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Abstract In the field of tissue engineering, achieving precise spatiotemporal control over engineered cells is critical for sculpting functional 2D cell cultures into intricate morphological shapes. In this study, we engineer light-responsive mammalian cells and target them with dynamic light patterns to realize 2D cell culture patterning control. To achieve this, we developed μPatternScope (μPS), a modular framework for software-controlled projection of high-resolution light patterns onto microscope samples. μPS comprises hardware and software suite governing pattern projection and microscope maneuvers. Together with a 2D culture of the engineered cells, we utilize μPS for controlled spatiotemporal induction of apoptosis to generate desired 2D shapes. Furthermore, we introduce interactive closed-loop patterning, enabling a dynamic feedback mechanism between the measured cell culture patterns and the light illumination profiles to achieve the desired target patterning trends. Our work offers innovative tools for advanced tissue engineering applications through seamless fusion of optogenetics, optical engineering, and cybernetics.

Abstract Recent advances in tissue engineering have been remarkable, yet the precise control of cellular behavior in 2D and 3D cultures remains challenging. One approach to address this limitation is to genomically engineer optogenetic control of cellular processes into tissues using gene switches that can operate with only a few genomic copies. Here, we implement blue and red light-responsive gene switches to engineer genomically stable two- and three-dimensional mammalian tissue models. Notably, we achieve precise control of cell death and morphogen-directed patterning in 2D and 3D tissues by optogenetically regulating cell necroptosis and synthetic WNT3A signaling at high spatiotemporal resolution. This is accomplished using custom-built patterned LED systems, including digital mirrors and photomasks, as well as laser techniques. These advancements demonstrate the capability of precise spatiotemporal modulation in tissue engineering and open up new avenues for developing programmable 3D tissue and organ models, with significant implications for biomedical research and therapeutic applications.

Abstract Introduction Cervical cancer continues to pose a major public health challenge in low-income countries. Cervical cancer screening programs enable early detection and effectively reduce the incidence of cervical cancer as well as late-stage diagnosis and mortality. However, screening uptake remains suboptimal in Uganda. This study assessed correlates of intention to screen for cervical cancer among women in the Kyotera district of Central Uganda. Methods We analyzed cross-sectional data collected to determine the effectiveness of community audio towers (CATs) as a modality of health communication to support cervical cancer prevention. Women (n = 430) aged 21–60 years without a prior history of cervical cancer screening were surveyed about demographics, sources of health information and cervical cancer screening intentions in 2020. We used generalized linear modelling with modified Poisson regression and backwards variable elimination to identify adjusted prevalence ratios and 95% confidence intervals (CI) to determine factors associated with intention to screen for cervical cancer. Results Half (50.2%) of the participants had intentions to screen for cervical cancer within twelve months and 26.5% had moderate knowledge about cervical cancer. Nearly half (46.0%) considered themselves at risk of cervical cancer. Compared to residents who primarily received their health information from social media and radio, participants who received health information primarily from CATs (aPR:0.64, 95% CI:0.52–0.80, p

Abstract Background Interventions aimed at upstream factors contributing to late‐stage diagnoses could reduce disparities and improve breast cancer outcomes. This study examines the association between measures of housing stability and contemporary mortgage lending bias on breast cancer stage at diagnosis among older women in the United States. Methods We studied 67,588 women aged 66–90 from the SEER‐Medicare linked database (2010–2015). The primary outcome was breast cancer stage at diagnosis. Multinomial regression models adjusted for individual and neighborhood socio‐economic factors were performed using a three‐category outcome (stage 0, early stage, and late stage). Key census tract‐level independent variables were residence in the same house as the previous year, owner‐occupied homes, and an index of contemporary mortgage lending bias. Results In models adjusted for individual factors, higher levels of mortgage lending bias were associated with later stage diagnosis (RR = 1.10, 95% CI 1.02–1.20; RR = 1.31, 95% CI 1.16–1.49; RR = 1.41, 95% CI 1.24–1.60 for least to high, respectively). In models adjusted for individual and neighborhood socio‐economic factors, moderate and high levels of mortgage lending bias were associated with later stage diagnosis (RR = 1.16, 95% CI 1.02–1.33 for moderate and RR = 1.18, 95% CI 1.02–1.37 for high). Owner occupancy and tenure were not associated with later stage diagnosis in adjusted models. Conclusions Contemporary mortgage lending bias demonstrated a significant gradient relationship with later stage at diagnosis of breast cancer. Policy interventions aimed at reducing place‐based mortgage disinvestment and its impacts on local resources and opportunities should be considered as part of an overall strategy to decrease late‐stage breast cancer diagnosis and improve prognosis.

Abstract Background Intimate partner violence (IPV) remains a pervasive form of gender-based violence (GBV) that is largely undisclosed, especially among women seeking healthcare services in Uganda. Prioritizing survivor needs may improve IPV disclosure. This study explores healthcare worker experiences from provider-patient interactions with survivors seeking antenatal care services (ANC) in Uganda. Methods In-depth interviews were conducted among twenty-eight experienced healthcare providers in a rural and an urban-based ANC clinic in Eastern and Central Uganda. Providers were asked what they viewed as the needs and fears of women identified as having experienced any form of IPV. Iterative, inductive/deductive thematic analysis was conducted to discover themes regarding perceived needs, fears, and normalizing violence experienced by IPV survivors. Results According to healthcare providers, IPV survivors are unaware of available support services, and have need for support services. Providers reported that some survivors were afraid of the consequences of IPV disclosure namely, community stigma, worries about personal and their children’s safety, retaliatory abuse, fear of losing their marriage, and partners’ financial support. Women survivors also blamed themselves for IPV. Contextual factors underlying survivor concerns included the socio-economic environment that ‘normalizes’ violence, namely, some cultural norms condoning violence, and survivors’ unawareness of their human rights due to self-blame and shame for abuse. Conclusions We underscore a need to empower IPV survivors by prioritizing their needs. Results highlight opportunities to create a responsive healthcare environment that fosters IPV disclosure while addressing survivors’ immediate medical and psychosocial needs, and safety concerns. Our findings will inform GBV prevention and response strategies that integrate survivor-centered approaches in Uganda.

Abstract NADP(H) is a central metabolic hub providing reducing equivalents to multiple biosynthetic, regulatory and antioxidative pathways in all living organisms. While biosensors are available to determine NADP+ or NADPH levels in vivo, no probe exists to estimate the NADP(H) redox status, a determinant of the cell energy availability. We describe herein the design and characterization of a genetically-encoded ratiometric biosensor, termed NERNST, able to interact with NADP(H) and estimate E NADP(H). NERNST consists of a redox-sensitive green fluorescent protein (roGFP2) fused to an NADPH-thioredoxin reductase C module which selectively monitors NADP(H) redox states via oxido-reduction of the roGFP2 moiety. NERNST is functional in bacterial, plant and animal cells, and organelles such as chloroplasts and mitochondria. Using NERNST, we monitor NADP(H) dynamics during bacterial growth, environmental stresses in plants, metabolic challenges to mammalian cells, and wounding in zebrafish. NERNST estimates the NADP(H) redox poise in living organisms, with various potential applications in biochemical, biotechnological and biomedical research.

PurposePlace-based measures of structural racism have been associated with breast cancer mortality, which may be driven, in part, by epigenetic perturbations. We examined the association between contemporary redlining, a measure of structural racism at the neighborhood level, and DNA methylation in breast tumor tissue.MethodsWe identified 80 Black and White women diagnosed and treated for a first-primary breast cancer at Emory University Hospitals (2008–2017). Contemporary redlining was derived for census tracts using the Home Mortgage Disclosure Act database. Linear regression models were used to calculate the association between contemporary redlining and methylation in breast tumor tissue. We also examined epigenetic age acceleration for two different metrics, regressing β values for each cytosine-phosphate-guanine dinucleotide (CpG) site on redlining while adjusting for covariates. We employed multivariable Cox-proportional hazards models and 95% confidence intervals (CI) to estimate the association between aberrant methylation and mortality.ResultsContemporary redlining was associated with 5 CpG sites after adjustment for multiple comparisons (FDR

Understanding and forecasting human mobility in response to climatic and environmental changes has become a subject of substantial political, societal, and academic interest. Quantitative models exploring the relationship between climatic factors and migration patterns have been developed since the early 2000s; however, different models have produced results that are not always consistent with one another or robust enough to provide actionable insights into future dynamics. Here we examine weaknesses of classical methods and identify next-generation approaches with the potential to close existing knowledge gaps. We propose six priorities for the future of climate mobility modeling: (i) the use of non-linear machine-learning rather than linear methods, (ii) the prioritization of explaining the observed data rather than testing statistical significance of predictors, (iii) the consideration of relevant climate impacts rather than temperature- and precipitation-based metrics, (iv) the examination of heterogeneities, including across space and demographic groups rather than aggregated measures, (v) the investigation of temporal migration dynamics rather than essentially spatial patterns, (vi) the use of better calibration data, including disaggregated and within-country flows. Improving both methods and data to accommodate the high complexity and context-specificity of climate mobility will be crucial for establishing the scientific consensus on historical trends and future projections that has eluded the discipline thus far.

The objective of the study was to present a novel analytical approach using Nigerian trauma registry data to determine risks and of road traffic crash-related mortality for patients treated at National Trauma Centre, Abuja. Patient characteristics were compared between those who died at the hospital (n = 118) and those who survived (n = 2018). Multiple logistic regression and cluster analyses were used to identify risks of mortality and of high mortality and injury rates. The patient’s status as pedestrian (p

African paleoanthropological studies typically focus on regions of the continent such as Eastern, Southern and Northern Africa, which hold the highest density of Pleistocene archaeological sites. Nevertheless, lesser known areas such as West Africa also feature a high number of sites. Here, we present a high-resolution map synthesising all well contextualised Pleistocene archaeological sites present in Sub-Saharan West Africa. A detailed elevation and ecoregional map was developed and correlated with palaeoanthropological sites. This map is supplemented with 1,000- and 2000-year interval climate reconstructions over the last 120,000 years for three subregions of high archaeological interest. The presented archaeological sites were compiled by reviewing published literature, and selected based on: (1) documented archaeological stratification or >10 characteristic artefacts, (2) published coordinates, and (3) published chronometric ages or relative dating. The data presented here elucidates the current state of knowledge of Pleistocene West Africa, highlighting the regional potential for human evolutionary studies.

Abstract Background Uganda clinical guidelines recommend routine screening of pregnant women for intimate partner violence (IPV) during antenatal care (ANC). Healthcare providers play a critical role in identifying IPV during pregnancy in ANC clinics. This study explored facilitators and barriers for IPV screening during pregnancy (perinatal IPV screening) by ANC-based healthcare workers in Uganda. Methods We conducted qualitative in-depth interviews among twenty-eight purposively selected healthcare providers in one rural and an urban-based ANC health center in Eastern and Central Uganda respectively. Barriers and facilitators to IPV screening during ANC were identified iteratively using inductive-deductive thematic analysis. Results Participants had provided ANC services for a median (IQR) duration of 4.0 (0.1–19) years. Out of 28 healthcare providers, 11 routinely screened women attending ANC clinics for IPV and 10 had received IPV-related training. Barriers to routine IPV screening included limited staffing and space resources, lack of comprehensive gender-based violence (GBV) training and provider unawareness of the extent of IPV during pregnancy. Facilitators were availability of GBV protocols and providers who were aware of IPV (or GBV) tools tended to use them to routinely screen for IPV. Healthcare workers reported the need to establish patient trust and a safe ANC clinic environment for disclosure to occur. ANC clinicians suggested creation of opportunities for triage-level screening and modification of patients’ ANC cards used to document women’s medical history. Some providers expressed concerns of safety or retaliatory abuse if perpetrating partners were to see reported abuse. Conclusions Our findings can inform efforts to strengthen GBV interventions focused on increasing routine perinatal IPV screening by ANC-based clinicians. Implementation of initiatives to increase routine perinatal IPV screening should focus on task sharing, increasing comprehensive IPV training opportunities, including raising awareness of IPV severity, trauma-informed care and building trusting patient-physician relationships.

Optimising the spatial distribution of global croplands could substantially reduce carbon emissions and biodiversity loss associated with rain-fed crop production, according to a mathematical framework applied to environmental impact and crop yield data

Abstract The major social and economic impacts of international migration have led to a strong interest in better understanding the drivers of cross-border movement. Quantitative models have sought to explain global migration patterns in terms of economic, social, climatic, and other variables, and future projections of these variables are increasingly being used to forecast international migration flows. An important implicit assumption in the most widely used class of these approaches, so-called gravity models, is that their parameterisation based on panel data enables them to describe the effects of predictor variables on migration flows across both space and time, i.e., that they explain flow variation both across country pairs at a given time and across time for a given country pair. Here we show that this assumption does not hold. Whilst gravity models describe spatial patterns of international migration very well, they fail to capture even basic temporal dynamics, indeed, often worse than even the time-invariant average of the historical flows. We show that standard validation techniques have been unable to detect this important limitation of gravity models due to the different orders of magnitude of migration flows across spatial corridors, on the one hand, and over time, on the other hand. Our analysis suggests that gravity-model-based inferences about the effects that certain variables have had, or will have, on international migration over time may in reality represent statistical artefacts rather than true mechanisms. We argue that future predictions based on gravity models lack statistical support and that, in its current form, this class of models is not suited for informing policy makers about migration trajectories in the coming years and decades.

Significance Globally, cardiovascular disease (CVD) and cancer are leading causes of morbidity and mortality. While having different etiologies, CVD and cancer are linked by multiple shared risk factors, the presence of which exacerbate adverse outcomes for individuals with either disease. For both pathologies, factors such as poverty, lack of physical activity (PA), poor dietary intake, and climate change increase risk of adverse outcomes. Prior research has shown that greenspaces and other nature-based interventions (NBIs) contribute to improved health outcomes and climate change resilience. Objective To summarize evidence on the impact of greenspaces or NBIs on cardiovascular health and/or cancer-related outcomes and identify knowledge gaps to inform future research. Methods Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 and Peer Review of Electronic Search Strategies (PRESS) guidelines, we searched five databases: Web of Science, Scopus, Medline, PsycINFO and GreenFile. Two blinded reviewers used Rayyan AI and a predefined criteria for article inclusion and exclusion. The risk of bias was assessed using a modified version of the Newcastle–Ottawa Scale (NOS). This review is registered with PROSPERO, ID # CRD42021231619. Results & discussion Of 2565 articles retrieved, 31 articles met the inclusion criteria, and overall had a low risk of bias. 26 articles studied cardiovascular related outcomes and 5 studied cancer-related outcomes. Interventions were coded into 4 categories: forest bathing, green exercise, gardening, and nature viewing. Outcomes included blood pressure (BP), cancer-related quality of life (QoL) and (more infrequently) biomarkers of CVD risk. Descriptions of findings are presented as well as visual presentations of trends across the findings using RAW graphs. Overall studies included have a low risk of bias; and alluvial chart trends indicated that NBIs may have beneficial effects on CVD and cancer-related outcomes. Conclusions & implications (1)Clinical implication: Healthcare providers should consider the promotion of nature-based programs to improve health outcomes. (2)Policy implication: There is a need for investment in equitable greenspaces to improve health outcomes and build climate resilient neighborhoods. (3)Research or academic implication: Research partnerships with community-based organizations for a comprehensive study of benefits associated with NBIs should be encouraged to reduce health disparities and ensure intergenerational health equity. There is a need for investigation of the mechanisms by which NBIs impact CVD and exploration of the role of CVD biological markers of inflammation among cancer survivors.

It is still unclear when and by which route modern humans expanded out of Africa. Here, Beyer et al. use paleoclimate reconstructions and estimates of human precipitation requirements to evaluate the survivability of spatial and temporal migration corridors to Eurasia over the last 300,000 years.

Measurement(s) temperature • precipitation • cloud cover • net primary productivity Technology Type(s) computational modeling technique Factor Type(s) atmospheric carbon dioxide • orbital parameters • land surface type (ocean, land, land ice) Sample Characteristic - Environment climate system Sample Characteristic - Location Earth (planet) Machine-accessible metadata file describing the reported data: https://doi.org/10.6084/m9.figshare.15073479

We examined the use of “edge of flood” habitats by larval silver carp during the extensive flooding that occurred in Pool 26 of the Upper Mississippi River near Alton, IL, USA and St. Louis, MO, USA during the summer of 2015. We captured over 12,700 individual fishes including eight taxa, over 12,000 of which (> 95%) were larval silver carp between 5 and 21 mm standard length. Peak catch rates occurred near the confluence of the Illinois and Mississippi rivers. These findings suggest that larval silver carp have an affinity for edge of the flood habitat and further study is needed to better understand how this affects the dynamics of this invasive species in the Mississippi River. The high catch rates observed at the confluence of the Illinois and Mississippi rivers suggests that the Illinois River is an important source of larval silver carp to the Upper Mississippi River.

Long-term dynamics of species’ range sizes play a crucial role in determining extinction risks. Here the authors simulate global vegetation cover and scenarios of anthropogenic land cover change to estimate habitat range sizes of thousands of mammal, bird, and amphibian species since 1700, and project trajectories up to 2100 under four emission scenarios and five socio-economic pathways.

The number of ecohydrological studies involving water stable isotope measurements has been increasing steadily due to technological (e.g., field-deployable laser spectroscopy and cheaper instruments) and methodological (i.e., tracer approaches or improvements in root water uptake models) advances in recent years. This enables researchers from a broad scientific background to incorporate water-isotope-based methods into their studies. Several isotope effects are currently not fully understood but might be essential when investigating root water uptake depths of vegetation and separating isotope processes in the soil–vegetation–atmosphere continuum. Different viewpoints exist on (i) extraction methods for soil and plant water and methodological artifacts potentially introduced by them, (ii) the pools of water (mobile vs. immobile) measured with those methods, and (iii) spatial variability and temporal dynamics of the water isotope composition of different compartments in terrestrial ecosystems. In situ methods have been proposed as an innovative and necessary way to address these issues and are required in order to disentangle isotope effects and take them into account when studying root water uptake depths of plants and for studying soil–plant–atmosphere interaction based on water stable isotopes. Herein, we review the current status of in situ measurements of water stable isotopes in soils and plants, point out current issues and highlight the potential for future research. Moreover, we put a strong focus and incorporate practical aspects into this review in order to provide a guideline for researchers with limited previous experience with in situ methods. We also include a section on opportunities for incorporating data obtained with described in situ methods into existing isotope-enabled ecohydrological models and provide examples illustrating potential benefits of doing so. Finally, we propose an integrated methodology for measuring both soil and plant water isotopes in situ when carrying out studies at the soil–vegetation–atmosphere continuum. Several authors have shown that reliable data can be generated in the field using in situ methods for measuring the soil water isotope composition. For transpiration, reliable methods also exist but are not common in ecohydrological field studies due to the required effort. Little attention has been paid to in situ xylem water isotope measurements. Research needs to focus on improving and further developing those methods. There is a need for a consistent and combined (soils and plants) methodology for ecohydrological studies. Such systems should be designed and adapted to the environment to be studied. We further conclude that many studies currently might not rely on in situ methods extensively because of the technical difficulty and existing methodological uncertainties. Future research needs to aim on developing a simplified approach that provides a reasonable trade-off between practicability and precision and accuracy.

Background: The burden of cancer in Africa is growing. Although cancer outcomes are understood as the product of influences at multiple socioecological levels, population-based studies of geographical factors and cancer outcomes in Africa are scarce. The objective of this study was to identify parish-level social factors associated with cervical cancer incidence in the Kampala Cancer Registry catchment area, using a novel linkage between population-based cancer registry data and small-area census data from Uganda. Methods: Kampala Cancer Registry cervical cancer records (2008–15) were augmented to add the parish of residence at diagnosis. Parish-level population and housing profile data (2014) were obtained from the Uganda Bureau of Statistics and linked to Kampala Cancer Registry records. Stepwise forward Poisson regression modelling was used to estimate incidence ratios (IR) assessing associations between social factors and incidence. Housing tenure, infrastructure, gender equality, economic status, and employment were examined, controlling for population density. The significance level was set at α=0·05. Findings: Factors related to higher incidence included a higher girl-to-boy ratio of 6–12-year-olds not attending school (IR 1·33 [95% CI 1·15–1·54]; p

Inteins catalyze their removal from a host protein through protein splicing. Inteins that contain an additional site-specific endonuclease domain display genetic mobility via a process termed “homing” and thereby act as selfish DNA elements. We elucidated the crystal structures of two archaeal inteins associated with an active or inactive homing endonuclease domain. This analysis illustrated structural diversity in the accessory domains (ACDs) associated with the homing endonuclease domain. To augment homing endonucleases with highly specific DNA cleaving activity using the intein scaffold, we engineered the ACDs and characterized their homing site recognition. Protein engineering of the ACDs in the inteins illuminated a possible strategy for how inteins could avoid their extinction but spread via the acquisition of a diverse accessory domain.

It is unclear whether bird migration patterns are restricted to interglacial periods or are maintained during glacial maxima. Somveille et al. apply a global migration simulation model to climate reconstruction to show that the prevalence of this phenomenon has likely been largely maintained up to 50,000 years ago.

Abstract Objective The objective of this study was to measure flow‐mediated dilation (FMD) prior to and following transient increases in intraluminal pressure (IILP) in resistance arterioles isolated from subjects with and without coronary artery disease (CAD) (CAD and non‐CAD) and non‐CAD subjects with hypertension. Methods Arterioles were isolated from discarded surgical tissues (adipose and atrial) from patients without coronary artery disease (non‐CAD; ≤1 risk factor, excluding hypertension), with CAD, and non‐CAD patients with hypertension (hypertension as the only risk factor). To simulate transient hypertension, increased IILP was generated (150 mmHg, 30 min) by gravity. Arterioles were constricted with endothelin‐1, followed by FMD and endothelial‐independent dilation prior to and following exposure to IILP. Results IILP reduced FMD in non‐CAD and CAD arterioles relative to pre‐IILP (p

The growing understanding of partially unfolded proteins increasingly points to their biological relevance in allosteric regulation, complex formation, and protein design. However, the structural characterization of disordered proteins remains challenging. NMR methods can access both the dynamics and structures of such proteins, yet suffering from a high degeneracy of NMR signals. Here, we overcame this bottleneck utilizing a salt-inducible split intein to produce segmentally isotope-labeled samples with the native sequence, including the ligation junction. With this technique, we investigated the NMR structure and conformational dynamics of TonB from Helicobacter pylori in the presence of a proline-rich low complexity region. Spin relaxation experiments suggest that the several nano-second time scale dynamics of the C-terminal domain (CTD) is almost independent of the faster pico-to-nanosecond dynamics of the low complexity central region (LCCR). Our results demonstrate the utility of segmental isotopic labeling for proteins with heterogenous dynamics such as TonB and could advance NMR studies of other partially unfolded proteins.

In this commentary, we summarize and build upon discussions that emerged during the workshop Isotope-based studies of water partitioning and plant–soil interactions in forested and agricultural environments held in San Casciano in Val di Pesa, Italy, in September 2017. Quantifying and understanding how water cycles through the Earth's critical zone is important to provide society and policymakers with the scientific background to manage water resources sustainably, especially considering the ever-increasing worldwide concern about water scarcity. Stable isotopes of hydrogen and oxygen in water have proven to be a powerful tool for tracking water fluxes in the critical zone. However, both mechanistic complexities (e.g. mixing and fractionation processes, heterogeneity of natural systems) and methodological issues (e.g. lack of standard protocols to sample specific compartments, such as soil water and xylem water) limit the application of stable water isotopes in critical-zone science. In this commentary, we examine some of the opportunities and critical challenges of isotope-based ecohydrological applications and outline new perspectives focused on interdisciplinary research opportunities for this important tool in water and environmental science.

The rural population in parts of the Cuvelai-Etosha Basin (CEB) in Namibia depends on groundwater as a source for drinking and livestock watering. The aim of this study is to investigate spatial-temporal patterns for understanding water origins and recharge processes of perched aquifers in the CEB. The study uses hydrochemical data and water stable isotope signatures (18O and 2H) of samples collected during (10) ten field campaigns over a three-year period (from 2014 to 2016) originating from two regions within the basin (Ohangwena and Omusati). A clear distinction between the shallow groundwater in the Omusati and Ohangwena regions documented by TDS values, hydrochemical water types as well as the isotopic compositions of water. These differences are influenced mainly by their location in the landscape, depth to water table, the type of well infrastructure and aquifer material. Spatial and temporal variations indicate that even though these perched aquifers are both within the same basin, they are chemically different because of the rock materials in which their flow. Therefore, these differences in the water hydrochemical composition as well as the processes governing perched aquifers must be taken into account when planning groundwater management in the basin.

We recently discovered a new groundwater age tracer, Halon-1301, which can be used to date groundwater recharged after the 1970s. In a previous study, we showed that Halon-1301 reliably inferred groundwater age at the majority of groundwater sites studied. At those sites, ages inferred from Halon-1301 agreed with those inferred from SF6 and tritium, two reliable widely applied groundwater age tracers. A few samples, however, showed reduced concentrations of Halon-1301, preventing meaningful age interpretation from its concentration. These reduced concentrations were likely a result of degradation or retardation of Halon-1301 in the aquifer. However, we could not provide full evidence for this due to the limited number of groundwater samples analysed (18 in total). In this study, we assess the potential of Halon-1301 as a groundwater age tracer for a larger dataset of groundwater samples under specific groundwater conditions, including highly anoxic young groundwater which can significantly degrade Halon-1301, to gain more information on the magnitude of occurrence and the causes of reduced Halon-1301 concentrations. In this study, we analysed 302 groundwater samples for Halon-1301, SF6, tritium and the CFCs CFC-11, CFC-12 and CFC-113. Comparison of age information inferred from the concentrations of these tracers allows assessment of the performance of Halon-1301 compared to other well established and widely used age tracers. The samples are taken from different groundwater environments in New Zealand and include anoxic and oxic waters with mean residence times ranging from The majority of assessed samples have reduced or elevated concentrations of CFCs, which makes it impossible to infer a reliable age using the CFCs for these samples. Halon-1301, however, reliably infers ages for CFC-contaminated waters. Three other groundwater samples were found to have elevated SF6 concentrations (contaminated). Again, at these SF6-contaminated sites, ages inferred from Halon-1301 agree with ages inferred from tritium. A few samples (14 sites) exhibit reduced concentrations of Halon-1301, which result in elevated inferred Halon-1301 ages in comparison to those inferred from SF6, tritium and/or CFC-113. Assessment of the groundwater environment at these sites gives further insight into the potential causes of Halon-1301 reduction in groundwater. Overall, Halon-1301 gives age information that matches ages inferred from SF6 and/or tritium for the majority (97 %) of the assessed groundwater sites. These findings suggest that Halon-1301 is a reasonably reliable groundwater age tracer, and is in particular significantly more reliable than the CFCs, which may have contamination and degradation problems. Halon-1301 thus has potential to become a useful groundwater age tracer where SF6 and the CFCs are compromised, and where additional independent tracers are needed to constrain complex mixing models.

Introduction: Elevated levels of mitochondrial reactive oxygen species (ROS) contribute to the development of numerous cardiovascular diseases. TERT, the catalytic subunit of telomerase, has been shown to translocate to mitochondria to suppress ROS while promoting ATP production. Acute overexpression of TERT increases survival and decreases infarct size in a mouse model of myocardial infarct, while decreased telomerase activity predisposes to mitochondrial defects and heart failure. In the present study, we examined the role of TERT on cardiac structure and function under basal conditions and conditions of acute or prolonged stress in a novel rat model of TERT deficiency.Methods: Cardiac structure and function were evaluated via transthoracic echocardiogram. Langendorff preparations were used to test the effects of acute global ischemia reperfusion injury on cardiac function and infarction. Coronary flow and left ventricular pressure were measured during and after ischemia/reperfusion (I/R). Mitochondrial DNA integrity was measured by PCR and mitochondrial respiration was assessed in isolated mitochondria using an Oxygraph. Angiotensin II infusion was used as an established model of systemic stress.Results: No structural changes (echocardiogram) or coronary flow/left ventricle pressure (isolated hearts) were observed in TERT−/− rats at baseline; however, after I/R, coronary flow was significantly reduced in TERT−/− compared to wild type (WT) rats, while diastolic Left Ventricle Pressure was significantly elevated (n = 6 in each group; p < 0.05) in the TERT−/−. Interestingly, infarct size was less in TERT−/− rats compared to WT rats, while mitochondrial respiratory control index decreased and mitochondrial DNA lesions increased in TERT−/− compared to WT. Angiotensin II treatment did not alter cardiac structure or function; however, it augmented the infarct size significantly more in TERT−/− compared to the WT.Conclusion: Absence of TERT activity increases susceptibility to stress like cardiac injury. These results suggest a critical role of telomerase in chronic heart disease.

Inteins are prevalent among extremophiles. Mini-inteins with robust splicing properties are of particular interest for biotechnological applications due to their small size. However, biochemical and structural characterization has still been limited to a small number of inteins, and only a few serve as widely used tools in protein engineering. We determined the crystal structure of a naturally occurring Pol-II mini-intein from Pyrococcus horikoshii and compared all three mini-inteins found in the genome of P. horikoshii. Despite their similar sizes, the comparison revealed distinct differences in the insertions and deletions, implying specific evolutionary pathways from distinct ancestral origins. Our studies suggest that sporadically distributed mini-inteins might be more promising for further protein engineering applications than highly conserved mini-inteins. Structural investigations of additional inteins could guide the shortest path to finding novel robust mini-inteins suitable for various protein engineering purposes.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Stable isotopes (deuterium, 2H, and oxygen-18, 18O) of soil water were measured in the field using a liquid water isotope analyzer (tunable off-axis integrated cavity output spectroscope, OA-ICOS, LGR) and commercially available soil gas probes (BGL-30, UMS, Munich) in the semi-arid Cuvelai–Etosha Basin (CEB), Namibia. Results support the applicability of an in situ measurement system for the determination of stable isotopes in soil pore water. High spatial and temporal resolution was achieved in the study area with reasonable accuracy and measurements were in agreement with laboratory-based cryogenic vacuum extraction and subsequent cavity ring-down laser spectroscopic isotope analysis (CRDS, L2120-i, Picarro Inc.). After drift and span correction of the in situ isotope data, precision for over 140 measurements taken during two consecutive field campaigns (June and November 2014) was 1.8 and 0.48 ‰ for δ2H and δ18O, respectively. Mean measurement trueness is determined using quality check standards and was 5 and 0.3 ‰ for δ2H and δ18O, respectively. The isotope depth profiles are used quantitatively to calculate a soil water balance. The contribution of transpiration to total evapotranspiration ranged between 72 and 92 %. Shortly after a rain event, the contribution of transpiration was much lower, at 35 to 50 %. Potential limitations of such an in situ system are related to environmental conditions which could be minimized by using a temperature-controlled chamber for the laser spectrometer. Further, the applicability of the system using previously oven-dried soil material might be limited by physicochemical soil properties (i.e., clay minerals). Uncertainty in the in situ system is suggested to be reduced by improving the calibration procedure and further studying fractionation effects influencing the isotope ratios in the soil water, especially at low water contents. Furthermore, the influence of soil-respired CO2 on isotope values within the root zone could not be deduced from the data.