment test for extremely low-coverage whole-genome sequence data.

Genomic assignment tests can provide important diagnostic biological characteristics, such as population of origin or ecotype. Yet, assignment tests often rely on moderate- to high-coverage sequence data that can be difficult to obtain for fields such as molecular ecology and ancient DNA. We have developed a novel approach that efficiently assigns biologically relevant information (i.e., population identity or structural variants such as inversions) in extremely low-coverage sequence data. First, we generate databases from existing reference data using a subset of diagnostic single nucleotide polymorphisms (SNPs) associated with a biological characteristic. Low-coverage alignment files are subsequently compared to these databases to ascertain allelic state, yielding a joint probability for each association. To assess the efficacy of this approach, we assigned haplotypes and population identity in Heliconius butterflies, Atlantic herring, and Atlantic cod using chromosomal inversion sites and whole-genome data. We scored both modern and ancient specimens, including the first whole-genome sequence data recovered from ancient Atlantic herring bones. The method accurately assigns biological characteristics, including population membership, using extremely low-coverage data (as low as 0.0001x) based on genome-wide SNPs. This approach will therefore increase the number of samples in evolutionary, ecological and archaeological research for which relevant biological information can be obtained

Hybridization of Atlantic puffins in the Arctic coincides with 20th-century climate change

The Arctic is experiencingthe fastest rates of globalwarming,leadingto shiftsin the distributionof its biotaandincreasingthe potentialfor hybridization. However, genomicevidenceof recenthybridization events in theArctic remainsunexpectedlyrare. Here, we use whole-genomesequencingof contemporary and 122-year-oldhistoricalspecimensto investigate the originof an Arctic hybridpopulation of Atlanticpuffins(Fr aterculaarctica)on Bjørnøya, Norway. We show that the hybridization between the High Arctic, large-bodiedsubspeciesF. a. naumanniand the temperate, smaller-sizedsubspeciesF. a. arcticabeganas recentlyas six generationsagodue to an unexpectedsouthward rangeexpansionofF. a. naumanni.Moreover, we find a significanttemporalloss of geneticdiversityacross Arctic and temperate puffinpopulations.Our observationsprovide compellinggenomicevidenceof the impacts of recentdistributionalshiftsand loss of diversityin Arctic communitiesduringthe 20th century.publishedVersio

A high urea-to-creatinine ratio predicts long-term mortality independent of acute kidney injury among patients hospitalized with an infection

Acute kidney injury (AKI) occurs frequently in patients with sepsis. Persistent AKI is, in contrast to transient AKI, associated with reduced long-term survival after sepsis, while the effect of AKI on survival after non-septic infections remains unknown. As prerenal azotaemia is a common cause of transient AKI that might be identified by an increased urea-to-creatinine ratio, we hypothesized that the urea-to-creatinine ratio may predict the course of AKI with relevance to long-term mortality risk. We studied the association between the urea-to-creatinine ratio, AKI and long-term mortality among 665 patients presented with an infection to the ED with known pre-existent renal function. Long-term survival was reduced in patients with persistent AKI. The urea-to-creatinine ratio was not associated with the incidence of either transient or non-recovered AKI. In contrast, stratification according to the urea-to-creatinine-ratio identifies a group of patients with a similar long-term mortality risk as patients with persistent AKI. Non-recovered AKI is strongly associated with all-cause long-term mortality after hospitalization for an infection. The urea-to-creatinine ratio should not be employed to predict prerenal azotaemia, but identifies a group of patients that is at increased risk for long-term mortality after infections, independent of AKI and sepsis

Combining bleach and mild predigestion improves ancient DNA recovery from bones.

The feasibility of genome-scale studies from archaeological material remains critically dependent on the ability to access endogenous, authentic DNA. In the majority of cases, this represents a few per cent of the DNA extract, at most. A number of specific pre-extraction protocols for bone powder aimed to improve ancient DNA recovery before library amplification have recently been developed. Here, we test the effects of combining two of such protocols, a bleach wash and a predigestion step, on 12 bone samples of Atlantic cod and domestic horse aged 750-1350 cal. years before present. Using high-throughput sequencing, we show that combined together, bleach wash and predigestion consistently yield DNA libraries with higher endogenous content than either of these methods alone. Additionally, the molecular complexity of these libraries is improved and endogenous DNA templates show larger size distributions. Other library characteristics, such as DNA damage profiles or the composition of microbial communities, are little affected by the pre-extraction protocols. Application of the combined protocol presented in this study will facilitate the genetic analysis of an increasing number of ancient remains and will reduce the cost of whole-genome sequencing

Association between oxidized nucleobases and mitochondrial DNA damage with long-term mortality in patients with sepsis

BACKGROUND: Sepsis not only leads to short-term mortality during hospitalization, but is also associated with increased mortality during long-term follow-up after hospital discharge. Metabolic stress during sepsis may cause oxidative damage to both nuclear and mitochondrial DNA (mtDNA) and RNA, which could affect long-term health and life span. Therefore, the aim of this study was to assess the association of sepsis with oxidized nucleobases and (mt)DNA damage and long-term all-cause mortality in septic patients. METHODS: 91 patients with sepsis who visited the emergency department (ED) of the University Medical Center Groningen between August 2012 and June 2013 were included. Urine and plasma were collected during the ED visit. Septic patients were matched with 91 healthy controls. Death rate was obtained until June 2020.The degree of oxidation of DNA, RNA and free nucleobases were assessed in urine by mass-spectrometry. Lipid peroxidation was assessed in plasma using a TBAR assay. Additionally, plasma levels of mtDNA and damage to mtDNA were determined by qPCR. RESULTS: Sepsis patients denoted higher levels of oxidated DNA, RNA, free nucleobases and lipid peroxidation than controls (all p < 0.01). Further, sepsis patients displayed an increase in plasma mtDNA with an increase in mtDNA damage compared to matched controls (p < 0.01). Kaplan meier survival analyses revealed that high degrees of RNA- and nucleobase oxidation were associated with higher long-term all-cause mortality after sepsis (p < 0.01 and p = 0.01 respectively). Of these two, high RNA oxidation was associated with long-term all-cause mortality, independent of adjustment for age, medical history and sepsis severity (HR 1.29 [(1.17-1.41, 95% CI] p < 0.01). CONCLUSIONS: Sepsis is accompanied with oxidation of nuclear and mitochondrial DNA and RNA, where RNA oxidation is an independent predictor of long-term all-cause mortality. In addition, sepsis causes mtDNA damage and an increase in cell free mtDNA in plasma