bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of CEACAM1 in Endothelial Cells Contributes to the Development of Cardiac Fibrosis
Authors: MUTURI HARRISON T. , GHADIEH HILDA E. , KHUDER SAJA S. , JASH SUKANTA , ABU HELAL RAGHD , DANIELS NIGEL A. , LIU JAMES , PURI VISHWAJEET , VAZQUEZ GUILLERMO , GUPTA RAJESH , NAJJAR SONIA M. 
Published: June 2018

Carcinoembryonic Antigen-related Cell Adhesion Molecule 1 (CEACAM1) has been implicated in various physiological processes in the vascular systems, including blood vessel formation, and endothelial barrier function. Consistent with a role for hepatic CEACAM1 in promoting insulin sensitivity by mediating insulin clearance and maintaining physiologic plasma insulin levels, we have recently identified a role for hepatic CEACAM1 in preventing cardiac hypertrophy and dysfunction. The aim of this study was to elucidate the contribution of endothelial cell CEACAM1 in cardiac function and fibrosis that has remained elusive. To this end, we generated an endothelial cell-specific Ceacam1 knockout mouse line (VECadCc1−/−). Gomori Trichrome Blue Staining revealed a significant perivascular and interstitial fibrosis in the heart of VECadCc1−/− mice compared to their control littermates. Consistently, the mRNA levels of markers of fibrosis [collagen 1A1, α-SMA and fibroblast-specific protein1 (FSP1)] were elevated with a reciprocal decrease of endothelial cell makers (CD31), indicating endothelial mesenchymal transition (EndMT) in these mice. Cardiac fibrosis was associated with increased activation of the TGF-β1-Smad2/3 signaling pathway. Immunofluorescence staining co-localized CD31 with α-SMA in the myocardial perivascular section of VECadCc1−/− hearts. Moreover, mRNA level of α-SMA, FSP1, SNAIL and SLUG were up-regulated while that of CD31 was down-regulated in mouse heart endothelial cells (MHEC) isolated from VECadCc1−/− mice. The Akt/eNOS pathway was basally inactivated, leading to lower nitric oxide (NO) level in MHEC isolated from VECadCc1−/− mice. Moreover, loss of endothelial CEACAM1 caused reduced association with SHP-2 phosphatase and consequently, reduction in insulin-stimulated IRS-1/Akt/eNOS activation.These studies identified an important role for the loss of CEACAM1 in endothelial cells as a mechanism underlying cardiac fibrosis.

Disclosure
H.T. Muturi: None. H.E. Ghadieh: None. S.S. Khuder: None. S. Jash: None. R. Abu Helal: None. N.A. Daniels: None. J. Liu: None. V. Puri: None. G. Vazquez: None. R. Gupta: None. S.M. Najjar: None.
Hepatic Overexpression of Retinoic Acid Receptor ß Downregulates Mitochondrial Pyruvate Carrier 1 and Protects from Metabolic Dysregulation in ob/ob Mice
Authors: KEYHANI NEJAD FARNAZ , LEE DONGHWAN , CHEN HANQING , LIANG ZERONG , MUSI NICOLAS , ZANG MENGWEI 
Published: June 2018

Our recent studies identify that retinoic acid receptor β (RARβ), a vitamin A-related nuclear receptor, regulates mitochondrial fatty acid oxidation in hepatocytes and in vivo in a fasting-inducible manner, which contributes to the metabolic adaptation to fasting. Mitochondrial pyruvate carrier (MPC) has been recently identified to control gluconeogenesis in hepatocytes, which contributes to the pathogenesis of diabetes. The present study aims to determine whether RARβ is linked to the control of glucose metabolism. We observed hepatic downregulation of RARβ occurred in high fat, high sucrose (HFHS) diet-fed mice and in genetically obese ob/ob mice. Treatment with all-trans-retinoic acid (RA), a natural ligand of RARβ and an approved drug for anticancer treatment, at doses of 2.5- 5.0 mg/kg/day for 4 weeks lowered fasting glucose levels and ameliorated glucose intolerance and insulin resistance in HFHS-fed mice, which was associated with increased hepatic gluconeogenic gene expression. Moreover, adenovirus-mediated overexpression of RARβ restored hepatic impairment of RARβ and systemic glucose intolerance and decreased hepatic gluconeogenesis by obesity. Mechanistically, overexpression of RARβ decreased obesity-induced expression of MPC1 and MPC2, key regulators of gluconeogenesis from pyruvate. Increasing RARβ in ob/ob mice also inhibited the induction of alanine transaminase (ALT), a key enzyme that converts alanine into pyruvate for glucose production. Moreover, overexpression of RARβ downregulated lipogenic gene expression, upregulated fatty acid oxidation, and improved hepatic steatosis and hyperlipidemia in ob/ob mice. Hepatic and systemic insulin resistance were also reduced by overexpressing RARβ. In sum, hepatic defective RARβ may contribute to the upregulation of MPC/ALT and gluconeogenesis in diabetes and RARβ can be a novel drug target to normalize metabolic dysregulation in diabetes.

Disclosure
F. Keyhani Nejad: None. D. Lee: None. H. Chen: None. Z. Liang: None. N. Musi: None. M. Zang: None.
Cognitive Function and Brain Plasticity in Obese Patients—The Impact of Bariatric Surgery
Authors: DARDANO ANGELA , DANIELE GIUSEPPE , LUNGHI CLAUDIA , CICCARONE ANNAMARIA , SANTINI FERRUCCIO , CECCARINI GIOVANNI , MORETTO CARLO , PENNO GIUSEPPE , MICCOLI ROBERTO , MORRONE MARIA CONCETTA , DEL PRATO STEFANO 
Published: June 2018

Background and Aim: Obesity adversely affects brain function, but the effect of bariatric surgery on cognitive function is still poorly evaluated. We assessed metabolic parameters and cognitive function in obese patients before and after bariatric surgery (RYGB).
Methods: A 75g OGTT was performed in 10 nondiabetic obese patients (BMI 46.0±5.4 kg/m2; age 43.6±10.5 years; HbA1c 43.3±4.6 mmol/mol) before and 6-months after RYGB for measurements of metabolic parameters. Mini Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were administered; neuronal plasticity (NP) of the visual cortex was measured as change in ocular dominance after 120 min monocular deprivation. No subject with drugs or alcohol abuse, psychiatric illness, head injury, brain tumor, dementia, epilepsy, learning disorder, developmental disability, and impaired sensory function were included.
Results: At baseline median MMSE score was 28.5 (range 25-30) and median MoCA score 26.5 (range 21-30), indicating early cognitive abnormalities (MoCA&lt;26 points). Six months after RYGB, BMI decreased to 35.2±6.1 kg/m2 (p=0.004) and glucose tolerance did not change. HOMA-IR (from 5.1±2.3 to 1.6±1.1) and disposition index (from 3.3±4.6 to 35.0±41.0) all improved (p&lt;0.05); post-OGTT GLP-1 increased from 5336.6±2262.6 to 11132.3±3412.2 pmol/lx120min (p&lt;0.05) and fasting plasma leptin decreased from 80.0±46.7 to 13.1±6.6 pmol/l (p&lt;0.008). NP increased from 0.03±0.1 to 0.11±0.14 (p&lt;0.01). Cognitive performance improved in all subjects (MMSE: 30, range 28-30; MoCA: 28.5, range 24-30; both p=0.03 or less vs. baseline). There was no correlation between MoCA and metabolic changes though it was positively correlated with NP post-RYGB (p=0.04).
Conclusion: Morbid obese subjects have subclinical cognitive impairments and brain plasticity alterations that significantly improve after RYGB. While no correlation was found between metabolic and hormonal changes, NP was positively associated with MoCA score.

Disclosure
A. Dardano: None. G. Daniele: None. C. Lunghi: None. A. Ciccarone: None. F. Santini: None. G. Ceccarini: None. C. Moretto: None. G. Penno: None. R. Miccoli: None. M. Morrone: None. S. Del Prato: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, GlaxoSmithKline plc., Intarcia Therapeutics, Inc., Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Servier, Sanofi, Takeda Pharmaceuticals U.S.A., Inc.. Research Support; Self; Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca. Speaker's Bureau; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharmaceuticals Corporation, Takeda Pharmaceuticals U.S.A., Inc.. Advisory Panel; Self; Janssen Biotech, Inc., Abbott.
Glucagon-Like Peptide-1 (GLP-1) Increases Skeletal and Cardiac Muscle, Microvascular Perfusion, and Improves Metabolic Insulin Action in Obese Humans
Authors: WANG NASUI , KIT TAN ALVIN WAI , JAHN LINDA , HARTLINE LEE , AYLOR KEVIN W. , BARRETT EUGENE , LIU ZHENQI 
Published: June 2018

Obesity is associated with metabolic and microvascular insulin resistance. The latter is characterized by impaired insulin-mediated microvascular recruitment. GLP-1 engenders vasodilation in addition to its glycemic actions. At physiological concentrations, GLP-1 acutely recruits skeletal and cardiac muscle microvasculature in healthy humans and this action is preserved in the insulin resistant rodents.
To examine whether GLP-1 recruits microvasculature and improves insulin’s action in obese humans, 15 obese adults (age 27 ± 2.2, BMI 33.7 ± 1.2) were studied thrice at random order after an overnight fast. Subjects received an intravenous infusion of GLP-1 (1.2 pmol/kg/min) for 150 mins or an intravenous infusion of either normal saline or GLP-1 (1.2 pmol/kg/min) for 150 mins with a euglycemic insulin clamp (1 mU/kg/min) superimposed over the last 120 mins. Skeletal and cardiac muscle microvascular blood volume (MBV), flow velocity (MFV) and blood flow (MBF) were determined at 0, 30 and 150 min. Steady-state glucose infusion rate (GIR) was calculated.
Insulin infusion alone did not change MBV or MBF in either skeletal or cardiac muscle, confirming the presence of microvascular insulin resistance. GLP-1 infusion alone increased MBV by ∼30% and ∼40% in skeletal and cardiac muscle respectively (p&lt;0.05) without affecting MFV, leading to a significant increase in MBF in both skeletal and cardiac muscle. Superimposition of insulin infusion to GLP-1 infusion did not further increase MBV or MBF in either skeletal or cardiac muscle, but raised the steady-state GIR from 4.1± 0.4 to 5.0 ± 0.5 mg/kg/min (p&lt;0.05).
We conclude that in obese humans with microvascular insulin resistance GLP-1's vasodilatory actions are preserved in both skeletal and cardiac muscle microvasculature. This may contribute to improving metabolic responses to insulin and decreasing cardiovascular morbidity and mortality associated with obesity and diabetes.

Disclosure
N. Wang: None. A. Tan: None. L. Jahn: None. L. Hartline: None. K.W. Aylor: None. E. Barrett: None. Z. Liu: None.
Functional and Developmental Heterogeneity in Human Adipose Tissue Depots
Authors: JOHNSON JAMES W. , CERO CHERYL , O'MARA ALANA , LINDERMAN JOYCE D. , BASKIN ALISON S. , CYPESS AARON 
Published: June 2018

Obesity results from the accumulation of excess white adipose tissue, and its increasing rates worldwide pose a significant risk to overall health due to its multiple comorbidities, like type 2 diabetes. A novel treatment strategy is to increase energy expenditure through activation of brown and beige adipocytes, which can use uncoupling protein 1 (UCP1) for thermogenesis. In addition to having different developmental lineages, brown and beige adipocytes differ functionally regarding the additional mechanisms by which they generate heat. Most human fat depots are not easily accessible, so little is known about their developmental and functional characteristics. In this study, we collected adipose tissue from 11 patients (age 16-84, 4 female/7 male) who had undergone autopsy. We sampled from 7 anatomically distinct fat depots: subcutaneous (sc), omental (om), retroperitoneal (rp), pericardial (pc), periadrenal (pa), paraspinal (ps), and supraclavicular (sv). We measured mRNA levels of lineage and functional genes associated with brown, beige, and white adipocytes. Energy storing white adipocytes were defined by high leptin expression, while energy expending brown and beige cells were defined by high UCP1; these were further distinguished as having a brown lineage using Zic1 and beige via Tbx1. Principal component analysis showed three different clusters of genes consistent with the brown, beige, and white lineages. Functionally, based on expression of leptin and UCP1 for white and brown fat, respectively, the sc and om were white (P&lt;0.05), while rp, pc, pa, ps, and sv were mostly brown (P&lt;0.05). Regarding lineage of the UCP1+ depots, ps had both beige and brown cells, while rp, pa, ps, and sv were predominantly brown.
In summary, human adipose tissue is not merely white or brown but instead is composed of many depots with distinct developmental lineages and functional capacities. These differences will be important when trying to use brown and beige adipocyte thermogenesis to treat obesity and diabetes.

Disclosure
J.W. Johnson: None. C. Cero: None. A. O'Mara: None. J.D. Linderman: None. A.S. Baskin: None. A. Cypess: None.
A Study of APOC3 Gene Polymorphism in Patients of Diabetes Mellitus with Hypertriglyceridemia
Authors: KALITA BIPUL C. , KOTOKEY RATAN K. , DAS SUBHALAKSHMI , CHAKRAVORTY AVISEK , SONI HARISH 
Published: June 2018

Aims and Objectives: To find out the presence of Sst I polymorphism in APOC3 gene in patients of diabetes mellitus with Hypertriglyceridemia.
To find out the correlation between serum triglyceride levels and Sst I polymorphism.
To find out the correlation between serum levels of various lipoprotein fractions and Sst I polymorphism.
Methods: The present study is a hospital-based observational case-control study, carried out in 300 patients of diabetes mellitus. Of these 300 subjects, 150 were hypertriglyceridemic and served as cases, while the other 150 who were normotriglyceridemic served as controls. PCR for APOC3 gene was done in all subjects.
Result: In a total of 300 subjects, 26.33% (n=79) had shown presence of Sst I polymorphism in APOC3 gene. Polymorphic gene was present in 35.33% of cases while only in 17.33% of controls. The mean cholesterol level in the group with Sst I polymorphism was significantly higher than in group with S1 allele only. The mean triglyceride in the group with S1 allele only was 152.97±45.86 mg/dl and that in the group with Sst I polymorphism was 212.84±103.07 mg/dl, and this increased mean triglyceride level was statistically significant as determined by chi-square test. Mean HDL level was also significantly lower in the group with Sst I polymorphism as compared to that with S1 allele only. The odds ratio of developing hypertriglyceridemia was 2.6059 in those carrying the Sst I polymorphism as compared to those with only S1 allele. The association between increasing levels of triglyceride and the increasing percentage of Sst I polymorphism was also statistically significant.
Conclusion: This is the first study from North-East India to determine the Sst I polymorphism in APOC3 gene, and demonstrate a clear association between the polymorphism and serum levels of various lipoprotein fractions, including triglycerides. A prospective study comprising of a wide population group will help us in forming a definite conclusion.

Disclosure
B.C. Kalita: None. R.K. Kotokey: None. S. Das: None. A. Chakravorty: None. H. Soni: None.
Sustained Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor-1 (FGF-1) Requires Activation of Integrin Receptor aνß3
Authors: SCARLETT JARRAD , BROWN JENNY M. , PHAN BAO ANH N. , KHAV EDDIE L. , MATSEN MILES E. , ACHARYA NIKHIL K. , NGUYEN HONG T. , SECHER ANNA , JORGENSEN RASMUS , MORTON GREGORY J. , SCHWARTZ MICHAEL W. 
Published: June 2018

The brain is implicated in the antidiabetic effects of members of the fibroblast growth factor (FGF) peptide family. Previously, we reported that a single intracerebroventricular (icv) injection of FGF-1 induces remission of hyperglycemia that is sustained for weeks in rodent models of type 2 diabetes (T2D). The integrin receptor ανβ3 plays a key role in FGF-1 signaling, and crosstalk between ανβ3 and the FGF-1-FGF receptor (FGF-R) complex is necessary for increased DNA synthesis and proliferation in cultured cells. These cellular responses downstream of FGF-1-FGFR-ανβ3 result from sustained (e.g., lasting for 5 hours) activation of the extracellular signal-regulated kinase (ERK) signal transduction pathway. In cultured fibroblasts, prolonged ERK activation induced by FGF-1 is not replicated with a mutant FGF-1 peptide (R50E) that activates FGFR normally, but does not activate ανβ3 signaling. Similarly, we report that whereas icv FGF-1 (3 µg) induces phosphorylation of ERK1/2 at both early (20 minute) and late (5 hr) time points in hypothalamic punches obtained from wild type mice, icv injection of the same dose of R50E elicited this effect only at the early time point. Thus, R50E fails to activate hypothalamic FGF-1-FGFR-ανβ3 signaling in vivo. To investigate whether activation of ανβ3 is required for the prolonged antidiabetic effect of icv FGF-1, diabetic ob/ob mice received a single injection of either vehicle (Veh), recombinant FGF-1 (3 µg), or R50E (3 µg). Whereas R50E mimicked the effect of native FGF-1 to reduce food intake, body weight and blood glucose for the first week after icv injection (P&lt;0.001 vs. Veh), the mutant peptide failed to elicit the sustained diabetes remission observed in animals receiving icv FGF-1. We conclude that signaling by the full FGF-1-FGFR-ανβ3 complex is necessary for the effect of FGF-1 signaling in the brain to elicit sustained remission of diabetic hyperglycemia in rodent models of T2D.

Disclosure
J. Scarlett: None. J.M. Brown: None. B.N. Phan: None. E.L. Khav: None. M.E. Matsen: None. N.K. Acharya: None. H.T. Nguyen: None. A. Secher: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Spouse/Partner; Gubra. R. Jorgensen: Employee; Self; Novo Nordisk Inc.. G.J. Morton: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.
A Single-Cell Transcriptomics Roadmap to Investigate Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor-1 (FGF-1)
Authors: BENTSEN MARIE A. , RAUSCH DYLAN , SCARLETT JARRAD , ALONGE KIMBERLY M. , TIMSHEL PASCAL N. , MIRZADEH ZAMAN , SECHER ANNA , JORGENSEN RASMUS , PERS TUNE , SCHWARTZ MICHAEL W. 
Published: June 2018

Recently, lasting remission of hyperglycemia was achieved in rodent models of type 2 diabetes (T2D) by a single intracerebroventricular (icv) injection of FGF-1. While the mechanism underlying this effect is unknown, the lack of association with changes of body fat mass or risk of hypoglycaemia raises the possibility that icv FGF-1 normalizes the defended level of glycemia, rather than simply lowering blood glucose levels. Recent work from our lab has identified gluco-regulatory neurocircuits in the mediobasal hypothalamus (MBH) as targets for this FGF-1 effect. To investigate the mechanism underlying FGF-1 action in this brain area, we used large-scale single cell RNA-sequencing to identify and characterize FGF-1-responsive cells in mouse MBH. Based on &gt;70,000 single cell MBH transcriptomes from diabetic ob/ob mice harvested 5d after a single icv injection of either FGF-1 or vehicle, we identified &gt;20 cell clusters and &gt;900 cell type-specific differentially expressed genes (p&lt;0.01). Among these potential targets of the MBH response to FGF-1 in diabetic mice, we found that FGF-1 treatment robustly increased the proportion of newly formed oligodendrocytes, a response that can be predicted to yield new mature oligodendrocytes and associated myelination of axons. These preliminary results demonstrate that transcriptional identification and characterisation of icv FGF-1-responsive cell clusters may both advance our understanding of brain control of glucose homeostasis and identify novel targets for diabetes prevention and treatment.

Disclosure
M.A. Bentsen: None. D. Rausch: None. J. Scarlett: None. K.M. Alonge: None. P.N. Timshel: None. Z. Mirzadeh: None. A. Secher: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Spouse/Partner; Gubra. R. Jorgensen: Employee; Self; Novo Nordisk Inc.. T. Pers: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.
The Cell Junction Protein Afadin Negatively Regulates Insulin Action and Modulates Adipose Tissue Function
Authors: LUNDH MORTEN , ISIDOR MARIE S. , PLUCINSKA KAJA , SHAMSI FARNAZ , PETERSEN PATRICIA , TSENG YU-HUA , EMANUELLI BRICE 
Published: June 2018

Adipose tissue dysfunction is one of the main culprits triggering metabolic imbalance associated with obesity. We identified the cell junction protein Afadin as one of the most highly phosphorylated proteins in response to insulin/IGF-1 stimulation using quantitative phospho-proteomic analysis in pre-adipocytes, and investigated the role of Afadin in mediating insulin action and in adipocyte biology. We found that phosphorylation of Afadin occurred in response to insulin in both pre- and mature adipocytes obtained from mice and human subjects. In addition, Afadin phosphorylation was observed in mouse adipose tissues following insulin injection, and after re-feeding, indicating physiological relevance. In obese mice, however, brown fat insulin-induced phosphorylation of Afadin was abolished. Surprisingly, glucose and insulin tolerance were improved in Fat-specific Afadin knock-out (KO) mice on a high fat diet, despite similar body weight gain and adipose mass as wild type controls. Furthermore, ablation of Afadin in vitro did not affect adipogenesis. Yet, prolonged insulin-induced phosphorylation of Akt was observed in KO adipocytes, and was associated with increased insulin-mediated fatty-acid uptake. Finally, phosphorylation of Afadin regulates the docking of several proteins, which may be involved in the regulation of insulin action and adipocyte metabolism. Thus, Afadin is a novel bona fide substrate of the insulin signaling pathway, and is involved in the regulation of insulin action and metabolism.

Disclosure
M. Lundh: None. M.S. Isidor: None. K. Plucinska: None. F. Shamsi: None. P. Petersen: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.. Research Support; Self; MedImmune. B. Emanuelli: None.
Impact of Aging on Risk of Hypoglycemia in Patients with Type 1 Diabetes
Authors: MUNSHI MEDHA , SLYNE CHRISTINE , ATAKOV-CASTILLO ASTRID , GREENBERG JORDAN , GREAVES TORI , CARL SAM P. , TOSCHI ELENA 
Published: June 2018

Background: As T1D patients routinely reach older age, more information is needed to understand the impact of aging on the risk of hypoglycemia and its interaction with other age-related issues.
Methods: We evaluated 2 groups of patients with T1D: older (age &gt;65 years) and younger (age 18-35 years). All patients were subjected to either blinded CGM for 2 weeks or assessment of personal CGM. A hypoglycemia fear survey (HFS II), and a Clarke hypoglycemia unawareness survey were completed. Patients were also assessed for cognitive function by the Montreal Cognitive Assessment (MoCA). Glycemic control was measured by A1C.
Results: We evaluated 56 patients with T1D; 23 in the older and 33 in the younger group. The average age was 70 vs. 28 years; diabetes duration 36 vs. 14 years; A1C 8.0% vs. 7.9%, respectively. Clinically significant hypoglycemia (glucose ?54 mg/dL for &gt; 15 mins/episode) occurred equally in 91% of older (mean 28 minutes/day) and 88% of younger (mean 23 mins/day) patients. Older patients reported hypoglycemia unawareness more (39% vs. 30%), with similar scores on HSF II (34 vs. 33). However, 61% of older patients had cognitive impairment (MoCA score &lt;26) compared to 12% in younger adults. There was a higher comorbidity burden in older patients (average number of comorbidities 4 vs. 1), with a higher number of daily medications (10 vs. 4).
Conclusion: The risk of hypoglycemia is similar in older and younger patients with T1D. However, higher incidence of comorbidities including cognitive dysfunction and polypharmacy may increase the risk of poor outcomes due to hypoglycemia.

Disclosure
M. Munshi: Consultant; Self; Sanofi. C. Slyne: None. A. Atakov-Castillo: None. J. Greenberg: None. T. Greaves: None. S.P. Carl: None. E. Toschi: None.