Microbial Ecology

Phenotyping Thermal Responses of Yeasts and Yeast-like Microorganisms at the Individual and Population Levels: Proof-of-Concept, Development and Application of an Experimental Framework to a Plant Pathogen Abstract Deciphering the responses of microbial populations to spatiotemporal changes in their thermal environment is instrumental in improving our understanding of their eco-evolutionary dynamics. Recent studies have shown that current phenotyping protocols do not adequately address all dimensions of phenotype expression. Therefore, these methods can give biased assessments of sensitivity to temperature, leading to misunderstandings concerning the ecological processes underlying thermal plasticity. We describe here a new robust and versatile experimental framework for the accurate investigation of thermal performance and phenotypic diversity in yeasts and yeast-like microorganisms, at the individual and population levels. In addition to proof-of-concept, the application of this framework to the fungal wheat pathogen Zymoseptoria tritici resulted in detailed characterisations for this yeast-like microorganism of (i) the patterns of temperature-dependent changes in performance for four fitness traits; (ii) the consistency in thermal sensitivity rankings of strains between in plantaand in vitro growth assessments; (iii) significant interindividual variation in thermal responses, with four principal thermotypes detected in a sample of 66 strains; and (iv) the ecological consequences of this diversity for population-level processes through pairwise competition experiments highlighting temperature-dependent outcomes. These findings extend our knowledge and ability to quantify and categorise the phenotypic heterogeneity of thermal responses. As such, they lay the foundations for further studies elucidating local adaptation patterns and the effects of temperature variations on eco-evolutionary and epidemiological processes.

Prokaryotic Diversity and Distribution in Different Habitats of an Alpine Rock Glacier-Pond System Abstract Rock glaciers (RG) are assumed to influence the biogeochemistry of downstream ecosystems because of the high ratio of rock:water in those systems, but no studies have considered the effects of a RG inflow on the microbial ecology of sediments in a downstream pond. An alpine RG-pond system, located in the NW Italian Alps has been chosen as a model, and Bacteria and Archaea 16S rRNA genes abundance, distribution and diversity have been assessed by qPCR and Illumina sequencing, coupled with geochemical analyses on sediments collected along a distance gradient from the RG inflow. RG surface material and neighbouring soil have been included in the analysis to better elucidate relationships among different habitats. Our results showed that different habitats harboured different, well-separated microbial assemblages. Across the pond, the main variations in community composition (e.g. Thaumarchaeota and Cyanobacteria relative abundance) and porewater geochemistry (pH, DOC, TDN and NH4+) were not directly linked to RG proximity, but to differences in water depth. Some microbial markers potentially linked to the presence of meltwater inputs from the RG have been recognised, although the RG seems to have a greater influence on the pond microbial communities due to its contribution in terms of sedimentary material.

Divergence in Diversity and Composition of Root-Associated Fungi Between Greenhouse and Field Studies in a Semiarid Grassland Abstract Investigations of plant-soil feedbacks (PSF) and plant-microbe interactions often rely exclusively on greenhouse experiments, yet we have little understanding of how, and when, results can be extrapolated to explain phenomena in nature. A systematic comparison of microbial communities using the same host species across study environments can inform the generalizability of such experiments. We used Illumina MiSeq sequencing to characterize the root-associated fungi of two foundation grasses from a greenhouse PSF experiment, a field PSF experiment, field monoculture stands, and naturally occurring resident plants in the field. A core community consisting < 10% of total fungal OTU richness but > 50% of total sequence abundance occurred in plants from all study types, demonstrating the ability of field and greenhouse experiments to capture the dominant component of natural communities. Fungal communities were plant species-specific across the study types, with the core community showing stronger host specificity than peripheral taxa. Roots from the greenhouse and field PSF experiments had lower among sample variability in community composition and higher diversity than those from naturally occurring, or planted monoculture plants from the field. Core and total fungal composition differed substantially across study types, and dissimilarity between fungal communities did not predict plant-soil feedbacks measured in experiments. These results suggest that rhizobiome assembly mechanisms in nature differ from the dynamics of short-term, inoculation studies. Our results validate the efficacy of common PSF experiment designs to test soil inoculum effects, and highlight the challenges of scaling the underlying microbial mechanisms of plant responses from whole-community inoculation experiments to natural ecosystems.

Adaptation to Fasting in Crucian Carp ( Carassius auratus ): Gut Microbiota and Its Correlative Relationship with Immune Function Abstract Fasting influences the overall physiology of fish, and the knowledge how the gut microbiota, growth performances, and immune function in response to intermittent and long-term fasting is still insufficient. Here, we characterized the effects of fasting on the host-gut microbiota in crucian carp, which would enhance our insight into physiological adaptation to fasting. To achieve this, we investigated the gut microbial communities of crucian carp with different fasting stress, and corresponding immune and growth parameters. The gut microbial communities were structured into four clusters according to different fasting stress, namely one control group (feed regularly), two intermittent fasting groups (fasting period and re-feeding period, respectively), and one long-term fasting group. Intermittent fasting significantly improved the activity of superoxide dismutase (SOD) and lysozyme (LZM) (ANOVA, p < 0.05) and significantly increased alpha diversity and ecosystem stability of gut microbiota (ANOVA, p < 0.05). Gut length (GL) and condition factor (CF) showed no significant difference between the control group (CG) and intermittent fasting group under re-feeding period (RIF) (ANOVA, p = 0.11), but relative gut length (RGL) in group RIF was higher than that in the CG (ANOVA, p = 0.00). The bacterial genera Bacteroides, Akkermansia, and Erysipelotrichaceae were enriched in fishes under intermittent fasting. Two Bacteroides OTUs (OTU50 and OTU1292) correlated positively with immune (SOD, complement, and LZM) and growth (GL and RGL) parameters. These results highlight the possible interplay between growth performances, immune function, and gut microbiota in response to fasting.

The Influence of Habitat and Phylogeny on the Skin Microbiome of Amphibians in Guatemala and Mexico Abstract Microbial symbionts are increasingly recognized as playing a critical role in organismal health across a wide range of hosts. Amphibians are unique hosts in that their skin helps to regulate the exchange of water, ions, and gases, and it plays an active role in defense against pathogens through the synthesis of anti-microbial peptides. The microbiome of amphibian skin includes a diverse community of bacteria known to defend against pathogens, including the global pandemic lineage of Batrachochytrium dendrobatidis associated with mass amphibian die-offs. The relative influence of host phylogeny and environment in determining the composition of the amphibian skin microbiome remains poorly understood. We collected skin swabs from montane amphibians in Mexico and Guatemala, focusing on two genera of plethodontid salamanders and one genus of frogs. We used high throughput sequencing to characterize the skin bacterial microbiome and tested the impact of phylogeny and habitat on bacterial diversity. Our results show that phylogenetic history strongly influences the diversity and community structure of the total bacterial microbiome at higher taxonomic levels (between orders), but on lower scales (within genera and species), the effect of habitat predominates. These results add to a growing consensus that habitat exerts a strong effect on microbiome structure and composition, particularly at shallow phylogenetic scales.

Comamonadaceae OTU as a Remnant of an Ancient Microbial Community in Sulfidic Waters Abstract Intraterrestrial waters harbor microbial communities being extensively studied to understand microbial processes underlying subsurface ecosystem functioning. This paper provides the results of an investigation on the microbiomes of unique, subsurface sulfidic waters associated with Upper Jurassic, Cretaceous, and Miocene sediments. We used high-throughput 16S rDNA amplicon sequencing to reveal the structure of bacterial and archaeal communities in water samples differing in sulfide content (20–960 mg/dm3), salinity (1.3–3.2%), and depth of extraction (60–660 m below ground level). Composition of the bacterial communities strongly varied across the samples; however, the bacteria participating in the sulfur cycle were common in all sulfidic waters. The shallowest borehole water (60 m bgl) was dominated by sulfur-oxidizing Epsilonproteobacteria(Sulfurimonas, Sulfurovum). In the waters collected from greater depths (148–300 m bgl), the prevalence of Betaproteobacteria (Comamonadaceae) and sulfate/sulfur-reducing Deltaproteobacteria (Desulfopila, Desulfomicrobium, MSBL7) was observed. Sulfate reducers (members of Clostridia: Candidatus Desulforudis) were the most abundant bacteria in the deepest borehole water (660 m bgl). Out of 850 bacterial OTUs, only one, affiliated with the Comamonadaceae family, was found abundant (> 1% of total bacterial sequences) in all samples. Contribution of Archaea to the whole microbial communities was lower than 0.5%. Archaeal communities did not differ across the samples and they consisted of Halobacteriaceae. Out of 372 archaeal OTUs, five, belonging to the four genera Natronomonas, Halorubrum, Halobellus, and Halorhabdus, were the most numerous.

Fungal, Bacterial, and Archaeal Diversity in Soils Beneath Native and Introduced Plants in Fiji, South Pacific Abstract The Fiji Islands is an archipelago of more than 330 islands located in the tropics of the South Pacific Ocean. Microbial diversity and biogeography in this region is still not understood. Here, we present the first molecular characterization of fungal, bacterial, and archaeal communities in soils from different habitats within the largest Fijian island, Viti Levu. Soil samples were collected from under native vegetation in maritime-, forest-, stream-, grassland-, and casuarina-dominated habitats, as well as from under the introduced agricultural crops sugarcane, cassava, pine, and mahogany. Soil microbial diversity was analyzed through MiSeq amplicon sequencing of 16S (for prokaryotes), ITS, LSU ribosomal DNA (for fungi). Prokaryotic communities were dominated by Proteobacteria (~ 25%), Acidobacteria (~ 19%), and Actinobacteria (~ 17%), and there were no indicator species associated with particular habitats. ITS and LSU were congruent in β-diversity patterns of fungi, and fungal communities were dominated by Ascomycota (~ 57–64%), followed by Basidiomycota (~ 20–23%) and Mucoromycota (~ 10%) according to ITS, or Chytridiomycota (~ 9%) according to LSU. Indicator species analysis of fungi found statistical associations of Cenococcum, Wilcoxina, and Rhizopogon to Pinus caribaea. We hypothesize these obligate biotrophic fungi were co-introduced with their host plant. Entoloma was statistically associated with grassland soils, and Fusarium and Lecythophora with soils under cassava. Observed richness varied from 65 (casuarina) to 404 OTUs (cassava) for fungi according to ITS region, and from 1268 (pine) to 2931 OTUs (cassava) for bacteria and archaea. A major finding of this research is that nearly 25% of the fungal OTUs are poorly classified, indicative of novel biodiversity in this region. This preliminary survey provides important baseline data on fungal, bacterial, and archaeal diversity and biogeography in the Fiji Islands.

Detecting Associations Between Ciliated Protists and Prokaryotes with Culture-Independent Single-Cell Microbiomics: a Proof-of-Concept Study Abstract Symbioses between prokaryotes and microbial eukaryotes, particularly ciliated protists, have been studied for a long time. Nevertheless, researchers have focused only on a few host genera and species, mainly due to difficulties in cultivating the hosts, and usually have considered a single symbiont at a time. Here, we present a pilot study using a single-cell microbiomic approach to circumvent these issues. Unicellular ciliate isolation followed by simultaneous amplification of eukaryotic and prokaryotic markers was used. Our preliminary test gave reliable and satisfactory results both on samples collected from different habitats (marine and freshwater) and on ciliates belonging to different taxonomic groups. Results suggest that, as already assessed for many macro-organisms like plants and metazoans, ciliated protists harbor distinct microbiomes. The applied approach detected new potential symbionts as well as new hosts for previously described ones, with relatively low time and cost effort and without culturing. When further developed, single-cell microbiomics for ciliates could be applied to a large number of studies aiming to unravel the evolutionary and ecological meaning of these symbiotic systems.

Variations in Gut Microbiota of Siberian Flying Squirrels Correspond to Seasonal Phenological Changes in Their Hokkaido Subarctic Forest Ecosystem Abstract Gut microbial communities of animals are influenced by diet and seasonal weather changes. Since foraging strategies of wild animals are affected by phenological changes, gut microbial communities would differ among seasons. However, interactions of plant-animal-microbiota with seasonal changes have not been well characterized. Here, we surveyed gut microbial diversity of Siberian flying squirrels (Pteromys volans orii) from a natural forest in Hokkaido during spring and summer of 2013 and 2014. Additionally, we compared microbial diversity to temperature changes and normalized difference vegetation index (NDVI). Changes in both seasonal temperature and phenology were significantly associated with alterations in gut microbiota. There were two clusters of OTUs, below and above 20 °C that were significantly correlated with low and high temperatures, respectively. Low-temperature cluster OTUs belonged to various phyla, whereas the high-temperature cluster was only constituted by Firmicutes. In conclusion, gut microbiota of Siberian flying squirrels varied with environmental changes on an ecological scale.

Mangrove Sediment Microbiome: Adaptive Microbial Assemblages and Their Routed Biogeochemical Processes in Yunxiao Mangrove National Nature Reserve, China Abstract Microorganisms play important roles in mangrove ecosystems. However, we know little about the ecological implications of mangrove microbiomes for high productivity and the efficient circulation of elements in mangrove ecosystems. Here, we focused on mangrove sediments located at the Yunxiao National Mangrove Reserve in southeast China, uncovering the mangrove microbiome using the 16S rRNA gene and shotgun metagenome sequencing approaches. Physicochemical assays characterized the Yunxiao mangrove sediments as carbon (C)-rich, sulfur (S)-rich, and nitrogen (N)-limited environment. Then phylogenetic analysis profiling a distinctive microbiome with an unexpected high frequency of Chloroflexi and Nitrospirae appeared to be an adaptive characteristic of microbial structure in S-rich habitat. Metagenome sequencing analysis revealed that the metabolic pathways of N and S cycling at the community-level were routed through ammonification and dissimilatory nitrate reduction to ammonium for N conservation in this N-limited habitat, and dissimilatory sulfate reduction along with polysulfide formation for generating bioavailable S resource avoiding the biotoxicity of sulfide in mangrove sediments. In addition, methane metabolism acted as a bridge to connect C cycling to N and S cycling. Further identification of possible biogeochemical linkers suggested Syntrophobacter, Sulfurovum, Nitrospira, and Anaerolinea potentially drive the coupling of C, N, and S cycling. These results highlighting the adaptive routed metabolism flow, a previously undescribed property of mangrove sediment microbiome, appears to be a defining characteristic of this habitat and may significantly contribute to the high productivity of mangrove ecosystems, which could be used as indicators for the health and biodiversity of mangrove ecosystems.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Diabetologia

Correction to: Bone marrow pericyte dysfunction in individuals with type 2 diabetes Unfortunately, three errors were made in the conversion of HbA1c to per cent values.

The Integrated Islet Distribution Program answers the call for improved human islet phenotyping and reporting of human islet characteristics in research articles

Soluble receptor for AGE in diabetic nephropathy and its progression in Finnish individuals with type 1 diabetes Abstract Aims/hypothesis Activation of the receptor for AGE (RAGE) has been shown to be associated with diabetic nephropathy. The soluble isoform of RAGE (sRAGE) is considered to function as a decoy receptor for RAGE ligands and thereby protects against diabetic complications. A possible association between sRAGE and diabetic nephropathy is still, however, controversial and a more comprehensive analysis of sRAGE with respect to diabetic nephropathy in type 1 diabetes is therefore warranted. Methods sRAGE was measured in baseline serum samples from 3647 participants with type 1 diabetes from the nationwide multicentre Finnish Diabetic Nephropathy (FinnDiane) Study. Associations between sRAGE and diabetic nephropathy, as well as sRAGE and diabetic nephropathy progression, were evaluated by regression, competing risks and receiver operating characteristic curve analyses. The non-synonymous SNP rs2070600 (G82S) was used to test causality in the Mendelian randomisation analysis. Results Baseline sRAGE concentrations were highest in participants with diabetic nephropathy, compared with participants with a normal AER or those with microalbuminuria. Baseline sRAGE was associated with progression from macroalbuminuria to end-stage renal disease (ESRD) in the competing risks analyses, but this association disappeared when eGFR was entered into the model. The SNP rs2070600 was strongly associated with sRAGE concentrations and with progression from macroalbuminuria to ESRD. However, Mendelian randomisation analysis did not support a causal role for sRAGE in progression to ESRD. Conclusions/interpretation sRAGE is associated with progression from macroalbuminuria to ESRD, but does not add predictive value on top of conventional risk factors. Although sRAGE is a biomarker of diabetic nephropathy, in light of the Mendelian randomisation analysis it does not seem to be causally related to progression from macroalbuminuria to ESRD.

Continuous glucose monitoring targets in type 1 diabetes pregnancy: every 5% time in range matters Abstract With randomised trial data confirming that continuous glucose monitoring (CGM) is associated with improvements in maternal glucose control and neonatal health outcomes, CGM is increasingly used in antenatal care. Across pregnancy, the ambition is to increase the CGM time in range (TIR), while reducing time above range (TAR), time below range (TBR) and glycaemic variability measures. Pregnant women with type 1 diabetes currently spend, on average, 50% (12 h), 55% (13 h) and 60% (14 h) in the target range of 3.5–7.8 mmol/l (63–140 mg/dl) during the first, second and third trimesters, respectively. Hyperglycaemia, as measured by TAR, reduces from 40% (10 h) to 33% (8 h) during the first to third trimester. A TIR of >70% (16 h, 48 min) and a TAR of <25% (6 h) is achieved only in the final weeks of pregnancy. CGM TBR data are particularly sensor dependent, but regardless of the threshold used for individual patients, spending ≥4% of time (1 h) below 3.5 mmol/l or ≥1% of time (15 min) below 3.0 mmol/l is not recommended. While maternal hyperglycaemia is a well-established risk factor for obstetric and neonatal complications, CGM-based risk factors are emerging. A 5% lower TIR and 5% higher TAR during the second and third trimesters is associated with increased risk of large for gestational age infants, neonatal hypoglycaemia and neonatal intensive care unit admissions. For optimal neonatal outcomes, women and clinicians should aim for a TIR of >70% (16 h, 48 min) and a TAR of <25% (6 h), from as early as possible during pregnancy.

Metformin increases endogenous glucose production in non-diabetic individuals and individuals with recent-onset type 2 diabetes Abstract Aims/hypothesis Metformin is the endorsed first-line glucose-lowering drug for treating patients with type 2 diabetes but despite more than 50 years of use, no consensus has been reached on its mechanisms of action. In this study, we investigated the glucose-lowering effects of metformin in individuals with type 2 diabetes and non-diabetic individuals. Methods We performed a randomised, placebo-controlled trial in 24 individuals with recent-onset type 2 diabetes (diabetes duration 50 [48] months) who had good glycaemic control (HbA1c 48 mmol/mmol [6.5%]). The studies were conducted at Aarhus University Hospital between 2013 and 2016. Participants were randomised to receive either metformin (2000 mg/day, n = 12, MET group) or placebo (n = 12, PLA group) for 90 days, using block randomisation set up by an unblinded pharmacist. Two participants withdrew from the study prior to completion and were replaced with two new participants receiving the same treatment. In addition, we recruited a group of non-diabetic individuals with similar age and BMI (n = 12, CONT group), who were all treated with 2000 mg metformin daily. Before and after treatment all individuals underwent studies of whole-body glucose metabolism by non-steady-state [3-3H]glucose kinetics, hyperinsulinaemic–euglycaemic clamping, indirect calorimetry, metabolomics, dual x-ray absorptiometry and muscle biopsies. The primary study endpoint was the effect of metformin treatment on lipid kinetics as well as glucose rate of disappearance (Rd) and endogenous glucose production (EGP). Results One participant from the CONT group withdrew due to intolerable gastrointestinal side-effects and was excluded from analysis. As expected, metformin treatment lowered fasting plasma glucose (FPG) in the MET group (~1.5 mmol/l, p < 0.01), whereas no effect was observed in the PLA and CONT groups. Body weight and composition did not change in any of the groups. In both of the metformin-treated groups (MET and CONT), basal glucose Rd, EGP and glucagon levels increased by ~30% (p < 0.05) whereas this was not the case in the PLA group. Conclusions/interpretation Ninety days of metformin treatment resulted in similar increases in EGP and glucose Rd in individuals with recent-onset type 2 diabetes and in non-diabetic control individuals. These results challenge the existing paradigm that metformin primarily acts in the liver by inhibiting EGP, at least in individuals with type 2 diabetes of short duration and who have discretely affected glycaemic status. Whether metformin increases basal glucose Rd by facilitating glucose uptake in other tissues such as the intestines remains to be further clarified. Trial registration ClinicalTrials.gov NCT01729156 Funding This study was supported by grants from The Danish Council for Independent Research | Medical Sciences, Aase Danielsen Fund, the Novo Nordisk Foundation, the Danish Diabetes Association and the Danish Diabetes Academy supported by the Novo Nordisk Foundation.

Type 2 diabetes: a multifaceted disease Abstract Type 2 diabetes is a complex disease usually diagnosed with little regard to aetiology. In the broader sense, it is a mix of different clearly defined aetiologies, such as monogenic diabetes, that we need to be better at identifying as this has major implications for treatment and patient management. Beyond this, however, type 2 diabetes is a highly heterogeneous polygenic disease. This review outlines the recent developments that recognise this heterogeneity by deconvoluting the aetiology of type 2 diabetes into pathophysiological processes, either by measuring physiological variables (such as beta cell function or insulin resistance) or using partitioned polygenic scores, and addresses recent work that clusters type 2 diabetes into distinct subgroups. Increasing evidence suggests that considering the aetiological components of type 2 diabetes matters, in terms of progression rates, treatment response and complications. In other words, clinicians need to recognise that type 2 diabetes is multifaceted and that its characteristics are important for how patients are managed.

Type 1 diabetes defined by severe insulin deficiency occurs after 30 years of age and is commonly treated as type 2 diabetes Abstract Aims/hypothesis Late-onset type 1 diabetes can be difficult to identify. Measurement of endogenous insulin secretion using C-peptide provides a gold standard classification of diabetes type in longstanding diabetes that closely relates to treatment requirements. We aimed to determine the prevalence and characteristics of type 1 diabetes defined by severe endogenous insulin deficiency after age 30 and assess whether these individuals are identified and managed as having type 1 diabetes in clinical practice. Methods We assessed the characteristics of type 1 diabetes defined by rapid insulin requirement (within 3 years of diagnosis) and severe endogenous insulin deficiency (non-fasting C-peptide <200 pmol/l) in 583 participants with insulin-treated diabetes, diagnosed after age 30, from the Diabetes Alliance for Research in England (DARE) population cohort. We compared characteristics with participants with retained endogenous insulin secretion (>600 pmol/l) and 220 participants with severe insulin deficiency who were diagnosed under age 30. Results Twenty-one per cent of participants with insulin-treated diabetes who were diagnosed after age 30 met the study criteria for type 1 diabetes. Of these participants, 38% did not receive insulin at diagnosis, of whom 47% self-reported type 2 diabetes. Rapid insulin requirement was highly predictive of severe endogenous insulin deficiency: 85% required insulin within 1 year of diagnosis, and 47% of all those initially treated without insulin who progressed to insulin treatment within 3 years of diagnosis had severe endogenous insulin deficiency. Participants with late-onset type 1 diabetes defined by development of severe insulin deficiency had similar clinical characteristics to those with young-onset type 1 diabetes. However, those with later onset type 1 diabetes had a modestly lower type 1 diabetes genetic risk score (0.268 vs 0.279; p < 0.001 [expected type 2 diabetes population median, 0.231]), a higher islet autoantibody prevalence (GAD-, islet antigen 2 [IA2]- or zinc transporter protein 8 [ZnT8]-positive) of 78% at 13 years vs 62% at 26 years of diabetes duration; (p = 0.02), and were less likely to identify as having type 1 diabetes (79% vs 100%; p < 0.001) vs those with young-onset disease. Conclusions/interpretation Type 1 diabetes diagnosed over 30 years of age, defined by severe insulin deficiency, has similar clinical and biological characteristics to that occurring at younger ages, but is frequently not identified. Clinicians should be aware that patients progressing to insulin within 3 years of diagnosis have a high likelihood of type 1 diabetes, regardless of initial diagnosis.

Up front

Human gut microbiota transferred to germ-free NOD mice modulate the progression towards type 1 diabetes regardless of the pace of beta cell function loss in the donor Abstract Aims/hypothesis This study aimed to assess the ability of human gut microbiota to delay the onset of type 1 diabetes when transferred into germ-free NOD mice. Methods Two children with rapid and three children with slow beta cell function loss (as assessed by C-peptide AUC change in the mixed-meal tolerance tests performed 1 and 12 months after type 1 diabetes onset), participating in an ongoing trial with gluten-free diet, donated faeces, which were transferred into germ-free NOD mice. The mice were subsequently followed for diabetes incidence. Results The bacterial profiles of bacteriome-humanised mice had significantly (p < 10−5) lower alpha diversity than the donor material, with marked shifts in ratios between the main phyla. Diabetes onset was significantly delayed in all bacteriome-humanised colonies vs germ-free NOD mice, but the pace of beta cell loss was not transferable to the mouse model. Conclusions/interpretation Germ-free NOD mice colonised with human gut microbiome are able to adopt a large proportion of transferred bacterial content, although the ratios of main phyla are reproduced only suboptimally. The recipient mice did not replicate the phenotype of the stool donor in relation to the pace towards type 1 diabetes. Trial registration ClinicalTrials.gov NCT02867436

Genome-wide association study of type 2 diabetes in Africa Abstract Aims/hypothesis Genome-wide association studies (GWAS) for type 2 diabetes have uncovered >400 risk loci, primarily in populations of European and Asian ancestry. Here, we aimed to discover additional type 2 diabetes risk loci (including African-specific variants) and fine-map association signals by performing genetic analysis in African populations. Methods We conducted two type 2 diabetes genome-wide association studies in 4347 Africans from South Africa, Nigeria, Ghana and Kenya and meta-analysed both studies together. Likely causal variants were identified using fine-mapping approaches. Results The most significantly associated variants mapped to the widely replicated type 2 diabetes risk locus near TCF7L2 (p = 5.3 × 10−13). Fine-mapping of the TCF7L2 locus suggested one type 2 diabetes association signal shared between Europeans and Africans (indexed by rs7903146) and a distinct African-specific signal (indexed by rs17746147). We also detected one novel signal, rs73284431, near AGMO (p = 5.2 × 10−9, minor allele frequency [MAF] = 0.095; monomorphic in most non-African populations), distinct from previously reported signals in the region. In analyses focused on 100 published type 2 diabetes risk loci, we identified 21 with shared causal variants in African and non-African populations. Conclusions/interpretation These results demonstrate the value of performing GWAS in Africans, provide a resource to larger consortia for further discovery and fine-mapping and indicate that additional large-scale efforts in Africa are warranted to gain further insight in to the genetic architecture of type 2 diabetes.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Mental Health

Individualised or Standardised Outcome Measures: A Co-habitation? Abstract Mental health outcome measurement is conflicted between two different schools of thought which underlie the division between standardised (nomothetic) and individualised or patient-generated (idiographic) measures. The underpinning philosophies of both approaches have very different starting points in terms of how we understand the world. And yet the strengths of both may contribute something useful for patients and mental health services. We suggest a convergence of approaches with new thinking on options for co-habitation.

Working with a Severe Mental Illness: Estimating the Causal Effects of Employment on Mental Health Status and Total Mental Health Costs Abstract Employment is an important goal for persons who have a severe mental illness (SMI). The current literature finds some evidence for a positive relationship between employment and measures of mental health (MH) status, however study design issues have prevented a causal interpretation. This study aims to measure the causal effect of employment on MH status and total MH costs for persons with SMI. In a quasi-experimental prospective design, self-reported data measured at baseline, 6-months, and 12-months, on MH status and employment are paired with Public Mental Health System (PMHS) claims data. The study population (N = 5162) is composed of persons with a SMI who received PMHS services for a year or more. Outcome variables are MH status symptom scores from the BASIS-24 instrument and total MH costs. The estimation method is full information maximum likelihood, which allows for tests of employment endogeneity. Outcomes with an insignificant test of endogeneity are estimated using tobit or ordinary least square (OLS). Employment has modest but meaningful effects on MH status (including overall MH status, functioning, and relationships) and reduces total mental health costs on average by $538 in a 6-month period. Tests of endogeneity were largely insignificant, except for the depression score that tested marginally statistically significant. Interaction terms between baseline MH scores and employment indicated larger employment effects for individuals with worse baseline scores. This study demonstrates the non-vocational benefits of employment for individuals with SMI. Results have high generalizability and should be of interest to federal and state governments in setting appropriate disability policy and funding vocational programs. From a methodological perspective, future research should still be concerned with potential endogeneity problems, especially if employment status and MH outcomes are simultaneously measured and/or baseline measures of MH are not adequately controlled for future research should continue to examine the multi-dimensional nature of MH status and costs. Our analyses also demonstrate the practical use of a state-wide outcomes measurement program in assessing the factors that influence the recovery trajectories of individuals with SMI.

State Approaches to Funding Home and Community-Based Mental Health Care for Non-Medicaid Youth: Alternatives to Medicaid Waivers Abstract Home and Community-Based Services (HCBS) Medicaid waivers for children increase the availability of public funding for HCBS by waiving or expanding the means tests for parents' income, basing child eligibility for Medicaid coverage primarily on clinical need. But many states provide mechanisms apart from HCBS waivers to increase coverage for youth with significant mental health needs. Through interviews with public mental health officials from 37 states, this study identifies and explains non-waiver funding strategies for HCBS services for otherwise ineligible youth. Results demonstrate that states expand Medicaid-eligibility through CHIP or use state general revenue funds to pay for medically necessary HCBS for non-Medicaid youth.

Relative Technical Efficiency Assessment of Mental Health Services: A Systematic Review Abstract The current prevalence of mental disorders demands improved ways of the management and planning of mental health (MH) services. Relative technical efficiency (RTE) is an appropriate and robust indicator to support decision-making in health care, but it has not been applied significantly in MH. This article systematically reviews the empirical background of RTE in MH services following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Finally, 13 studies were included, and the findings provide new standard classifications of RTE variables, efficiency determinants and strategies to improve MH management and planning.

Effects of Fidelity-Focused Consultation on Clinicians' Implementation: An Exploratory Multiple Baseline Design Abstract Identification of effective consultation models could inform implementation efforts. This study examined the effects of a fidelity-focused consultation model among community-based clinicians implementing Attachment and Biobehavioral Catch-up. Fidelity data from 1217 sessions from 7 clinicians were examined in a multiple baseline design. In fidelity-focused consultation, clinicians received feedback from consultants' fidelity coding, and also coded their own fidelity. Clinicians' fidelity increased after fidelity-focused consultation began, but did not increase during other training periods. Fidelity was sustained for 30 months after consultation ended. Findings suggest that consultation procedures involving fidelity coding feedback and self-monitoring of fidelity may promote implementation outcomes.

Impact of Serious Mental Illness on Medicaid and Other Public Healthcare Costs in Texas Abstract Medicaid-enrolled adults with serious mental illness may be dually-enrolled in Medicare, and may receive health care services from other state and local programs. To understand cross-program costs of care, we linked 2012 payment data across Medicaid, Medicare, state, and local programs. Average costs were calculated according to presence/absence of SMI, Medicare coverage, SSI coverage, medical comorbidities, and other characteristics. Costs for Medicaid adults with SMI were 57.4% greater than adults without SMI, but only 23.6% of costs were SMI-related. Greater costs were associated with Medicaid-Medicare dual-eligibility, multiple SMI diagnoses, and medical comorbidities. The results support cross-program efforts such as joint Medicaid-Medicare managed care and integrated care.

Impact of Supervisory Support on Turnover Intention: The Mediating Role of Burnout and Job Satisfaction in a Longitudinal Study Abstract High rates of provider turnover are problematic for our mental health system. Research indicates that supervisory support could alleviate some turnover intention by decreasing emotional exhaustion (a key component of burnout) as well as by increasing job satisfaction. However, the potential mediation mechanisms have not been rigorously tested. Longitudinal data collected from 195 direct clinical care providers at two community mental health centers identified positive effects of supervisory support on reduced turnover intention through reduced emotional exhaustion. Job satisfaction was not a significant mediator. Supervisory support may help mitigate turnover intention through work-related stress reduction.

Measuring and Predicting Service Providers' Use of an Evidence-Based Parenting Program Abstract This study addressed the predictors of service providers' use of a multi-level evidence-based program (EBP). Of the 92 trained providers participating in the study, 67 (72.8%) used the EBP at least once. A multidimensional index of the amount of usage (MUI) was created using three indicators. Providers' self-efficacy and the amount of training they had received predicted their amount of usage. The community to which the providers belonged was also associated with their amount of usage. The findings underline the importance of studying many indicators of usage in implementation research and considering both provider-level and broader contextual variables as determinants of the use of EBPs.

Observed Outcomes: An Approach to Calculate the Optimum Number of Psychiatric Beds Abstract The number of psychiatric beds, in most developed countries, has decreased progressively since the late 1950s. Many clinicians believe that this reduction has gone too far. But how can we determine the number of psychiatric beds a mental health system needs? While the population health approach has advantages over the normative approach, it makes assumptions about optimal and minimum duration of hospitalization required for various psychiatric disorders. In this paper, we describe a naturalistic approach that estimates the required number of psychiatric beds by comparing the bed levels at which negative outcomes develop in different jurisdictions. We hypothesize that there will be a threshold below which negative outcomes will be seen across jurisdictions. We predict that hospital key performance indices will be more sensitive to bed reductions than the clinical and social outcomes of patients. The observed outcome approach can complement other approaches to determining bed numbers at the national and local levels, and should be a priority for future health services research.

Predicting Youth Improvement in Community-Based Residential Settings with Practices Derived from the Evidence-Base Abstract The current investigation conducted descriptive analyses on key variables in community-based residential (CBR) settings and investigated the extent to which disruptive youth between the ages of 13 and 17 years improved based on therapists' reported alignment with using practices derived from the evidence-base (PDEBs). Results from both the descriptive analyses and multilevel modeling suggested that therapists are using practices that both do and do not align with the evidence-base for disruptive youth. In addition, both PDEBs and practices with minimal evidence-support predicted or marginally predicted final average progress rating for these youth. Findings are discussed as they relate to the importance of continued exploration of treatment outcomes for CBR youth.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Transplantation

Clostridium difficile: a frequent infection in children after intestinal transplantation Background: Organ transplantation is a risk factor for Clostridium difficile infection (CDI). After intestinal transplantation (ITx), few data are available on the impact of this graft infection and the possible induction of rejection. Methods: We included retrospectively all children after ITx in our unit, with at least one year of graft survival. All samples positive for Clostridium difficile and its toxin were considered. Results: Among the 57 ITx recipients (60 transplantations), 22 children (39%) developed culture-proven CDI, 12 after isolated small bowel Tx, 9 after liver-small bowel Tx, one after multivisceral Tx. Twenty patients had diarrhea, 8 bloody stools, 4 fever, one hypothermia. Nine were hospitalized for an average of 6.5 days (2-20), four with severe dehydration. Nine (40%) had received antibiotics for an average of 19 days (7-60) before CDI. Two patients were asymptomatic. CDI was treated with metronidazole in 12 children, vancomycin in 6 and both of them in 3. Three children presented mild to severe rejections. Two patients presented concomitantly CDI and rejection. The third patient presented a rejection with severe complications 4 years after CDI. Recurrence of toxinogenic CD was observed in 9 children, in 7 associated with clinical symptoms. During the last follow-up, the stool number was the same as before CDI except for one patient with ongoing infection. Conclusion: CDI is more prevalent in children after ITx compared with other organ Tx; it is most often symptomatic but mildly or moderately severe. Standard antibiotics efficiently control the symptoms. Induction of rejection is a rare event. RDL, FL, JB and LN participated, in the writing of the paper and in data analysis, AF, CC and OG participated in research design and in the writing of the paper. Disclosure: "The authors declare no conflict of interest." Financial disclosure:"The authors declare no financial disclosure." Corresponding Author: Dr Rémi Duclaux-Loras, Hôpital Femme Mère Enfant, 59 boulevard Pinel 69677 BRON, remi.duclaux-loras@chu-lyon.fr Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Role of preimplantation biopsies in kidney donors with acute kidney injury. No abstract available

The Forgotten Member of Patient Selection Committees: The U.S. Constitution No abstract available

The outcomes of portal vein reconstruction with vein graft interposition in pediatric liver transplantation for small children with biliary atresia Background: Several technical modifications in portal vein (PV) reconstruction have shown excellent outcomes in pediatric liver transplantation (LT); however, which procedure is the best for PV reconstruction of the hypoplastic PV in pediatric LT remains unclear. Methods: One hundred sixteen pediatric patients aged 1 or younger with BA undergoing living donor liver transplantation (LDLT) at our center were enrolled in the present study to investigate the outcomes of the different types of PV reconstruction. We compared the results between patients with and without VG interposition (VG group, n = 33; non-VG group, n = 83) to analyze the risk factors for PV complications (PVCs). The median follow-up period was 4.7 years. Results: PVCs occurred in 10 cases (7.2%), including 5 cases in the non-VG group and 5 in the VG group. Stenosis and thrombosis occurred in 10 and 3 cases, respectively, and 3 cases suffered from both. Three patients were surgically treated for PVCs and two underwent stent insertion to treat short-term recurrence after the initial treatment. The incidence of PVCs in the two groups did not differ to a statistically significant extent. Although retrograde PV flow was one of significant risk factors in a univariate analysis, a multivariate analysis revealed that early transplant era was the only independent risk factor for PVCs. Conclusions: VG interposition for PV reconstruction in LDLT appears to be a feasible alternative option with acceptable outcomes for patients with BA. Achieving sufficient PV flow is essential to preventing PVCs after LDLT. Authorship: S. Sakamoto, H. Uchida and T. Kitajima participated in study design, data analysis, and the writing of the paper, S. Shimizu critically reviewed the statistical analysis. S. Yoshimura, M. Takeda, Y. Hirata and A. Fukuda collected the clinical data, M. Kasahara critically reviewed the article. Conflict of interest: None Corresponding Author:Seisuke Sakamoto MD, PhD, 2-10-1 Okura, Setagaya, Tokyo, 157-8535, Japan. E-mail: sakamoto-si@ncchd.go.jp, Phone: +81-3-3416-0181. Fax: +81-3-3416-222 Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Long-term outcomes following kidney transplantation from donors with acute kidney injury Background: Kidneys from deceased donors with acute kidney injury (AKI) are more likely to be discarded because of concerns for poor outcomes after transplantation. The aim of this study was to determine the long-term outcomes of a large cohort of patients transplanted utilizing kidneys from deceased donors with acute kidney injury. Methods: All patients receiving a deceased donor kidney transplant during a recent 10-year period were included. Acute Kidney Injury Network (AKIN) criteria were used to classify the donors. Donor kidneys with more than 10% cortical necrosis or more than mild chronic changes were discarded. The primary outcome is the combined endpoint of death or graft loss. Results: The cohort included 1,313 kidneys from 974 donors, AKIN stage 0 (no AKI) in 319 (24.3%), stage 1 in 370 (28.2%), stage 2 in 177 (13.5) and stage 3 in 447 (34.0%). Estimated 5-year graft survival (95% CI) was 78.5% (72.5-84.5), 77.8% (72.8-82.1), 83.8% (76.8-88.9) and 84.6% (79.5-88.7) for AKIN donor stage 0 to 3, respectively (log-rank p=0.10). After adjusting for baseline differences, the HR (95% CI) for the combined endpoint for the AKIN stage 3 group (relative to AKIN 0 group) was 0.70 (0.45-1.10). DGF occurred in 44.6% and 75.4% of AKIN 2 and 3 groups, as compared to 33.9% and 33.5% in AKIN 0 and 1 (p<.001). Conclusion We conclude that transplanting selected kidneys from deceased donors with AKI with preimplantation biopsy showing less than 10% cortical necrosis and no more than mild chronic changes have excellent long-term graft survival. Disclosures: The authors declare no conflicts of interest. Conclusion: We conclude that transplanting selected kidneys from deceased donors with AKI with pre-implantation biopsy showing less than 10% cortical necrosis and no more than mild chronic changes have excellent long-term graft survival. Corresponding Author: Raymond L. Heilman, MD, 5777 East Mayo Boulevard, Phoenix, AZ 85054, Email Heilman.raymond@mayo.edu Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Performance of creatinine clearance and estimated GFR in assessing kidney function in living donor candidates Background: Glomerular filtration rate (GFR) assessment is a key aspect in the evaluation of living kidney donor candidates; however, data on performance of commonly used methods are limited. Methods: We examined 769 living kidney donor candidates with 24-hour urine collections assessed as accurate by comparing measured creatinine excretion rate (CER) to CER estimated using a four-variable equation previously developed and validated using robust methodology. Results: Of all collections, 42.6% would have been deemed inaccurate, mostly under-collections, using the conventional weight- and gender-based CER estimation. Creatinine clearance (CrCl) overestimated 125I-iothalamate GFR (iGFR), estimated GFR (eGFR) underestimated iGFR, and their average [Avg (CrCl and eGFR)] essentially eliminated the GFR bias (median bias = +2.2, -5.4 and -1.0 ml/min/1.73m2, respectively; p<0.001). This held true for all subgroups except blacks, where all three measures overestimated iGFR. Avg (CrCl and eGFR) also offered modestly improved accuracy compared with CrCl alone, as measured by the proportion of values falling within 10% (50.7% vs. 45.3%; p=0.009) and 30% of iGFR (94.5% vs. 89.3%; p<0.001). Conclusions: When measured GFR is unavailable, the Avg (CrCl and eGFR) provides a better estimate of kidney function in kidney donor candidates than either measure alone, although in blacks the estimates are neither better nor worse. The authors declare no conflicts of interest. Disclosures: The authors of this manuscript have no conflicts of interest to disclose as described by the American Journal of Transplantation. * Authors contributed equally to the study Corresponding author: Emilio D. Poggio, M.D. Department of Nephrology and Hypertension, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, Ohio 44195, E-mail: poggioe@ccf.org, Tel: 216-659-2734, Fax: 216-444-9378 Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Cytotoxic Effects of Rabbit Anti-Thymocyte Globulin preparations on Primary Human Thymic Epithelial Cells Background: Graft-versus-host disease (GvHD) presents a major cause for morbidity and mortality after allogeneic hematopoietic stem cell transplantation (alloHSCT). Rabbit anti-thymocyte globulin (rATG) treatment reduces the incidence of GvHD after alloHSCT. However, delayed immune reconstitution following rATG treatment, partly due to hampered thymic function, is being discussed. The present study aimed at elucidating possible cytotoxic effects of two commonly used rATG preparations on cultured human thymic stroma, especially thymic epithelial cells (TECs). Methods: A primary thymic epithelial cell culture was established and the binding and cytotoxicity of two rATG preparations to the aforementioned cells were assessed by flow cytometry and immunofluorescence analyses. The release of several cytokines by cultured TSCs in response to rATG was analyzed via multiplex ELISA. Results: Both preparations showed a comparable dose-dependent binding to TECs and exerted a similar complement-independent, dose-dependent cytotoxicity. rATG exposure further resulted in hampered secretion of IL-7, IL-15 and IL-6, cytokines being involved in thymic T cell development and proliferation. Pretreatment with keratinocyte growth factor (KGF) diminished rATG-induced cytotoxicity of TECs and restored their IL-7 and IL-15 secretion. Conclusions: Cytotoxic effects on TECs link a rATG-induced thymic damage to the delayed T cell reconstitution witnessed after rATG treatment. Our data support a combination treatment of rATG and thymus-protective strategies such as KGF in order to simultaneously offer sufficient GvHD prophylaxis and overcome delayed T cell reconstitution due to thymic damage. Funding and disclosure: The authors of this manuscript have conflicts of interest to disclose as described by Transplantation. The studies reported in this publication were supported by a grant from Neovii Biotech GmbH. Although a financial conflict of interest was identified for management based on the overall scope of the project and its potential benefit to Neovii Biotech GmbH, the research findings included in this publication may not be necessarily related to the interests of Neovii Biotech GmbH. The terms of this arrangement have been reviewed and approved by Charité Universitätsmedizin Berlin in accordance with its policy on objectivity in research. Corresponding author: Univ.-Prof. Dr. med. Il-Kang Na, Charité - Campus Virchow-Klinikum, Hematology, Oncology and Tumor Immunology, Augustenburger Platz 1, 13353 Berlin, Germany. Phone: +49 30 450-540155/ -653598, E-mail: il-kang.na@charite.de Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Long-Term Effects of Hypothermic Ex-Situ Perfusion on Skeletal Muscle Metabolism, Structure, and Force Generation After Transplantation Background: Hypothermic ex-situ perfusion (HESP) systems are used to prolong allograft survival in solid organ transplantations and have been shown to be superior to static cold storage (SCS) methods. However, the effect of this preservation method on limb allograft survival and long-term function has not yet been tested. In this study, we investigated the long-term effects of the HESP on skeletal muscle metabolism, structure, and force generation and compared it with the current standard of preservation . Methods: Forty male Lewis rats (250 ± 25 gr) were divided into 5 groups including naive control, sciatic nerve transection/repair, immediate transplantation, SCS, and HESP. For the SCS group, limbs were preserved at 4˚C for 6 hours. In the HESP group, limbs were continuously perfused with oxygenated Histidine-Tryptophan-Ketoglutarate (HTK) solution at 10-15˚C for 6 hours. Hemodynamic and biochemical parameters of perfusion were recorded throughout the experiment. At 12 weeks, electromyographyand muscle force measurements (maximum twitch and tetanic forces) were obtained along with muscle samples for histology and metabolomics analysis. Results: Histology demonstrated 48% myocyte injury in the HESP group compared to 49% in immediate transplantation (p=0.96), and 74% in the SCS groups (p<0.05). The maximum twitch force measurement revealed a significantly higher force in the HESP group compared to the SCS group (p=0.029). Essential amino acid levels of the gastrocnemius muscle did not reach significance, with the exception of higher proline levels in the HESP group. Conclusion: Hypothermic ex-situ perfusion using HTK protects viability of the limb but fails to restore muscle force in the long-term. Authorship: Study design: EG, and KO. Acquisition of data: EG, CK, EG, MP, and MJH. Analysis and interpretation: EG, CK, EG, and MJH. Manuscript drafted by EG. Revision: EG, AR, SK, RHB and KO. Disclosures The authors declare no conflicts of interest. Acknowledgment: This study was partially supported by the American Foundation for Surgery of Hand (Award Number 1429) and Michigan Regional Comprehensive Metabolomics Resource Core Grant U24 DK097153. * Corresponding Author: Kagan Ozer, MD, Clinical Professor, Surgical Director for Hand Transplantation Program, Hand and Upper Extremity, Department of Orthopedic Surgery, University of Michigan Hospitals, 2098 South Main St. , Ann Arbor, MI 48103 USA. Phone: +1 (734) 998-6541, E-mail: kozer@med.umich.edu Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

An outpatient hospital-based exercise training programme for patients with cirrhotic liver disease awaiting transplantation: a feasibility trial Background: Time spent on the waiting list prior to liver transplantation (LT) provides an opportunity to optimise recipient fitness through prehabilitation; potentially reducing the physiological impact of major surgery. We assessed the feasibility and effectiveness of a six-week exercise programme in patients with cirrhotic liver disease awaiting LT. Methods: This single centre, prospective cohort, feasibility study, enrolled patients awaiting LT to a six week period of thrice weekly, supervised exercise on a static bike. Cardiopulmonary exercise testing (CPET) was used to objectively assess cardiopulmonary fitness at baseline and after six weeks of exercise. A follow-up CPET was performed at 12 weeks. CPET-derived measures were used to guide prescription of the training programme. A non-randomised control cohort of LT patients were selected to match the exercise group based on specific demographic data. Allocation to study arms was primarily based on the distance participants lived from the hospital where training occurred. Both groups received structured nutritional advice. Results: The exercise programme was feasible, with 9/16 (56%) patients completing the full programme of six weeks. Peak oxygen consumption (VO2peak) in the exercise group rose from a mean (SD) of 16.2 (± 3.4) ml/kg/min at baseline to 18.5 (±4.6) ml/kg/min at week 6 (p=0.02). In the control group VO2peak decreased from a mean (SD) of 19.0 (± 6.1) ml/kg/min to 17.1 (±6.0) at week 6 (p=0.03). Conclusion: We have demonstrated that it is feasible to engage patients awaiting LT in an intensive aerobic exercise programme with a signal of improvement in fitness being detected. Grants and financial support: None Conflicts of interest: None declared Acknowledgments: We would like to thank all the patients that contributed to this study whilst awaiting their liver transplants. Corresponding author: Dr Clare Morkane MBBCh, MRCP, FRCA, FFICM, Royal Free Perioperative Research Group (RoFPoR), Department of Anaesthesia, Royal Free Hospital, Pond Street, London. c.morkane@nhs.net, 07725269398 Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Differential impact of delayed graft function in deceased donor renal transplant recipients with and without donor-specific HLA-antibodies Background: Delayed graft function (DGF) and pre-transplant donor-specific HLA-antibodies (DSA) are both regarded as risk factors for rejection and lower graft survival. However, the combined impact of DGF and DSA has not been studied in detail. Methods: We investigated 375 deceased donor kidney transplantations, which had DSA assignment by single-antigen bead technology and which had surveillance biopsies at 3/6 months. Median follow-up time was 6.1 years. Results: DGF occurred in 137/375 patients (37%), and DSA were present in 85/375 patients (23%). The incidence of DGF was similar in DSApos-patients and DSAneg-patients (40% vs 36%; p=0.45). In DSAneg-patients, five-year graft survival was not different with/without DGF (81% vs 83%; p=0.48). By contrast, in DSApos-patients, five-year graft survival was significantly lower with DGF (64% vs 79%; p=0.01). Moreover, DSApos-patients with DGF had a higher one-year incidence of (sub)clinical rejection, which were mostly antibody-mediated or mixed rejection phenotypes. Graft loss due to rejection was significantly more frequent in DSApos-patients with DGF (5/34; 15%) compared to DSApos-patients without DGF (2/51; 4%), and DSAneg-patients with/without DGF (3/103; 3% and 4/187; 2%, respectively) (p=0.005). In a multivariate Cox model, DSA with DGF was an independent predictor for graft (HR=2.84 [95%-CI 1.54-5.06]; p=0.001) and death-censored graft loss (HR=4.65 [95%-CI 1.83-11.51]; p=0.002). Conclusions: DGF has a much more detrimental impact in DSApos-patients than in DSAneg-patients, which is likely related to a higher incidence of antibody-mediated rejection. If possible, the combined risks of DGF and DSA should be avoided. Disclosure: None Funding: SS is supported by the Swiss National Foundation (grant # 32003B_169310). Acknowledgments: We thank the nurses in the outpatient clinic for their outstanding help in the management of the patients. Corresponding Author: Stefan Schaub, MD MSc, Clinic for Transplantation Immunology and Nephrology, University Hospital Basel, Petersgraben 4, 4031 Basel, Switzerland. Tel: 0041 61 265 45 33, Fax: 0041 61 265 24 10, e-mail: stefan.schaub@usb.ch, ORCID: 0000-0002-9170-1341 Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Sex Roles

Pursuing Medicina [Medicine]: Latina Physicians and Parental Messages on Gendered Career Choices Abstract Prior research underscores that college-educated Latinas are hyper-segregated into highly feminized occupations and that Latino parents socialize their daughters to seek out these careers. Despite this trend, a small and growing number of Latinas are steadily entering prestigious non-traditional careers in medicine. Drawing on in-depth interviews with 20 Latina physicians of various generational backgrounds in California, I examine how Latina doctors explain the role parental messages played in their occupational trajectories. I document the fluid and often contradicting gendered expectations Latina physicians received from parents and identify three primary patterns in these messages, including: (a) fathers' contradictory gendered expectations for their wives versus their U.S. born/raised daughters, (b) fathers and mothers' differing messages about procuring financial and social independence from men, and (c) parental messages about sexuality. The messages that young Latinas receive vary and reflect tensions between gendered expectations for women in the immigrant parents' home country and those available in the United States. The messages of support and resistance that Latina physicians receive from their parents regarding the pursuit of a career in medicine show how changing structural contexts—such as access and entry into prestigious non-traditional fields—rearranges gendered dynamics within Latino families.

The Impact of Gendered Stereotypes on Perceptions of Violence: A Commentary Abstract The present commentary explores the impact of gender role stereotypes on perceptions of both intimate partner violence (IPV) and sexual violence. Two papers published in this issue of Sex Roles explored the influence of gender stereotypes on both IPV (Bates et al. 2019) and rape myths (Klement et al. 2019). An overarching theme of these papers is how gender stereotypes may influence incorrect beliefs in how we view and approach interventions to these two types of violence. Reflecting on this convergence, we have come together as authors to consider how influential and damaging these stereotypes can be to victims of both partner violence and sexual violence. Our paper considers the nature of these stereotypes, who is harmed by them considering both gender and sexuality, and also the impact they have in societal and service responses to violence, as well as policy and practice development.

Reducing a Male Bias in Language? Establishing the Efficiency of Three Different Gender-Fair Language Strategies Abstract Different strategies of gender-fair language have been applied to reduce a male bias, which means the implicit belief that a word describing an undefined person describes a man. This male bias might be caused by the words themselves in terms of generic masculine or masculine forms or by androcentrism (the conflation of men with humanity). In two experiments, we tested how different gender-fair strategies used as labels of an unknown social target (an applicant in a recruitment situation) could eliminate the male bias. The three types of gender-fair strategies tested were: (a) paired forms (he/she), (b) traditional neutral words (e.g., singular they, "the applicant"), or (c) gender-neutral third-person pronouns actively created to challenge the binary gender system (ze, Swedish hen). The two experiments were performed in Swedish with 417 undergraduates in Sweden and in English with 411 U.S. participants recruited online. In Swedish, the third-person gender-neutral pronoun singular (hen) was used. In English, several forms of such gender-neutral pronouns have been suggested (e.g., ze). In both experiments, results indicated that paired forms and actively created gender-neutral pronouns eliminated the male bias, whereas traditional neutral words contained a male bias. Thus, gender-fair language strategies should avoid using traditional words. Consequences of using paired forms and creating new gender-neutral words are discussed. We argue that an actively created gender-neutral pronoun is of highest value because it is more inclusive.

When Seeing Is Not Believing: An Examination of the Mechanisms Accounting for the Protective Effect of Media Literacy on Body Image Abstract The present study aimed to explore the moderating role of three dimensions of media literacy on the relationship between media exposure and body dissatisfaction, mediated by thin-ideal internalization and appearance comparison among early female adolescents. A sample of 284 Australian female adolescents in single-sex schools (Mage = 13.15 years, range 11–16) reported on their media exposure, thin-ideal internalization, appearance comparison, body dissatisfaction, and three dimensions of media literacy: realism scepticism (scepticism regarding the extent to which media images portray reality, similarity scepticism (scepticism regarding the extent to which images portray a reality that is compatible with one's personal experience), and critical thinking (with regard to the intention of the message, its meaning, and influence). Moderated mediation analyses were conducted. Findings revealed different patterns of relationships for the different dimensions of media literacy, with similarity scepticism moderating the mediated relationship between media exposure and body dissatisfaction via both thin-ideal internalization and appearance comparison. In contrast, reality scepticism and critical thinking revealed negative associations with body dissatisfaction but were not found to serve as moderators. Findings suggest that the mechanisms of action may vary for different dimensions of media literacy, and they highlight the importance of targeting media literacy in intervention and prevention efforts.

Body Surveillance Predicts Men's and Women's Perceived Loneliness: A Serial Mediation Model Abstract Previous research on self-objectification mainly focuses on its influences on intrapersonal psychological distress whereas our study examined whether self-objectification would influence interpersonal distress (i.e., loneliness) and its corresponding mechanisms in a sample of American women and men recruited with MTurk. Participants' self-objectification was indexed by their level of body surveillance, and we proposed that body surveillance would increase women's and men's tendency to experience shame about their body and decrease their general self-esteem, which would in turn predict their level of loneliness. A total of 373 Americans (235 women; Mdnage = 33 years-old, range = 18–77) participated in the present study, and the results provided support for the proposed theoretical model. Specifically, we found that body surveillance positively predicted people's body shame, and body shame negatively predicted self-esteem, which in turn predicted people's loneliness. Moreover, this mediational model was not different between men and women. These results expand the scope of investigation by incorporating male samples, and they suggest that in addition to intrapersonal consequences, self-objectification can also influence people's interpersonal well-being. Implications were discussed.

Accusers Lie and Other Myths: Rape Myth Acceptance Predicts Judgments Made About Accusers and Accused Perpetrators in a Rape Case Abstract Previous research results have yielded a consistent link between rape myth acceptance and sexual assault victim blaming: Individuals reporting higher levels of rape myth acceptance also report higher levels of victim blaming. In four studies we explored whether the presentation of rape-myth confirming information or rape-myth debunking information might moderate these tendencies. In these studies, U.S. undergraduates (97 in Study 1, 84 in Study 2, 98 in Study 3, and 116 in Study 4) read scenarios of a heterosexual sexual assault case and were randomly assigned to a control condition, a rape myth confirmation condition, or a rape myth debunking condition; they also reported the extent to which they endorsed or accepted rape myths. Rape myth acceptance robustly correlated with judgments made about accusers and accused rapists regardless whether the accuser/accused pairing was female/male (Studies 1 and 2) or male/female (Studies 3 and 4). For example, those who most strongly endorsed rape myths were also likely to disbelieve accusers. There were few instances indicating that the presentation of rape myth confirming information or rape myth debunking information moderated these effects. This lack of moderation occurred regardless of whether the information came from trial lawyers or from expert witnesses in the case. The relative impotence of the information presentations could be due to several factors (e.g., entrenched nature of rape myth acceptance, psychological reactance, timing and strength of manipulation), and we suggest ideas for how to overcome this relative impotence in future research.

Do Women in the Newsroom Make a Difference? Coverage Sentiment toward Women and Men as a Function of Newsroom Composition Abstract Positive or negative media coverage may have important consequences for individuals' lives and ability to succeed. One potential factor that may affect the tone of coverage, in particular for women, is the gender of newsroom managers. Some scholars have suggested that women in key editorial and managerial roles should have a positive effect on the overall coverage of issues in the news, and specifically on the coverage of women. We used fixed effects regression to analyze panel data on the coverage sentiment of 212 U.S. newspapers from various cities and states between 2004 and 2009 to examine the effects of the gendered composition of newsrooms on coverage tone for both men and women. Our results showed that individuals with female names receive more positive coverage than those with male names do in every section of the newspaper. We also found that increases in female representation on newspapers' editorial boards resulted in coverage for women that is moderately more positive. However, there is no evidence that under female executive editorship coverage sentiment favors women. Our findings are consistent with the work of gender sociologists and media scholars who have highlighted the media's rigid gender structures and their resistance to change.

How Societal Changes Have Influenced German Children's Gender Representations as Expressed in Human Figure Drawings in 1977 and 2015 Abstract We investigated German first graders' gender representations in human figure drawings done in 1977 and 2015. We hypothesized that increasing gender-status equality in society as well as growing gender differentiation in childcare and marketing are reflected in depictions of the human figure. Drawings were collected from a total of 376 children between 5 years 10 months and 8 years of age. Overall, the results are in accordance with the hypotheses: In contrast to 1977, the proportion of male and female figures was more balanced in 2015. In 2015, more girls drew a figure of their own gender and the femininity of female figures was higher than in 1977. Unexpectedly, the masculinity of male figures did not increase over this time. These results provide some insight into dynamic changes of children's view of gender roles reflecting societal conditions. Drawings as a nonverbal approach to children's gender representations proved sensitive in research but may also serve as a starting point in social and pedagogical work addressing gender issues. Considering gender status equality and gender specification as independent aspects of gender representations contributes to a better understanding by researchers as well as by practice professionals.

What about the Male Victims? Exploring the Impact of Gender Stereotyping on Implicit Attitudes and Behavioural Intentions Associated with Intimate Partner Violence Abstract Although intimate partner violence (IPV) is considered stereotypically as a gendered phenomenon, empirical evidence contradicts such gender asymmetry in reported rates of victimisation and perpetration. The current research explored the impact of stereotype priming on implicit attitudes associated with IPV victimisation (Study 1) and perpetration (Study 2), and further examined behavioural intentions associated with hypothetical gendered scenarios of IPV. Participants recruited in the United Kingdom were primed with either stereotype congruent, incongruent or no information about IPV victimisation (Study 1, n = 122) or perpetration rates (Study 2, n = 101). They then completed an Implicit Association Test and reported their subjective norms, self-efficacy, behavioural intentions, and outcome expectancies pertaining to different scenarios depicting gendered IPV. Findings indicate that priming an incongruent stereotype did not impact significantly on implicit or explicit attitudes toward IPV. Gendered scenarios were found to be influential on explicit attitudes, with IPV less likely to be identified toward male victims and considered more acceptable compared to when the victim was female. Moreover, individuals reported feeling more capable and likely to intervene in an act of IPV when the victim was female compared to male, were more likely to report such an incident, and anticipated greater outcomes. These findings highlight the need for an inclusive research approach that recognises men's victimisation.

Does Traditional Stereotyping of Career as Male Affect College Women's, but Not College Men's, Career Decision Self-Efficacy and Ultimately Their Career Adaptability? Abstract In South Korea, strong beliefs about traditional gender roles in accordance with Confucian and patriarchic atmosphere still strongly influence daily life and the career development process. Cultural and contextual factors impact the development of gender role socialization, which influences an individual's career decision self-efficacy (CDSE) and adaptability to manage the challenging career decision-making process. In our study, we recruited 291 South Korean undergraduate students (138 women, 153 men) and investigated how an implicit gender-career stereotyping impacts career adaptability via CDSE and whether there is a gender difference on the direct and indirect effects of implicit gender-career stereotyping on career adaptability. The Implicit Association Test (IAT) was implemented to measure implicit gender-career stereotyping. By testing a moderated mediation model, we found a mediated effect of CDSE in the link between implicit gender-career stereotyping and career adaptability in the female students. Moreover, the direct relationship between implicit gender-career stereotyping and CDSE was significant only for female students. Given the findings, practitioners and educators who work with South Korean women need to explore the degree of clients' traditional gender role stereotyping and provide tailored interventions to increase their level of career adaptability by minimizing the negative impacts of gender role stereotyping and by increasing CDSE.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com