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Cases of death from chloroform during anesthesia or immediately after are well known. But even Casper in 1850 claimed that harmful effects after prolonged chloroform anesthesia could affect later (Nachwirkungen) - "to a certain extent, chronic chloroform poisoning".

Transferrin-bound iron (TBI), the physiological circulating iron form, is acquired by cells through the transferrin receptor (TfR1) by endocytosis. In erythroid cells, most of the acquired iron is incorporated into heme in the mitochondria. Cellular trafficking of heme is indispensable for erythropoiesis and many other essential biological processes. Comprehensive elucidation of molecular pathways governing and regulating cellular iron acquisition and heme trafficking is required to better understand physiological and pathological processes affecting erythropoiesis. Here, we report the first genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screens in human erythroid cells to identify determinants of iron and heme uptake, as well as heme-mediated erythroid differentiation. We identified several candidate modulators of TBI acquisition including TfR1, indicating that our approach effectively revealed players mechanistically relevant to the process. Interestingly, components of the endocytic pathway were also revealed as potential determinants of transferrin acquisition. We deciphered a role for the vacuolar-type H+ - ATPase (V- ATPase) assembly factor coiled-coil domain containing 115 (CCDC115) in TBI uptake and validated this role in CCDC115 deficient K562 cells. Our screen in hemin-treated cells revealed perturbations leading to cellular adaptation to heme, including those corresponding to trafficking mechanisms and transcription factors potentiating erythroid differentiation. Pathway analysis indicated that endocytosis and vesicle acidification are key processes for heme trafficking in erythroid precursors. Furthermore, we provided evidence that CCDC115, which we identified as required for TBI uptake, is also involved in cellular heme distribution. This work demonstrates a previously unappreciated common intersection in trafficking of transferrin iron and heme in the endocytic pathway of erythroid cells.

Minority-carrier lifetimes and diffusion lengths have been deduced from a comparison of band structure simulations and experimental measurements on mid-wave infrared InAsSb XBn and long-wave infrared InAs/GaSb type II superlattice (T2SL) XBp barrier detectors with low diffusion-limited dark current close to mercury cadmium telluride Rule 07 and high quantum efficiency. For the XBn devices, a lifetime of 1.9 μs was observed with a corresponding diffusion length of 14.5 μm. In contrast, the T2SL exhibited a much shorter lifetime of 7.5 ns, but the diffusion length of ∼ 7 μm was long enough to ensure that almost 90% of the photocarriers are collected. The lifetime appears to be Auger limited in the case of n-type InAsSb, but for the p-type T2SL, Shockley–Read–Hall (SRH) traps appear to dominate. In the second case, possible scenarios for the dominance of SRH recombination are discussed to identify pathways for further performance optimization.

Background Short-term treatment for people with type 2 diabetes using a low dose of the selective endothelin A receptor antagonist atrasentan reduces albuminuria without causing significant sodium retention. We report the long-term effects of treatment with atrasentan on major renal outcomes.Methods We did this double-blind, randomised, placebo-controlled trial at 689 sites in 41 countries. We enrolled adults aged 18-85 years with type 2 diabetes, estimated glomerular filtration rate (eGFR) 25-75 mL/min per 1.73 m(2) of body surface area, and a urine albumin-to-creatinine ratio (UACR) of 300-5000 mg/g who had received maximum labelled or tolerated renin-angiotensin system inhibition for at least 4 weeks. Participants were given atrasentan 0.75 mg orally daily during an enrichment period before random group assignment. Those with a UACR decrease of at least 30% with no substantial fluid retention during the enrichment period (responders) were included in the double-blind treatment period. Responders were randomly assigned to receive either atrasentan 0.75 mg orally daily or placebo. All patients and investigators were masked to treatment assignment. The primary endpoint was a composite of doubling of serum creatinine (sustained for >= 30 days) or end-stage kidney disease (eGFR = 90 days, chronic dialysis for >= 90 days, kidney transplantation, or death from kidney failure) in the intention-to-treat population of all responders. Safety was assessed in all patients who received at least one dose of their assigned study treatment. The study is registered with ClinicalTrials. gov, number NCT01858532.Findings Between May 17, 2013, and July 13, 2017, 11 087 patients were screened; 5117 entered the enrichment period, and 4711 completed the enrichment period. Of these, 2648 patients were responders and were randomly assigned to the atrasentan group (n=1325) or placebo group (n=1323). Median follow-up was 2.2 years (IQR 1.4-2.9). 79 (6.0%) of 1325 patients in the atrasentan group and 105 (7.9%) of 1323 in the placebo group had a primary composite renal endpoint event (hazard ratio [HR] 0.65 [95% CI 0.49-0.88]; p=0.0047). Fluid retention and anaemia adverse events, which have been previously attributed to endothelin receptor antagonists, were more frequent in the atrasentan group than in the placebo group. Hospital admission for heart failure occurred in 47 (3.5%) of 1325 patients in the atrasentan group and 34 (2.6%) of 1323 patients in the placebo group (HR 1.33 [95% CI 0.85-2.07]; p=0.208). 58 (4.4%) patients in the atrasentan group and 52 (3.9%) in the placebo group died (HR 1.09 [95% CI 0.75-1.59]; p=0.65).Interpretation Atrasentan reduced the risk of renal events in patients with diabetes and chronic kidney disease who were selected to optimise efficacy and safety. These data support a potential role for selective endothelin receptor antagonists in protecting renal function in patients with type 2 diabetes at high risk of developing end-stage kidney disease. Copyright (C) 2019 Elsevier Ltd. All rights reserved.

The equivalent conductivity of an ion in solution is derived by multiplying the equivalent conductivity of the entire electrolyte, Λ, by the ion’s transport number. Λ is obtained directly from the specific conductivity, S , but a better way of converting S to Λ is through σi, the specific conductivity of ion i. Redefined to include the contribution of the counterion to the mobility of ion i, using a new concept of “ionic power”, σi is theoretically developed here and its concentration dependence is demonstrated and discussed for select electrolytes of different valence families.

The physical essence of the limiting equivalent ionic conductivity in solution, λ0i, has been a continuing challenge over almost a century. Here I briefly present an ab initio theoretical treatment providing (1) a new insight into the nature of λ0i, and (2) a mathematical formula for computing λ0i. In the new treatment, one assumes that any chosen ion i is surrounded by a spherical body of oriented solvent dipoles carrying the charge of the counterion, and the bulk solvent is a continuum with no molecular detail. λ0i is thus the result of the tandem operation, at hydrodynamic equilibrium, of the dipole body's electrophoretic and relaxation forces exerted on the drifting ion. λ0i is found to be proportional to the radius of ion i, and independent of the ionic charge. From experimental λ0i's, the ion radius can be computed as ‘electric radius.’ An electric ion-radius scale so derived compares well with other ion-size scales. The current theory expresses λ0i using only universal constants and unitary...

XBn or XBp barrier detectors exhibit diffusion-limited dark currents comparable with mercury cadmium telluride Rule-07 and high quantum efficiencies. In 2011, SemiConductor Devices (SCD) introduced “HOT Pelican D”, a 640 × 512/15-μm pitch InAsSb/AlSbAs XBn mid-wave infrared (MWIR) detector with a 4.2-μm cut-off and an operating temperature of ∼150 K. Its low power (∼3 W), high pixel operability (>99.5%) and long mean time to failure make HOT Pelican D a highly reliable integrated detector-cooler product with a low size, weight and power. More recently, “HOT Hercules” was launched with a 1280 × 1024/15-μm format and similar advantages. A 3-megapixel, 10-μm pitch version (“HOT Blackbird”) is currently completing development. For long-wave infrared applications, SCD’s 640 × 512/15-μm pitch “Pelican-D LW” XBp type II superlattice (T2SL) detector has a ∼9.3-μm cut-off wavelength. The detector contains InAs/GaSb and InAs/AlSb T2SLs, and is fabricated into focal plane array (FPA) detectors using standard production processes including hybridization to a digital silicon read-out integrated circuit (ROIC), glue underfill and substrate thinning. The ROIC has been designed so that the complete detector closely follows the interfaces of SCD’s MWIR Pelican-D detector family. The Pelican-D LW FPA has a quantum efficiency of ∼50%, and operates at 77 K with a pixel operability of >99% and noise equivalent temperature difference of 13 mK at 30 Hz and F/2.7.

The variation of the activity coefficient of aqueous sulfuric acid with acid concentration, solution's temperature, and solvent's (water) dielectric constant, clearly fits a typical strong 1–3 electrolyte behaviour [Fraenkel, J. Phys. Chem. B. 116, 11662 (2012)]. Therefore, the aqueous acid was proposed to have the molecular formula H4SO5, and to dissociate to 3H+ (3H3O+) and 1HSO53−. When the above study was reported, electric conductivity data of the aqueous acid could not be theoretically interpreted to prove or disprove the 1–3 electrolyte behaviour, due to the lack of an appropriate conductance theory. The improved Debye–Hückel–Onsager (DHO) theory, DHO–SiS, recently reported [Fraenkel, Phys. Chem. Chem. Phys. 20, 29896 (2018)], allows, unlike its parent theory, a quantitative analysis of strong 1–1, 1–2 and 1–3 electrolytes. A good-to-excellent theory–experiment agreement is achieved for dilute aqueous H2SO4 at 0–50°C, but only if the acid is considered, as before, as a strong 1–3 electrolyte; the new analysis thus corroborates the previous activity-based findings. In contrast, the conductance of dilute sulfuric acid in pure methanol follows the conventional H2SO4 dissociation pattern, exhibiting a ‘mixed 1–1/1–2’ electrolyte behaviour.

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Small molecule screens are widely used to prioritize pharmaceutical development. However, determining the pathways targeted by these molecules is challenging, since the compounds are often promiscuous. We present a network strategy that takes into account the polypharmacology of small molecules in order to generate hypotheses for their broader mode of action. We report a screen for kinase inhibitors that increase the efficacy of gemcitabine, the first-line chemotherapy for pancreatic cancer. Eight kinase inhibitors emerge that are known to affect 201 kinases, of which only three kinases have been previously identified as modifiers of gemcitabine toxicity. In this work, we use the SAMNet algorithm to identify pathways linking these kinases and genetic modifiers of gemcitabine toxicity with transcriptional and epigenetic changes induced by gemcitabine that we measure using DNaseI-seq and RNA-seq. SAMNet uses a constrained optimization algorithm to connect genes from these complementary datasets through a small set of protein-protein and protein-DNA interactions. The resulting network recapitulates known pathways including DNA repair, cell proliferation and the epithelial-to-mesenchymal transition. We use the network to predict genes with important roles in the gemcitabine response, including six that have already been shown to modify gemcitabine efficacy in pancreatic cancer and ten novel candidates. Our work reveals the important role of polypharmacology in the activity of these chemosensitiz-ing agents., National Institutes of Health (U.S.) (Grant R01-GM089903), National Institutes of Health (U.S.) (Grant U01-CA184898)

Seasonal variations of 25-hydroxyvitamin D, PTH and calcium levels are not well characterized in primary hyperparathyroidism (PHPT). Our objectives were to characterize seasonal changes in these parameters in PHPT patients, and to assess whether these seasonal changes affect clinical decision making.This is a retrospective study based on the electronic medical records of Clalit Health service in the south of Israel between 2000 and 2012. Patients 18years and older with PHPT (PTHupper limit of norm (ULN) and serum calcium10.5mg%) were included. Patients with renal failure or on Thiazide diuretics were excluded. All serum levels of calcium, PTH and 25-hydroxyvitamin D were collected and then stratified according to season.792 patients were classified as PHPT (72.2% female) and had a total of 2659 PTH tests, 1395 25-hydroxyvitamin D tests and 7426 calcium test. Fifty six percent of 25-hydroxyvitamin D levels were50nmol/L. Seasonality was demonstrated in all three parameters: mean 25-hydroxyvitamin D was 13% higher in the summer compared to the winter (P0.001), median PTH values showed opposite trend with a fall of about 8.4% in summer compared to winter (P0.001). Calcium levels were higher during the autumn with a rise of about 0.2mg/dL in the mean calcium levels compared to spring and summer (P0.001). The odds ratio of calcium level above 11.5mg/dL is highest in the autumn (OR=1.275, P=0.018).We show seasonal variation in serum 25-hydroxyvitamin D, PTH, and calcium levels in patients with PHPT. These seasonal variations cause transition to pathological values that may influence diagnosis and treatment of PHPT patients.

BACKGROUND AND AIMS While there is broad recognition of the high societal costs of substance use disorders (SUD), treatment rates are low. We examined whether, in the United States, participants with substance or alcohol use disorder would report a greater willingness to enter SUD treatment located in a primary care setting (primary care) or more commonly found specialty care setting in the United States (usual care). DESIGN Randomized survey-embedded experiment. SETTING US web-based research panel in which participants were randomized to read one-paragraph vignettes describing treatment in usual care (specialty drug or alcohol treatment center), primary care or collaborative care within a primary care setting. PARTICIPANTS A total of 42 451 panelists aged 18+ were screened for substance or alcohol use disorder using validated diagnostic criteria. Participants included 344 with a substance use disorder and 634 with an alcohol use disorder not in treatment with no prior treatment history. MEASURES Willingness to enter treatment across vignettes by condition. FINDINGS Among participants with a substance use disorder, 24.6% of those randomized to usual care reported being willing to enter drug treatment compared with 37.2% for primary care [12.6 percentage point difference; 95% confidence interval (CI) = 0.8, 24.4) and 34.0% for collaborative care (9.4 percentage point difference; 95% CI = -2.0, 20.8). Among participants with an alcohol use disorder, 17.6% of those randomized to usual care reported being willing to enter alcohol treatment compared with 20.3% for primary care (2.6 percentage point difference; 95% CI = -4.9, 10.1) and 20.8% for collaborative care (3.1 percentage point difference; 95% CI = -4.3, 10.6). The most common reason for not being willing to enter drug (63%) and alcohol (78%) treatment was the belief that treatment was not needed. CONCLUSIONS In the United States, people diagnosed with substance or alcohol use disorders appear to be more willing to enter treatment in a primary care setting than in a specialty drug treatment center. Expanding availability of primary care-based substance use disorder treatment could increase treatment rates in the United States.

Altered cellular metabolism is believed to be an important contributor to pathogenesis of the neurodegenerative disorder Huntington’s disease (HD). Research has primarily focused on mitochondrial toxicity, which can cause death of the vulnerable striatal neurons, but other aspects of metabolism have also been implicated. Most previous studies have been carried out using postmortem human brain or non-human cells. Here, we studied bioenergetics in an induced pluripotent stem cell-based model of the disease. We found decreased adenosine triphosphate (ATP) levels in HD cells compared to controls across differentiation stages and protocols. Proteomics data and multiomics network analysis revealed normal or increased levels of mitochondrial messages and proteins, but lowered expression of glycolytic enzymes. Metabolic experiments showed decreased spare glycolytic capacity in HD neurons, while maximal and spare respiratory capacities driven by oxidative phosphorylation were largely unchanged. ATP levels in HD neurons could be rescued with addition of pyruvate or late glycolytic metabolites, but not earlier glycolytic metabolites, suggesting a role for glycolytic deficits as part of the metabolic disturbance in HD neurons. Pyruvate or other related metabolic supplements could have therapeutic benefit in HD.

Newly recorded Raman spectra of aqueous sulfuric acid provide scattering shifts of very high sensitivity hence signal-to-noise ratios, at a broad concentration range, up to 17 M. I interpret the spectra as (a) providing no evidence for incomplete H2SO4 ionization at low concentration, (b) reflecting only one dissociation event below ∼5 M, (c) indicating a gradual ion association at ∼5–12.5 M, and (d) exhibiting further structural changes of the sulfate above 12.5 M. The analysis of the Raman shifts supports the postulated presence in solution of the para-bisulfate anion HSO53− as a sole sulfate ion up to ∼5 M, as concluded before (D. Fraenkel, J. Phys. Chem. B, 2012, 116, 11662; D. Fraenkel, J. Phys. Chem. B, 2012, 116, 11678; D. Fraenkel, J. Chem. Thermodyn., 2014, 78, 215), an ion association producing para-sulfuric acid, H4SO5 between 5 and 12.5 M, and a dehydration of H4SO5 to H2SO4 above 12.5 M. These conclusions are rationalized and corroborated by (1) a correlation of the Raman spectra with well-known physicochemical properties of aqueous sulfuric acid, and (2) structural analogies between the proposed para-sulfates and related compounds and anions.

Numerous genes and molecular pathways are implicated in neurodegenerative proteinopathies, but their inter-relationships are poorly understood. We systematically mapped molecular pathways underlying the toxicity of alpha-synuclein (α-syn), a protein central to Parkinson's disease. Genome-wide screens in yeast identified 332 genes that impact α-syn toxicity. To “humanize” this molecular network, we developed a computational method, TransposeNet. This integrates a Steiner prize-collecting approach with homology assignment through sequence, structure, and interaction topology. TransposeNet linked α-syn to multiple parkinsonism genes and druggable targets through perturbed protein trafficking and ER quality control as well as mRNA metabolism and translation. A calcium signaling hub linked these processes to perturbed mitochondrial quality control and function, metal ion transport, transcriptional regulation, and signal transduction. Parkinsonism gene interaction profiles spatially opposed in the network (ATP13A2/PARK9 and VPS35/PARK17) were highly distinct, and network relationships for specific genes (LRRK2/PARK8, ATXN2, and EIF4G1/PARK18) were confirmed in patient induced pluripotent stem cell (iPSC)-derived neurons. This cross-species platform connected diverse neurodegenerative genes to proteinopathy through specific mechanisms and may facilitate patient stratification for targeted therapy. Keywords: alpha-synuclein; iPS cell; Parkinson’s disease; stem cell; mRNA translation; RNA-binding protein; LRRK2; VPS35; vesicle trafficking; yeast

Uncovering the molecular context of dysregulated metabolites is crucial to understand pathogenic pathways. However, their system-level analysis has been limited owing to challenges in global metabolite identification. Most metabolite features detected by untargeted metabolomics carried out by liquid-chromatography-mass spectrometry cannot be uniquely identified without additional, time-consuming experiments. We report a network-based approach, prize-collecting Steiner forest algorithm for integrative analysis of untargeted metabolomics (PIUMet), that infers molecular pathways and components via integrative analysis of metabolite features, without requiring their identification. We demonstrated PIUMet by analyzing changes in metabolism of sphingolipids, fatty acids and steroids in a Huntington's disease model. Additionally, PIUMet enabled us to elucidate putative identities of altered metabolite features in diseased cells, and infer experimentally undetected, disease-associated metabolites and dysregulated proteins. Finally, we established PIUMet's ability for integrative analysis of untargeted metabolomics data with proteomics data, demonstrating that this approach elicits disease-associated metabolites and proteins that cannot be inferred by individual analysis of these data., National Institutes of Health (U.S.) (grant R01-GM089903), National Institutes of Health (U.S.) (grant U54-NS091046), National Institutes of Health (U.S.) (grant U01-CA184898), National Cancer Institute (U.S.) (grant U54 CA112967), National Cancer Institute (U.S.) (grant P30 CA014051), Searle Scholars Program

Iron overload causes the generation of reactive oxygen species that can lead to lasting damage to the liver and other organs. The goal of this study was to identify genes that modify the toxicity of iron overload. We studied the effect of iron overload on the hepatic transcriptional and metabolomic profile in mouse models using a dietary model of iron overload and a genetic model, the hemojuvelin knockout mouse. We then evaluated the correlation of nicotinamide N-methyltransferase (NNMT) expression with body iron stores in human patients and the effect of NNMT knockdown on gene expression and viability in primary mouse hepatocytes. We found that iron overload induced significant changes in the expression of genes and metabolites involved in glucose and nicotinamide metabolism and that NNMT, an enzyme that methylates nicotinamide and regulates hepatic glucose and cholesterol metabolism, is one of the most strongly down-regulated genes in the liver in both genetic and dietary iron overload. We found that hepatic NNMT expression is inversely correlated with serum ferritin levels and serum transferrin saturation in patients who are obese, suggesting that body iron stores regulate human liver NNMT expression. Furthermore, we demonstrated that adenoviral knockdown of NNMT in primary mouse hepatocytes exacerbates iron-induced hepatocyte toxicity and increases expression of transcriptional markers of oxidative and endoplasmic reticulum stress, while overexpression of NNMT partially reversed these effects. Conclusion: Iron overload alters glucose and nicotinamide transcriptional and metabolic pathways in mouse hepatocytes and decreases NNMT expression, while NNMT deficiency worsens the toxic effect of iron overload. For these reasons, NNMT may be a drug target for the prevention of iron-induced hepatotoxicity. (Hepatology Communications 2017;1:803-815).

The calculation of thermodynamic properties of many strong electrolytes in solution, including aqueous sulfuric acid, has been performed over the past four decades using so-called thermodynamic models, such as the well-known Pitzer model. I have recently pointed out (Fraenkel, 2012) [15,16] that H 2 SO 4 in water appears to follow the mean ionic activity pattern of a strong 1–3 electrolyte, and postulated that this H 3 A acid may be H 4 SO 5 fully ionizing to 3H + (3H 3 O + ) and HSO 5 3 - . This contrasts with the traditional view of the aqueous acid – claimed to be supported by thermodynamic models – according to which H 2 SO 4 retains its molecular structure in water and dissociates primarily to H + and HSO 4 - , and at HSO 4 - dissociates further to H + and SO 4 2 - . I now show that a good fit of Pitzer model with the activity coefficients reported by Hamer and Harned can be obtained for the “1–3 H 2 SO 4 ” even by using the simple 3-parameter equation of the model; the best-fit Pitzer parameters are β (0) = 0.240, β (1) = 4.30 and C MX = −0.0134, and the standard deviation, σ is 0.0152. With the corrected activity coefficients as proposed in the first reference above, the best-fit parameters are β (0) = 0.230, β (1) = 3.60 and C MX = −0.0120, and σ = 0.0081. σ of the analysis of the “1–3 acid” is in both cases considerably lower than that of the “1–2 acid” ( σ = 0.049) that provides a best-fit β (1) value of −3.000; a negative β (1) is inappropriate since it is parallel to a negative ion–ion distance of closest approach in Debye–Huckel-type expressions of the activity coefficient.

Structural studies of the N-(2,4-dinitrophenyl) derivative of a DielsAlder-cyclized 1,2-dihydropyridine both unequivocally established the polycyclic framework and revealed interesting distortions of aromatic structure and unique dimeric clustering of the aromatic entities in the solid state.

Hepcidin, a peptide hormone that decreases intestinal iron absorption and macrophage iron release, is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. Endogenous stimulants of Hepcidin transcription include bone morphogenic protein 6 (BMP6) and interleukin-6 (IL-6) by effects on mothers against decapentaplegic homolog (Smad)4 or signal transducer and activator of transcription (Stat)3, respectively. We conducted a small-scale chemical screen in zebrafish embryos to identify small molecules that modulate hepcidin expression. We found that treatment with the isoflavone, genistein, from 28-52 hours postfertilization in zebrafish embryos enhanced Hepcidin transcript levels, as assessed by whole-mount in situ hybridization and quantitative real-time reverse-transcriptase polymerase chain reaction. Genistein's stimulatory effect was conserved in human hepatocytes: Genistein treatment of HepG2 cells increased both Hepcidin transcript levels and promoter activity. We found that genistein's effect on Hepcidin expression did not depend on estrogen receptor signaling or increased cellular iron uptake, but was impaired by mutation of either BMP response elements or the Stat3-binding site in the Hepcidin promoter. RNA sequencing of transcripts from genistein-treated hepatocytes indicated that genistein up-regulated 68% of the transcripts that were up-regulated by BMP6; however, genistein raised levels of several transcripts involved in Stat3 signaling that were not up-regulated by BMP6. Chromatin immunoprecipitation and ELISA experiments revealed that genistein enhanced Stat3 binding to the Hepcidin promoter and increased phosphorylation of Stat3 in HepG2 cells. Conclusion: Genistein is the first small-molecule experimental drug that stimulates Hepcidin expression in vivo and in vitro. These experiments demonstrate the feasibility of identifying and characterizing small molecules that increase Hepcidin expression. Genistein and other candidate molecules may subsequently be developed into new therapies for iron overload syndromes., United States. Army Research Office (Institute for Collaborative Biotechnologies Grant W911NF-09-001)

The Raman spectrum of DMAPP in the solid, colloid and solution phases is presented for the first time. Its suitability to follow the dissociation reactions of DMAPP (thermal and photochemical) under different conditions is confirmed.

We construct a pathway de novo for microenvironment-specific genetic regulators of acute lymphoblastic leukemia using RNAi screening, and mRNA data., Data integration stands to improve interpretation of RNAi screens which, as a result of off-target effects, typically yield numerous gene hits of which only a few validate. These off-target effects can result from seed matches to unintended gene targets (reagent-based) or cellular pathways, which can compensate for gene perturbations (biology-based). We focus on the biology-based effects and use network modeling tools to discover pathways de novo around RNAi hits. By looking at hits in a functional context, we can uncover novel biology not identified from any individual ‘omics measurement. We leverage multiple ‘omic measurements using the Simultaneous Analysis of Multiple Networks (SAMNet) computational framework to model a genome scale shRNA screen investigating Acute Lymphoblastic Leukemia (ALL) progression in vivo. Our network model is enriched for cellular processes associated with hematopoietic differentiation and homeostasis even though none of the individual ‘omic sets showed this enrichment. The model identifies genes associated with the TGF-beta pathway and predicts a role in ALL progression for many genes without this functional annotation. We further experimentally validate the hidden genes – Wwp1, a ubiquitin ligase, and Hgs, a multi-vesicular body associated protein – for their role in ALL progression. Our ALL pathway model includes genes with roles in multiple types of leukemia and roles in hematological development. We identify a tumor suppressor role for Wwp1 in ALL progression. This work demonstrates that network integration approaches can compensate for off-target effects, and that these methods can uncover novel biology retroactively on existing screening data. We anticipate that this framework will be valuable to multiple functional genomic technologies – siRNA, shRNA, and CRISPR – generally, and will improve the utility of functional genomic studies.

SCD has developed a range of advanced infrared detectors based on III-V semiconductor heterostructures grown on GaSb. The XBn/XBp family of barrier detectors enables diffusion limited dark currents, comparable with MCT Rule-07, and high quantum efficiencies. This work describes some of the technical challenges that were overcome, and the ultimate performance that was finally achieved, for SCD’s new 15 μm pitch “Pelican-D LW” type II superlattice (T2SL) XBp array detector. This detector is the first of SCD's line of high performance two dimensional arrays working in the LWIR spectral range, and was designed with a ~9.3 micron cut-off wavelength and a format of 640 x 512 pixels. It contains InAs/GaSb and InAs/AlSb T2SLs, engineered using k • p modeling of the energy bands and photo-response. The wafers are grown by molecular beam epitaxy and are fabricated into Focal Plane Array (FPA) detectors using standard FPA processes, including wet and dry etching, indium bump hybridization, under-fill, and back-side polishing. The FPA has a quantum efficiency of nearly 50%, and operates at 77 K and F/2.7 with background limited performance. The pixel operability of the FPA is above 99% and it exhibits a stable residual non uniformity (RNU) of better than 0.04% of the dynamic range. The FPA uses a new digital read-out integrated circuit (ROIC), and the complete detector closely follows the interfaces of SCD’s MWIR Pelican-D detector. The Pelican- D LW detector is now in the final stages of qualification and transfer to production, with first prototypes already integrated into new electro-optical systems.

Hepcidin, a peptide produced in the liver, decreases intestinal iron absorption and macrophage iron release by causing degradation of the iron exporter, ferroportin. Because its levels are inappropriately low in patients with iron overload syndromes, hepcidin is a potential drug target. We previously conducted a chemical screen that revealed ipriflavone, an orally available small molecule, as a potent inducer of hepcidin expression. To evaluate ipriflavone's effect on iron homeostasis, we placed groups of 5-week old wild type or thalassemia intermedia (HbbTh3+/−) mice on a soy-free, iron-sufficient diet, AIN-93G containing 220 mg iron and 0-750 mg ipriflavone per kg of food for 50 days. Ipriflavone 500 mg/kg significantly reduced liver iron stores and intestinal ferroportin expression in WT mice, while increasing the ratio of hepcidin transcript levels to liver iron stores. Ipriflavone supplementation in HbbTh3+/− mice failed to alleviate iron overload and was associated with a milder reduction in intestinal ferroportin and a failure to alter the ratio of hepcidin transcript levels to liver iron stores or splenic expression of the hepcidin-regulatory hormone, erythroferrone. These data suggest that dietary supplementation with ipriflavone alone would not be sufficient to treat iron overload in thalassemia intermedia.

OBJECTIVE To examine longitudinal change in vitamin D binding protein (DBP) levels during the first year after initiation of tenofovir disoproxil fumarate (TDF)/lamivudine/efavirenz and compare these findings with concurrent changes in markers of skeletal metabolism. DESIGN Secondary analysis of plasma samples collected from an ongoing multicenter clinical trial. METHODS Plasma samples collected at 0, 24, and 48 weeks after initiation of TDF + lamivudine + efavirenz from 134 adult participants enrolled in a multicenter randomized trial were analyzed. Data regarding sociodemographic and clinical characteristics were obtained as part of the parent study. Laboratory analyses included plasma DBP, intact parathyroid hormone, total 25-hydroxy vitamin D, phosphorus, the bone resorption marker collagen type 1 cross-linked C-telopeptide, and the bone formation marker total procollagen type 1 N-terminal propeptide. Repeated measures analysis of variance was used to measure changes in biomarkers over time. RESULTS Our sample included 108 men and 26 women (mean age 33.6 ± 9.6 years). Median levels of DBP increased significantly from baseline to 48 weeks [154 (91.8-257.4) versus 198.3 (119.6-351.9) μg/ml, P

In a one-pot process without isolation of intermediates, (but-3-en-1-yl)pyridine (13) is treated sequentially with dicyclohexylborane, trimethylaluminium, and ethyl carbonochloridate yielding ethyl 1,4-dihydro-4,4-(tetramethylene)pyridine-1-carboxylate (=ethyl 8-azaspiro[4.5]deca-6,9-diene-8-carboxylate; 2) in 46% yield based on starting alkenylpyridine 13 (Scheme 5).