Metabolic Evaluation Research Articles

P0317EARLY MARKERS OF RENAL INJURY AND ENDOTHELIAL DYSFUNCTION IN PATIENTS WITH METABOLIC SYNDROME

Abstract Background and Aims The definition of subclinical renal damage markers in patients with metabolic syndrome (MS) and evaluation the relations between the indicators of subclinical renal injury and endothelial dysfunction. Method The study included 62 patients with MS, 32 (51.6%) men and 30 (48.4%) women aged 51.2±5.4 years. The control group included 25 practically healthy individuals, 13 (52.0%) men and 12 (48.0%) women, aged 47.3±6.1 years. All patients were underwent clinical and laboratory examination, calculation of glomerular filtration rate (GFR) using the formula CKD-EPI, ultrasound of the carotid arteries, determination of the insulin resistance index (IR) (HOMA-IR), as well as determination of the content of endothelin-1 (ET-1) in serum and albumin excretion in a single portion of urine by IFA. Results When calculating GFR, hyperfiltration was detected in 10 (16.1%), optimal filtration - in 23 (37.1%), slightly reduced - in 29 (46.8%). At the same time, in patients with MS, a statistically significant excess of urinary albumin excretion (UAE) in a single dose was revealed compared with the control group (26.6±5.8 mcg/ml and 9.2±3.4 mcg/ml, respectively; p <0.001). Based on the increase in level (ET-1) in patients of the main group, compared with the control (6.3±3.2 fmol/ml and 3.4±1.0 fmol/ml, respectively; p=0.03) may indicate the development in patients of the main group of endothelial dysfunction. A correlation analysis revealed a direct correlation between the level of UAE and ET-1 (R = 0.45; p = 0.005). A direct correlation was found between UAE and IR-HOMA (R = 0.68; p <0.007), interventricular septum thickness (R = 0.71; p <0.001), and intima–media thickness (IMT) (R = 0.49; p <0.008) and systolic blood pressure (R = 0.47; p = 0.026). Similar associations were noted also for the ET-1 level: IR-HOMA2 (R = 0.78; p <0.001), with the thickness of the interventricular septum (R = 0.76; p = 0.001), IMT (R = 0.46; p <0.033) and systolic blood pressure (R = 0.45; p = 0.035). Conclusion In patients with metabolic syndrome, in addition to the well-known manifestations, in the early stages, may be a violation of renal function and endothelium, manifested by the change of glomerular filtration rate, increased levels of UAE and increased concentration of ET-1 in the blood. Direct correlation of the level of UAE and ET-1 level, and the coincidence of the correlation relationships of these indicators with a number of clinical and laboratory indicators of the metabolic syndrome will make it possible to consider an increasing of the level of UAE as one of the markers of endothelial dysfunction.

363-OR: Comprehensive Assessment of Glycemic Control after Pancreas Transplantation

Introduction: Limited data are available about comprehensive metabolic evaluation after Pancreas transplantation (PT) for type 1 diabetes (T1D). Methods: After IRB approval, we prospectively studied 15 PT recipients at least 1 year post transplant. After a screening visit, subjects underwent Dexcom G4 CGM for one week and Mixed Meal Test (MMT) according to standard procedure. Results: PT recipient were 53.1 ± 10.6 years old, F/M 12/3, BMI (Kg/m2) 30.8 ± 6 and mean duration of diabetes was 33 ± 12.3 years with 6 ± 5 (1-17) years mean interval from PT to the study. Twelve patients underwent pancreas transplant alone and three underwent simultaneous pancreas kidney transplantation. Baseline values were Hemoglobin 12.3 ± 1.4 (g/dl), HbA1c 5.4 ± 0.6 %, Albumin 4.0 ± 0.3 g/dl, Fructosamine 233 ± 27 (µmol/L), Fasting Glucose 94.3 ± 17.5 mg/dl, C-peptide 2.5 ± 0.9 (ng/ml), eGFR 56.3 ± 25.5 ml/min and AST 23.4 ± 8.5 U/L. MMT revealed Glucose concentration increased from 90.9 ± 9.6 mg/dl at baseline to 140.6 ± 41.5 mg/dl at 30min followed by 123.2 ± 48.0 mg/dl and 115.2 ± 34.6 mg/dl at 60mins and 120 mins respectively. C-peptide was 2.4 ± 0.7 ng/ml at baseline, peaked to 7.5 ± 5.4 ng/ml at 60 mins and was 6.0 ± 3.6 mins and 5.9 ± 2.4 at 30 and 120 mins respectively with insulin peaking from 13.5 ± 5.5 uIU/ml at baseline to 107 ± 81.6 uIU/ml at 30 mins and 85.8 ± 78 uIU/ml and 42.2 ± 23.2 uIU/ml at 60 mins and 120 mins respectively. CGM analysis revealed mean CGM glucose was 120.4 ± 14.8 with SD 26.2 ± 7.7. Glucose control was excellent with 94 ± 9.2% of readings within 70-180mg/dl and 79.8 ± 17.6% within 70-140 mg/dl. Percentage time < 70 mg/dl and < 54 mg/dl were 1.1 ± 1.1% and 0.1 ± 0.4% respectively. Percentage time > 180 and > 250 mg/dl were 4.8 ± 9.3% and 0.4 ± 0.7% respectively. HBGI, LBGI and ADRR were 1.3 ± 1.6, 0.7 ± 0.5 and 17.4 ± 8.9 respectively. Conclusion: PT recipients demonstrate excellent glucose control on comprehensive assessment one or more years after the procedure. CGM and MMT data are different from normal people. Disclosure R. Kaur: None. C. Reid: None. P. Thapa: None. S. Rizvi: None. S.K. McCrady-Spitzer: None. S. Batthula: None. B.H. Smith: None. P. Dean: None. W.K. Kremers: Research Support; Self; AstraZeneca. A. Kukla: None. M.D. Stegall: None. Y.C. Kudva: Research Support; Self; Dexcom, Inc., Roche Diabetes Care. Other Relationship; Self; Abbott.

Pearls and Pitfalls of Metabolic Liver Magnetic Resonance Imaging in the Pediatric Population

Recent advances in magnetic resonance imaging (MRI) technology have moved imaging beyond anatomical assessment to characterization of tissue composition. There are now clinically validated MRI-based quantitative techniques for assessing liver fat, iron, and fibrosis, and MRI is now routinely used in metabolic liver disease evaluation in both pediatric and adult patients. These MRI techniques provide noninvasive quantitation of liver metabolic biomarkers that are increasingly relied upon in the clinical management of pediatric patients with nonalcoholic fatty liver disease, metabolic syndrome, and hemochromatosis and/or hemosiderosis. This article provides a review of the clinical indications and technical parameters for performing metabolic liver MRI in the pediatric population, along with common pearls and pitfalls encountered during its performance.

Metabolic, cardiovascular, and substance use evaluation of living kidney donor candidates: US practices in 2017.

We surveyed US transplant centers to assess practices regarding the evaluation and selection of living kidney donors based on metabolic, cardiovascular, and substance use risk factors. Our companion article describes renal aspects of the evaluation. Response rate was 31%. Compared with 2005, programs have become more accepting of hypertensive candidates: 65% in 2017% vs 41% in 2005 consider candidates with hypertension well controlled with 1 medication. One notable exception is black hypertensive candidates, who are frequently excluded regardless of severity. The most common body mass index (BMI) cutoff remains 35kg/m2 , and fewer programs now consider candidates with BMI >40kg/m2 . A 2-hour oral glucose tolerance test of ≥140mg/dL remains the most common criterion for exclusion of prediabetic candidates. One quarter to one third of programs exclude based on isolated cardiac abnormalities, such as mild aortic stenosis; a similar proportion consider these candidates only if older than 50years. Cigarette or marijuana smoking are infrequently criteria for exclusion, although 45% and 37% programs, respectively, require cessation 4weeks prior to surgery. In addition to providing an overview of current practices in living kidney donor evaluation, our study highlights the importance of research evaluating outcomes with various comorbidities to guide practice.

SLIPS-LAB-A bioinspired bioanalysis system for metabolic evaluation of urinary stone disease.

Urinary stone disease is among the most common medical conditions. Standard evaluation of urinary stone disease involves a metabolic workup of stone formers based on measurement of minerals and solutes excreted in 24-hour urine samples. Nevertheless, 24-hour urine testing is slow, expensive, and inconvenient for patients, which has hindered widespread adoption in clinical practice. Here, we demonstrate SLIPS-LAB (Slippery Liquid-Infused Porous Surface Laboratory), a droplet-based bioanalysis system, for rapid measurement of urinary stone-associated analytes. The ultra-repellent and antifouling properties of SLIPS, which is a biologically inspired surface technology, allow autonomous liquid handling and manipulation of physiological samples without complicated sample preparation procedures and supporting equipment. We pilot a study that examines key urinary analytes in clinical samples from patients with urinary stone. The simplicity and speed of SLIPS-LAB hold the potential to provide actionable diagnostic information for patients with urinary stone disease and rapid feedback for responses to dietary and pharmacologic treatments.

A Pharmacokinetic and Pharmacodynamic Evaluation of the Anti-Hepatocellular Carcinoma Compound 4-N-Carbobenzoxy-gemcitabine (Cbz-dFdC).

Gemcitabine (dFdC) demonstrates significant effectiveness against solid tumors in vitro and in vivo; however, its clinical application is limited because it tends to easily undergo deamination metabolism. Therefore, we synthesized 4-N-carbobenzoxy-gemcitabine (Cbz-dFdC) as a lead prodrug and conducted a detailed pharmacokinetic, metabolic, and pharmacodynamic evaluation. After intragastric Cbz-dFdC administration, the Cmax of Cbz-dFdC and dFdC was 451.1 ± 106.7 and 1656.3 ± 431.5 ng/mL, respectively. The Tmax of Cbz-dFdC and dFdC was 2 and 4 h, respectively. After intragastric administration of Cbz-dFdC, this compound was mainly distributed in the intestine due to low carboxylesterase-1 (CES1) activity. Cbz-dFdC is activated by CES1 in both humans and rats. The enzyme kinetic curves were well fitted by the Michaelis–Menten equation in rats’ blood, plasma, and tissue homogenates and S9 of the liver and kidney, as well as human liver S9 and CES1 recombinase. The pharmacodynamic results showed that the Cbz-dFdC have a good antitumor effect in the HepG2 cell and in tumor-bearing mice, respectively. In general, Cbz-dFdC has good pharmaceutical characteristics and is therefore a good candidate for a potential prodrug.

OR17-02 Changes in Hepatokines and Apolipoproteins Are Associated with Metabolic Response to Metreleptin in Partial Lipodystrophy

Introduction Metreleptin treatment may improve the metabolic aspects of partial lipodystrophy; however, the treatment response is heterogeneous. This study aimed to explore changes in circulating apolipoprotein concentrations, as well as ANGPLT3, ANGPLT4, and IGF-1 levels in patients treated with Metreleptin as part of a clinical study investigating the efficacy of Metreleptin in nonalcoholic steatohepatitis (NASH) associated with partial lipodystrophy (ClinicalTrials.gov identifier: NCT01679197). Methods Serum samples of 18 patients with partial lipodystrophy who underwent a full metabolic evaluation and paired liver biopsies before and after Metreleptin were studied. Patients were tested at baseline, month (M) 3, M6, and M12. Glycemic response was defined as “more than 1% HbA1c reduction from baseline”. Lipid response was defined as “more than 30% decrease in triglycerides from baseline”. The hepatic response was defined as “a decrease of 2 points or more from baseline in NASH score, without an increase in fibrosis”. Patients with “any 2 of 3 above” at M12 were defined as metabolic responders. Results Metreleptin treatment resulted in significant reductions in triglycerides (346 mg/dL vs. 253 mg/dL; F: 8.474; p < 0.001), apo B (145.24 mg/dL vs. 111.09 mg/dL; F: 9.266: p < 0.001), apo CII (18.65 mg/dL vs. 15.95 mg/dL; F: 6.663: p = 0.001), apo CIII (62.95 mg/dL vs. 49.33 mg/dL; F: 5.640, p = 0.002), apo E (8.16 mg/dL vs. 6.52 mg/dL; F: 11.056, p < 0.001), and ANGPLT3 (14.36 ng/mL vs. 12.00 mg/dL; F: 4.348; p = 0.008) over time. IGF-1 levels significantly increased at M3 (134 ng/mL vs. 139 ng/mL; p = 0.001), however the difference was not significant over time. Metabolic responders had lower baseline leptin (12.4 ng/mL vs. 27.8 ng/mL; p = 0.024) and IGF-1 (95 ng/ml vs. 151 ng/mL; p = 0.008), and higher apo CII (39.06 mg/dL vs. 17.90 mg/dL; p = 0.011), apo CIII (173.57 mg/dL vs. 51.51 mg/dL; p = 0.015), apo E (18.41 mg/dL vs. 5.89 mg/dL; p = 0.002), and ANGPLT3 (17.33 ng/mL vs. 10.06 ng/mL; p = 0.04). Metabolic responders had a significant increase in IGF-1 (95 ng/mL vs. 134 ng/mL; p = 0.019), which was statistically distinguished from non-responders (p = 0.004). Responders also had a greater reduction in apo CII (20.51 mg/dL vs. -1.84 mg/dL; p = 0.001), apo CIII (32.59 mg/dL vs. -7.83 mg/dL; p = 0.007), apo E (8.17 mg/dL vs. 0.22 mg/dL; p = 0.001), and ANGPLT3 (6.08 ng/mL vs. -0.16 ng/mL; p = 0.005) early after treatment at M3. Conclusions Metreleptin treatment lowers levels of apolipoproteins associated with triglyceride metabolism as well as ANGPLT3 in patients with partial lipodystrophy. Metabolic response to Metreleptin appears to be correlated with early changes in these factors and an increase in IGF-1 levels.

Targeting Casein kinase 2 with quercetin or enzymatically modified isoquercitrin as a strategy for boosting the type 1 interferon response to viruses and promoting cardiovascular health

The serine/threonine kinase CK2 has been shown to down-regulate the production of type 1 interferons in response to viral infections by conferring an inhibitory phosphorylation on RIG-I, which functions to detect double-stranded RNA generated during replication of RNA viruses. Quercetin and certain other planar flavones/flavonols can inhibit CK2 in high nanomolar concentrations; this may explain quercetin’s ability to slow the proliferation of RNA viruses in cell cultures and in mice. Limited clinical evidence suggests that supplemental quercetin may decrease risk for upper respiratory infections in humans. Quercetin and enzymatically-modified isoquercitrin (EMIQ – a food additive/nutraceutical that upon oral administration achieves far higher plasma concentrations of quercetin than quercetin per se) also have exerted a range of vascular-protective effects clinically and in rodents – improving endothelial function, warding off atherosclerosis, lowering blood pressure, decreasing C-reactive protein, aiding glycemic control, stabilizing platelets - that might also, at least in part, reflect CK2 inhibition. The utility of quercetin, EMIQ, and other clinically feasible CK2 inhibitors for aiding control of viral infections and promoting vascular and metabolic health merits further evaluation.

Association between dietary pattern and metabolic disorders in children and adolescents with urolithiasis

ObjectiveTo describe the dietary patterns and occurrence of metabolic disorders in children and adolescents with urolithiasis treatment at a referral hospital in southern Brazil in order to learn the features of urolithiasis in this population to better develop preventive actions. MethodsDescriptive study conducted between 2016 and 2017 in a tertiary care referral hospital. Fourty patients aged 2–19 years old with urolithiasis proven by imaging were included. Clinical and dietary data were obtained through interviews and medical records. For statistical analyses, the chi‐squared test was performed. Results40 individuals were analyzed. Mean age at diagnosis was 7.2±4 years. 25% were overweight or obese. 95% had metabolic disorders, hypocitraturia being the predominant type. Protein intake was adequate in all participants and carbohydrate intake, in 70% of them; 37.5% had lipid intake above recommended and 65% had low fiber intake. The mean daily sodium intake was 2.64g (±1.74), with 55% of participants ingesting more than the recommended amount. A total of 52.5% had low potassium intake, with a mean of 4.79g/day (±2.49). Calcium intake was adequate in 27.5%. No significant differences were identified in relation to mean daily consumption among participants with or without the various metabolic disorders. ConclusionPediatric urolithiasis is often accompanied by metabolic disorders; therefore, metabolic evaluation should be part of the diagnostic process and subsequent analysis of these patients’ dietary patterns, helping to optimize treatment and prevent recurrences and complications.

Genetic IGF1R defects: new cases expand the spectrum of clinical features.

We aimed to identify the phenotypic variability of IGF1R defects in a cohort of short children with normal GH secretion gathered through the last decade. Fifty children (25 girls) with short stature and a basal/stimulated growth hormone (GH) over 10ng/ml having either a low birth weight or microcephaly were enrolled. MLPA and then Sanger sequence analysis were performed to detect IGF1R defects. The auxological and metabolic evaluation were carried out in index cases and their first degree family members whenever available. A total of seven (14%) IGF1R defects were detected. Two IGF1R deletions and five heterozygous variants (one frameshift, four missense) were identified. Three (likely) pathogenic, one VUS and one likely benign were classified by using ACMG. All children with IGF1R defects had a height < - 2.5SDS, birth weight < - 1.4SDS, and head circumference < - 1.36SDS. IGF-1 ranged from - 2.44 to 2.13 SDS. One child with a 15q terminal deletion had a normal phenotype and intelligence, whereas low IQ is a finding in a case with missense variant. Two parents who carried IGF1R mutations had diabetes mellitus, hypertension and hyperlipidemia, one of whom also had hypergonadotropic hypogonadism. We found a deletion or variant in IGF1R in 14% of short children. Birth weight, head circumference, intelligence, dysmorphic features, IGF-1 levels and even height are not consistent among patients. Additionally, metabolic and gonadal complications may appear during adulthood, suggesting that patients should be followed into adulthood to monitor for these late complications.

Isolation, Characterization and Targeted Metabolic Evaluation of Endophytic Fungi Harbored in 14 Seed-Derived Hypericum Species.

Medicinal plants of the genus Hypericum are rich sources of bioactive naphthodianthrones, which are unique in the plant kingdom, but quite common in fungal endophytes. Cultivable endophytic fungi were isolated from 14 different Hypericum spp. originating from seeds grown under in vitro conditions and further acclimated to outdoor conditions. Among 37 fungal isolates yielded from the aerial and underground plant organs, 25 were identified at the species level by the fungal barcode marker internal transcribed spacer rDNA and protein-coding gene region of tef1α. Ten of them were isolated from Hypericum spp. for the first time. The axenic cultures of the isolated endophytes were screened for the production of extracellular enzymes, as well as bioactive naphthodianthrones and their putative precursors by Bornträger's test and HPLC-HRMS. Traces of naphthodianthrones and their intermediates, emodin, emodin anthrone, skyrin, or pseudohypericin, were detected in the fungal mycelia of Acremonium sclerotigenum and Plectosphaerella cucumerina isolated from Hypericum perforatum and Hypericum maculatum, respectively. Traces of emodin, hypericin, and pseudohypericin were released in the broth by Scedosporium apiospermum, P. cucumerina, and Fusarium oxysporum during submerged fermentation. These endophytes were isolated from several hypericin-producing Hypericum spp. Taken together, our results reveal the biosynthetic potential of cultivable endophytic fungi harbored in Hypericum plants as well as evidence of the existence of remarkable plant-endophyte relationships in selected non-native ecological niches. A possible role of the extracellular enzymes in plant secondary metabolism is discussed.

CARACTERÍSTICAS METABÓLICAS DE LOS PACIENTES CON APNEAS OBSTRUCTIVAS DEL SUEÑO

Introducción: la apnea obstructiva del sueño (AOS) es una patología de alta prevalencia asociada al sobrepeso y la obesidad. La relación entre el síndrome metabólico (SM) y la AOS no ha sido comunicada en nuestro medio.Materiales y métodos: estudio retrospectivo en adultos seleccionados para poligrafía respiratoria y evaluación metabólica clasificados según la presencia de SM.Resultados: incluimos 302 pacientes. La prevalencia de obesidad fue de 66,88% y de SM de 62,58%. El 19% presentó síntomas de somnolencia diurna y el 48,3% mostró cinco o más componentes del cuestionario STOP-BANG.Los pacientes con AOS fueron mayormente del sexo masculino, con mayor edad; peso corporal, circunferencia de cintura y cuello comparados con el grupo sin AOS (57 vs 49 años, p&lt;0,001; 93,89 vs 85 kg, p&lt;0,05; 108 vs 100 cm, p&lt;0,001 respectivamente). Mostraron mayores valores de triglicéridos, presión arterial sistólica, obesidad e índice de desaturación (134 vs 99 mg/dl, p&lt;0,001; 134 vs 128 mmHg, p&lt;0,05; 69,2 vs 52,3%, p&lt;0,05 y 14,6 vs 2 ev/h, p&lt;0,001 respectivamente). La cantidad de componentes de STOP-BANG fue mayor en los pacientes con AOS (5 vs 3; p&lt;0,001).Conclusiones: AOS y SM están frecuentemente relacionados y permanecen subdiagnosticados. La utilización de cuestionarios validados facilita la identificación de candidatos a estudios de sueño. Se necesita la implementación de programas de gestión de hábitos saludables para prevenir las complicaciones de ambas patologías.

V04-06 ENCRUSTATED METALLIC URETERAL STENT TREATED BY MINI NEPHROLITHOTOMY

INTRODUCTION AND OBJECTIVE: The use of polymeric double J stents for the management of ureteral strictures is effective for a short period of time, although they have limited use due to the risk of obstruction and to the need for periodic stent changes. Metal stents have been developed with the aim of maintaining greater ureteric patency, alleviating symptoms, and, especially, allowing less frequent exchanges. Although metallic stent changes are approved for 12 months, there are no concrete data defining a safe dwell-time to prevent clinically significant encrustations. Likewise, there are no reports of surgeries performed to treat metal stent encrustations. Our objective with this video is to describe a case of a large metallic ureteral stent encrustation successfully treated by Mini Percutaneous Nephrolithotomy (MiniPerc) in an obese patient. METHODS: A 48-year-old female was admitted at our hospital for treatment. The patient had previously undergone a metallic DJ stent implantation for the treatment of a chronic ureteral stricture but hasn’t had a stent exchange in the last 18 months due to loss of follow-up. Radiological exams revealed a large encrustation at the right renal pelvis (2,5x1,6cm). After the attempt of removing the stent by cystoscopy failed, a mini percutaneous nephrolithotomy was performed to treat the kidney stone and to implant another stent. RESULTS: After the procedure, the patient became stone-free and had a silicone DJ stent implanted. Metabolic evaluation of the urine did not reveal hypercalciuria or any other contributing factor for this severe complication. CONCLUSIONS: Large encrustations on a metallic ureteral stent are rare complications and may require surgical treatment. Mini percutaneous nephrolithotomy seems to provide a safe and feasible treatment option for these cases, even in obese patients. Source of Funding: Authors

Revisiting an Old Concept in a New Era: 36 Hour Lung Preservation Using 10ºC Static Cold Storage

Despite remote reports indicating 10°C to be the optimal temperature for cold static organ preservation (CSP), preservation of lungs in an ice cooler at 4°C remains the standard strategy in clinical transplantation. Here, we used a commercially available device that can keep a donor lung at 10°C conveniently and compared 10°C vs. 4°C during a prolonged period of CSP followed by functional and biological assessment of lung grafts. Lungs were procured from Yorkshire pigs (28-35kg), flushed with a low-potassium dextran solution and randomized into two groups (n=5 each): CSP at 4°C vs. CSP at 10°C (MyTemp Mini Incubator, Benchmark Scientific). After 36h of CSP, lungs were subjected to 12h of normothermic ex vivo lung perfusion (EVLP) with hourly functional assessment. Lung biopsies and perfusate samples were taken for biological and metabolic evaluation. During 12h of EVLP, lungs stored at 10°C presented superior physiological parameters compared to 4°C (Fig 1A). Reduced edema formation was seen as represented by a lower lung weight gain during EVLP (30 ± 34.1 vs. 201 ± 33.2 g, p = 0.0159). Metabolic profiles of the EVLP perfusate were significantly more favorable in the 10°C group compared to 4°C (Fig 1B). Furthermore, the 10°C group had significantly less inflammation demonstrated by lower levels of pro-inflammatory cytokines IL-1β (p = <0.0001, Fig 1C), and IL-8 (p = <0.0001, Fig 1C) in the EVLP perfusate. The levels of IL-6 were significantly lower in both EVLP perfusate (p = <0.0001, Fig 1C) and post-perfusion lung tissue (p = 0.0317). Immunohistochemical analysis for 8-Hydroxyguanosine demonstrated that lungs stored at 10°C tended to have less oxidative stress during CSP (p = 0.0952). With simple controllable refrigerators, 10°C preservation is easily achievable and provides significantly superior CSP in comparison to conventional 4°C storage. This could potentially remarkably improve the clinical and transportation logistics in lung transplantation.

Cytotoxic Effects of Zoom® Whitening Product in Human Fibroblasts.

Tooth whitening procedures are increasing; however, side effects can occur, such as damage to pulp cells, by the whitening products. This study aims to assess the cellular effects promoted by a whitening product, namely, the oxidative stress fostered by the active agent hydrogen peroxide, with and without photoactivation. Additionally, if cellular recovery occurred, we intended to determine the time point where cells recover from the tooth whitening induced damage. Human fibroblasts were exposed to hydrogen peroxide, Zoom®, Zoom® + irradiation, and irradiation alone. The following analysis was performed: metabolic activity evaluation by the MTT assay; cell viability, mitochondrial membrane potential, peroxides production, superoxide radical production, and reduced glutathione expression by flow cytometry. We determined the IC50 value for all groups, and a dose-dependent cytotoxic effect was verified. At the times analyzed, hydrogen peroxide groups showed no metabolic activity recovery while a cell recovery was observed after 24 h (Zoom®) and 48 h (Zoom® + irradiation). Cell death was seen in hydrogen peroxide and Zoom® + irradiation groups, mainly by apoptosis, and the irradiation had a cytotoxic effect per se. This in vitro study supports that whitening products with moderate hydrogen peroxide (HP) concentration have a temporary effect on cells, allowing a cellular recovery.

Linking 24-h urines to clinical phenotypes: what alternatives does the future bring?

The 24-h urine test is recommended as part of the metabolic evaluation for patients with nephrolithiasis to guide preventive interventions. However, this test may be challenging to interpret and has limits in its predictive ability. In this review, we summarize and discuss the most recent research on the opportunities and challenges for utilizing urinary biomarkers for kidney stone prevention. Contemporary studies utilizing the 24-h urine test have improved our understanding of how to better administer testing and interpret test results. Beyond the standard panel of 24-h urine parameters, recent applications of proteomics and metabolomics have identified protein and metabolic profiles of stone formers. These profiles can be assayed in future studies as potential biomarkers for risk stratification and prediction. Broad collaborative efforts to create large datasets and biobanks from kidney stone formers will be invaluable for kidney stone research. Recent advances in our understanding of kidney stone risk have opened opportunities to improve metabolic testing for kidney stone formers. These strategies do not appear to be mutually exclusive of 24-h urine testing but instead complementary in their approach. Finally, large clinical datasets hold promise to be leveraged to identify new avenues for stone prevention.

Analysis of urine composition from split 24-h samples: use of 12-h overnight samples to evaluate risk factors for calcium stones in healthy and stone-forming children

The analysis of 24-h urine is the gold standard to diagnose metabolic abnormalities in the stone-forming patient. However, urinary composition changes throughout the dayand analyzing the whole 24-h urine may mask peaks of increased risk of crystallization. To examine variations of stone-promoting and stone-inhibiting factors in urine using split 24-h samples from healthy and stone-forming children. Urine was collected from 87 healthy and 26 stone-forming children using a split collection procedure (12-h daytime urine and 12-h overnight urine). Urine volume, pH, calcium (Ca), magnesium (Mg), phosphate (P), citrate (Cit), uric acid (Ur), and oxalate (Ox) were determined, and the Ca/Cit ratio was calculated. The overnight urine samples in both groups had higher levels of P and Mg, lower volume, lower pH, and less citrate and uric acid. As can be seen in the table, higher percentages of healthy and stone-forming children had altered 12-h night urine than 24-h urine with regards to Ca/Cr, Cit/Cr and Ca/Cit ratios. All healthy subjects and all stone-forming children (except one) with altered Cit/Cr ratios or Ca/Cit ratios in the 24-h sample also had altered ratios in the 12-h overnight sample. This study indicates that urine composition changes throughout the day, and that there is daily variability in most of the parameters related to kidney stone formation. Furthermore, 12-h overnight samples seem to be more sensitive than 24-h samples in detecting the most common urinary abnormalities. The main limitation of this study is the relative low sample size of stone-forming children, owing to the low prevalence of nephrolithiasis in childhood. We observed a higher excretion of stone-promoting substances and a lower citrate in urine at night. However, the study results do not provide enough evidence to conclude that the use of a 12-h overnight sample collection can replace 24h urine analysis in the metabolic evaluation of children with lithiasis.

Metabolic Profile and Evaluation of Biological Activities of Extracts from the Stems of Cissus trifoliata

Cissus trifoliata (L.) L belongs to the Vitaceae family and is an important medicinal plant used in Mexico for the management of infectious diseases and tumors. The present study aimed to evaluate the metabolic profile of the stems of C. trifoliata and to correlate the results with their antibacterial and cytotoxic activities. The hexane extract was analyzed using gas chromatography coupled with mass spectrometry (GC-MS) and the CHCl3-MeOH and aqueous extracts by ultraperformance liquid chromatography quadrupole time of fly mass spectrometry (UPLC-QTOF-MS). The antibacterial activity was determined by broth microdilution and the cytotoxicity was evaluated using MTS cell proliferation assay. Forty-six metabolites were putatively identified from the three extracts. Overall, terpenes, flavonoids and stilbenes characterize the metabolic profile. No antibacterial activity was found in any extract against the fifteen bacteria strains tested (MIC >500 µg/mL). However, high cytotoxic activity (IC50 ≤ 30 µg/mL) was found in the hexane and aqueous extracts against hepatocarcinoma and breast cancer cells (Hep3B, HepG2 and MCF7). This is the first report of the bioactive compounds of C. trifoliata stems and their antibacterial and cytotoxic properties. The metabolic profile rich in anticancer compounds correlate with the cytotoxic activity of the extracts from the stems of C. trifoliata. This study shows the antitumor effects of this plant used in the traditional medicine and justifies further research of its anticancer activity.

Metabolic Profiling-Based Evaluation of the Fermentative Behavior of Aspergillus oryzae and Bacillus subtilis for Soybean Residues Treated at Different Temperatures.

Soybean processing, e.g., by soaking, heating, and fermentation, typically results in diverse metabolic changes. Herein, multivariate analysis-based metabolic profiling was employed to investigate the effects of fermentation by Aspergillus oryzae or Bacillus subtilis on soybean substrates extracted at 4, 25, or 55 °C. As metabolic changes for both A. oryzae and B. subtilis were most pronounced for substrates extracted at 55 °C, this temperature was selected to compare the two microbial fermentation strategies, which were shown to be markedly different. Specifically, fermentation by A. oryzae increased the levels of most organic acids, γ-aminobutyric acid, and glutamine, which were ascribed to carbohydrate metabolism and conversion of glutamic acid into GABA and glutamine. In contrast, fermentation by B. subtilis increased the levels of most amino acids and isoflavones, which indicated the high activity of proteases and β-glucosidase. Overall, the obtained results were concluded to be useful for the optimization of processing steps in terms of nutritional preferences.

Integration of metabolic evaluation and exome sequencing for the diagnosis of children with urolithiasis

Metabolic evaluation and exome sequencing are both important etiological diagnostic methods for pediatric urolithiasis. There is a lack of integrated analysis of metabolic evaluation and exome sequencing in a pediatric urolithiasis cohort. A retrospective analysis of both metabolic and genomic data was performed for the first 105 pediatric urolithiasis patients who underwent exome sequencing at our hospital from February 2016 to October 2018. Measurements included the diagnostic yield of exome sequencing and the conformity between molecular diagnoses and metabolic evaluation. Exploratory data analyses were performed to compare the phenotypic differences between cases with positive and negative genetic findings. The present study involved a cohort of 105 pediatric patients with urolithiasis, from which diagnostic variants were identified in 37 patients (35%). In subgroup analyses, 39% hyperoxaluria cases are primary hyperoxaluria caused by monogenic defects, and 100% of the causes of cystine stones can be explained by monogenic defects. However, no appropriate inherited causes were identified for hypocitraturia, hypercalciuria, and hyperuricosuria in the cohort. A high conformity (100%) between molecular diagnoses and metabolic evaluation was obtained. Compared with negative cases, cases with positive genetic findings were correlated with an early onset age (median difference: -1.25 yr; 95% CI: -2.33 to -0.33, P < 0.001), a lower eGFR (median difference: -25 ml/min/1.73 m2; 95% CI: -41 to -9, P = 0.002), and a more severe stone burden (odds ratio: 10.7; 95% CI: 4.13 to 27.7, P < 0.001). Exome sequencing is an effective and reliable diagnostic method for inherited urolithiasis in children, and the molecular diagnostic yield varies greatly across different metabolic abnormalities. Early onset, lower eGFR, or severe stone burden are “red flag” conditions, suggestive of inherited urolithiasis.