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Plant productivity varies due to environmental heterogeneity, and theory suggests that plant diversity can reduce this variation. While there is strong evidence of diversity effects on temporal variability of productivity, whether this mechanism extends to variability across space remains elusive. Here we determine the relationship between plant diversity and spatial variability of productivity in 83 grasslands, and quantify the effect of experimentally increased spatial heterogeneity in environmental conditions on this relationship. We found that communities with higher plant species richness (alpha and gamma diversity) have lower spatial variability of productivity as reduced abundance of some species can be compensated for by increased abundance of other species. In contrast, high species dissimilarity among local communities (beta diversity) is positively associated with spatial variability of productivity, suggesting that changes in species composition can scale up to affect productivity. Experimentally increased spatial environmental heterogeneity weakens the effect of plant alpha and gamma diversity, and reveals that beta diversity can simultaneously decrease and increase spatial variability of productivity. Our findings unveil the generality of the diversity-stability theory across space, and suggest that reduced local diversity and biotic homogenization can affect the spatial reliability of key ecosystem functions.

Loss of fatty acid β-oxidation (FAO) in the proximal tubule is a critical mediator of acute kidney injury and eventual fibrosis. However, transcriptional mediators of FAO in proximal tubule injury remain understudied. Krüppel-like factor 15 (KLF15), a highly enriched zinc-finger transcription factor in the proximal tubule, was significantly reduced in proximal tubule cells after aristolochic acid I (AAI) treatment, a proximal tubule-specific injury model. Proximal tubule specific knockout of Klf15 exacerbated proximal tubule injury and kidney function decline compared to control mice during the active phase of AAI treatment, and after ischemia-reperfusion injury. Furthermore, along with worsening proximal tubule injury and kidney function decline, knockout mice exhibited increased kidney fibrosis as compared to control mice during the remodeling phase after AAI treatment. RNA-sequencing of kidney cortex demonstrated increased transcripts involved in immune system and integrin signaling pathways and decreased transcripts encompassing metabolic pathways, specifically FAO, and PPARα signaling, in knockout versus control mice after AAI treatment. In silico and experimental chromatin immunoprecipitation studies collectively demonstrated that KLF15 occupied the promoter region of key FAO genes, CPT1A and ACAA2, in close proximity to transcription factor PPARα binding sites. While the loss of Klf15 reduced the expression of Cpt1a and Acaa2 and led to compromised FAO, induction of KLF15 partially rescued loss of FAO in AAI-treated cells. Klf15, Ppara, Cpt1a, and Acaa2 expression was also decreased in other mouse kidney injury models. Tubulointerstitial KLF15 independently correlated with eGFR, PPARA and CPT1A appearance in expression arrays from human kidney biopsies. Thus, proximal tubule-specific loss of Klf15 exacerbates acute kidney injury and fibrosis, likely due to loss of interaction with PPARα leading to loss of FAO gene transcription.

Heat and drought affect plant chemical defenses and thereby plant susceptibility to pests and pathogens. Monoterpenes are of particular importance for conifers as they play critical roles in defense against bark beetles. To date, work seeking to understand the impacts of heat and drought on monoterpenes has primarily focused on young potted seedlings, leaving it unclear how older age classes that are more vulnerable to bark beetles might respond to stress. Furthermore, we lack a clear picture of what carbon resources might be prioritized to support monoterpene synthesis under drought stress. To address this, we measured needle and woody tissue monoterpene concentrations and physiological variables simultaneously from mature piñon pines (Pinus edulis) from a unique temperature and drought manipulation field experiment. While heat had no effect on total monoterpene concentrations, trees under combined heat and drought stress exhibited ~ 85% and 35% increases in needle and woody tissue, respectively, over multiple years. Plant physiological variables like maximum photosynthesis each explained less than 10% of the variation in total monoterpenes for both tissue types while starch and glucose + fructose measured 1-month prior explained ~ 45% and 60% of the variation in woody tissue total monoterpene concentrations. Although total monoterpenes increased under combined stress, some key monoterpenes with known roles in bark beetle ecology decreased. These shifts may make trees more favorable for bark beetle attack rather than well defended, which one might conclude if only considering total monoterpene concentrations. Our results point to cumulative and synergistic effects of heat and drought that may reprioritize carbon allocation of specific non-structural carbohydrates toward defense. Supplementary Information The online version contains supplementary material available at 10.1007/s00442-021-05058-8.

Inflammatory factors have been considered to contribute to keratoconus (KC) pathogenesis. This study aims to determine the immune cells subsets and soluble inflammatory factor profile on the ocular surface of KC patients. 32 KC subjects (51 eyes) across different grades of severity and 15 healthy controls (23 eyes) were included in the study. Keratometry and pachymetry measurements were recorded. Ocular surface immune cells (collected by ocular surface wash) immunophenotyped using flow cytometry include leukocytes, neutrophils, macrophages, natural killer (NK) cells, pan-T cells, gamma delta T (gamma delta T) cells and NKT cells. Tear fluid collected using Schirmer's strip was used to measure 50 soluble factors by multiplex ELISA. Proportions of activated neutrophils, NK cells and gamma delta T cells were significantly increased in KC patients. Significantly higher levels of tear fluid IL-1 beta, IL-6, LIF, IL-17A, TNF alpha, IFN alpha/beta/gamma, EPO, TGF beta 1, PDGF-BB, sVCAM, sL-selectin, granzyme-B, perforin, MMP2, sFasL and IgE, along with significantly lower levels of IL-1 alpha and IL-9 were observed in KC patients. Alterations observed in few of the immuno-inflammatory parameters correlated with grades of disease, allergy, eye rubbing and keratometry or pachymetry measurements. The observation implies a distinct immuno-inflammatory component in KC pathogenesis and its potential as an additional therapeutic target in KC management.

Context Invasive species are a major cause of biodiversity loss across much of the world, and a key threat to Australia’s diverse reptile fauna. There has been no previous comprehensive analysis of the potential impact of the introduced European red fox, Vulpes vulpes, on Australian reptiles. Aims We seek to provide an inventory of all Australian reptile species known to be consumed by the fox, and identify characteristics of squamate species associated with such predation. We also compare these tallies and characteristics with reptile species known to be consumed by the domestic cat, Felis catus, to examine whether predation by these two introduced species is compounded (i.e. affecting much the same set of species) or complementary (affecting different groups of species). Methods We collated records of Australian reptiles consumed by foxes in Australia, with most records deriving from fox dietary studies (tallying >35 000 samples). We modelled presence or absence of fox predation records against a set of biological and other traits, and population trends, for squamate species. Key results In total, 108 reptile species (~11% of Australia’s terrestrial reptile fauna) have been recorded as consumed by foxes, fewer than that reported for cats (263 species). Eighty-six species have been reported to be eaten by both predators. More Australian turtle species have been reported as consumed by foxes than by cats, including many that suffer high levels of predation on egg clutches. Twenty threatened reptile species have been reported as consumed by foxes, and 15 by cats. Squamate species consumed by foxes are more likely to be undergoing population decline than those not known to be consumed by foxes. The likelihood of predation by foxes increased with squamate species’ adult body mass, in contrast to the relationship for predation by cats, which peaked at ~217 g. Foxes, but not cats, were also less likely to consume venomous snakes. Conclusions The two introduced, and now widespread, predators have both compounding and complementary impacts on the Australian reptile fauna. Implications Enhanced and integrated management of the two introduced predators is likely to provide substantial conservation benefits to much of the Australian reptile fauna.

Background Plant-derived cysteine proteinases of the papain family (CPs) attack nematodes by digesting the cuticle, leading to rupture and death of the worm. The nematode cuticle is composed of collagens and cuticlins, but the specific molecular target(s) for the proteinases have yet to be identified. Methods This study followed the course of nematode cuticle disruption using immunohistochemistry, scanning electron microscopy and proteomics, using a free-living nematode, Caenorhabditis elegans and the murine GI nematode Heligmosomoides bakeri (H. polygyrus) as target organisms. Results Immunohistochemistry indicated that DPY-7 collagen is a target for CPs on the cuticle of C. elegans. The time course of loss of DPY-7 from the cuticle allowed us to use it to visualise the process of cuticle disruption. There was a marked difference in the time course of damage to the cuticles of the two species of nematode, with H. bakeri being more rapidly hydrolysed. In general, the CPs’ mode of attack on the nematode cuticle was by degrading the structural proteins, leading to loss of integrity of the cuticle, and finally death of the nematode. Proteomic analysis failed conclusively to identify structural targets for CPs, but preliminary data suggested that COL-87 and CUT-19 may be important targets for the CPs, the digestion of which may contribute to cuticle disruption and death of the worm. Cuticle globin was also identified as a cuticular target. The presence of more than one target protein may slow the development of resistance against this new class of anthelmintic. Conclusions Scanning electron microscopy and immunohistochemistry allowed the process of disruption of the cuticle to be followed with time. Cuticle collagens and cuticlins are molecular targets for plant cysteine proteinases. However, the presence of tyrosine cross-links in nematode cuticle proteins seriously impeded protein identification by proteomic analyses. Multiple cuticle targets exist, probably making resistance to this new anthelmintic slow to develop. Graphical Abstract

The red fox Vulpes vulpes is one of the world’s most widespread carnivores. A key to its success has been its broad, opportunistic diet. The fox was introduced to Australia about 150 years ago, and within 30 years of its introduction was already recognised as a threat to livestock and native wildlife. We reviewed 85 fox diet studies (totalling 31693 samples) from throughout the species’ geographic range within Australia. Mammals were a major component of fox diet, being present in 70 ± 19% of samples across n = 160 locations. Invertebrates (38 ± 26% n = 130) and plant material (26 ± 25% n = 123) were also both staple foods and often the dominant food category recorded. Birds (13 ± 11% n = 137) and reptiles (10 ± 15% n = 132) were also commonly reported, while frogs were scarcely represented (1.6 ± 3.6% n = 111) in fox diet studies. Biogeographical differences reveal factors that likely determine prey availability. Diet composition varied with ecosystem, level of vegetation clearing and condition, and climate zone. Sample type (i.e. stomach versus scat samples) also significantly influenced reporting of diet composition. Livestock and frogs were underrepresented in records based on analysis of scats, whereas small mammals (native rodents, dasyurid marsupials, and bats) were more likely to be recorded in studies of scats than in studies of stomach contents. Diet varied seasonally, reflecting activity patterns of prey species and food availability. This synthesis also captures temporal shifts in fox diet over 70 years (1951–2020), as foxes have switched to consuming more native species in the wake of successful broadscale biological control of the invasive European rabbit Oryctolagus cuniculus. Diet analyses, such as those summarised in this review, capture the evidence required to motivate for greater control of foxes in Australia. This synthesis also highlights the importance of integrated pest species management to meet biodiversity conservation outcomes.

Determining the factors that drive the distributions of threatened species is often critical for informing effective conservation management actions. Species distribution models can be used to distinguish common habitat features shared by limited historical records and identify other areas where a species might persist. In this study, we built a species distribution model for the Endangered and cryptic Kangaroo Island dunnart (Sminthopsis fuliginosus aitkeni). We fitted generalised linear models using incidental records and presence-absence data from surveys between 1969 and 2018. In the models we included the variables rainfall, percentage native vegetation in the surrounding 2 km2, and post-fire vegetation age. The modelling suggested that rainfall and to a lesser extent post-fire vegetation age are good predictors of dunnart occurrence, with dunnart occurrence greatest in areas of high rainfall (>600 mm) and vegetation age classes

Mg-protoporphyrin IX monomethyl ester (MgPME) cyclase catalyses the formation of the isocyclic ring, producing protochlorophyllide a and contributing substantially to the absorption properties of chlorophylls and bacteriochlorophylls. The O2-dependent cyclase is found in both oxygenic phototrophs and some purple bacteria. We overproduced the simplest form of the cyclase, AcsF, from Rubrivivax gelatinosus, in Escherichia coli. In biochemical assays the di-iron cluster within AcsF is reduced by ferredoxin furnished by NADPH and ferredoxin:NADP+ reductase, or by direct coupling to Photosystem I photochemistry, linking cyclase to the photosynthetic electron transport chain. Kinetic analyses yielded a turnover number of 0.9 min−1, a Michaelis–Menten constant of 7.0 µM for MgPME and a dissociation constant for MgPME of 0.16 µM. Mass spectrometry identified 131-hydroxy-MgPME and 131-keto-MgPME as cyclase reaction intermediates, revealing the steps that form the isocyclic ring and completing the work originated by Sam Granick in 1950. Mg-protoporphyrin IX monomethyl ester cyclase catalyses the formation of the isocyclic ring in the synthesis of chlorophyll. Kinetic analyses and mass spectrometry identified intermediates in the reaction.

Research on the African cheetah (Acinonyx jubatus) exhibits strong geographical biases, with most studies taking place within a few, well‐studied populations. Here, we provide the first insights into the status and distribution of a globally important cheetah population in the 50,000 km2 Ruaha‐Rungwa landscape in southern Tanzania. We employed data from four methods (systematic camera trap surveys, sign surveys, community camera trapping, and observations by photo‐tourism guides) to improve knowledge of this understudied population. Our findings indicate that cheetah are widespread across the landscape, although they appear to exist at relatively low population densities, likely primarily due to biotic factors. Our surveys revealed an extension of confirmed geographical range of the species and provide some of the first evidence that miombo woodlands may be an important habitat for cheetah across its eastern African range. We employ these findings to identify research priorities for the species elsewhere in the region. Community camera trapping revealed that cheetah are using unprotected areas, although rarely. Finally, we show that collaborations with tourism operators can be employed to monitor cheetah populations, but also identify limitations of this method. Our results have implications for conservation of the species both in southern Tanzania and across its African range.

Homeostatic signaling systems are fundamental forms of biological regulation that maintain stable functionality in a changing environment. In the nervous system, synapses are crucial substrates for homeostatic modulation, serving to establish, maintain, and modify the balance of excitation and inhibition. Synapses must be sufficiently flexible to enable the plasticity required for learning and memory but also endowed with the stability to last a lifetime. In response to the processes of development, growth, remodeling, aging, and disease that challenge synapses, latent forms of adaptive plasticity become activated to maintain synaptic stability. In recent years, new insights into the homeostatic control of synaptic function have been achieved using the powerful Drosophila neuromuscular junction (NMJ). This review will focus on work over the past 10 years that has illuminated the cellular and molecular mechanisms of five homeostats that operate at the fly NMJ. These homeostats adapt to loss of postsynaptic neurotransmitter receptor functionality, glutamate imbalance, axonal injury, as well as aberrant synaptic growth and target innervation. These diverse homeostats work independently yet can be simultaneously expressed to balance neurotransmission. Growing evidence from this model glutamatergic synapse suggests these ancient homeostatic signaling systems emerged early in evolution and are fundamental forms of plasticity that also function to stabilize mammalian cholinergic NMJs and glutamatergic central synapses.

Negative interactions with humans resulting from livestock predation is a major factor influencing the decline of African lion Panthera leo populations across Africa. Here we investigate lion depredation within two Maasai communities in southern Kenya where people and lions coexist in the absence of any formal protected areas. We explore the factors that increase the frequency and severity of lion attacks on pastoralists and their livestock and assess the effectiveness of livestock guarding to reduce damage. Finally, we examine in which circumstances lion depredation triggers retaliation by people. Over a period of 26 months, lions attacked livestock 29 times, resulting in 41 livestock deaths and 19 injuries. There were also two attacks on people. Lions preferred cattle over the more numerous sheep and goats. Attacks on livestock occurred mostly during the dry season and were not affected by changes in prey density or variation in pastoral settlement that brought livestock into closer proximity with lions. Livestock were guarded during 48.2% of lion attacks. Active guarding at pasture disrupted the majority of lion attacks, resulting in lower mortality rates. Passive guarding in corrals at night also disrupted attacks but did not lead to lower livestock mortality.

Significance Statement Developmental acclimation to light intensity in plants involves changes in the organisation and regulation of photosynthetic electron transfer chain components in the chloroplast thylakoid membrane. Here, using mass spectrometry, we quantified changes in the thylakoid proteome and correlated these with key photosynthetic parameters to gain insight into the process., Summary Photosynthetic acclimation, the ability to adjust the composition of the thylakoid membrane to optimise the efficiency of electron transfer to the prevailing light conditions, is crucial to plant fitness in the field. While much is known about photosynthetic acclimation in Arabidopsis, to date there has been no study that combines both quantitative label‐free proteomics and photosynthetic analysis by gas exchange, chlorophyll fluorescence and P700 absorption spectroscopy. Using these methods we investigated how the levels of 402 thylakoid proteins, including many regulatory proteins not previously quantified, varied upon long‐term (weeks) acclimation of Arabidopsis to low (LL), moderate (ML) and high (HL) growth light intensity and correlated these with key photosynthetic parameters. We show that changes in the relative abundance of cytb 6 f, ATP synthase, FNR2, TIC62 and PGR6 positively correlate with changes in estimated PSII electron transfer rate and CO2 assimilation. Improved photosynthetic capacity in HL grown plants is paralleled by increased cyclic electron transport, which positively correlated with NDH, PGRL1, FNR1, FNR2 and TIC62, although not PGR5 abundance. The photoprotective acclimation strategy was also contrasting, with LL plants favouring slowly reversible non‐photochemical quenching (qI), which positively correlated with LCNP, while HL plants favoured rapidly reversible quenching (qE), which positively correlated with PSBS. The long‐term adjustment of thylakoid membrane grana diameter positively correlated with LHCII levels, while grana stacking negatively correlated with CURT1 and RIQ protein abundance. The data provide insights into how Arabidopsis tunes photosynthetic electron transfer and its regulation during developmental acclimation to light intensity.

Ancestral dingoes arrived in Australia at some time, or times, during the Holocene, heralding a period of long and uneasy coexistence with the human inhabitants of the continent. For the first Australians, dingoes became a valued and integral part of the culture but also exacted diverse social and economic costs. For early Europeans and later arrivals, dingoes were alternately revered for their strength and intelligence or feared and reviled for their attacks on livestock. These disparate views have scarcely changed in 232 years; if anything, the schism in perspectives about the dingo has widened as more has been discovered about this divisive and still enigmatic animal. Here, we show that current arguments about the dingo have deep origins by tracing the history of debate about the taxon’s name, when dingoes arrived in Australia, whether they are native or introduced, the early effects of dingoes on native fauna, and their current impacts as a ‘biodiversity regulator’ and destroyer of livestock. We suggest that some debates concerning the dingo will be resolved when more evidence is gained or new discoveries are made, whereas other debates will progress only when proponents and protagonists are able to agree on a research agenda and on thresholds for interpretation of the results that the agenda produces. Such new evidence, and new collaborative thinking, should provide a more robust underpinning for when, where and how dingoes are conserved and managed in future.

Public opposition has shaped management of wild animals in Australia, but public interest in dingo control has been minimal. We hypothesised that this is due to lack of awareness of dingo management practices, in part because using the term “wild dogs” to describe management renders “dingoes” invisible, framing the issue as one of control of introduced pests rather than control of an iconic Australian animal. We distributed an online questionnaire survey to the Australian public (N = 811) to measure how the public perceived dingoes and their management, how these views compared with other animals managed as pests in Australia, and whether the term “wild dogs” has shaped views and knowledge of dingo management. Most respondents (84.6%) considered dingoes to be native to Australia and there was low approval of lethal control methods, except when justification was provided (e.g., to protect livestock or endangered native species). Only 19.1% were aware that “wild dog” management included dingoes, and attitudes towards “wild dogs” were more negative than those towards dingoes. If public awareness about dingo management increases, pressure from the public may result and shape future management actions, including restricting the use of lethal control practices like poison baiting on public lands. As such, public attitudes should be incorporated into decision-making, and appropriate communication strategies need to be employed to prevent backlash.

Pathogens and other adverse environmental conditions can trigger endoplasmic reticulum (ER) stress. ER stress signaling increases the expression of cytoprotective ER-chaperones. The inositol-requiring enzyme (IRE1) is one ER stress sensor that is activated to splice the bZIP60 mRNA that produces a truncated transcription factor that activates gene expression in the nucleus. The IRE1/bZIP60 pathway is associated with restricting potyvirus and potexvirus infection. This study shows that the Plantago asiatica mosaic virus (PlAMV) triple gene block 3 (TGB3) and the Turnip mosaic virus (TuMV) 6K2 proteins activate alternative transcription pathways involving the bZIP17, bZIP28, BAG7, NAC089 and NAC103 factors in Arabidopsis thaliana. Using the corresponding knockout mutant lines, we show that bZIP17, bZIP60, BAG7 and NAC089 are factors in reducing PlAMV infection, whereas bZIP28 and bZIP60 are factors in reducing TuMV infection. We propose a model in which bZIP60 and bZIP17 synergistically induce genes restricting PlAMV infection, while bZIP60 and bZIP28 independently induce genes supporting PlAMV infection. Regarding TuMV-green fluorescent protein (GFP) infection, bZIP60 and bZIP28 serve to repress local and systemic infection. Finally, tauroursodeoxycholic acid treatments were used to demonstrate that the protein folding capacity significantly influences PlAMV accumulation.

Recurrent upper tract urothelial carcinomas (UTUCs) arise in the context of nephropathy linked to exposure to the herbal carcinogen aristolochic acid (AA). Here we delineated the molecular programs underlying UTUC tumorigenesis in patients from endemic aristolochic acid nephropathy (AAN) regions in Southern Europe. We applied an integrative multiomics analysis of UTUCs, corresponding unaffected tissues and of patient urines. Quantitative microRNA (miRNA) and messenger ribonucleic acid (mRNA) expression profiling, immunohistochemical analysis by tissue microarrays and exome and transcriptome sequencing were performed in UTUC and nontumor tissues. Urinary miRNAs of cases undergoing surgery were profiled before and after tumor resection. Ribonucleic acid (RNA) and protein levels were analyzed using appropriate statistical tests and trend assessment. Dedicated bioinformatic tools were used for analysis of pathways, mutational signatures and result visualization. The results delineate UTUC-specific miRNA:mRNA networks comprising 89 miRNAs associated with 1, 862 target mRNAs, involving deregulation of cell cycle, deoxyribonucleic acid (DNA) damage response, DNA repair, bladder cancer, oncogenes, tumor suppressors, chromatin structure regulators and developmental signaling pathways. Key UTUC- specific transcripts were confirmed at the protein level. Exome and transcriptome sequencing of UTUCs revealed AA-specific mutational signature SBS22, with 68% to 76% AA-specific, deleterious mutations propagated at the transcript level, a possible basis for neoantigen formation and immunotherapy targeting. We next identified a signature of UTUC- specific miRNAs consistently more abundant in the patients' urine prior to tumor resection, thereby defining biomarkers of tumor presence. The complex gene regulation programs of AAN-associated UTUC tumors involve regulatory miRNAs prospectively applicable to noninvasive urine-based screening of AAN patients for cancer presence and recurrence.

The cornea is the clear window that lets light into the eye. It is composed of five layers: epithelium, Bowman’s layer, stroma, Descemet’s membrane and endothelium. The maintenance of its structure and transparency are determined by the functions of the different cell types populating each layer. Attempts to regenerate corneal tissue and understand disease conditions requires knowledge of how cell profiles vary across this heterogeneous tissue. We performed a single cell transcriptomic profiling of 19,472 cells isolated from eight healthy donor corneas. Our analysis delineates the heterogeneity of the corneal layers by identifying cell populations and revealing cell states that contribute in preserving corneal homeostasis. We identified expression of CAV1, HOMER3 and CPVL in the corneal epithelial limbal stem cell niche, CKS2, STMN1 and UBE2C were exclusively expressed in highly proliferative transit amplifying cells, CXCL14 was expressed exclusively in the suprabasal/superficial limbus, and NNMT was exclusively expressed by stromal keratocytes. Overall, this research provides a basis to improve current primary cell expansion protocols, for future profiling of corneal disease states, to help guide pluripotent stem cells into different corneal lineages, and to understand how engineered substrates affect corneal cells to improve regenerative therapies.

We combine a chemically-synthesized, voltage-sensitive fluorophore with a genetically encoded, self-labeling enzyme to enable voltage imaging in Drosophila melanogaster. Previously, we showed that a rhodamine voltage reporter (RhoVR) combined with the HaloTag self-labeling enzyme could be used to monitor membrane potential changes from mammalian neurons in culture and brain slice. Here, we apply this hybrid RhoVR-Halo approach in vivo to achieve selective neuron labeling in intact fly brains. We generate a Drosophila UAS-HaloTag reporter line in which the HaloTag enzyme is expressed on the surface of cells. We validate the voltage sensitivity of this new construct in cell culture before driving expression of HaloTag in specific brain neurons in flies. We show that selective labeling of synapses, cells, and brain regions can be achieved with RhoVR-Halo in either larval neuromuscular junction (NMJ) or in whole adult brains. Finally, we validate the voltage sensitivity of RhoVR-Halo in fly tissue via dual-electrode/imaging at the NMJ, show the efficacy of this approach for measuring synaptic excitatory post-synaptic potentials (EPSPs) in muscle cells, and perform voltage imaging of carbachol-evoked depolarization and osmolarity-evoked hyperpolarization in projection neurons and in interoceptive subesophageal zone neurons in fly brain explants following in vivo labeling. We envision the turn-on response to depolarizations, fast response kinetics, and two-photon compatibility of chemical indicators, coupled with the cellular and synaptic specificity of genetically-encoded enzymes, will make RhoVR-Halo a powerful complement to neurobiological imaging in Drosophila.

Mutations not only alter allele frequencies in a genetic pool but may also determine the fate of an evolutionary process. Here we study which allele fixes in a one-step, one-way model including the wild type and two adaptive mutations. We study the effect of the four basic evolutionary mechanisms---genetic drift, natural selection, mutation, and gene flow---on mutant fixation and its kinetics. Determining which allele is more likely to fix is not simply a question of comparing fitnesses and mutation rates. For instance, if the allele of interest is less fit than the other, then not only must it have a greater mutation rate, but also its mutation rate must exceed a specific threshold for it to prevail. We find exact expressions for such conditions. Our conclusions are based on the mathematical description of two extreme but important regimes, as well as on simulations.

Significance Disentangling the effects of rainfall timing and magnitude on animal and plant populations is essential to reveal the biological consequence of diverse climate change scenarios around the world. We conducted a 10-y, large-scale, manipulative experiment to examine the bottom-up effects of changes in rainfall regime on the population dynamics of Brandt’s voles in the steppe grassland of Inner Mongolia, China. We found that a moderate rainfall increase during the early growing season could produce marked increases in vole population size by increasing the biomass of preferred plant species, whereas large increases in rainfall produced no additional increase in vole population growth. Our study highlights the importance of rainfall magnitude and timing on the nonlinear population dynamics of herbivores., Climate change–induced shifts in species phenology differ widely across trophic levels, which may lead to consumer–resource mismatches with cascading population and ecosystem consequences. Here, we examined the effects of different rainfall patterns (i.e., timing and amount) on the phenological asynchrony of population of a generalist herbivore and their food sources in semiarid steppe grassland in Inner Mongolia. We conducted a 10-y (2010 to 2019) rainfall manipulation experiment in 12 0.48-ha field enclosures and found that moderate rainfall increases during the early rather than late growing season advanced the timing of peak reproduction and drove marked increases in population size through increasing the biomass of preferred plant species. By contrast, greatly increased rainfall produced no further increases in vole population growth due to the potential negative effect of the flooding of burrows. The increases in vole population size were more coupled with increased reproduction of overwintered voles and increased body mass of young-of-year than with better survival. Our results provide experimental evidence for the fitness consequences of phenological mismatches at the population level and highlight the importance of rainfall timing on the population dynamics of small herbivores in the steppe grassland environment.

Predators are fundamentally important in regulating many global ecosystems, and perturbations of predator populations through impacts by invasive species are frequently implicated in the degradation of these ecosystems. In considering recent declines of native critical weight range (35–5500 g) mammals in northern Australia, most attention has focused on predator–prey interactions between a mammalian invader, the feral cat (Felis catus), and these mammals. Little consideration has been given to the possible implications of changed reptilian predator assemblages resulting from invasion by another vertebrate, the toxic cane toad (Rhinella marina). We used reptile removal records from licenced reptile catchers in three widely spaced towns in the savannas of northern Australia to explore potential impacts of toads on apex and meso-predatory snakes and large lizards. In addition, simultaneous fauna survey data from one town with reptile removal records were used to identify cascading faunal impacts associated with toad invasion. Intervention analyses revealed empirical linkages between toad invasion, apex predator declines, meso-predator increases and declines of critical weight range mammals and other prey groups. Based on the timing and strength of intervention we postulate a novel hypothesis and supporting conceptual model linking recent mammal declines with trophic cascades following toad invasion. The conceptual model is discussed in relation to the prevailing feral cat focussed hypotheses regarding northern Australia’s dramatic small mammal declines. Future studies will need to test putative interactions and their importance so that appropriate management can be implemented to stem ongoing mammal losses.

Disturbances such as fire reduce the structural complexity of terrestrial habitats, increasing the risk of predation for small prey species. The postfire effect of predation has especially deleterious effects in Australian habitats owing to the presence of invasive mammalian predators, the red fox (Vulpes vulpes) and feral cat (Felis catus), that rapidly exploit burned habitats. Here, we investigated whether the provision of artificial shelter could alleviate the risk of predation perceived by two species of small marsupial, the dunnarts Sminthopsis hirtipes and S. youngsoni, in open postfire habitat in the sandridge system of the Simpson Desert, central Australia. We installed artificial shelters constructed from wire mesh that allowed passage of the dunnarts but not of their predators at one site, and measured and compared the perceived risk of predation by the dunnarts there with those on a control site using optimal patch-use theory (giving-up densities, GUDs). GUDs were lower near artificial shelters than away from them, and near dune crests where dunnarts typically forage, suggesting that the shelters acted as corridors for dunnarts to move up to the crests from burrows in the swales. Foraging was lower near the crest in the control plot. Two-day foraging bouts were observed in dunnart activity, with recruitment to GUD stations occurring a day earlier in the augmented shelter plot. Despite these results, the effects of the shelters were localized and not evident at the landscape scale, with GUDs reduced also in proximity to sparse natural cover in the form of regenerating spinifex grass hummocks. Mapping dunnart habitat use using the landscape of fear (LOF) framework confirmed that animals perceived safety near shelter and risk away from it. We concluded that the LOF framework can usefully assess real-time behavioral responses of animals to management interventions in situations where demographic responses take longer to occur.

Introduction: Suppression of gastric acid secretion with proton-pump inhibitors (PPI) is an integral part of the treatment of Helicobacter pylori infection. Esomeprazole has been shown to be superior to other PPIs when used in the context of triple therapy; however, comparative data for PPI efficacy in quadruple therapy are lacking. Current guidelines recommend H. pylori eradication with quadruple therapy in areas with high clarithromycin resistance. Objective: To determine whether esomeprazole is more effective than other PPIs in the context of quadruple therapy for H. pylori eradication. Methods: We retrospectively identified 25- to 60-year-old subjects with a positive 13C-urea breath test and no prior laboratory or endoscopic test for H. pylori infection. Pharmacy dispensation data were retrieved. Results: A total of 7,896 subjects including 2,856 (36.2%) males, aged 40.4 ± 10.6 years, were identified. Of those, 78.1% received omeprazole, 20.1% received lansoprazole, 1.5% received esomeprazole, and 0.34% received pantoprazole together with antibiotics for H. pylori eradication. Esomeprazole was associated with a greater proportion of successful eradication (85.0 vs. 77.5%, esomeprazole vs. omeprazole, OR 1.64; 95% CI 0.99–2.72; p = 0.05). A nonsignificant trend favored esomeprazole over omeprazole among subjects receiving quadruple therapy (90.0 vs. 82.0%, respectively, OR 1.98; 95% CI 0.68–5.72; p = 0.16). Independent predictors of treatment success included older age and quadruple therapy. Conclusion: Esomeprazole is more beneficial than other PPIs for H. pylori eradication. Studies with larger subgroups are necessary to confirm our findings among subjects receiving quadruple therapy.

The membrane-embedded FtsH proteases found in bacteria, chloroplasts, and mitochondria are involved in diverse cellular processes including protein quality control and regulation. The genome of the model cyanobacterium Synechocystis sp PCC 6803 encodes four FtsH homologs designated FtsH1 to FtsH4. The FtsH3 homolog is present in two hetero-oligomeric complexes: FtsH2/3, which is responsible for photosystem II quality control, and the essential FtsH1/3 complex, which helps maintain Fe homeostasis by regulating the level of the transcription factor Fur. To gain a more comprehensive insight into the physiological roles of FtsH hetero-complexes, we performed genome-wide expression profiling and global proteomic analyses of Synechocystis mutants conditionally depleted of FtsH3 or FtsH1 grown under various nutrient conditions. We show that the lack of FtsH1/3 leads to a drastic reduction in the transcriptional response to nutrient stress of not only Fur but also the Pho, NdhR, and NtcA regulons. In addition, this effect is accompanied by the accumulation of the respective transcription factors. Thus, the FtsH1/3 complex is of critical importance for acclimation to iron, phosphate, carbon, and nitrogen starvation in Synechocystis. Germany Federal Ministry of Education and Research [031L0106B] Grant Agency of the Czech RepublicGrant Agency of the Czech Republic [P501-12-G055] Czech Ministry of Education Ministry of Education, Youth & Sports - Czech Republic [LO1416] Portuguese Fundacao para a Ciencia e a Tecnologia (Foundation for Science and Technology) [PTDC/BIA-MIC/4418/2012, IF/00881/2013, UID/Multi/04326/2013] United Kingdom Biotechnology and Biological Sciences Research Council (BBSRC)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/M012166/1, BB/M000265/1] European Research CouncilEuropean Research Council (ERC) [338895]

Apex predators play a critical role in maintaining the health of ecosystems but are highly susceptible to habitat degradation and loss caused by land-use changes, and to anthropogenic mortality. The leopard Panthera pardus is the last free-roaming large carnivore in the Western Cape province, South Africa. During 2011–2015, we carried out a camera-trap survey across three regions covering c. 30,000 km2 of the Western Cape. Our survey comprised 151 camera sites sampling nearly 14,000 camera-trap nights, resulting in the identification of 71 individuals. We used two spatially explicit capture–recapture methods (R programmes secr and SPACECAP) to provide a comprehensive density analysis capable of incorporating environmental and anthropogenic factors. Leopard density was estimated to be 0.35 and 1.18 leopards/100 km2, using secr and SPACECAP, respectively. Leopard population size was predicted to be 102–345 individuals for our three study regions. With these estimates and the predicted available leopard habitat for the province, we extrapolated that the Western Cape supports an estimated 175–588 individuals. Providing a comprehensive baseline population density estimate is critical to understanding population dynamics across a mixed landscape and helping to determine the most appropriate conservation actions. Spatially explicit capture–recapture methods are unbiased by edge effects and superior to traditional capture–mark–recapture methods when estimating animal densities. We therefore recommend further utilization of robust spatial methods as they continue to be advanced.

African lion (Panthera leo) populations have been reduced by almost half in the past two decades, with national parks and game reserves maintaining vital source populations, particularly in East Africa. However, much of the habitats necessary to support lion populations occur in unprotected lands surrounding protected areas. There is an ongoing need for understanding the ecological determinants of lion occurrence in these unprotected habitats, where lions are most vulnerable to extinction. This study evaluated variations in lion site use along a gradient of anthropogenic pressure encompassing the Ruaha National Park, Pawaga‐Idodi Wildlife Management Area (WMA) and unprotected village lands via camera‐trapping. We collected lion occurrence data in the dry seasons of 2014 and 2015, and modelled lion site use as a function of environmental and anthropogenic variables under a Bayesian framework. We recorded 143 lion detections within the national park, 14 in the WMA and no detections in village lands. This result does not imply that lions never use the village lands, but rather that we did not detect them in our surveys during the dry season. Our findings suggest that lion site use was primarily associated with high seasonal wild prey biomass in protected areas. Thus, we infer that human‐induced prey depletion and lion mortality are compromising lion site use of village lands. Seasonal prey movements, and a corresponding concentration inside the park during sampling, could also play an important role in lion site use. These findings reinforce the need to secure large‐bodied prey base to conserve lions, and the importance of protected areas as key refugia for the species.

Predation by cats (Felis catus) is implicated in the decline and extinction of many Australian mammal species. We estimate the number of mammals killed by cats across Australia through meta-analysis of data on the frequency of mammals in cat diet samples from 107 studies. For feral cats in largely natural landscapes, the spatially-weighted mean frequency of mammals in diet samples was 70% (44% for native species, 34% for introduced species). Frequency was significantly higher on the mainland, and in areas of low temperature and topographic ruggedness. Geographic patterns varied markedly between native and introduced mammals, with native mammals most frequent in northern Australia. We estimate that: (i) 815 million individuals yr-1 are killed by feral cats in natural landscapes, 56% of which are native species; (ii) 149 million individuals yr-1 are killed by unowned cats in highly modified landscapes; and (iii) 180 million individuals yr-1 are killed by pet cats. For the latter two components, mainly introduced species are killed. Collectively, across the three components of the cat population, 1,144 million individuals yr-1 are killed by cats, of which, at least 40% (459 million individuals yr-1) are native species. It remains challenging to interpret this tally in terms of its impact on population viability for Australian mammals, because demographic information is not available for most species. However, our estimate of annual mammal mortality due to cat predation is substantially higher than that due to another key threatening process, land clearing.

Mammals comprise the bulk of the diet of free-ranging domestic cats Felis catus (defined as including outdoor pet cats, strays, and feral cats) in most parts of their global range. In Australia, predation by introduced feral cats has been implicated in the extinction of many mammal species, and in the ongoing decline of many extant species. Here, we collate a wide range of records of predation by cats (including feral and pet cats) on Australian mammals and model traits of extant, terrestrial, native mammal species associated with the relative likelihood of cat predation. We explicitly seek to overcome biases in such a continental-scale compilation by excluding possible carrion records for larger species and accounting for differences in the distribution and abundance of potential prey species, as well as study effort, throughout each species’ range. For non-volant species, the relative likelihood of predation by cats was greatest for species in an intermediate weight range (peaking at ca. 400 g), in lower rainfall areas and not dwelling in rocky habitats. Previous studies have shown the greatest rates of decline and extinction in Australian mammals to be associated with these traits. As such, we provide the first continental-scale link between mammal decline and cat predation through quantitative analysis. Our compilation of cat predation records for most extant, terrestrial, native mammal species (151 species, or 52% of the Australian species’ complement) is substantially greater than previously reported (88 species) and includes 50 species listed as threatened by the IUCN or under Australian legislation (57% of Australia's 87 threatened terrestrial mammal species). We identify the Australian mammal species most likely to be threatened by predation by cats (mulgaras Dasycercus spp., kowari Dasyuroides byrnei, many smaller dasyurids and medium-sized to large rodents, among others) and hence most likely to benefit from enhanced mitigation of cat impacts, such as translocations to predator-free islands, the establishment of predator-proof fenced exclosures, and broad-scale cat poison baiting.

Magnesium chelatase initiates chlorophyll biosynthesis, catalysing the MgATP2−-dependent insertion of a Mg2+ ion into protoporphyrin IX. The catalytic core of this large enzyme complex consists of three subunits: Bch/ChlI, Bch/ChlD and Bch/ChlH (in bacteriochlorophyll and chlorophyll producing species, respectively). The D and I subunits are members of the AAA+ (ATPases associated with various cellular activities) superfamily of enzymes, and they form a complex that binds to H, the site of metal ion insertion. In order to investigate the physical coupling between ChlID and ChlH in vivo and in vitro, ChlD was FLAG-tagged in the cyanobacterium Synechocystis sp. PCC 6803 and co-immunoprecipitation experiments showed interactions with both ChlI and ChlH. Co-production of recombinant ChlD and ChlH in Escherichia coli yielded a ChlDH complex. Quantitative analysis using microscale thermophoresis showed magnesium-dependent binding (Kd 331 ± 58 nM) between ChlD and H. The physical basis for a ChlD–H interaction was investigated using chemical cross-linking coupled with mass spectrometry (XL–MS), together with modifications that either truncate ChlD or modify single residues. We found that the C-terminal integrin I domain of ChlD governs association with ChlH, the Mg2+ dependence of which also mediates the cooperative response of the Synechocystis chelatase to magnesium. The interaction site between the AAA+ motor and the chelatase domain of magnesium chelatase will be essential for understanding how free energy from the hydrolysis of ATP on the AAA+ ChlI subunit is transmitted via the bridging subunit ChlD to the active site on ChlH.

As climatic conditions change, species will be forced to move or adapt to avoid extinction. Exacerbated by ongoing climate change, California recently experienced a severe and exceptional drought from 2011 to 2017. To investigate whether an adaptive response occurred during this event, we conducted a “resurrection” study of the cutleaf monkeyflower (Mimulus laciniatus), an annual plant, by comparing trait means and variances of ancestral seed collections (“pre‐drought”) with contemporary descendant collections (“drought”). Plants were grown under common conditions to test whether this geographically restricted species has the capacity to respond evolutionarily to climate stress across its range. We examined if traits shifted in response to the recent, severe drought and included populations across an elevation gradient, including populations at the low‐ and high‐elevation edges of the species range. We found that time to seedling emergence in the drought generation was significantly earlier than in the pre‐drought generation, a response consistent with drought adaptation. Additionally, trait variation in days to emergence was reduced in the drought generation, which suggests selection or bottleneck events. Days to first flower increased significantly by elevation, consistent with climate adaptation across the species range. Drought generation plants were larger and had greater reproduction, which was likely a carryover effect of earlier germination. These results demonstrate that rapid shifts in trait means and variances consistent with climate adaptation are occurring within populations, including peripheral populations at warm and cold climate limits, of a plant species with a relatively restricted range that has so far not shifted its elevation distribution during contemporary climate change. Thus, rapid evolution may mitigate, at least temporarily, range shifts under global climate change. This study highlights the need for better understanding rapid adaptation as a means for plant communities to cope with extraordinary climate events.

OBJECTIVES Helicobacter pylori (H. pylori) guidelines, including the recent ACG clinical guideline, recommend avoiding clarithromycin-based triple therapy (TT-C) among patients with past macrolide exposure. Data to support this recommendation are scarce, and the impact of macrolide exposure on quadruple therapies is unclear. We aimed to determine the impact of macrolide exposure on the efficacy of H. pylori treatment in our region. METHODS We searched the Clalit Health Services database to identify subjects aged 25-60 years who underwent the first-ever C-urea breath test between 2010 and 2015. Patients who underwent a previous H. pylori stool antigen test or gastroscopy were excluded. Pharmacy dispensation data were retrieved. RESULTS We identified 7,842 subjects (36.1% male individuals, age: 40.3 ± 10.5 years), including 3,062 (39.0%) with previous macrolide exposure. The efficacy of TT-C was 74.3% and 82.4% among subjects with and without macrolide exposure, respectively (odds ratio (OR), 0.62; 95% confidence interval (CI), 0.55-0.70; P < 0.0001). TT success was adversely affected by exposure to clarithromycin (55.5%; OR, 0.31; 95% CI, 0.24-0.39; P < 0.0001), roxythromycin (74.4%; OR, 0.65; 95% CI, 0.58-0.74; P < 0.0001), and erythromycin (73.9%; OR, 0.72; 95% CI, 0.57-0.89; P < 0.01) but not by exposure to azithromycin. A greater time elapsed because exposure to clarithromycin and roxythromycin was associated with higher eradication (OR, 1.007; 95% CI, 1.002-1.012; P < 0.01 and OR, 1.004; 95% CI, 1.002-1.006; P < 0.0001). A higher dose of clarithromycin and roxythromycin was associated with a lower likelihood of successful eradication (OR, 0.99988; 95% CI, 0.99982-0.99996; P < 0.01 and OR, 0.99981; 95% CI, 0.99971-0.99992; P < 0.001). The efficacies of sequential and concomitant therapies were 82.7% and 81.3%, respectively, and were not significantly affected by macrolide exposure. CONCLUSIONS TT-C is adversely affected by previous exposure to macrolide antibiotics. Sequential, concomitant, and bismuth-based treatment may be preferred in this setting.

The chromosome complement of the human fungal pathogen Candida albicans is unusually unstable, suggesting that the process of nuclear division is error prone. The Cdc14 phosphatase plays a key role in organising the intricate choreography of mitosis and cell division. In order to understand the role of Cdc14 in C. albicans we used quantitative proteomics to identify proteins that physically interact with Cdc14. To distinguish genuine Cdc14-interactors from proteins that bound non-specifically to the affinity matrix, we used a substrate trapping mutant combined with mass spectrometry analysis using Stable Isotope Labelling with Amino Acids in Cell Culture (SILAC). The results identified 126 proteins that interact with Cdc14 of which 80% have not previously been identified as Cdc14 interactors in C. albicans or S. cerevisiae. In this set, 55 proteins are known from previous research in S. cerevisiae and S. pombe to play roles in the cell cycle, regulating the attachment of the mitotic spindle to kinetochores, mitotic exit, cytokinesis, licensing of DNA replication by re-activating pre-replication complexes, and DNA repair. Five Cdc14-interacting proteins with previously unknown functions localised to the Spindle Pole Bodies (SPBs). Thus, we have greatly increased the number of proteins that physically interact with Cdc14 in C. albicans.

PGC-1α plays a central role in maintaining the mitochondrial and energy metabolism homeostasis, linking external stimuli to the transcriptional co-activation of genes involved in adaptive and age-related pathways. The carboxyl-terminus encodes a serine/arginine-rich (RS) region and a putative RNA recognition motif, however potential RNA-processing role(s) have remained elusive for the past 20 years. Here, we show that the RS domain of human PGC-1α directly interacts with RNA and the nuclear RNA export factor NXF1. Inducible depletion of endogenous PGC-1α and expression of RNAi-resistant RS-deleted PGC-1α further demonstrate that the RNA-binding activity is required for nuclear export of co-activated transcripts and mitochondrial homeostasis. Moreover, a quantitative proteomics approach confirmed PGC-1α-dependent RNA transport and mitochondrial-related functions, identifying also novel mRNA nuclear export targets in age-related telomere maintenance. Discovering a novel function for a major cellular homeostasis regulator provides new directions to further elucidate the roles of PGC-1α in gene expression, metabolic disorders, ageing and neurodegenerative diseases.

Africa is home to some of the world’s most functionally diverse guilds of large carnivores. However, they are increasingly under threat from anthropogenic pressures that may exacerbate already intense intra-guild competition. Understanding the coexistence mechanisms employed by these species in human-impacted landscapes could help shed light on some of the more subtle ways in which humans may impact wildlife populations, and inform multi-species conservation planning. We used camera trap data from Tanzania’s Ruaha-Rungwa landscape to explore temporal and spatiotemporal associations between members of an intact East African large carnivore guild, and determine how these varied across gradients of anthropogenic impact and protection. All large carnivores except African wild dog (Lycaon pictus) exhibited predominantly nocturnal road-travel behaviour. Leopard (Panthera pardus) appeared to employ minor temporal avoidance of lion (Panthera leo) in all sites except those where human impacts were highest, suggesting that leopard may have been freed up from avoidance of lion in areas where the dominant competitor was less abundant, or that the need for leopard to avoid humans outweighed the need to avoid sympatric competitors. Lion appeared to modify their activity patterns to avoid humans in the most impacted areas. We also found evidence of avoidance and attraction among large carnivores: lion and spotted hyaena (Crocuta crocuta) followed leopard; leopard avoided lion; spotted hyaena followed lion; and lion avoided spotted hyaena. Our findings suggest that large carnivores in Ruaha-Rungwa employ fine-scale partitioning mechanisms to facilitate coexistence with both sympatric species and humans, and that growing human pressures may interfere with these behaviours.

Rates at which predators encounter, hunt, and kill prey are influenced by, among other things, the intrinsic condition of prey. Diseases can considerably compromise body condition, potentially weakening ability of afflicted prey to avoid predation. Understanding predator-prey dynamics is particularly important when both species are threatened, as is the case with lions (Panthera leo) and giraffes (Giraffa camelopardalis). Importantly, an emergent disease called giraffe skin disease (GSD) may affect predatory interactions of lions and giraffes. Hypotheses suggest GSD may negatively affect the likelihood of giraffes surviving lion attacks. We evaluated giraffe-lion interactions in Ruaha National Park, Tanzania, where 85% of the giraffe population has GSD. We monitored lion hunting behavior and estimated proportion of the giraffe population with GSD and evidence of ‘lion marks’ from assumed previous lion predation attempts (i.e. claw marks, bite marks, and missing tails). Although we recorded lions hunting and feeding on 16 different prey species, giraffes represented the largest prey category (27%; n = 171 of 641). For age and sex cohorts combined, 26% (n = 140 of 548) of encountered giraffes displayed evidence of previous lion predation attempts. Occurrence of lion marks was higher for adults and males in the giraffe population, suggesting that these individuals were more likely to survive lion attacks. We also found marginal evidence of a positive relationship between giraffes with severe GSD and occurrence of lion marks. Our results identify giraffes as important prey species for lions in Ruaha National Park and suggest that GSD severity plays a minor role in likelihood of surviving a lion attack. This is the first study to explore connections between lion predation and GSD. We explore the ecological implications of disease ecology on predator-prey interactions and consider opportunities for future research on causal links between GSD and giraffe vulnerability to lion predation.

We combine a chemically-synthesized, voltage-sensitive fluorophore with a genetically encoded, self-labeling enzyme to enable voltage imaging in Drosophila melanogaster. Previously, we showed that a rhodamine voltage reporter (RhoVR) combined with the HaloTag self-labeling enzyme could be used to monitor membrane potential changes from mammalian neurons in culture and brain slice. Here, we apply this hybrid RhoVR-Halo approach in vivo to achieve selective neuron labeling in intact fly brains. We generate a Drosophila UAS-HaloTag reporter line in which the HaloTag enzyme is expressed on the surface of cells. We validate the voltage sensitivity of this new construct in cell culture before driving expression of HaloTag in specific brain neurons in flies. We show that selective labeling of synapses, cells, and brain regions can be achieved with RhoVR-Halo in either larval neuromuscular junction (NMJ) or in whole adult brains. Finally, we validate the voltage sensitivity of RhoVR-Halo in fly tissue via dual-electrode/imaging at the NMJ, show the efficacy of this approach for measuring synaptic excitatory post-synaptic potentials (EPSPs) in muscle cells, and perform voltage imaging of carbachol-evoked depolarization and osmolarity-evoked hyperpolarization in projection neurons and in interoceptive subesophageal zone neurons in fly brain explants following in vivo labeling. We envision the turn-on response to depolarizations, fast response kinetics, and two-photon compatibility of chemical indicators, coupled with the cellular and synaptic specificity of genetically-encoded enzymes, will make RhoVR-Halo a powerful complement to neurobiological imaging in Drosophila.Significance StatementVoltage imaging is a powerful method for interrogating neurobiology. Chemical indicators possess fast response kinetics, turn-on responses to membrane depolarization, and can be compatible with two-photon excitation. However, selective cell labeling in intact tissues and in vivo remains a challenge for completely synthetic fluorophores. Here, we show that a chemical – genetic hybrid approach in Drosophila enables cell-specific staining in vivo and voltage imaging in whole-brain explants and at neuromuscular junction synapses.

Short-term plant respiration (R) increases exponentially with rising temperature, but drought could reduce respiration by reducing growth and metabolism. Acclimation may alter these responses. We examined if species with different drought responses would differ in foliar R response to +4.8°C temperature and -45% precipitation in a field experiment with mature pinon and juniper trees, and if any differences between species were related to differences in photosynthesis rates, shoot growth and non-structural carbohydrates (NSCs). Short-term foliar R had a Q10 of 1.6 for pinon and 2.6 for juniper. Pinon foliar R did not respond to the +4.8°C temperatures, but R increased 1.4x for juniper. Across treatments, pinon foliage had higher growth, lower NSC content, 29% lower photosynthesis rates, and 44% lower R than juniper. Removing 45% precipitation had little impact on R for either species. Species differences in the response of R under elevated temperature were related to substrate availability and stomatal response to leaf water potential. Despite not acclimating to the higher temperature and having higher R than pinon, greater substrate availability in juniper suggests it could supply respiratory demand for much longer than pinon. Species responses will be critical in ecosystem response to a warmer climate. This article is protected by copyright. All rights reserved.

The cornea is the clear window that lets light into the eye. It is composed of five layers: epithelium, Bowman’s layer, stroma, Descemet’s membrane and endothelium. The maintenance of its structure and transparency are determined by the functions of the different cell types populating each layer. Attempts to regenerate corneal tissue and understand disease conditions requires knowledge of how cell profiles vary across this heterogeneous tissue. We performed a single cell transcriptomic profiling of 19,472 cells isolated from eight healthy donor corneas. Our analysis delineates the heterogeneity of the corneal layers by identifying cell populations and revealing cell states that contribute in preserving corneal homeostasis. We identified that the expression of CAV1, CXCL14, HOMER3 and CPVL were exclusive to the corneal epithelial limbal stem cell niche, CKS2, STMN1 and UBE2C were exclusively expressed in highly proliferative transit amplifying cells, and NNMT was exclusively expressed by stromal keratocytes. Overall, this research provides a basis to improve current primary cell expansion protocols, for future profiling of corneal disease states, to help guide pluripotent stem cells into different corneal lineages, and to understand how engineered substrates affect corneal cells to improve regenerative therapies.

Feral individuals of the cat Felis catus are recognized internationally as a threat to biodiversity. Open, non-insular systems support a large proportion of the world's biodiversity, but the population-level impacts of feral cats in these systems are rarely elucidated. This limits prioritization and assessment of the effectiveness of management interventions. We quantified the predatory impact of feral cats on small mammals in open, non-insular forest systems in Tasmania, Australia in the context of other factors hypothesized to affect small mammal densities and survival, namely the density of a native carnivore, co-occurring small mammals, and rainfall. Change in feral cat density was the most important determinant of small mammal density and survival. We calculated that, on average, a 50% reduction in feral cat density could result in 25% and 10% increases in the density of the swamp rat Rattus lutreolus and long-tailed mouse Pseudomys higginsi, respectively. Low-level culling of feral cats that we conducted on two of our four study sites to experimentally alter feral cat densities revealed that swamp rat survival was highest when feral cat densities were stable. We conclude that feral cats exert downward pressure on populations of indigenous small mammals in temperate forest systems. However, alleviating this downward pressure on prey by culling a large proportion of the feral cat population is difficult as current methods for reducing feral cat populations in cool temperate forest systems are ineffective, and potentially even counterproductive. We suggest using an adaptive approach that regularly and robustly monitors how feral cats and small mammals respond to management interventions that are intended to conserve vulnerable prey species.

Common reed (Phragmites) interacts with a large suite of other organisms including cryptogams attached to live or dead culm bases. We report an unusual observation of at least ten taxa of lichens and hepatics attached to persistent reed stubble on the swampy bank of freshwater Cypress Creek north of Tampa, Hillsborough County, Florida. This bryoid material included the minute, leafy hepatics Microlejeunea globosa, Microlejeunea cf. ulicina, Myriocoleopsis minutissima, and Frullania cf. inflata; and the lichens Arthonia subdiffusa, Chrysothrix xanthina, Opegrapha viridis, Phaeographis sp., Physcia sp., and an unidentified species of Parmeliaceae. These generally corticolous or epiphyllous taxa have not previously been reported from common reed, and suggest unrecognized complexity in both cryptogamic microhabitats and reed epiphytes.

Epigenetic modifications regulate gene expression in the host response to a diverse range of pathogens. The extent and consequences of epigenetic modification during macrophage responses to Streptococcus pneumoniae, and the role of pneumolysin, a key Streptococcus pneumoniae virulence factor, in influencing these responses, are currently unknown. To investigate this, we infected human monocyte derived macrophages (MDMs) with Streptococcus pneumoniae and addressed whether pneumolysin altered the epigenetic landscape and the associated acute macrophage transcriptional response using a combined transcriptomic and proteomic approach. Transcriptomic analysis identified 503 genes that were differentially expressed in a pneumolysin-dependent manner in these samples. Pathway analysis highlighted the involvement of transcriptional responses to core innate responses to pneumococci including modules associated with metabolic pathways activated in response to infection, oxidative stress responses and NFκB, NOD-like receptor and TNF signalling pathways. Quantitative proteomic analysis confirmed pneumolysin-regulated protein expression, early after bacterial challenge, in representative transcriptional modules associated with innate immune responses. In parallel, quantitative mass spectrometry identified global changes in the relative abundance of histone post translational modifications (PTMs) upon pneumococcal challenge. We identified an increase in the relative abundance of H3K4me1, H4K16ac and a decrease in H3K9me2 and H3K79me2 in a PLY-dependent fashion. We confirmed that pneumolysin blunted early transcriptional responses involving TNF-α and IL-6 expression. Vorinostat, a histone deacetylase inhibitor, similarly downregulated TNF production, reprising the pattern observed with pneumolysin. In conclusion, widespread changes in the macrophage transcriptional response are regulated by pneumolysin and are associated with global changes in histone PTMs. Modulating histone PTMs can reverse pneumolysin-associated transcriptional changes influencing innate immune responses, suggesting that epigenetic modification by pneumolysin plays a role in dampening the innate responses to pneumococci.Author summaryPneumolysin is a toxin that contributes to how Streptococcus pneumoniae, the leading cause of pneumonia, causes disease. In this study, the toxin alters gene expression in immune cells called macrophages, one of the first lines of defence against bacteria at sites of infection. Modulation involved multiple immune responses, including generation of chemical signals coordinating responses in immune cells termed cytokines. In addition, changes were observed in histone proteins that are involved in controlling gene expression in the cell. Pneumolysin reduced early production of the cytokine TNF-α and a medicine vorinostat that modifies these ‘epigenetic’ histone modifications had a similar affect, suggesting epigenetic mechanisms contribute to the ability of pneumolysin to reduce immune responses.

This investigation aimed to determine the effect of a multi-ingredient pre-workout supplement (MIPS) on heart rate (HR), perceived exertion (RPE), lactate concentration, and time to fatigue (TTF) during a running task to volitional exhaustion. Eleven NCAA Division I cross-country runners (20 ± 2 year, height: 171 ± 14 cm, weight: 63.5 ± 9.1 kg) participated in this randomized, double-blind, placebo-controlled cross-over study. Bayesian statistical methods were utilized, and parameter estimates were interpreted as statistically significant if the 95% highest-density intervals (HDIs) did not include zero. TTF was increased in the MIPS condition with a posterior Meandiff = 154 ± 4.2 s (95% HDI: −167, 465) and a 0.84 posterior probability that the supplement would increase TTF relative to PL. Blood lactate concentration immediately post-exercise was also higher in the MIPS condition compared to PL with an estimated posterior Meandiff = 3.99 ± 2.1 mmol (95% HDI: −0.16, 7.68). There were no differences in HR or RPE between trials. These findings suggest that a MIPS ingested prior to sustained running at lactate threshold has an 84% chance of increasing TTF in highly trained runners and may allow athletes to handle a higher level of circulating lactate before reaching exhaustion.

Our knowledge of the conservation status of reptiles, the most diverse class of terrestrial vertebrates, has improved dramatically over the past decade, but still lags behind that of the other tetrapod groups. Here, we conduct the first comprehensive evaluation (~92% of the world’s ~1,714 described species) of the conservation status of skinks (Scincidae), a speciose reptile family with a worldwide distribution. Using International Union for Conservation of Nature (IUCN) criteria, we report that ~20% of species are threatened with extinction, and nine species are Extinct or Extinct in the Wild. The highest levels of threat are evident in Madagascar and the Neotropics, and in the subfamilies Mabuyinae, Eugongylinae and Scincinae. The vast majority of threatened skink species were listed based primarily on their small geographic ranges (Criterion B, 83%; Criterion D2, 13%). Although the population trend of 42% of species was stable, 14% have declining populations. The key threats to skinks are habitat loss due to agriculture, invasive species, and biological resource use (e.g., hunting, timber harvesting). The distributions of 61% of species do not overlap with protected areas. Despite our improved knowledge of the conservation status of the world’s skinks, 8% of species remain to be assessed, and 14% are listed as Data Deficient. The conservation status of almost a quarter of the world’s skink species thus remains unknown. We use our updated knowledge of the conservation status of the group to develop and outline the priorities for the conservation assessment and management of the world’s skink species.

Made available in DSpace on 2021-06-25T15:08:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-09 Wellcome Trust From 1985 to 2016, the prevalence of underweight decreased, and that of obesity and severe obesity increased, in most regions, with significant variation in the magnitude of these changes across regions. We investigated how much change in mean body mass index (BMI) explains changes in the prevalence of underweight, obesity, and severe obesity in different regions using data from 2896 population-based studies with 187 million participants. Changes in the prevalence of underweight and total obesity, and to a lesser extent severe obesity, are largely driven by shifts in the distribution of BMI, with smaller contributions from changes in the shape of the distribution. In East and Southeast Asia and sub-Saharan Africa, the underweight tail of the BMI distribution was left behind as the distribution shifted. There is a need for policies that address all forms of malnutrition by making healthy foods accessible and affordable, while restricting unhealthy foods through fiscal and regulatory restrictions. Imperial Coll London, London, England Harvard TH Chan Sch Publ Hlth, Boston, MA USA Middlesex Univ, London, England World Hlth Org, Geneva, Switzerland Minist Hlth, Victoria, Seychelles Univ Lausanne, Lausanne, Switzerland Eduardo Mondlane Univ, Maputo, Mozambique Victor Babes Univ Med & Pharm Timisoara, Timisoara, Romania Univ Sydney, Sydney, NSW, Australia Natl Inst Biomed Innovat Hlth & Nutr, Tokyo, Japan Univ Auckland, Auckland, New Zealand Seoul Natl Univ, Seoul, South Korea ICMR Natl Inst Nutr, Hyderabad, India Capital Med Univ, Beijing An Zhen Hosp, Beijing, Peoples R China Univ Peruana Cayetano Heredia, Lima, Peru Univ Tunis El Manar, Tunis, Tunisia Univ Oulu, Oulu, Finland Univ Zagreb, Zagreb, Croatia Univ Ljubljana, Ljubljana, Slovenia Caja Costarricense Seguro Social, San Jose, Costa Rica Al Quds Univ, East Jerusalem, State Of Palest, Palestine Natl Ctr Publ Healthcare, Nur Sultan, Kazakhstan Minist Hlth, Kuala Lumpur, Malaysia Qatar Univ, Doha, Qatar Birzeit Univ, Birzeit, State Of Palest, Israel Usmanu Danfodiyo Univ Teaching Hosp, Sokoto, Nigeria Inst Mexicano Seguro Social, Mexico City, DF, Mexico Flinders Univ S Australia, Adelaide, SA, Australia Mahidol Univ, Salaya, Nakhon Pathom, Thailand BRAC James P Grant Sch Publ Hlth, Dhaka, Bangladesh Univ Copenhagen, Copenhagen, Denmark Copenhagen Univ Hosp, Copenhagen, Denmark Food & Nutr Res Inst, Taguig, Philippines Urmia Univ Med Sci, Orumiyeh, Iran Inst Nacl Ciencias Med & Nut, Mexico City, DF, Mexico Univ Amsterdam, Amsterdam, Netherlands Shahrekord Univ Med Sci, Shahrekord, Iran Noncommunicable Dis Res Ctr, Tehran, Iran Univ Oslo, Oslo, Norway Univ Bremen, Bremen, Germany Republican Ctr Hlth Promot, Bishkek, Kyrgyzstan Natl Ctr Diabet Endocrinol & Genet, Amman, Jordan Princess Nourah Bint Abdulrahman Univ, Riyadh, Saudi Arabia Kuwait Inst Sci Res, Kuwait, Kuwait King Abdulaziz Univ, Jeddah, Saudi Arabia Dasman Diabet Inst, Kuwait, Kuwait Luxembourg Inst Hlth, Strassen, Luxembourg Aldara Hosp & Med Ctr, Riyadh, Saudi Arabia King Abdullah Int Med Res Ctr, Riyadh, Saudi Arabia Bispebjerg & Frederiksberg Hosp, Copenhagen, Denmark Barcelona Inst Global Hlth CIBERESP, Barcelona, Spain World Hlth Org Reg Off Eastern Mediterranean, Cairo, Egypt Bombay Hosp & Med Res Ctr, Mumbai, Maharashtra, India Res Ctr Social Determinants Hlth, Tehran, Iran UMR CNRS MNHN 7206 Ecoanthropologie, Paris, France Univ Lille, Lille, France Lille Univ Hosp, Lille, France Western Norway Univ Appl Sci, Sogndal, Norway Norwegian Sch Sport Sci, Oslo, Norway Madras Diabet Res Fdn, Chennai, Tamil Nadu, India Zahedan Univ Med Sci, Zahedan, Iran Natl Inst Publ Hlth, Tunis, Tunisia Univ Porto, Inst Publ Hlth, Porto, Portugal Norwegian Inst Publ Hlth, Oslo, Norway Univ Massachusetts, Amherst, MA 01003 USA Abt Associates Inc, Kathmandu, Nepal Univ Iceland, Reykjavik, Iceland Univ Yaounde I, Yaounde, Cameroon Univ Fed Pelotas, Pelotas, RS, Brazil Univ Med 1, Yangon, Myanmar Banska Bystrica Reg Author Publ Hlth, Banska Bystrica, Slovakia Tel Aviv Univ, Tel Aviv, Israel Hebrew Univ Jerusalem, Jerusalem, Israel Univ Porto, Med Sch, Porto, Portugal Neyshabur Univ Med Sci, Neyshabur, Iran Res Inst Endocrine Sci, Tehran, Iran Indian Council Med Res, New Delhi, India Natl Inst Publ Hlth, Copenhagen, Denmark Univ Sci & Technol, Sanaa, Yemen Med Univ Lodz, Lodz, Poland Med Univ Gdansk, Gdansk, Poland Univ Autonoma Madrid CIBERESP, Madrid, Spain Univ Rzeszow, Rzeszow, Poland Univ Palermo, Palermo, Italy Pan Amer Hlth Org, Washington, DC USA Mohammed V Univ Rabat, Rabat, Morocco Univ Pernambuco, Recife, PE, Brazil Baqai Inst Diabetol & Endocrinol, Karachi, Pakistan Univ Fed Santa Catarina, Florianopolis, SC, Brazil Dalhousie Univ, Halifax, NS, Canada Jordan Univ Sci & Technol, Irbid, Jordan Univ Fed Maranhao, Sao Luis, Brazil Farabi Kazakh Natl Univ, Alma Ata, Kazakhstan Univ Coll Dublin, Dublin, Ireland Christian Med Coll & Hosp, Vellore, Tamil Nadu, India Fed Minist Social Affairs Hlth Care & Consumer Pr, Vienna, Austria Cafam Univ Fdn, Bogota, Colombia Kazakh Natl Med Univ, Alma Ata, Kazakhstan Lithuanian Univ Hlth Sci, Kaunas, Lithuania Univ Sao Paulo, Sao Paulo, Brazil BJ Med Coll, Ahmadabad, Gujarat, India Chirayu Med Coll, New Delhi, India Sunder Lal Jain Hosp, Delhi, India Hosp Sick Children, Toronto, ON, Canada Shandong Univ Tradit Chinese Med, Jinan, Peoples R China Shanghai Jiao Tong Univ, Sch Med, Shanghai, Peoples R China Univ Republica, Montevideo, Uruguay Inst Med Res & Med Plant Studies, Yaounde, Cameroon Ufa Eye Res Inst, Ufa, Russia Nepal Hlth Res Council, Kathmandu, Nepal Univ Montenegro, Niksic, Montenegro Univ Southern Denmark, Copenhagen, Denmark Univ Gothenburg, Gothenburg, Sweden Univ Fed Rio de Janeiro, Rio De Janeiro, Brazil Natl Inst Publ Hlth & Environm, Bilthoven, Netherlands Univ Turin, Turin, Italy UCL, London, England Liverpool John Moores Univ, Liverpool, Merseyside, England Nanyang Technol Univ Singapore, Singapore, Singapore German Inst Human Nutr, Potsdam, Germany Endocrinol Res Ctr, Moscow, Russia Ctr Educ Med & Invest Clin, Buenos Aires, DF, Argentina IRCCS Neuromed, Pozzilli, Italy Toulouse Univ, Sch Med, Toulouse, France Univ Zurich, Zurich, Switzerland Univ Hosp KU Leuven, Leuven, Belgium Flemish Agcy Care & Hlth, Brussels, Belgium Univ Ghent, Ghent, Belgium FrieslandCampina, Amersfoort, Netherlands Univ Cent Venezuela, Caracas, Venezuela Bielefeld Univ, Bielefeld, Germany World Hlth Org Reg Off Europe, Moscow, Russia German Canc Res Ctr, Heidelberg, Germany Fred Hollows Fdn, Auckland, New Zealand Univ Med & Pharm Bucharest, Bucharest, Romania Swansea Univ, Swansea, W Glam, Wales Univ Coll Copenhagen, Copenhagen, Denmark Inst Publ Hlth, Tirana, Albania Munster Technol Univ, Cork, Ireland Univ La Laguna, Tenerife, Spain Univ Malta, Msida, Malta Vanderbilt Univ, 221 Kirkland Hall, Nashville, TN 37235 USA Minist Hlth, Tongatapu, Tonga Canadian Fitness & Lifestyle Res Inst, Ottawa, ON, Canada Hosp Santa Maria, Lisbon, Portugal Istanbul Univ Cerrahpasa, Istanbul, Turkey Univ Fed Juiz de Fora, Juiz De Fora, Brazil Minist Publ Hlth, Asuncion, Paraguay Natl Inst Publ Hlth, Prague, Czech Republic Gaetano Fucito Hosp, Mercato San Severino, Italy Univ Groningen, Groningen, Netherlands Karolinska Inst, Huddinge, Sweden Ctr Estudios Nutr Infantil, Buenos Aires, DF, Argentina Univ Porto, Porto, Portugal Univ Zaragoza, Zaragoza, Spain Santiago de Compostela Univ, Santiago De Compostela, Spain Minist Hlth, Ankara, Turkey Council Agr Res & Econ, Rome, Italy Fed Univ Rio Grande, Rio Grande, Brazil India Diabet Res Fdn, Chennai, Tamil Nadu, India Duke NUS Med Sch, Singapore, Singapore ICMR Natl Inst Med Stat, New Delhi, India Singapore Eye Res Inst, Singapore, Singapore Acad Sinica, Taipei, Taiwan Capital Inst Pediat, Beijing, Peoples R China Duke Univ, Durham, NH USA Kailuan Gen Hosp, Tangshan, Peoples R China Univ Oxford, Oxford, England Ahvaz Jundishapur Univ Med Sci, Ahvaz, Iran Gertner Inst Epidemiol & Hlth Policy Res, Ramat Gan, Israel Natl Ctr Publ Hlth & Anal, Sofia, Bulgaria Univ Fribourg, Fribourg, Switzerland Minist Hlth & Welf, Taipei, Taiwan CIBER Epidemiol & Salud Publ, Murcia, Spain Univ Southern Denmark, Odense, Denmark Univ Estadual Paulista, Presidente Prudente, Brazil Med Univ Silesia, Katowice, Poland Charles Univ Prague, Prague, Czech Republic Primary Hlth Care, Floriana, Malta Carol Davila Univ Med & Pharm, Bucharest, Romania Katholieke Univ Leuven, Leuven, Belgium STAT Canada, Ottawa, ON, Canada UMR CNRS MNHN 7206 Ecoanthropol, Marseille, France Agcy Prevent & Social Med, Bregenz, Austria Univ Southampton, Southampton, Hants, England Inst Pasteur, Lille, France Malawi Epidemiol & Intervent Res Unit, Lilongwe, Malawi CIBEROBN, Madrid, Spain Univ Debrecen, Debrecen, Hungary Univ Med & Pharm Carol Davila, Bucharest, Romania Univ Fed Rio Grande do Sul, Porto Alegre, RS, Brazil CNR, Reggio Di Calabria, Italy Eftimie Murgu Univ Resita, Resita, Romania Spanish Agcy Food Safety & Nutr, Madrid, Spain Indian Stat Inst, Kolkata, India Tabriz Hlth Serv Management Res Ctr, Tabriz, Iran Geneva Univ Hosp, Geneva, Switzerland Sciensano, Brussels, Belgium Univ Med Ctr Groningen, Groningen, Netherlands Natl Res Ctr Prevent Med, Moscow, Russia Innovating Hlth Int, Port Au Prince, Haiti Univ Montreal, Montreal, PQ, Canada French Natl Res Inst Sustainable Dev, Montpellier, France French Publ Hlth Agcy, St Maurice, France Mediterranea Cardioctr, Naples, Italy Univ Vale Rio dos Sinos, Sao Leopoldo, Brazil Natl Inst Hyg Epidemiol & Microbiol, Havana, Cuba Natl Council Sci & Tech Res, Buenos Aires, DF, Argentina Minist Hlth & Med Educ, Tehran, Iran Univ Novi Sad, Novi Sad, Serbia Natl Inst Nutr, Hanoi, Vietnam Univ Queensland, Brisbane, Qld, Australia Ist Super Sanita, Rome, Italy Univ Cuenca, Cuenca, Ecuador Helmholtz Zentrum Munchen, Munich, Germany Univ Tehran Med Sci, Tehran, Iran Minist Sante & Hyg Publ, Abidjan, Cote Ivoire Univ Hosp Dusseldorf, Dusseldorf, Germany Natl Inst Cardiol, Warsaw, Poland Beijing Ctr Dis Prevent & Control, Beijing, Peoples R China UMI 3189 ESS, Marseille, France Beth Israel Deaconess Med Ctr, Boston, MA 02215 USA Scuola Super Sant Anna, Pisa, Italy Univ Latvia, Riga, Latvia Minist Hlth & Med Serv, Gizo, Solomon Islands Hormozgan Univ Med Sci, Bandar Abbas, Iran Univ Benin, Benin, Nigeria Univ Skovde, Skovde, Sweden Natl Inst Nutr & Food Technol, Tunis, Tunisia Univ West Indies, Kingston, Jamaica Univ Calif Davis, Davis, CA 95616 USA Univ Stellenbosch, Cape Town, South Africa Univ Duisburg Essen, Duisburg, Germany Karadeniz Tech Univ, Trabzon, Turkey Univ Helsinki, Helsinki, Finland Mashhad Univ Med Sci, Mashhad, Razavi Khorasan, Iran Rafsanjan Univ Med Sci, Rafsanjan, Iran Queens Univ Belfast, Belfast, Antrim, North Ireland Tabriz Univ Med Sci, Tabriz, Iran Fasa Univ Med Sci, Fasa, Iran Shiraz Univ Med Sci, Shiraz, Iran Baqai Med Univ, Karachi, Pakistan Ctr Salud Villanueva Norte, Badajoz, Spain Hosp Don Benito Villanueva Serena, Badajoz, Spain Minist Hlth, Buenos Aires, DF, Argentina Univ Insubria, Varese, Italy Pontificia Univ Catolica Chile, Santiago, Chile Inst Mother & Child Hlth, Warsaw, Poland Swiss Trop & Publ Hlth Inst, Basel, Switzerland Univ Tartu, Tartu, Estonia Univ Sains Malaysia, Kota Baharu, Kelantan, Malaysia Umea Univ, Umea, Sweden Univ Fed Sao Paulo, Sao Paulo, Brazil Hosp Univ Son Espases, Palma De Mallorca, Spain Hosp Clin Porto Alegre, Porto Alegre, RS, Brazil Univ Sumatera Utara, Medan, Indonesia Kindai Univ, Osaka, Japan Kyoto Univ, Kyoto, Japan Med Univ Warsaw, Warsaw, Poland Jagiellonian Univ, Med Coll, Krakow, Poland Minist Salute DG Prevenz Sanit, Rome, Italy Univ Catania, Catania, Italy CIBER Epidemiol & Salud Publ, Alicante, Spain Africa Hlth Res Inst, Mtubatuba, South Africa Univ Geneva, Med Sch, Geneva, Switzerland CIBER Epidemiol & Salud Publ, Barcelona, Spain Univ Fed Minas Gerais, Belo Horizonte, MG, Brazil Univ Utrecht, Utrecht, Netherlands Wageningen Univ, Wageningen, Netherlands Univ Adelaide, Adelaide, SA, Australia Lund Univ, Lund, Sweden Aristotle Univ Thessaloniki, Thessaloniki, Greece Inst Natl Sante & Rech Med, Villejuif, France Minist Hlth, Jerusalem, Israel McGill Univ, Montreal, PQ, Canada Gasol Fdn, Barcelona, Spain PASs Hirszfeld Inst Immunol & Expt Therapy, Wroclaw, Poland Univ Politecn Madrid, Madrid, Spain St Annes Univ Hosp, Brno, Czech Republic Natl Inst Publ Hlth, Cuernavaca, Morelos, Mexico Ctr Estudios Diabet AC, Mexico City, DF, Mexico Univ Autonoma Santo Domingo, Santo Domingo, Dominican Rep Minist Hlth, Lisbon, Portugal Inst Clin & Expt Med, Prague, Czech Republic Childrens Mem Hlth Inst, Warsaw, Poland Natl Ctr Cardiovasc Dis, Beijing, Peoples R China Univ Ferrara, Ferrara, Italy Author Sanitaria San Marino, San Marino, CA USA Iceland Heart Assoc, Kopavogur, Iceland Univ Icesi, Cali, Colombia Univ Estadual Montes Claros, Montes Claros, MG, Brazil Kings Coll London, London, England Int Agcy Res Canc, Lyon, France Capital Med Univ, Beijing, Peoples R China Capital Med Univ, Beijing Tongren Hosp, Beijing, Peoples R China Healis Sekhsaria Inst Publ Hlth, Navi Mumbai, India Eternal Heart Care Ctr & Res Inst, Jaipur, Rajasthan, India Univ Ibadan, Ibadan, Nigeria Inst Clin Effectiveness & Hlth Policy, Buenos Aires, DF, Argentina Natl Hlth Insurance Serv, Wonju, South Korea Prevent Metab Disorders Res Ctr, Tehran, Iran Res & Educ Inst Child Hlth, Nicosia, Cyprus Danish Canc Soc Res Ctr, Copenhagen, Denmark Univ West Indies, Cave Hill, Barbados Kermanshah Univ Med Sci, Kermanshah, Iran Univ Eastern Finland, Kuopio, Finland Beijing Inst Ophthalmol, Beijing, Peoples R China Int Hellen Univ, Thessaloniki, Greece Kyushu Univ, Fukuoka, Japan Univ Bergen, Bergen, Norway Tulane Univ, New Orleans, LA USA Natl Res Inst Hlth & Family Planning, Beijing, Peoples R China Chinese Ctr Dis Control & Prevent, Beijing, Peoples R China Univ Pecs, Pecs, Hungary Danish Hlth Author, Copenhagen, Denmark Joep Lange Inst, Amsterdam, Netherlands Univ Fed Pernambuco, Recife, PE, Brazil Oulu Univ Hosp, Oulu, Finland Univ Autonoma Bucaramanga, Bucaramanga, Colombia Swiss Fed Inst Technol, Zurich, Switzerland Chron Dis Res Ctr, Tehran, Iran Univ Hong Kong, Hong Kong, Peoples R China Chinese Univ Hong Kong, Hong Kong, Peoples R China Univ Western Australia, Perth, WA, Australia Kingston Hlth Sci Ctr, Kingston, ON, Canada Fdn Oftalmol Santander, Bucaramanga, Colombia Univ Oran 1, Oran, Algeria Nay Pyi Taw, Independent Publ Hlth Specialist, Nay Pyi Taw, Myanmar Minist Hlth & Sports, Nay Pyi Taw, Myanmar Peking Univ, Beijing, Peoples R China Natl Inst Publ Hlth, Bucharest, Romania Vrije Univ Amsterdam Med Ctr, Amsterdam, Netherlands Amer Univ Beirut, Beirut, Lebanon CIBER Epidemiol & Salud Publ, San Sebastian, Spain Cairo Univ, Cairo, Egypt Erasmus MC, Rotterdam, Netherlands Med Univ Varna, Varna, Bulgaria Univ Tokyo, Tokyo, Japan Deakin Univ, Geelong, Vic, Australia Tokyo Metropolitan Inst Gerontol, Tokyo, Japan Hadassah Univ Med Ctr, Jerusalem, Israel Norwegian Univ Sci & Technol, Trondheim, Norway Univ Melbourne, Melbourne, Vic, Australia Sports Univ Tirana, Tirana, Albania Univ Hosp Ctr Zagreb, Zagreb, Croatia Univ Zagreb, Sch Med, Zagreb, Croatia Heart Fdn, Melbourne, Vic, Australia Guangzhou 12th Hosp, Guangzhou, Peoples R China Univ Eugenio Maria Hostos, Santo Domingo, Dominican Rep Simon Fraser Univ, Burnaby, BC, Canada Inst Mol & Clin Ophthalmol Basel, Basel, Switzerland Univ New South Wales, Sydney, NSW, Australia World Hlth Org Country Off, Delhi, India Guilan Univ Med Sci, Rasht, Iran Inst Publ Hlth, Belgrade, Serbia Univ Opole, Opole, Poland Finnish Inst Hlth & Welf, Helsinki, Finland Gulu Univ, Gulu, Uganda Univ Crete, Iraklion, Greece Sri Venkateswara Univ, Tirupati, Andhra Pradesh, India Sree Chitra Tirunal Inst Med Sci & Technol, Trivandrum, Kerala, India Hellen Med Assoc Obes, Athens, Greece Natl & Kapodistrian Univ Athens, Athens, Greece Maharajgunj Med Campus, Kathmandu, Nepal Aarhus Univ, Aarhus, Denmark Johns Hopkins Bloomberg Sch Publ Hlth, Baltimore, MD USA Pennington Biomed Res Ctr, Baton Rouge, LA USA Natl Inst Epidemiol, Chennai, Tamil Nadu, India Univ Munster, Munster, Germany Israel Ctr Dis Control, Ramat Gan, Israel Res Inst Primordial Prevent Noncommunicable Dis, Esfahan, Iran Amsterdam UMC Publ Hlth Res Inst, Amsterdam, Netherlands South African Med Res Council, Cape Town, South Africa Kyrgyz State Med Acad, Bishkek, Kyrgyzstan Res Inst Child Nutr, Dortmund, Germany Shahid Beheshti Univ Med Sci, Tehran, Iran Univ Cambridge, Cambridge, England Mazandaran Univ Med Sci, Sari, Iran Hypertens Res Ctr, Esfahan, Iran Med Univ Innsbruck, Innsbruck, Austria Muhimbili Univ Hlth & Allied Sci, Dar Es Salaam, Tanzania Yonsei Univ, Coll Med, Seoul, South Korea Natl Canc Ctr, Goyang Si, South Korea Univ Coll South Denmark, Haderslev, Denmark Stat Austria, Vienna, Austria BP Koirala Inst Hlth Sci, Dharan, Nepal Univ Vienna, Vienna, Austria Tartu Univ Clin, Tartu, Estonia Kansai Med Univ, Hirakata, Osaka, Japan Hungarian Sch Sport Federat, Budapest, Hungary Minist Hlth & Qual Life, Port Louis, Mauritius Univ Hosp Ulm, Ulm, Germany North West Univ, Potchefstroom, South Africa Univ Jyvaskyla, Jyvaskyla, Finland Inst Publ Hlth, Podgorica, Montenegro Amrita Inst Med Sci, Cochin, Kerala, India Inst Endocrinol, Prague, Czech Republic All India Inst Med Sci, New Delhi, India African Populat & Hlth Res Ctr, Nairobi, Kenya Hanoi Univ Publ Hlth, Hanoi, Vietnam Hassan First Univ Settat, Settat, Morocco Minist Hlth, Algiers, Algeria Harokopio Univ, Athens, Greece Sahlgrens Acad, Gothenburg, Sweden Endocrinol & Metab Res Ctr, Tehran, Iran Univ Publ Hlth, Yangon, Myanmar Int Food Policy Res Inst, Dakar, Senegal Natl Univ Singapore, Singapore, Singapore Hong Kong Polytech Univ, Hong Kong, Peoples R China Tampere Univ Hosp, Tampere, Finland Univ Douala, Douala, Cameroon Univ Cape Town, Cape Town, South Africa Natl Inst Hlth Dev, Tallinn, Estonia West Virginia Univ, Morgantown, WV 26506 USA Oswaldo Cruz Fdn Rene Rachou Res Inst, Belo Horizonte, MG, Brazil Natl Taiwan Univ, Taipei, Taiwan Univ Chinese Acad Sci, Shanghai, Peoples R China Uppsala Univ, Uppsala, Sweden Taipei Med Univ, Taipei, Taiwan Sports Med Ctr Minho, Braga, Portugal Univ San Martin Porres, Lima, Peru Consejeria Sanidad Junta Castilla & Leon, Valladolid, Spain Univ Fed Ciencias Saude Porto Alegre, Porto Alegre, RS, Brazil Norrbotten Cty Council, Lulea, Sweden Ilembula Lutheran Hosp, Ilembula, Iran Univ Fed Ouro Preto, Ouro Preto, Brazil Univ Coimbra, Coimbra, Portugal Coimbra Univ Hosp Ctr, Coimbra, Portugal Inst Neurosci Natl Res Council, Padua, Italy Baker Heart & Diabet Inst, Melbourne, Vic, Australia Agr Univ Athens, Athens, Greece Hosp Israelita Albert Einstein, Sao Paulo, Brazil SB RAS Fed Res Ctr Inst Cytol & Genet, Novosibirsk, Russia Univ Otago, Dunedin, New Zealand Univ Padua, Padua, Italy Hellen Mediterranean Univ, Siteia, Greece Loughborough Univ, Loughborough, Leics, England Minist Hlth, Nicosia, Cyprus Lausanne Univ Hosp, Lausanne, Switzerland Escola Nacl Saude Publ Sergio Arouca, Rio De Janeiro, Brazil CIBERCV, Madrid, Spain Mary Immaculate Coll, Limerick, Ireland Emory Univ, Atlanta, GA 30322 USA Hungarian Soc Sports Med, Budapest, Hungary Paracelsus Med Univ, Salzburg, Austria Univ Estadual Centro Oeste, Guarapuava, Brazil Sherikashmir Inst Med Sci, Srinagar, India UiT Arctic Univ Norway, Tromso, Norway ICMR Natl Ctr Dis Informat & Res, Bengaluru, India Cape Peninsula Univ Technol, Cape Town, South Africa Brown Univ, Providence, RI 02912 USA London Sch Hyg & Trop Med, London, England Univ Edinburgh, Edinburgh, Midlothian, Scotland Shahid Sadoughi Univ Med Sci, Tehran, Iran Inst Natl Sante & Rech Med, Lille, France Robert Koch Inst, Berlin, Germany Univ Lisbon, Lisbon, Portugal Womens Social & Hlth Studies Fdn, Trivandrum, Kerala, India Democritus Univ, Alexandroupolis, Greece Grigore T Popa Univ Med & Pharm, Iasi, Romania Univ Firenze, Florence, Italy Ain Shams Univ, Cairo, Egypt Isfahan Cardiovasc Res Ctr, Esfahan, Iran Univ Strasbourg, Strasbourg, France Mulago Hosp, Kampala, Uganda Univ San Carlos, Guatemala City, Guatemala Univ Limpopo, Sovenga, South Africa Univ Med Sci Cienfuegos, Cienfuegos, Cuba Royal Coll Surgeons Ireland, Dublin, Ireland Trobe Univ, Melbourne, Vic, Australia Int Inst Mol & Cell Biol, Warsaw, Poland Inst Conmemorat Gorgas Estudios Salud, Panama City, Panama World Hlth Org Country Off, Lilongwe, Malawi Dept Publ Hlth, Nay Pyi Taw, Myanmar Univ Brescia, Brescia, Italy Minist Hlth & Social Protect, Dushanbe, Tajikistan Univ Limerick, Limerick, Ireland Croatian Inst Publ Hlth, Zagreb, Croatia Univ Kebangsaan Malaysia, Kuala Lumpur, Malaysia Bushehr Univ Med Sci, Bushehr, Iran Ulm Univ, Ulm, Germany Dept Stat, Kuala Lumpur, Malaysia Kobe Univ, Kobe, Hyogo, Japan Suraj Eye Inst, Nagpur, Maharashtra, India UNICEF, Yaounde, Cameroon Univ Med Greifswald, Greifswald, Germany Karolinska Inst, Stockholm, Sweden Natl Inst Hyg & Epidemiol, Hanoi, Vietnam Univ Med & Pharm Ho Chi Minh City, Ho Chi Minh City, Vietnam Hanoi Med Univ, Hanoi, Vietnam LifeDoc Hlth, Memphis, TN USA Heartfile, Islamabad, Pakistan Eastern Mediterranean Publ Hlth Network, Amman, Jordan Univ Manchester, Manchester, Lancs, England State Univ Med & Pharm, Kishinev, Moldova Tachikawa Gen Hosp, Nagaoka, Niigata, Japan Korea Ctr Dis Control & Prevent, Cheongju, South Korea Japan Wildlife Res Ctr, Tokyo, Japan Istanbul Univ, Istanbul, Turkey Minist Hlth, Bandar Seri Begawan, Brunei Univ Fed Parana, Curitiba, Parana, Brazil Univ Madeira, Funchal, Portugal Univ Puerto Rico, San Juan, PR 00936 USA Ctr Clin Res & Prevent, Glostrup, Denmark MRC Lifecourse Epidemiol Unit, Southampton, Hants, England Kwame Nkrumah Univ Sci & Technol, Kumasi, Ghana UniSante, Lausanne, Switzerland Inst Canc Res, Prevent & Clin Network, Florence, Italy Univ Wisconsin, Madison, WI USA Heidelberg Univ, Heidelberg, Germany IRCCS Ente Osped Specializzato Gastroenterol S de, Bari, Italy Zayed Univ, Abu Dhabi, U Arab Emirates Catholic Univ Daegu, Daegu, South Korea Univ Med Pharm Sci & Technol Targu Mures, Targu Mures, Romania Jivandeep Hosp, Anand, Gujarat, India South African Med Res Council, Durban, South Africa Natl Dent Care Ctr Singapore, Singapore, Singapore Univ Malaga, Malaga, Spain Univ Hosp Varese, Varese, Italy Vietnam Natl Heart Inst, Hanoi, Vietnam Clin Med Avanzada Dr Abel Gonzalez, Santo Domingo, Dominican Rep Leibniz Inst Prevent Res & Epidemiol BIPS, Bremen, Germany Univ Sarajevo, Sarajevo, Bosnia & Herceg Cardiovasc Prevent Ctr Udine, Udine, Italy Pisa Univ Hosp, Pisa, Italy Minist Hlth & Med Serv, Honiara, Solomon Islands Publ Hlth Agcy Catalonia, Barcelona, Spain Inst Hosp del Mar Invest Med, Barcelona, Spain Ardabil Univ Med Sci, Ardebil, Iran Ctr Dis Prevent & Control, Riga, Latvia Alborz Univ Med Sci, Karaj, Iran Minist Hlth, Hanoi, Vietnam Pure Earth, Dhaka, Bangladesh Inst Epidemiol Dis Control & Res, Dhaka, Bangladesh Univ Turku, Turku, Finland Inst Univ Invest Atenc Primaria Jordi Gol, Girona, Spain Univ Putra Malaysia, Serdang, Malaysia Univ Malaya, Kuala Lumpur, Malaysia Sotiria Hosp, Athens, Greece Univ Valencia, Valencia, Spain Univ Philippines, Manila, Philippines Slovak Acad Sci, Bratislava, Slovakia Nutr Res Fdn, Barcelona, Spain Minas Gerais State Secretariat Hlth, Belo Horizonte, MG, Brazil CS S Agustin Ibsalut, Palma De Mallorca, Spain Amsterdam Inst Global Hlth & Dev, Amsterdam, Netherlands Natl Inst Hlth Doutor Ricardo Jorge, Lisbon, Portugal Univ Nove Julho, Sao Paulo, Brazil Publ Hlth Agcy Canada, Ottawa, ON, Canada Canarian Hlth Serv, Tenerife, Spain Univ Ind Santander, Bucaramanga, Colombia Assoc Calabrese Epatol, Reggio Di Calabria, Italy Univ Minho, Braga, Portugal Fiji Natl Univ, Suva, Fiji Spanish Nutr Fdn, Madrid, Spain Publ Hlth Agcy Sweden, Stockholm, Sweden Inst Food Sci Natl Res Council, Avellino, Italy Sitaram Bhartia Inst Sci & Res, New Delhi, India Kindergarten Avlonari, Evia, Greece Natl Inst Hlth, Lima, Peru Minist Hlth, Jakarta, Indonesia Catalan Dept Hlth, Barcelona, Spain Biodonostia Hlth Res Inst, San Sebastian, Spain Inst Saude Ambiental, Lisbon, Portugal South Karelia Social & Hlth Care Dist, Lappeenranta, Finland McMaster Univ, Hamilton, ON, Canada Natl Canc Ctr, Tokyo, Japan Univ Sao Paulo Clin Hosp, Sao Paulo, Brazil Hosp Italiano Buenos Aires, Buenos Aires, DF, Argentina Rigshosp, Copenhagen, Denmark Acad Med Ctr Univ Amsterdam, Amsterdam, Netherlands Ctr Oral Hlth Serv & Res Midnorway, Trondheim, Norway Lagos State Univ, Coll Med, Lagos, Nigeria Univ Las Palmas Gran Canaria, Las Palmas Gran Canaria, Spain Comenius Univ, Bratislava, Slovakia Natl Ctr Dis Control & Publ Hlth, Tbilisi, Georgia Nippon Med Sch, Tokyo, Japan Sungkyunkwan Univ, Seoul, South Korea Finnish Inst Occupat Hlth, Helsinki, Finland Publ Hlth Promot & Dev Org, Kathmandu, Nepal St Vincents Hosp, Sydney, NSW, Australia Nes Municipal, Aarnes, Norway Hlth Polytech Jakarta II Inst, Jakarta, Indonesia Diponegoro Univ, Semarang, Indonesia Univ Bari, Bari, Italy Inst Reg Sante Publ, Ouidah, Benin Univ Bordeaux, Bordeaux, France Oslo Metropolitan Univ, Oslo, Norway Inst Publ Hlth, Skopje, North Macedonia Univ Leuven, Leuven, Belgium Lamprecht & Stamm Sozialforsch & Beratung AG, Zurich, Switzerland Inst Natl Sante & Rech Med, Nancy, France Univ Bonn, Bonn, Germany Sotiria Hosp, Sotiria, Greece Kalina Malina Kindergarten, Pazardjik, Bulgaria Jikei Univ, Sch Med, Tokyo, Japan Fu Jen Catholic Univ, Taipei, Taiwan Natl Stat Off, Praia, Cape Verde Natl Inst Publ Hlth Natl Inst Hyg, Warsaw, Poland Deakin Univ, Sydney, NSW, Australia Sci Res Inst Maternal & Child Hlth, Ashkhabad, Turkmenistan Univ Lincoln, Lincoln, England Minist Hlth, Amman, Jordan UNICEF, Niamey, Niger Cent Univ Kerala, Kasaragod, India Amsterdam Publ Hlth Res Inst, Amsterdam, Netherlands Hlth Serv Murcia, Murcia, Spain Inst Invest Sanitaria Illes Balears, Menorca, Spain Univ Bologna, Bologna, Italy Hellen Hlth Fdn, Athens, Greece Govt Med Coll, Bhavnagar, Gujarat, India Sefako Makgatho Hlth Sci Univ, Ga Rankuwa, South Africa Addis Ababa Univ, Addis Ababa, Ethiopia Minist Hlth, Wellington, New Zealand Univ Ctr Occidental Lisandro Alvarado, Barquisimeto, Venezuela Meharry Med Coll, Nashville, TN 37208 USA Dokuz Eylul Univ, Izmir, Turkey Univ Tampere, Tays Eye Ctr, Tampere, Finland Sabiha Gokcen Ilkokulu, Ankara, Turkey Polytech Inst Porto, Porto, Portugal Icahn Sch Med Mt Sinai, New York, NY 10029 USA Univ CEU San Pablo, Madrid, Spain CNR, Pisa, Italy Univ Miguel Hernandez, Alicante, Spain Vrije Univ Amsterdam, Amsterdam, Netherlands Sunflower Nursery Sch, Craiova, Romania North Karelia Ctr Publ Hlth, Joensuu, Finland Univ Witwatersrand, Johannesburg, South Africa Med Univ Vienna, Vienna, Austria Cork Inst Technol, Cork, Ireland Inst Med Res, Kuala Lumpur, Malaysia Xinjiang Med Univ, Urumqi, Peoples R China Shanghai Educ Dev Co Ltd, Shanghai, Peoples R China 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Compared to emblematic large carnivores, less charismatic or smaller carnivore species often receive little conservation attention, despite facing increasing anthropogenic pressure, and their status being poorly understood across much of their range. We employed systematic camera trapping and spatially explicit capture-recapture modelling to estimate variation in population density of serval, striped hyaena and aardwolf across the mixed-use Ruaha-Rungwa landscape in southern Tanzania. Three sites were selected as representative of different habitat types, management strategies, and levels of anthropogenic pressure: Ruaha National Park’s core tourist area, dominated by Acacia-Commiphora bushlands and thickets; the Park’s miombo woodland; and the neighbouring community-run MBOMIPA Wildlife Management Area, also covered in Acacia-Commiphora.Our results show that the Park’s miombo woodlands supported a higher serval density than the core tourist area and Wildlife Management Area, with 5.56 [Standard Error = ±2.45], 3.45 [±1.04] and 2.08 [±0.74] individuals per 100 km2, respectively. This variation is likely driven by precipitation, the abundance of apex predators, and the level of anthropogenic pressure.Striped hyaena were detected only in the Wildlife Management Area and at low density (1.36 [±0.5] individuals per 100 km2), potentially due to the location of the surveyed sites at the edge of the species’ global range, high densities of sympatric competitors and anthropogenic edge effects.Finally, aardwolf were captured in both the Park’s core tourist area and the Wildlife Management Area, with a higher density in the Wildlife Management Area (13.25 [±2.48] versus 9.19 [±1.66] individuals per 100 km2), possibly as a result of lower intraguild predation and late fire outbreaks in the area surveyed.By shedding light on three understudied African carnivore species, this study highlights the importance of miombo woodland conservation and community-managed conservation, as well as the value of camera trap data to improve ecological knowledge of lesser-studied carnivores.

Objectives Identification and validation of novel therapeutic targets is imperative to tackle the rise of drug resistance in tuberculosis. An essential Mur ligase-like gene (Rv3712), expected to be involved in cell-wall peptidoglycan (PG) biogenesis and conserved across mycobacteria, including the genetically depleted Mycobacterium leprae, was the primary focus of this study. Methods Biochemical analysis of Rv3712 was performed using inorganic phosphate release assays. The operon structure was identified using reverse-transcriptase PCR and a transcription/translation fusion vector. In vivo mycobacterial protein fragment complementation assays helped generate the interactome. Results Rv3712 was found to be an ATPase. Characterization of its operon revealed a mycobacteria-specific promoter driving the co-transcription of Rv3712 and Rv3713. The two gene products were found to interact with each other in vivo. Sequence-based functional assignments reveal that Rv3712 and Rv3713 are likely to be the mycobacterial PG precursor-modifying enzymes MurT and GatD, respectively. An in vivo network involving Mtb-MurT, regulatory proteins and cell division proteins was also identified. Conclusions Understanding the role of the enzyme complex in the context of PG metabolism and cell division, and the implications for antimicrobial resistance and host immune responses will facilitate the design of therapeutics that are targeted specifically to M. tuberculosis.