Fat Biopsies Research Articles

Abstract 12963: The Prevalence and Predictors of Transthyretin Amyloidosis in Patients Underwent Transcatheter Aortic Valve Implantation

Background: Several studies have recently reported that transthyretin (ATTR) amyloidosis is present in patients with aortic stenosis (AS). We aimed to 1) examine the prevalence of ATTR amyloidosis in AS patients undergoing transcatheter aortic valve implantation (TAVI), 2) compare clinical characteristics between AS patients with and without ATTR, and 3) identify predictors of ATTR amyloidosis in AS patients. Methods and Results: We prospectively analyzed consecutive 32 patients with severe AS who underwent screening tests for ATTR in prior to TAVI. To detect ATTR, we performed cardiac technetium-99m pyrophosphate ( 99m Tc-PYP) scintigraphy in 24 patients and biopsies of fat tissue from the puncture site in 24 patients. Of 32 AS patients who were screened for ATTR, 6 (18.8%) patients were diagnosed as ATTR by cardiac 99m Tc-PYP scintigraphy (n=2) and fat biopsy (n=5). Compared with non-ATTR patients, ATTR patients presented higher levels of B-type natriuretic peptide (BNP, 242.3 vs. 1101.4 ng/ml, P=0.004), lower left ventricular ejection fraction (LVEF, 62.3 vs. 45.5%, P=0.002) and lower mean pressure gradient of AS (50.2 vs. 32.0 mmHg, P=0.016). In contrast, age, sex, STS score, clinical frailty scale, prevalence of chronic kidney disease and atrial fibrillation, and levels of hemoglobin, albumin, estimated glomerular filtration rate and troponin I were comparable between the two groups. Other echocardiographic parameters including interventricular septal wall thickness, posterior wall thickness and aortic valve area did not differ between the two groups. To detect ATTR among AS patients, receiver operating characteristic (ROC) analysis demonstrated that area under the curve of BNP, LVEF, and mean pressure gradient was 0.814, 0.766 and 0.846, respectively. Although TAVI was successfully completed in all patients, and there was no procedural complication in both groups, one of ATTR patients re-hospitalized due to decompensated heart failure 3 weeks after TAVI. Conclusion: In AS patients undergoing TAVI, especially in cases of low-flow low-gradient pattern and high BNP levels, screening of ATTR should be considered. In this population, cardiac 99m Tc-PYP scintigraphy and fat biopsy are useful to detect latent ATTR non-invasively.

Abnormalities of the lymphatic system and impaired fluid clearance in patients with heart failure with preserved ejection fraction

Abstract Background Coronary and skeletal muscle microvascular dysfunction have been proposed as main factors in the pathogenesis of Heart Failure with Preserved Ejection Fraction (HFpEF). However, assessment of systemic arterial function has only been indirect thus far; most importantly, no direct link between systemic microvasculature and congestion, one of the core characteristics of the syndrome, has yet been investigated. Purpose To provide direct functional and anatomical characterisation of the systemic microvasculature and to explore in vivo parameters of capillary fluid extravasation and lymphatic clearance in HFpEF. Methods In 16 patients with HFpEF and 16 age- and sex-matched healthy controls (72±6 and 68±5 years, respectively) we determined peripheral microvascular filtration coefficient (proportional to vascular permeability and area) and isovolumetric pressure (above which lymphatic drainage cannot compensate for fluid extravasation) by venous occlusion plethysmography and collected a skin biopsy for vascular immunohistochemistry and gene expression analysis (TaqMan). Additionally, we measured brachial flow-mediated dilatation (FMD) and assessed by wire myography the vascular function of resistance arteries isolated from gluteal subcutaneous fat biopsies. Results Skin biopsies in patients with HFpEF showed rarefaction of small blood vessels (82±31 vs 112±21 vessels/mm2; p=0.003) and in ex-vivo analysis (n=6/group) we found defective relaxation of peripheral resistance arteries (p<0.001). Accordingly, post-ischaemic hyperaemic response (fold-change vs baseline, 4.6±1.6 vs 6.7±1.7; p=0.002) and FMD (3.9±2.1 vs 5.6±1.5%; p=0.014) were found to be reduced in patients with HFpEF compared to controls. In the skin of patients with HFpEF we also observed a reduced number (85±27 vs 130±60 vessels/mm2; p=0.012) but larger average diameter of lymphatic vessels (42±19 vs 26±9 μm2; p=0.007) compared to control subjects. These changes were paralleled by reduced expression of LYVE1 (p<0.05) and PROX1 (p<0.001), key determinants of lymphatic differentiation and function. Whilst patients with HFpEF had reduced peripheral capillary fluid extravasation compared to controls (microvascular filtration coefficient, leg 33.1±13.3 vs 48.4±15.2, p<0.01; trend for arm 49.9±20.5 vs 66.3±30.1, p=0.09), they had lower lymphatic clearance (isovolumetric pressure: leg 22±4 vs 16±4 mmHg, p<0.005; arm 25±5 vs 17±4 mmHg, p<0.001). Conclusions We provide direct evidence of systemic dysfunction and rarefaction of small blood vessels in patients with HFpEF. Despite a reduced microvascular filtration coefficient, which is in keeping with microvascular rarefaction, the clearance of extravasated fluid in HFpEF is limited by an anatomically and functionally defective lymphatic system. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): British Heart Foundation Centre of Research Excellence Award

Robotic Management of Pelvic Lipomatosis—Experience with Difficulties Encountered and the Techniques to Successful Outcomes

BackgroundPelvic lipomatosis (PL) is a rare condition characterized by diffuse pelvic overgrowth of nonmalignant but infiltrative adipose tissue in perivesical and perirectal space. ObjectiveTo share our robotic experience and difficulties encountered and suggested techniques to overcome them successfully. It is the first series from India. Design, setting, and participantsA prospective observational study was conducted. All consecutive patients diagnosed with PL between 2016 and 2019 underwent robotic-assisted wide bladder fat extirpation and bilateral ureteral reimplantation (extravesical refluxing type) with double J stenting and were evaluated prospectively. Outcome measurements and statistical analysisDemographics, serum creatinine level, radiographic features, postoperative complications, and patient-reported outcomes were evaluated. Continuous variables are presented as median and range, as the sample size is very small. Results and limitationsWe encountered a total of five patients with PL. The median console time was 126 (range 120–130) min, with a median estimated blood loss of 120 (range 100–150) ml. Postoperative complications were Clavien-Dindo grade I in three cases, and the median hospital stay was 2 d. Distal ureteric margins showed subepithelial edema with submucosal fibrosis, and bladder fat biopsies were reported as adipose tissue with hemorrhage. At initial 3-mo and annual follow-ups, all patients had normal serum creatinine and there was no evidence of disease recurrence. Limitations of our study are the very small sample size (a low incidence of PL) and short follow-up time period (the question of how long the surgical effect will be sustained due to abnormal proliferation of fat cells remains unanswered). ConclusionsRobotic management of wide bladder fat extirpation and bilateral ureteral reimplantation with double J stenting has a good success rate and good outcome in PL. Patient summaryPelvic lipomatosis is a very rare condition. Robotic management in treating the condition has good outcome for the patient. Here, we have discussed the difficulties encountered in treating the condition and the techniques used to overcome them.

Testosterone treatment is associated with reduced adipose tissue dysfunction and nonalcoholic fatty liver disease in obese hypogonadal men

PurposeIn both preclinical and clinical settings, testosterone treatment (TTh) of hypogonadism has shown beneficial effects on insulin sensitivity and visceral and liver fat accumulation. This prospective, observational study was aimed at assessing the change in markers of fat and liver functioning in obese men scheduled for bariatric surgery.MethodsHypogonadal patients with consistent symptoms (n = 15) undergoing 27.63 ± 3.64 weeks of TTh were compared to untreated eugonadal (n = 17) or asymptomatic hypogonadal (n = 46) men. A cross-sectional analysis among the different groups was also performed, especially for data derived from liver and fat biopsies. Preadipocytes isolated from adipose tissue biopsies were used to evaluate insulin sensitivity, adipogenic potential and mitochondrial function. NAFLD was evaluated by triglyceride assay and by calculating NAFLD activity score in liver biopsies.ResultsIn TTh-hypogonadal men, histopathological NAFLD activity and steatosis scores, as well as liver triglyceride content were lower than in untreated-hypogonadal men and comparable to eugonadal ones. TTh was also associated with a favorable hepatic expression of lipid handling-related genes. In visceral adipose tissue and preadipocytes, TTh was associated with an increased expression of lipid catabolism and mitochondrial bio-functionality markers. Preadipocytes from TTh men also exhibited a healthier morpho-functional phenotype of mitochondria and higher insulin-sensitivity compared to untreated-hypogonadal ones.ConclusionsThe present data suggest that TTh in severely obese, hypogonadal individuals induces metabolically healthier preadipocytes, improving insulin sensitivity, mitochondrial functioning and lipid handling. A potentially protective role for testosterone on the progression of NAFLD, improving hepatic steatosis and reducing intrahepatic triglyceride content, was also envisaged.Clinical Trial RegistrationClinicalTrials.gov Identifier: NCT02248467, September 25th 2014

Exploring the Paradoxical Relationship of a Creb 3 Regulatory Factor Missense Variant With Body Mass Index and Diabetes Among Samoans: Protocol for the Soifua Manuia (Good Health) Observational Cohort Study.

BackgroundThe prevalence of obesity and diabetes in Samoa, like many other Pacific Island nations, has reached epidemic proportions. Although the etiology of these conditions can be largely attributed to the rapidly changing economic and nutritional environment, a recently identified genetic variant, rs373863828 (CREB 3 regulatory factor, CREBRF: c.1370G>A p.[R457Q]) is associated with increased odds of obesity, but paradoxically, decreased odds of diabetes.ObjectiveThe overarching goal of the Soifua Manuia (Good Health) study was to precisely characterize the association of the CREBRF variant with metabolic (body composition and glucose homeostasis) and behavioral traits (dietary intake, physical activity, sleep, and weight control behaviors) that influence energy homeostasis in 500 adults.MethodsA cohort of adult Samoans who participated in a genome-wide association study of adiposity in Samoa in 2010 was followed up, based on the presence or absence of the CREBRF variant, between August 2017 and March 2019. Over a period of 7-10 days, each participant completed the main study protocol, which consisted of anthropometric measurements (weight, height, circumferences, and skinfolds), body composition assessment (bioelectrical impedance and dual-energy x-ray absorptiometry), point-of-care glycated hemoglobin measurement, a fasting blood draw and oral glucose tolerance test, urine collection, blood pressure measurement, hand grip strength measurement, objective physical activity and sleep apnea monitoring, and questionnaire measures (eg, health interview, cigarette and alcohol use, food frequency questionnaire, socioeconomic position, stress, social support, food and water insecurity, sleep, body image, and dietary preferences). In January 2019, a subsample of the study participants (n=118) completed a buttock fat biopsy procedure to collect subcutaneous adipose tissue samples.ResultsEnrollment of 519 participants was completed in March 2019. Data analyses are ongoing, with results expected in 2020 and 2021.ConclusionsWhile the genetic variant rs373863828, in CREBRF, has the largest known effect size of any identified common obesity gene, very little is currently understood about the mechanisms by which it confers increased odds of obesity but paradoxically lowered odds of type 2 diabetes. The results of this study will provide insights into how the gene functions on a whole-body level, which could provide novel targets to prevent or treat obesity, diabetes, and associated metabolic disorders. This study represents the human arm of a comprehensive and integrated approach involving humans as well as preclinical models that will provide novel insights into metabolic disease.International Registered Report Identifier (IRRID)RR1-10.2196/17329

Increased levels of VEGF-C and macrophage infiltration in lipedema patients without changes in lymphatic vascular morphology

Lipedema is a chronic adipose tissue disorder characterized by the disproportional subcutaneous deposition of fat and is commonly misdiagnosed as lymphedema or obesity. The molecular determinants of the lipedema remain largely unknown and only speculations exist regarding the lymphatic system involvement. The aim of the present study is to characterize the lymphatic vascular involvement in established lipedema. The histological and molecular characterization was conducted on anatomically-matched skin and fat biopsies as well as serum samples from eleven lipedema and ten BMI-matched healthy patients. Increased systemic levels of vascular endothelial growth factor (VEGF)-C (P = 0.02) were identified in the serum of lipedema patients. Surprisingly, despite the increased VEGF-C levels no morphological changes of the lymphatic vessels were observed. Importantly, expression analysis of lymphatic and blood vessel-related genes revealed a marked downregulation of Tie2 (P < 0.0001) and FLT4 (VEGFR-3) (P = 0.02) consistent with an increased macrophage infiltration (P = 0.009), without changes in the expression of other lymphatic markers. Interestingly, a distinct local cytokine milieu, with decreased VEGF-A (P = 0.04) and VEGF-D (P = 0.02) expression was identified. No apparent lymphatic anomaly underlies lipedema, providing evidence for the different disease nature in comparison to lymphedema. The changes in the lymphatic-related cytokine milieu might be related to a modified vascular permeability developed secondarily to lipedema progression.

CD5L, Macrophage Apoptosis Inhibitor, Was Identified in Epicardial Fat-Secretome and Regulated by Isoproterenol From Patients With Heart Failure.

ObjectivesNeurohormonal dysfunction, which can regulate epicardial fat activity, is one of the main promoters of atrial fibrillation (AF) in patients with heart failure (HF). Our aim was to study the epicardial fat mediators for AF in patients with HF and its catecholaminergic regulation.MethodsWe have included 29 patients with HF who underwent cardiac surgery and were followed up for 5 years. Released proteins by epicardial adipose tissue (EAT) after isoproterenol treatment were identified by nano-high-performance liquid chromatography (HPLC) and triple time-of-flight (TOF) analysis. Common and differential identified proteins in groups of patients with AF before and after surgery were determined by the FunRich tool. Plasma and epicardial fat biopsy proteins were quantified by western blot.ResultsOur results identified 17 common released proteins by EAT, after isoproterenol treatment, from HF patients who suffered AF or developed new-onset AF during follow-up. Mostly, they were involved on inflammatory response and extracellular matrix. One of them was CD5L, a macrophage apoptosis inhibitor. Its secretion by isoproterenol treatment was validated on western blot. The CD5L levels on epicardial fat were also higher in the group of male patients who present or develop AF (0.44 ± 0.05 vs. 0.18 ± 0.15; p < 0.016). However, there were no differences regarding plasma levels.ConclusionOur results suggest the role of epicardial fat CD5L as a mediator of AF and its possible paracrine effect by catecholaminergic activity.

Effect of increased potassium intake on the renin-angiotensin-aldosterone system and subcutaneous resistance arteries: a randomized crossover study.

Increased potassium intake lowers blood pressure (BP) in hypertensive patients. The underlying mechanism is not fully understood but must be complex because increased potassium intake elevates circulating concentrations of the BP-raising hormone aldosterone. In a randomized placebo-controlled crossover study in 25 normotensive men, we investigated the effect of 4 weeks of potassium supplement (90 mmol/day) compared with 4 weeks of placebo on the renin-angiotensin-aldosterone system (RAAS), urine composition and 24-h ambulatory BP. Vascular function was also assessed through wire myograph experiments on subcutaneous resistance arteries from gluteal fat biopsies. Higher potassium intake increased urinary potassium excretion (144.7 ± 28.7 versus 67.5 ± 25.5 mmol/24-h; P < 0.0001) and plasma concentrations of potassium (4.3 ± 0.2 versus 4.0 ± 0.2 mmol/L; P = 0.0002), renin {mean 16 [95% confidence interval (CI) 12-23] versus 11 [5-16] mIU/L; P = 0.0047}, angiotensin II [mean 10.0 (95% CI 6.2-13.0) versus 6.1 (4.0-10.0) pmol/L; P = 0.0025] and aldosterone [mean 440 (95% CI 336-521) versus 237 (173-386) pmol/L; P < 0.0001]. Despite RAAS activation, systolic BP (117.6 ± 5.8 versus 118.2 ± 5.2 mmHg; P = 0.48) and diastolic BP (70.8 ± 6.2 versus 70.8 ± 6.3 mmHg; P = 0.97) were unchanged. In the wire myograph experiments, higher potassium intake did not affect endothelial function as assessed by acetylcholine [logarithmically transformed half maximal effective concentration (pEC50): 7.66 ± 0.95 versus 7.59 ± 0.85; P = 0.86] and substance P (pEC50: 8.42 ± 0.77 versus 8.41 ± 0.89; P = 0.97) or vascular smooth muscle cell reactivity as assessed by angiotensin II (pEC50: 9.01 ± 0.86 versus 9.02 ± 0.59; P = 0.93) and sodium nitroprusside (pEC50: 7.85 ± 1.07 versus 8.25 ± 1.32; P = 0.25) but attenuated the vasodilatory response of retigabine (pEC50: 7.47 ± 1.16 versus 8.14 ± 0.90; P = 0.0084), an activator of Kv7 channels. Four weeks of increased potassium intake activates the RAAS in normotensive men without changing BP and this is not explained by improved vasodilatory responses ex vivo.

2313-PUB: Alterations in Basal State of Insulin and mTOR-Dependent Signalings Closely Related to Impaired Incretin Profile and Type 2 Diabetes in Subcutaneous Adipose Tissue of Obese Patients

Objective: In our study we have compared basal insulin and mTOR signaling in subcutaneous fat of obese T2DM vs. obese subjects with normal glucose tolerance (NGT). We have performed correlation analysis of phosphorylation’s parameters against clinical parameters of carbohydrate metabolism and incretin secretion profiles. Methods: 22 patients with long (&amp;gt;10 years) and morbid (BMI &amp;gt; 35 kg/m2) obesity, 12 of which had NGT and 10 had T2DM were enrolled in this study. Hyperinsulinemic-euglycemic clamp test and HOMA-IR were used to measure insulin resistance. Blood samples taken at 0, 30 and 120 min of food load test were used to assess incretin profile, insulin and glucose levels. Amount of total and visceral fat was determined by bioelectrical impedance analysis. Subcutaneous fat biopsies were obtained during bariatric surgery for all patients and signaling state was analyzed by western blots. Results: As assessed by western blots of insulin receptor substrate (IRS), Akt, Raptor, Rictor, mTOR and S6K1, the basal insulin signaling and mTORC activities were comparable in NGT and T2DM groups, whereas phosphorylation of AS160 (pAS160-S318) was significantly lower and serum and glucocorticoid-induced kinase (SGK; pSGK-S422) was significantly higher in T2DM group. Various correlations were found between the degree of insulin resistance and amount of visceral fat, changes in incretin profile, glucose metabolic parameters and phosphorylation level of AS160, incretin secretion profile and phosphorylated levels of AS160 or SGK1. Conclusions: Altered phosphorylation of AS160 and SGK1 is associated with obese T2DM phenotype and impaired incretin profile. We think that AS160 and SGK1 phosphorylations can be important markers of the prediabetes-to-diabetes transition. Disclosure I. Stafeev: None. I. Sklyanik: None. S. Michurina: None. E. Shestakova: Employee; Spouse/Partner; AstraZeneca. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharma K.K., Novo Nordisk A/S, Sanofi-Aventis, Takeda Pharmaceutical Company Limited. A. Yurasov: None. K. Yahyaev: None. A. Karmadonov: None. E. Ratner: None. M. Menshikov: None. A.V. Vorotnikov: None. Y.V. Parfyonova: None. M.V. Shestakova: None. Funding Russian Science Foundation (17-15-01435)

1703-P: Altered UCP1 Expression in ADSC-Derived Beige Adipocytes Determines Mitochondrial ROS Production in T2DM Patients

Background: Under normal physiological conditions, adipose derived stem cells (ADSC) maintain adipose tissue homeostasis by responding to metabolic loading through proliferation and differentiation. Nevertheless, in T2DM their properties might be compromised. ADSC give rise to thermogenic beige adipocytes that make a significant contribution into excess energy utilization in obesity. We compared ADSC response to beiging inductors in obese patients with type 2 diabetes mellitus (T2DM) and with normal glucose tolerance (NGT). Methods: We collected subcutaneous fat biopsies from 5 obese NGT (BMI&amp;gt;35kg/m2) and 5 obese T2DM patients during bariatric surgery. ADSC were enzymatically isolated and underwent white and beige adipogenic differentiation. UCP-1 level was evaluated by RT-PCR and Western blot. Expression of lipid metabolism and electron transport chain (ETC) genes were analyzed by RT-PCR. ROS production was evaluated by fluorescent microscopy. Results: UCP-1 expression was decreased in beige adipocytes from T2DM patients in mRNA and protein levels. Nevertheless, expression and phosphorylation of lipolytic proteins were equal in beige adipocytes of NGT and T2DM groups. Expression analysis of ETC genes also has not shown any statistically significant differences. All the while, we revealed increased mitochondrial ROS production in T2DM beige adipocytes during beige differentiation. Conclusions: These results indicate association of ADSC thermogenic potential and T2DM development. Decreased UCP1 level in complex with normal lipolysis and ETC activity in beige adipocytes from T2DM patients may be liable to elevated ROS level, known to initiate mitochondria damage and insulin resistance. Disclosure S. Michurina: None. I. Stafeev: None. I. Sklyanik: None. E. Shestakova: Employee; Spouse/Partner; AstraZeneca. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharma K.K., Novo Nordisk A/S, Sanofi-Aventis, Takeda Pharmaceutical Company Limited. A. Yurasov: None. K. Yahyaev: None. E. Ratner: None. M. Menshikov: None. M.V. Shestakova: None. Y.V. Parfyonova: None. Funding Russian Science Foundation (17-15-01435)

1987-P: Inflammation and Insulin Resistance in Adipose Tissue

Background: Adipose tissue inflammation is linked to adipose tissue insulin resistance. However, whether this is a causal relationship remains controversial. Therefore, we aimed to explore the relationship between measures of adipose tissue inflammation and adipose tissue insulin sensitivity over a wide range of body fat distribution and adipose tissue insulin sensitivity. Methods: We recruited a total of 87 volunteers (30 men and 57 women). Adipose tissue insulin sensitivity was quantified as the insulin concentration required for a 50% suppression of lipolysis (IC50). Lipolysis was measured as steady-state palmitate flux before and during a hyperinsulinemic-euglycemic clamp. Adipose tissue macrophage (ATM) was quantified by immunohistochemistry with antibodies against CD68 (total ATM), CD14 (pro-inflammatory ATM) and CD206 (anti-inflammatory ATM) and senescent preadipocyte content was quantified by β-galactosidase positivity using fluorescent microscopy in abdominal and femoral fat biopsies. Results: Total ATM content was only weakly associated with IC50 in abdominal fat (r=0.26, p=0.01) and no association was observed for femoral fat. No association was observed between IC50 and pro-inflammatory ATM content. Anti-inflammatory ATM content was associated with IC50 in abdominal (r=0.46, p&amp;lt;0.001) and femoral (r=0.33, p=0.02) fat. Senescent preadipocyte content was weakly associated with IC50 in femoral fat (r=0.36, p=0.01), but no association was observed in abdominal fat. Fat cell size was strongly associated with IC50 in both abdominal (r=0.6 p&amp;lt;0.001) and femoral (r=0.48, p&amp;lt;0.01) fat. Conclusions: ATM content is only weakly associated to adipose tissue insulin sensitivity. Only the content of anti-inflammatory, not pro-inflammatory macrophages, was associated with adipose tissue insulin sensitivity. This argues against pro-inflammatory macrophages as mediators of insulin resistance in adipose tissue. Disclosure E. Søndergaard: None. A.E. Espinosa De Ycaza: None. M. Morgan-Bathke: None. K. Lytle: None. D.A. Delivanis: None. B.G. Carranza Leon: None. M.D. Jensen: None. Funding National Institutes of Health (DK40404, DK45343)

P0307KIDNEY-HEMOPATHY CONSULTATION: A SINGLE-CENTER EXPERIENCE

Abstract Background and Aims Renal involvement in plasma cell dyscrasia has been widely described, including Monoclonal Gammopathy of Renal significance (MGRS). Chronic kidney disease (CKD) in hematological disorders carries major implications for management, prognosis and the potential for progressive renal injury and ESRD. Specific monthly Kidney-Hemopathy Consultation has been created in 2017 in the department of Nephrology. The aimed of this was to describe the clinical, biological and histological characteristics of patients with hematological disorders and CKD. Method From 01/05/2017 to 31/12/2019 adults with hematological disorders and CKD (eGFR&amp;lt;60ml/min and/or proteinuria or hematuria) were prospectively included. Results Fifty-six patients (27 men) were enrolled: Multiple Myeloma (n=30), MGUS (n=11), Lymphoma (n=5), Acute Leukemia (n=4), LMC (n=2), 1 LLC (n=2), Cryoglobulinemia (n=1), Refractory Anemia with Excess blasts (n=1), Bone marrow aplasia (n=1). This nephrology consultation was the first for 22 patients (39%), referred by a hematologist in 38 cases (68%). Renal injury was confirmed after the hematologic diagnosis in 34 patients (61%) and precedes the diagnosis of plasma cell dyscrasia for 8 patients (14%). The median eGFR was 54.1ml/min/1.73m (128-4): CKD stage II (n=16), stage III (n=28), stage IV (n=7) and stage V (n=2). Proteinuria was &amp;gt;0.5g/day for 22 (39.3%) patients including a Bence-Jones proteinuria for 15 patients (27%). Acute Kidney Injury (AKI) was confirmed in 34 patients (63%) and 8 (24%) requiring acute dialysis. Histologic diagnostic of kidney injury was confirmed for 21 patients (38%): 16 kidney biopsies and 5 AL renal amyloidosis confirmed by salivary glands or abdominal fat biopsy. MGRS was confirmed in 34 patients (61%): Cast nephropathy (n=10;29%), AL amyloidosis (n=7;20%); C3 nephropathy (n=3; 9%), Proliferative Glomerulonephritis with Monoclonal Immunoglobulin Deposits (n=3), Light Chain Proximal Tubulopathy (n=3), Randall-type monoclonal immunoglobulin deposition disease (n=2, 6%), 1 Minimal Change Disease (n=1; 3%), Glomerulonephritis with Organized Microtubular Monoclonal Ig Deposits (n=1) and exclusive tumoral renal infiltration (n=1). Associated tumoral infiltration or crystalline podocytopathy was confirmed in 2 and 1 biopsy respectively. Glomerular MGRS was highly suspected for 2 patients, even if no biopsy was performed. MGRS was excluded for 22 patients (39%): AKI due to antibiotics or sepsis (n=8; 36%), hypovolemia (n=4; 18%) or hypercalcemia (n=3; 14%); 2 renal artery stenosis due to tyrosine kinase inhibitors (n=2; 9%), nephroangiosclerosis (n=1), lithiasis (n=1), CKD post- nephrectomy CKD (n=1), and immunoallergic nephritis (n=1). Hematological treatment was initiated in 54 patients (96%). All patients with MGRS were treated, whom 17 (50%) completely or partially in the Nephrology department. Proteasome inhibitors, alkylant agents and IMIDs were the main molecule used in first line therapy (56%, 23% and 12% respectively). Eleven patients (20%) had stem cell transplantation. After a median follow up of 16 months (38-0), complete or very good partial hematologic response was obtained in 48 patients (89%); whom 30 with MGRS (88%). Median eGFR was 45ml/min/1.73m (142-4): CKD stage II (n=18), stage III (n=23) and stage IV (n=9). Only 1/3 CKD stade V patient needed chronic replacement therapy. Eight patients died (14%): 4 uncontrolled hematologic disease (50%) and 3 septic complications (38%). Conclusion Very closed collaboration between Nephrology and Hematology Department in our Center contributes to improve early management of patients with hematological disorders associated with CKD, and probably their survey. Hemato-Nephrology is a viable and emergent specialty.

SAT-618 Nattokinase to Improve Insulin Sensitivity and Weight Loss in Women with Obesity +/- Diabetes

Background: Diabetes (DM) and obesity are related health issues which are increasing in prevalence. But not all obesity is related to DM. Women suffering from Lipedema are categorized as gynoid obese. Nattokinase (Natto) is an enzyme supplement that has been shown to degrade fibrin. Patients with obesity tend to have elevated clotting factors which can lead to adipose tissue hypoxia, impaired insulin signaling, and lead to insulin resistance. Research in fat disorders noted that fat biopsies from women with Lipedema likely had micro-clots, and patients with Lipedema treated with Natto reported a decrease in clothing size and fat distribution. Objective: Determine the effect of Natto on participants with Obesity and DM and in patients with Lipedema without DM. Materials and Methods: Group 1: Involved subjects with Obesity and DM. This was a double blinded, randomized controlled clinical trial over 3 months. A total of 17 female patients were recruited from a rural clinic. Nine received Natto 2,000 FU daily and eight received an identical placebo capsule daily. Fasting labs, questionnaires, bioimpedance, and anthropometric measurements were completed at Baseline and 3 months. Group 2: 42 women with Lipedema seen at a Fat Disorder Clinic. 21 received Natto and 21 did not. We compared for weight only from the day Nattokinase was started until follow-up, which varied from 4 months to 1 year 8 months. Results: Group 1: After 3 months, there was no difference in weight loss in both groups. Per Bioimpedance, more subjects lost water weight in the Natto group (63%) compared to Placebo (33%). More subjects in the Natto group had a decrease in HbA1c (43%) compared to Placebo (22%), with average decrease in the Natto group of 0.9%. Also, more subjects in the Natto group had lower fasting insulin levels (75% vs 22%), lower fasting glucose level (50% vs 22%) and lower HOMA index (63% vs 22%). Group 2: 57% of patients in the Natto group lost weight compared to only 33% of patients not on Natto. Conclusions: In participants with obesity and DM, regardless of weight loss, metabolic health improved after taking Natto for 3 months. Higher percentage of subjects in the treated group had improved HbA1c, fasting Insulin, glucose, and HOMA score. We hypothesize that if treatment time was beyond 3 months, further metabolic improvement would be noted, indicating that Natto could have potential as an adjunct to DM care. The difference in weight loss between Metabolic Obesity and Lipedema reflects the difference in adipose tissue, likely differing in etiology and pathophysiology. Further studies are needed to evaluate long term benefits of Natto, including larger and longer randomized controlled trials, and assessment of clotting factors.

Thigh Adipocyte Size is Inversely Related to Energy Intake and Respiratory Quotient in Healthy Women.

The relationship between adipocyte size and ad libitum energy intake has not been previously examined. This study hypothesized an inverse relationship between adipocyte size and daily energy intake (DEI). Seventy healthy adults (39 men and 31 women; BMI 30.0 [SD6.3]) underwent dual-energy x-ray absorptiometry and subcutaneous fat biopsies from the abdomen and thigh. Osmium-fixed adipocytes were sized with a Coulter counter. Volunteers self-selected food from a vending machine paradigm as the only source of energy intake over 3 days as inpatients. Volunteersalso had 24-hour respiratory quotient (RQ) measured in a whole-room indirect calorimeter. In women, the large cell peak diameter of the thigh depot was greater than that of the abdominal depot (Δ = +15.8 μm; P < 0.0001). In women, thigh peak diameter was inversely associated with DEI (β = -264.7 kcal/d per 10-μm difference; P = 0.03) after adjusting for demographics and body composition. The thigh peak diameter in women was associated with 24-hour RQ (r = -0.47, P = 0.04) after adjusting for demographics, body composition, and 24-hour energy balance. These associations did not extend to men or the abdominal depot. In women, thigh adipocyte size was associated with reduced DEI and 24-hour RQ, indicating a special role for thigh fat in women. This depot-specific sexual dimorphism indicates common regulation of energy intake and adipocyte size in the thigh region of women.

Fat Distribution in Women Is Associated With Depot-Specific Transcriptomic Signatures and Chromatin Structure.

BackgroundPreferential accumulation of fat in the upper body (apple shape) is associated with higher risk of developing metabolic syndrome relative to lower body fat (pear shape). We previously discovered that chromatin openness partially defined the transcriptome of preadipocytes isolated from abdominal and gluteofemoral fat. However, the molecular mechanisms underlying interindividual variation in body shape are unknown.MethodsAdipocyte fraction was isolated from abdominal and gluteofemoral fat biopsies of premenopausal women (age and body mass index matched) segregated initially only by their waist-to-hip ratio. We evaluated transcriptomic and chromatin accessibility using RNA sequencing and assay for transposase-accessible chromatin using sequencing (ATAC-seq) along with key clinical parameters.ResultsOur data showed that higher lower body fat mass was associated with better lipid profile and free fatty acid decrease after glucose administration. Lipid and glucose metabolic pathways genes were expressed at higher levels in gluteofemoral adipocyte fraction in pears, whereas genes associated with inflammation were higher both in abdominal and gluteofemoral apple adipocyte fraction. Gluteofemoral adipocyte chromatin from pear-shaped women contained a significantly higher number of differentially open ATAC-seq peaks relative to chromatin from the apple-shaped gluteofemoral adipocytes. In contrast, abdominal adipocyte chromatin openness showed few differences between apple- and pear-shaped women. We revealed a correlation between gene transcription and open chromatin at the proximity of the transcriptional start site of some of the differentially expressed genes.ConclusionsIntegration of data from all 3 approaches suggests that chromatin openness partially governs the transcriptome of gluteofemoral adipocytes and may be involved in the early metabolic syndrome predisposition associated with body shape.

Resveratrol and metabolic health in COPD: A proof-of-concept randomized controlled trial.

Patients with COPD are often characterized by disturbed metabolic health which is reflected in altered body composition. Current studies in healthy subjects suggest that resveratrol improves metabolic health by enhancing muscle mitochondrial function and adipose tissue morphology. The primary objective was to investigate the effect of four weeks resveratrol supplementation on muscle mitochondrial function in patients with COPD. Secondary objectives were to investigate the effect of resveratrol on adipose tissue inflammatory and metabolic gene expression, systemic inflammation and body composition in patients with COPD. In a double-blind randomized placebo-controlled proof-of-concept study, 21 COPD patients (FEV1: 53±15% predicted; age: 67±9 years and BMI: 24.5±3.3kg/m2) received resveratrol (150mg/day) or placebo for four weeks. Before and after intervention, blood samples, quadriceps muscle and subcutaneous abdominal fat biopsies were obtained for metabolic and inflammatory profiling. Body composition was assessed by dual energy X-ray absorptiometry. Muscle mitochondrial biogenesis regulators AMPK, SIRT1 and PGC-1α as well as mitochondrial respiration, Oxphos complexes, oxidative enzyme activities and kynurenine aminotransferases were not improved by resveratrol. Plasma high-sensitive C-reactive protein and kynurenine did not change after resveratrol supplementation. Adipose tissue inflammatory markers were unaffected by resveratrol, while markers of glycolysis and lipolysis were significantly increased compared to placebo supplementation. Body weight decreased after resveratrol supplementation (resveratrol-0.95±1.01kg vs placebo-0.16±0.66kg, p=0.049) due to a reduction in lean mass (resveratrol-1.79±1.67kg vs 0.37±0.86kg, p=0.026). We do not confirm previously reported positive effects of resveratrol on skeletal muscle mitochondrial function in patients with COPD, but show an unexpected decline in lean mass. Clinicaltrials.gov NCT02245932.

Genetic determinism of boar taint and relationship with growth traits, meat quality and lesions

Breeding entire males is an alternative to surgical castration to improve their welfare. However, entire males may have a major quality defect called boar taint. Boar taint is partly due to the presence of androstenone in fat. In this study, we estimated the genetic parameters between androstenone and production traits to evaluate the consequences of selection against boar taint for traits of interest. We focused on growth traits, meat quality, lesions, hormone levels and computerised tomography measurements in purebred Piétrain (P) or Piétrain cross Large White (X) entire males. The number of measured animals varied from 670 P and 734 X for hormones concentrations to 553 P and 645 X for computerised tomography measurements. Skin lesions were measured on live pigs shortly after mixing, at the end of the fattening period, and on carcasses. Heritabilities of traits measured by tomography ranged from low to high: femur density (P: 0.34, X: 0.69), loin eye area (P: 0.53, X: 0.88) and loin eye density (P: 0.12, X: 0.18). The mean number of lesions at each stage was lower in purebred pigs than in crossbreds (entering the fattening stage 4.01 in P and 4.68 in X; before slaughter 3.72 in P and 4.22 in X; on carcass 4.50 in P and 4.96 in X). We also observed a decrease in the average number of lesions between the two stages in live pigs. We found high genetic correlations between stages in purebred pigs (0.74 to 0.76) but low correlations (−0.30 to 0.29) in crossbred pigs. Selection aiming to decrease fat androstenone is feasible (h2 = 0.57 in P and h2 = 0.71 in X). It would have overall positive effects on meat production and quality traits. Selection aiming to reduce plasma oestradiol would strongly reduce the level of fat androstenone (rg = 0.89 in P and rg = 0.84 in X). Selection against oestradiol is easier and less invasive since it would only require a blood sample rather than a fat biopsy in live animals.

Obesity and Insulin Resistance Are Inversely Associated with Serum and Adipose Tissue Carotenoid Concentrations in Adults

Background:Low tissue concentrations of carotenoids have been suggested to contribute to insulin resistance in obesity. Objectives:The objectives of the study were to 1) evaluate the relations of adipose tissue and serum carotenoids with body fat, abdominal fat distribution, muscle, adipose tissue and liver insulin resistance, and dietary intake; 2) evaluate the relations and distributions of carotenoids detected in adipose tissue and serum; and 3) compare serum carotenoids and retinol concentrations in subjects with and without obesity. Methods:Post hoc analysis of serum and adipose tissue carotenoids in individuals [n = 80; 31 men, 49 women; age (mean ± SEM): 51.4 α 1.1 y] who participated in 2 separate studies conducted at the Clinical Research Facility at the Garvan Institute of Medical Research (Sydney) between 2008 and 2013. Retinol, α-carotene, β-carotene, ζ-carotene, lutein, lycopene, phytoene, and phytofluene were measured using HPLC. Body composition was measured by dual-energy X-ray absorptiometry. Insulin resistance was measured by 2-step hyperinsulinemic-euglycemic clamps with deuterated glucose (n = 64), and subcutaneous and visceral abdominal volume and liver and pancreatic fat by MRI (n = 60). Periumbilical subcutaneous fat biopsy was performed and carotenoids and retinol measured in the tissue (n = 16). Results:We found that ζ-carotene, phytoene, and phytofluene were stored in considerable amounts in adipose tissue (25% of adipose tissue carotenoids). Carotenoid concentrations in adipose tissue and serum correlated significantly, but they followed different distributions: ζ-carotene was 3-fold higher in adipose tissue compared with serum, while lutein and lycopene made up 20% and 21% of serum carotenoids compared with 2% and 12% of adipose tissue carotenoids, respectively. Liver (P ≤ 0.028) and adipose tissue (P = 0.023), but not muscle (P ≥ 0.16), insulin resistance correlated inversely with many of the serum carotenoids. Conclusions:Multiple serum and adipose tissue carotenoids are associated with favorable metabolic traits, including insulin sensitivity in liver and adipose tissue in humans.

Testosterone Increases the Expression and Phosphorylation of AMP Kinase α in Men With Hypogonadism and Type 2 Diabetes.

Adenosine 5'-monophosphate-activated protein kinase-α (AMPKα) is a mediator of exercise-induced glucose uptake in skeletal muscle. We evaluated whether AMPKα expression and phosphorylation are reduced in skeletal muscle and adipose tissue of patients with hypogonadotropic hypogonadism (HH), and whether testosterone replacement therapy results in restoration of the expression and phosphorylation of AMPKα. This is a secondary analysis of a previously completed trial that showed an insulin-sensitizing effect of testosterone therapy in men with type 2 diabetes and HH. Clinical research center at university. Thirty-two men with HH and 32 eugonadal men were compared at baseline. Men with HH were treated with intramuscular injections of testosterone or placebo every 2 weeks for 22 weeks. Quadriceps muscle biopsies and subcutaneous abdominal fat biopsies were obtained before and after 4-hour euglycemic hyperinsulinemic clamp, prior to and after testosterone or placebo therapy. mRNA expression of AMPKα in hypogonadal men was lower by 37% in adipose tissue and 29% in skeletal muscle, respectively, compared with levels in eugonadal men, while phosphorylated AMPKα was lower by 22% and 28%, respectively. Following testosterone replacement, the expression of AMPKα did not alter in the fasting state but increased markedly by 41% and 46% in adipose tissue and muscle, respectively, after the clamp. In contrast, phosphorylated AMPKα increased by 69% in muscle after testosterone therapy but did not change following the clamp. Testosterone modulates the expression of AMPKα and phosphorylated AMPKα. These effects may contribute to the improved insulin sensitivity following testosterone therapy.

Metformin does not compromise energy status in human skeletal muscle at rest or during acute exercise: A randomised, crossover trial.

5´AMP–activated protein kinase (AMPK) is a mediator of a healthy metabolic phenotype in skeletal muscle. Metformin may exacerbate the energy disturbances observed during exercise leading to enhanced AMPK activation, and these disturbances may provoke early muscular fatigue. We studied acute (1 day) and short‐term (4 days) effects of metformin treatment on AMPK and its downstream signaling network, in healthy human skeletal muscle and adipose tissue at rest and during exercise, by applying a randomized blinded crossover study design in 10 lean men. Muscle and fat biopsies were obtained before and after the treatment period at rest and after a single bout of exercise. Metformin treat ment elicited peak plasma and muscle metformin concentrations of 31 μM and 11 μM, respectively. Neither of the treatments affected AMPK activity in skeletal muscle and adipose at rest or during exercise. In contrast, whole‐body stress during exercise was elevated as indicated by increased plasma lactate and adrenaline concentrations as well as increased heart rate and rate of perceived exertion. Also whole‐body insulin sensitivity was enhanced by 4 days metformin treatment, that is reduced fasting plasma insulin and HOMA‐IR. In conclusion, acute and short‐term metformin treatment does not affect energy homeostasis and AMPK activation at rest or during exercise in skeletal muscle and adipose tissue of healthy subjects. However, metformin treatment is accompanied by slightly enhanced perceived exertion and whole‐body stress which may provoke a lesser desire for physical activity in the metformin‐treated patients.