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Introduction Management of chronic kidney disease (CKD) entails high medical complexity and often results in high hospitalization burden. There are limited data on the associations of longitudinal hospital utilization patterns with adverse clinical outcomes in individuals with CKD. Methods We derived cumulative all-cause hospitalization trajectory groups using latent class trajectory analysis in 3012 participants of the Chronic Renal Insufficiency Cohort (CRIC) Study who were alive and did not reach end-stage kidney disease (ESKD) within 4 years of study entry. Cox proportional hazards models tested the associations between hospitalization trajectory groups and risks of ESKD and death prior to the onset of ESKD (ESKD-censored death). Results Within 4 years of study entry, there were 5658 hospitalizations among 3012 participants. We identified 3 distinct subgroups of individuals with CKD based on cumulative all-cause hospitalization trajectories over 4 years: low-utilizer (n = 1066), intermediate-utilizer (n = 1802), and high-utilizer (n = 144). High-utilizers represented a patient population of lower socioeconomic status who had a greater prevalence of comorbid conditions and lower kidney function compared with intermediate- and low-utilizers. After the 4-year ascertainment period to form the trajectory subgroups, there were 544 ESKD events and 437 ESKD-censored deaths during a median follow-up time of 5.1 years. Compared with low-utilizers, intermediate-utilizers and high-utilizers were at 1.49-fold (95% confidence interval [CI] 1.22–1.84) and 1.75-fold (95% CI 1.20–2.56) higher risk of ESKD in adjusted analyses, respectively. Compared with low-utilizers, intermediate-utilizers and high-utilizers were at 1.48-fold (95% CI 1.17–1.87) and 2.58-fold (95% CI 1.74–3.83) higher risk of ESKD-censored death in adjusted analyses, respectively. Conclusions Trajectories of cumulative all-cause hospitalization identify subgroups of individuals with CKD who are at high risk of ESKD and death., Graphical abstract

Three-dimensional (3D) body scanner technology for body composition assessment is expanding. The aim of this study was to assess the validity of a 3D body scanner. One hundred and ninety-four participants (43% male; age: 23.52 ± 5.47 years; body mass index: 23.98 ± 3.24 kg·m−2) were measured using 3D scanner and a 4-compartment (4C) model utilizing dual-energy X-ray absorptiometry (DXA), air displacement plethysmography, and bioelectrical impedance spectroscopy. Dependent t tests, validity statistics including total error (TE), standard error of the estimate, constant error, and Bland–Altman analyses were utilized. Compared with 4C, 3D scanner fat mass (FM) [mean difference (MD; 3D−4C): 2.66 kg ± 3.32 kg] and percent body fat (%BF) (MD: 4.13% ± 5.36%) were significantly (p < 0.001) over-predicted; fat free mass (FFM) was significantly underpredicted (MD: −3.15 kg ± 4.75 kg; p < 0.001). 3D demonstrated poor validity indicated by TE (%BF: 5.61%; FM: 4.50 kg; FFM: 5.69 kg). In contrast, there were no significant differences between 3D and DXA measures; 3D scanner demonstrated acceptable measurement for %BF (TE: 4.25%), FM (TE: 2.92 kg), and lean mass (TE: 3.86 kg). Compared with the 4C criterion, high TE values indicated 3D estimates were not valid. In contrast, 3D estimates produced acceptable measurement agreement when compared with DXA; an average overestimation of %BF by 5.31% (vs. 4C) and 4.20% (vs. DXA) may be expected. Novelty: 3D body composition estimates are not valid compared with the 4-C criterion model. 3D estimates appeared to be more valid in females, compared with males. When compared with DXA, 3D estimates were acceptable.

Basal cell carcinoma (BCC) of the prostate is a rare tumor. Compared with the more common acinar adenocarcinoma (AAC) of the prostate, BCCs show features of basal cell differentiation and are thought to be biologically distinct from AAC. The spectrum of molecular alterations of BCC has not been comprehensively described, and genomic studies are lacking. Herein, whole genome sequencing was performed on archival formalin-fixed, paraffin-embedded specimens of two cases with BCC. Prostatic BCCs were characterized by an overall low copy number and mutational burden. Recurrent copy number loss of chromosome 16 was observed. In addition, putative driver gene alterations in KIT, DENND3, PTPRU, MGA, and CYLD were identified. Mechanistically, depletion of the CYLD protein resulted in increased proliferation of prostatic basal cells in vitro. Collectively, these studies show that prostatic BCC displays distinct genomic alterations from AAC and highlight a potential role for loss of chromosome 16 in the pathogenesis of this rare tumor type.

The molecular mechanisms of SGLT2 inhibitors (SGLT2i) remain incompletely understood. Single-cell RNA sequencing and morphometrics data were collected from research kidney biopsies donated by participants with youth onset type 2 diabetes (T2D), aged 12-21 years of age, and healthy controls (HC) to study the effects of SGLT2i on kidney transcriptomics. Participants with T2D were more obese, had higher glomerular filtration rate, mesangial and glomerular volumes than HC. There were no clinically significant differences between participants prescribed SGLT2i (T2Di(+), n=10) and other T2D (T2Di(-), n=6). Transcriptional profiles showed SGLT2 expression exclusively in the proximal tubular (PT) cluster. Transcriptional alterations in T2Di(+) compared to T2Di(-) were seen across most nephron segments, most prominently in the distal nephron. SGLT2i treatment was associated with suppression of genes in the glycolysis, gluconeogenesis, tricarboxylic acid cycle pathways in PT, but enhanced expression in thick ascending limb. The energy sensitive mTOR signaling pathway transcripts were suppressed towards HC level in all nephron segments in T2Di(+). These transcriptional changes were confirmed in a diabetes mouse model treated with SGLT2i. Therefore, the beneficial effects of SGLT2i treatment to the kidneys might be from mitigating diabetes-induced metabolic perturbations via suppression of mTORC1 signaling across nephron segments, including those not expressing SGLT2.

Anthropometric variables will influence maximal respiratory pressure (MRP) values. Since significant variations exist in pulmonary nomograms amongst different races, it is important that tribe specific tables of normal maximal inspiratory pressures (MIP) and maximal expiratory pressures (MEP) be developed. To date, MRP prediction equations do not exist for Hopi children. PURPOSE: The purpose of this study was to develop MRP reference values and prediction equations for Hopi children in the ages 4–13 years. METHODS: A cross-sectional study was undertaken with 288 healthy children (125 male, 163 female), a 36% representative population of all the Hopi Native children attending Hopi Tribal Elementary Schools in Arizona. MIP and MEP values were measured. RESULTS: Age and the inverse of body mass were consistently significant predictors of the MRPs for both sexes. Predictions using the derived Hopi equations were significantly different (p≤0.001) than those using the equations for Navajo and Caucasian youth across both sexes, making it important for this population to have specific formulae to provide more accurate reference values. CONCLUSIONS: These data were collected from the children of Hopi ancestry resulting in MIP and MEP reference equations which should be used when measuring MIP and MEP in these children ages 4–13 years.

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Objective: We examined changes in the excretion of various amino acids and in glycolysis and ketogenesis-related metabolites, during and after diabetic ketoacidosis (DKA) diagnosis, in youth with known or new onset type 1 diabetes (T1D). Methods: Urine samples were collected from 40 youth with DKA (52% boys, mean age 11 ± 4 years, venous pH 7.2 ± 0.1, blood glucose 451 ± 163 mg/dL) at 3 time points: 0–8 h and 12–24 h after starting an insulin infusion, and 3 months after hospital discharge. Mixed-effects models evaluated the changes in amino acids and other metabolites in the urine. Results: Concentrations of urine histidine, threonine, tryptophan, and leucine per creatinine were highest at 0–8 h (148.8 ± 23.5, 59.5 ± 12.3, 15.4 ± 1.4, and 24.5 ± 2.4% of urine creatinine, respectively), and significantly decreased over 3 months (p = 0.028, p = 0.027, p = 0.019, and p < 0.0001, respectively). Urine histidine, threonine, tryptophan, and leucine per urine creatinine decreased by 10.6 ± 19.2, 0.7 ± 0.9, 1.3 ± 0.9, and 0.5 ± 0.3-fold, respectively, between 0 and 8 h and 3 months. Conclusions: In our study, DKA was associated with profound aminoaciduria, suggestive of proximal tubular dysfunction analogous to Fanconi syndrome.

TG concentrations across lipoprotein subclasses associate with diabetic kidney disease (DKD) in adults with T1D, but little is known about this relationship in youth with T1D. We evaluated cross-sectional relationships among TG concentrations across lipoprotein subclasses, intraglomerular hemodynamics, and renal oxygen availability (RO2) in youth with T1D. Glomerular filtration rate (GFR) , RO2, renal plasma flow (RPF) , afferent arteriolar resistance (RA) , efferent arteriolar resistance (RE) , intraglomerular pressure (PGLO) , and urine albumin-to-creatinine ratio (UACR) were assessed. Concentrations of lipid constituents from lipoprotein subclasses were quantified via targeted nuclear magnetic resonance spectroscopy (Nightingale Health Ltd., Helsinki, Finland) . Particle sizes for VLDL, LDL, intermediate-density lipoprotein, and HDL were measured. Fifty youth with T1D (age 16.0 ± 3.0 years, 50% female, HbA1c 8.7 ± 1.3%, T1D duration 5.7 ± 2.6 years) and 20 without T1D were included. VLDL-TG concentrations and small LDL-TG correlated with intraglomerular hemodynamic function parameters and RO2. To conclude, strong relationships were found among renal function markers and TG concentrations across lipoprotein subclasses in adolescents with T1D. Disclosure M.E.Pauley: None. K.L.Tommerdahl: None. C.Vinovskis: None. L.Pyle: None. R.Wadwa: Advisory Panel; Dompé, Consultant; Beta Bionics, Inc., Other Relationship; Tandem Diabetes Care, Inc., Research Support; Dexcom, Inc., Eli Lilly and Company, Tandem Diabetes Care, Inc. R.G.Nelson: None. K.J.Nadeau: None. P.Bjornstad: Advisory Panel; AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Horizon Therapeutics plc, LG Chem, Lilly, Novo Nordisk, Consultant; AstraZeneca, Bristol-Myers Squibb Company. Funding MEP supported by NIH (T32DK063687) ; KLT supported by NIH (K23 HL159292) , Children’s Hospital Colorado Research Institute Research Scholar Award, University of Colorado Diabetes Research Center (P30 DK116073) , Ludeman Family Center for Women’s Health Research at the University of Colorado, and Dept of Pediatrics, Section of Endocrinology at University of Colorado School of Medicine; PB supported by NIDDK (RDK129211, R21 DK129720, K23 DK116720, UC DK114886, and P30 DK116073) , JDRF (2-SRA-2019-845-S-B, 3-SRA-2017-424-M-B, 3-SRA-2022-1097-M-B) , Boettcher Foundation, American Heart Association (20IPA35260142) , Ludeman Family Center for Women’s Health Research at the University of Colorado, Department of Pediatrics, Section of Endocrinology and Barbara Davis Center for Diabetes at University of Colorado School of Medicine

Our objective was to compare renal hemodynamic function between obese youth with and without type 2 diabetes (T2D) and relate these measures to adipose insulin resistance (IR) . We assessed insulin sensitivity and kidney function in obese youth with (n=31, 15.8 ± 1.8 years, BMI 35.6 ± 6.6 kg/m2) , and without (n=20, 15.3 ± 2.1 years, BMI 38.2 ± 7.4 kg/m2) T2D. A hyperglycemic clamp was performed with 20% dextrose to maintain mild hyperglycemia for 240 minutes. Free fatty acids (FFA) were measured at baseline and every minutes during the steady state. FFA suppression (baseline FFA subtracted from steady state FFA) was used to estimate adipose IR. Iohexol and p-aminohippurate clearances were used to measure GFR and renal plasma flow, respectively. Renal hemodynamic parameters were calculated using Gomez equations. FFA suppression was attenuated in youth with T2D compared to obese controls (55.6% vs. 92.1%, p Youth with T2D had impaired FFA suppression compared to obese controls, indicating adipose IR. Impaired FFA suppression was associated with perturbed renal hemodynamics, indicating a potential role for adipose IR in the development of early diabetic kidney disease. Disclosure C.Platnick: None. P.V.Prasad: None. L.Li: None. L.Pyle: None. R.G.Nelson: None. D.Van raalte: Advisory Panel; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Merck & Co., Inc., Sanofi, Research Support; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Merck & Co., Inc., Sanofi. K.J.Nadeau: None. P.Bjornstad: Advisory Panel; AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Horizon Therapeutics plc, LG Chem, Lilly, Novo Nordisk, Consultant; AstraZeneca, Bristol-Myers Squibb Company.

Genetic studies in American Indians from Arizona previously identified a population specific Arg1420 His mutation in ABCC8 which causes hyperinsulinemic hypoglycemia in infancy. The His allele is carried by 3% of this population and associates with increased birthweight, suggesting increased insulin secretion in-utero, and doubles the risk of type 2 diabetes, a disease characterized by reduced insulin secretion. To model the contrary effects of this mutation, we used gene-edited induced pluripotent stem cell (iPSC) derived pancreatic islet-like cells. An iPS cell line created from an American Indian heterozygous for the mutation was gene-edited using CRISPR-CAS9 to generate iPSCs homozygous for the risk (His) and the non-risk (Arg) allele. Multiple iPSC lines for each genotype were differentiated towards pancreatic islet-like cells using a seven-stage differentiation protocol and the effect of the mutation on islet function were assessed. Higher insulin secretion was observed from the His-allele carrying islet-like cells when stimulated with low glucose (2mM) after 1 and 2 weeks of maturation, consistent with a clinical phenotype of hyperinsulinemia in His-allele carriers. Stimulation with 20mM glucose resulted in increased insulin secretion after 2 weeks of maturation only in the Arg-allele (non-risk) carrying islets while no increase was seen in the His-allele carrying islets, consistent with its role in T2D risk. Treatment with diazoxide significantly reduced insulin secretion with no difference between the genotypes. Single cell RNA sequencing of islet-like cells cultured for an extended time (4 weeks) identified many differentially expressed genes in cells with the mutant vs. reference allele that could potentially reveal the mechanism of the increased future T2D risk including G6PC2, a known negative regulator of glucose-stimulated insulin secretion, in beta-like cells and CRH, which has been linked to increased T2D risk, in alpha-like cells. Disclosure A.K.Nair: None. M.Traurig: None. J.Sutherland: Stock/Shareholder; Bristol-Myers Squibb Company, Johnson & Johnson, Pfizer Inc. R.G.Nelson: None. C.Bogardus: None. L.Baier: None. Funding NIH Intramural

In its broadest conceptualization, violence includes everything from physical assault to the creation and maintenance of institutional structures that are discriminatory and exclusionary. However, ...

An anthropological approach is needed to counter a rising chorus of biobabble about masculinities, maleness, and violence. Anthropological lenses enable us to examine what we know and what we do no...

Non-supplemental carotenoids and retinol may potentiate antioxidant and anti-inflammatory mechanisms. Chronic intraprostatic inflammation is linked to prostate carcinogenesis. We investigated the association of circulating carotenoids and retinol with intraprostatic inflammation in benign tissue. We included 235 men from the Prostate Cancer Prevention Trial placebo arm who had a negative end-of-study biopsy, most (92.8%) done without clinical indication. α-carotene, β-carotene, β-cryptoxanthin, lycopene, and retinol were assessed by high-performance liquid chromatography using pooled year 1 and 4 serum. Presence and extent of intraprostatic inflammation in benign tissue was assessed in 3 (of 6-10) biopsy cores. Logistic (any core with inflammation vs none) and polytomous logistic (some or all cores with inflammation vs none) regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) of intraprostatic inflammation by concentration tertile adjusting for age, race, prostate cancer family history, and serum cholesterol. None of the carotenoids or retinol was associated with intraprostatic inflammation, except β-cryptoxanthin, which appeared to be positively associated with any core with inflammation [vs none, T2: OR (95% CI) = 2.67 (1.19, 5.99); T3: 1.80 (0.84, 3.82), P-trend = 0.12]. These findings suggest that common circulating carotenoids and retinol are not useful dietary intervention targets for preventing prostate cancer via modulating intraprostatic inflammation.

Excess nutrients from agricultural settings contribute to surface water and groundwater impairment. Constructed wetlands have been widely used for water quality protection in various agricultural systems. We used a synthesis approach to document the performance of constructed wetlands for nutrient removal from a range of landscapes and geographic regions with the following objectives: (1) review the current use of constructed wetlands in agricultural applications, (2) summarize the nutrient removal efficiency of constructed wetlands, and (3) identify the geographic usage and costs associated with constructed wetlands. We reviewed over 130 publications and reports to characterize nutrient removal performance for the following types of agricultural effluents: cropland surface and subsurface drainage, and wastewater from livestock production, greenhouse, aquaculture, and hydroponic systems. Data from the reviewed studies indicate that constructed wetlands are efficient in protecting water quality in agricultural production settings. However, differences in constructed wetland characteristics reported by the studies suggest that standards are needed to ensure nutrient removal goals are met based on wetland design. Researchers should consider including basic performance parameters for constructed wetlands in published reports, including influent and effluent concentrations, hydraulic retention time, hydraulic loading rate, watershed to treatment wetland ratios, and plant species and relative cover. Future studies are needed to explore cost-benefit analyses to assess the feasibility and potential promotion of wetland incentive programs in various geographic regions and watershed nonpoint-source pollution goals for using these systems in agricultural settings. Keywords: Agricultural wastewater, Agricultural water quality, Aquaculture, Cropland runoff, Greenhouse, Hydroponic, Livestock, Review, Subsurface, Treatment wetland.

Epidemiology, clinical and experimental animal studies suggest high fructose diets are detrimental to metabolic status and may contribute to tumor development. This due to increased obesity and metabolic syndrome, known risk factors for many types of cancer. We compared tumor development in N-methyl-N-nitrosourea (MNU)-treated rats fed either a high (60%)-fructose diet (HFD) or a standard diet (SD). Female Sprague-Dawley rats at 43 days of age (DOA) were fed a SD or HFD followed by administration of MNU at 50 DOA. Rats were palpated weekly and sacrificed at 190 DOA. MNU-treated rats on HFD exhibited decreased tumor latency and roughly a two-fold increase in tumor multiplicity. RNA-Seq on frozen tumors (SD vs. HFD rats) showed altered expression of approximately 10% of genes (P < 0.05). When examined by Ingenuity Pathway Analysis, multiple highly significant pathways were identified including A) mechanisms of cancer, B) Wnt pathway, C) immune response (e.g., “Th1 and Th2 activation” and “antigen presentation”) and D) LXR/RXR nuclear receptor. These generalized pathways were indirectly confirmed by alterations of various interrelated disease pathways (epithelial cancers, T cell numbers and apoptosis). In a second study, serum was collected from rats on the HFD or SD pre-MNU and at the time of sacrifice. Metabolomics revealed that the HFD yielded: A) increased levels of fructose, B) increases of various monoglycerols, C) reduced levels of various diacylglycerols and oxygenated inflammatory lipids (9 and 13 HODE and 12,13 DHOME) and D) increased levels of secondary bile acids (hyodeoxycholate and 6-oxolithocholate), which may reflect microbiome changes. These metabolomic changes, which are distinct from those on a high-fat diet, may prove relevant when examining individuals who consume higher levels of fructose.

From a clinical, morphological and molecular perspective, prostate cancer is a heterogeneous disease. Primary prostate cancers are often multifocal, having topographically and morphologically distinct tumour foci. Sequencing studies have revealed that individual tumour foci can arise as clonally distinct lesions with no shared driver gene alterations. This finding demonstrates that multiple genomically and phenotypically distinct primary prostate cancers can be present in an individual patient. Lethal metastatic prostate cancer seems to arise from a single clone in the primary tumour but can exhibit subclonal heterogeneity at the genomic, epigenetic and phenotypic levels. Collectively, this complex heterogeneous constellation of molecular alterations poses obstacles for the diagnosis and treatment of prostate cancer. However, advances in our understanding of intra-tumoural heterogeneity and the development of novel technologies will allow us to navigate these challenges, refine approaches for translational research and ultimately improve patient care.

COVID-19 morbidity and mortality are increased in patients with diabetes and kidney disease via unknown mechanisms. SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) for entry into host cells. Since ACE2 is a susceptibility factor for infection, we investigated how diabetic kidney disease and medications alter ACE2 receptor expression in kidneys. Single cell RNA profiling of healthy living donor and kidney biopsies from patients with diabetic kidney disease revealed ACE2 expression primarily in proximal tubular epithelial cells. This cell specific localization was confirmed by in situ hybridization. ACE2 expression levels were unaltered by exposures to renin angiotensin aldosterone system inhibitors in diabetic kidney disease. Bayesian integrative analysis of a large compendium of public-omics datasets identified molecular network modules induced in ACE2-expressing proximal tubular epithelial cells in diabetic kidney disease (searchable at hb.flatironinstitute.org/covid-kidney) that were linked to viral entry, immune activation, endomembrane reorganization, and RNA processing. The diabetic kidney disease ACE2-positive proximal tubular epithelial cell module overlapped with expression patterns seen in SARS-CoV-2 infected cells. Similar cellular programs were seen in ACE2-positive proximal tubular epithelial cells obtained from urine samples of 13hospitalized patients with COVID-19, suggesting a consistent ACE2-coregulated proximal tubular epithelial cell expression program that may interact with the SARS-CoV-2 infection processes. Thus SARS-CoV-2 receptor networks can seed further research into risk stratification and therapeutic strategies for COVID-19 related kidney damage., Graphical abstract

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West Avalonia is a composite terrane that rifted from the supercontinent Gondwana in the Ordovician and accreted to Laurentia during the latest Silurian to Devonian Acadian orogeny. The nature and extent of West Avalonia are well constrained in Nova Scotia, New Brunswick, and Newfoundland, Canada, by U-Pb detrital zircon data and/or isotope geochemistry of (meta)sedimentary and igneous rocks. The southeastern New England Avalon terrane in eastern Massachusetts, Connecticut, and Rhode Island has generally been interpreted as an along-strike continuance of West Avalonia in Canada, but the ages and origins of metasedimentary units along the western boundary of the Avalon terrane in Massachusetts and Connecticut remain poorly constrained. In this study, new detrital zircon U-Pb and Lu-Hf laser-ablation–inductively coupled plasma–mass spectrometry (LA-ICP-MS) data from three samples of metasedimentary units along the western boundary of the southeastern New England Avalon terrane in Connecticut and Massachusetts were compared with existing data to test whether these metasedimentary units can be correlated along strike. The data were also compared with existing detrital zircon U-Pb and εHf data in New England and Canada in order to constrain the extent and provenance of West Avalonia. The maximum depositional age of two of the three detrital zircon samples analyzed in this study, based on the youngest single grain in each sample (600 ± 28 Ma, n = 1; 617 ± 28 Ma, n = 1) and consistency with existing analyses elsewhere in the southeastern New England Avalon terrane, is Ediacaran, while that of the third sample is Tonian (959 ± 40 Ma, n = 4). Detrital zircon analyses of all three samples from this study showed similar substantial Mesoproterozoic and lesser Paleoproterozoic and Archean populations. Other existing detrital zircon U-Pb data from quartzites in the southeastern New England Avalon terrane show similar Tonian populations with or without Ediacaran grains or populations. Most published detrital zircon U-Pb data from (meta)sedimentary rocks in West Avalonia in Canada yielded Ediacaran youngest detrital zircon age populations, except for a quartzite unit within the Gamble Brook Formation in the Cobequid Highlands of Nova Scotia, which showed a Tonian maximum depositional age, and otherwise a nearly identical detrital zircon signature with rocks from the southeastern New England Avalon terrane. All samples compiled from the southeastern New England Avalon terrane and West Avalonia in Canada show main age populations between ca. 2.0 Ga and ca. 1.0 Ga, with major peaks at ca. 1.95, ca. 1.50, ca. 1.20, and ca. 1.00 Ga, and minor ca. 3.1–3.0 Ga and ca. 2.8–2.6 Ga populations. The εHf(t) values from the three samples yielded similar results to those from West Avalonia in Canada, suggesting that both regions were derived from the same cratonic sources. The εHf(t) values of all West Avalonian samples overlap with both Amazonia and Baltica, suggesting that there is a mixed signature between cratonic sources, possibly as a result of previous collision and transfer of basement fragments between these cratons during the formation of supercontinent Rodinia, or during subsequent arc collisions.

Cultured cells are widely used in molecular biology despite poor understanding of how cell line genomes change in vitro over time. Previous work has shown that Drosophila cultured cells have a higher transposable element content than whole flies, but whether this increase in transposable element content resulted from an initial burst of transposition during cell line establishment or ongoing transposition in cell culture remains unclear. Here, we sequenced the genomes of 25 sublines of Drosophila S2 cells and show that transposable element insertions provide abundant markers for the phylogenetic reconstruction of diverse sublines in a model animal cell culture system. DNA copy number evolution across S2 sublines revealed dramatically different patterns of genome organization that support the overall evolutionary history reconstructed using transposable element insertions. Analysis of transposable element insertion site occupancy and ancestral states support a model of ongoing transposition dominated by episodic activity of a small number of retrotransposon families. Our work demonstrates that substantial genome evolution occurs during long-term Drosophila cell culture, which may impact the reproducibility of experiments that do not control for subline identity.

Objective: Understanding mechanisms underlying rapid estimated glomerular filtration rate (eGFR) decline is important to predict and treat kidney disease in type 1 diabetes (T1D). Research Design and Methods: We performed a case-control study nested within four T1D cohorts to identify urinary proteins associated with rapid eGFR decline. Cases and controls were defined by eGFR decline >3 and 2/year, respectively. We used targeted liquid chromatography-tandem mass spectrometry to measure 38 peptides from 20 proteins implicated in diabetic kidney disease. Significant proteins were investigated in complementary human cohorts and in mouse proximal tubular epithelial cell cultures. Results: The cohort study included 1270 participants followed a median 8 years. In the discovery set, only cathepsin D peptide and protein were significant on full adjustment for clinical and laboratory variables. In the validation set, associations of cathepsin D with eGFR decline were replicated in minimally-adjusted models but lost significance with adjustment for albuminuria. In a meta-analysis combining discovery and validation sets, the odds ratio for the association of cathepsin D with rapid eGFR decline was 1.29 per SD (95%CI 1.07-1.55). In complementary human cohorts, urine cathepsin D was associated with tubulointerstitial injury, and tubulointerstitial cathepsin D expression was associated with increased cortical interstitial fractional volume. In mouse proximal tubular epithelial cell cultures, advanced glycation end product-bovine serum albumin increased cathepsin D activity and inflammatory and tubular injury markers, which were further increased with cathepsin D siRNA. Conclusion: Urine cathepsin D is associated with rapid eGFR decline in T1D and reflects kidney tubulointerstitial injury.

Background Deoxycholic acid (DCA) is a secondary bile acid that may promote vascular calcification in experimental settings. Higher DCA levels were associated with prevalent coronary artery calcification (CAC) in a small group of individuals with advanced chronic kidney disease. Whether DCA levels are associated with CAC prevalence, incidence, and progression in a large and diverse population of individuals with chronic kidney disease stages 2 to 4 is unknown. Methods and Results In the CRIC (Chronic Renal Insufficiency Cohort) study, we evaluated cross‐sectional (n=1057) and longitudinal (n=672) associations between fasting serum DCA levels and computed tomographic CAC using multivariable‐adjusted regression models. The mean age was 57±12 years, 47% were women, and 41% were Black. At baseline, 64% had CAC (CAC score >0 Agatston units). In cross‐sectional analyses, models adjusted for demographics and clinical factors showed no association between DCA levels and CAC >0 compared with no CAC (prevalence ratio per 1‐SD higher log DCA, 1.08 [95% CI, 0.91–1.26). DCA was not associated with incident CAC (incidence per 1‐SD greater log DCA, 1.08 [95% CI, 0.85–1.39]) or CAC progression (risk for increase in ≥100 and ≥200 Agatston units per year per 1‐SD greater log DCA, 1.05 [95% CI, 0.84–1.31] and 1.26 [95% CI, 0.77–2.06], respectively). Conclusions Among CRIC study participants, DCA was not associated with prevalent, incident, or progression of CAC.

Refereed/Peer-reviewed Purpose: Pathways involving sex hormones and insulin-like growth factors (IGFs) have been proposed to explain, in part, the lower risk of prostate cancer among men with diabetes. To gain insights into potential biological mechanisms we explored differences in serum concentrations of sex hormones and IGFs across the trajectory from normoglycemia to prediabetes to poorly controlled diabetes. Methods: Using cross-sectional data from the National Health and Nutrition Examination Survey III we examined differences in levels of circulating sex hormones, sex hormone-binding globulin (SHBG), IGF-1, and IFG-binding protein 3 (IGFBP-3), according to diabetes status: no diabetes [n = 648], prediabetes [n = 578], undiagnosed diabetes [n = 106], well-controlled diabetes [n = 42], and poorly controlled diabetes [n = 56]. Adjusted geometric mean concentrations were derived using multivariable linear regression, adjusted for age, race, and other lifestyle factors. Results: Total testosterone concentrations were lower among prediabetics (4.89 ng/mL, 95% confidence interval (CI) 4.95–5.21) than men without prediabetes/diabetes (5.29 ng/mL, 95% CI 5.06–5.53) but did not reduce further across diabetes groups. Concentrations of estradiol, estimated free testosterone, SHGB, IGF-1, and IGFBP-3 did not differ. While the ratio of IGF-1 to IGFBP-3 was lower among men with prediabetics and undiagnosed diabetes than men without prediabetes/diabetes, there was no trend across groups. A positive trend for the ratio of estradiol-to-testosterone levels was observed across groups (p trend = 0.045). Conclusion: Our findings do not provide clear support for either an androgen driven or IGF-driven pathway for the inverse association between diabetes and prostate cancer risk.

Background: Early identification of youth with type 1 diabetes (T1D) at risk for diabetic kidney disease may improve clinical outcomes. We examined the cross-sectional relationship between kidney biomarkers neutrophil gelatinase–associated lipocalin (NGAL), copeptin, interleukin-18 (IL-18), kidney injury molecule-1 (KIM-1), chitinase-3-like protein-1 (YKL-40), and monocyte chemoattractant protein-1 (MCP-1) and intrarenal hemodynamic function in adolescents with T1D. Methods: Urine albumin-to-creatinine ratio (UACR), renal vascular resistance (RVR), glomerular filtration rate (GFR), intraglomerular pressure (P GLO), efferent arteriole resistance (R E), afferent arteriolar resistance (R A), and renal plasma flow (RPF), and the above indicated biomarkers were assessed in youth aged 12–21 years with and without T1D of < 10 years duration. Results: Fifty adolescents with T1D (16.1 ± 3.0 years, HbA1c 8.6 ± 1.2%) and 20 adolescents of comparable BMI without T1D (16.1 ± 2.9 years, HbA1c 5.2 ± 0.2%) were enrolled. Adolescents with T1D demonstrated significantly higher GFR, RPF, R E, and P GLO than controls (39%, 33%, 74%, and 29%, respectively, all p < 0.0001). Adolescents with T1D also exhibited significantly lower RVR and R A than controls (25% and 155%, respectively, both p < 0.0001). YKL-40 and KIM-1 concentrations, respectively, were positively associated with GFR (r: 0.43, p = 0.002; r: 0.41, p = 0.003), RPF (r: 0.29, p = 0.08; r: 0.34, p = 0.04), UACR (r: 0.33, p = 0.02; r: 0.50, p = 0.0002), and P GLO (r: 0.45, p = 0.006; r: 0.52, p = 0.001) in adolescents with T1D. Conclusions: Higher concentrations of biomarkers YKL-40 and KIM-1 may help define the risk for intraglomerular hemodynamic dysfunction in youth with T1D. Graphical abstract: A higher resolution version of the Graphical abstract is available as Supplementary information. [Figure not available: see fulltext.]

Defective cholesterol metabolism primarily linked to reduced ATP-binding cassette transporter A1 (ABCA1) expression is closely associated with the pathogenesis and progression of kidney diseases, including diabetic kidney disease and Alport Syndrome. However, whether the accumulation of free or esterified cholesterol contributes to progression in kidney disease remains unclear. Here, we demonstrate that inhibition of sterol-O-acyltransferase-1 (SOAT1), the enzyme at the endoplasmic reticulum that converts free cholesterol to cholesterol esters, which are then stored in lipid droplets, effectively reduced cholesterol ester and lipid droplet formation in human podocytes. Furthermore, we found that inhibition of SOAT1 in podocytes reduced lipotoxicity-mediated podocyte injury in diabetic kidney disease and Alport Syndrome in association with increased ABCA1 expression and ABCA1-mediated cholesterol efflux. In vivo, Soat1 deficient mice did not develop albuminuria or mesangial expansion at 10-12 months of age. However, Soat1 deficiency/inhibition in experimental models of diabetic kidney disease and Alport Syndrome reduced cholesterol ester content in kidney cortices and protected from disease progression. Thus, targeting SOAT1-mediated cholesterol metabolism may represent a new therapeutic strategy to treat kidney disease in patients with diabetic kidney disease and Alport Syndrome, like that suggested for Alzheimer's disease and cancer treatments.