Hybrid Machine Learning and Experimental Studies of Antiviral Potential of Ionic Liquids against P100, MS2, and Phi6.

Viruses are a group of widespread organisms that are often responsible for very dangerous diseases, as most of them follow a mechanism to multiply and infect their hosts as quickly as possible. Pathogen viruses also mutate regularly, with the result that measures to prevent virus transmission and recover from the disease caused are often limited. The development of new substances is very time-consuming and highly budgeted and requires the sacrifice of many living organisms. Computational chemistry methods allow faster analysis at a much lower cost and, most importantly, reduce the number of living organisms sacrificed experimentally to a minimum. Ionic liquids (ILs) are a group of chemical compounds that could potentially find a wide range of applications due to their potential virucidal activity. In our study, we conducted a complex computational analysis to predict the antiviral activity of ionic liquids against three surrogate viruses: two nonenveloped viruses, Listeria monocytogenes phage P100 and Escherichia coli phage MS2, and one enveloped virus, Pseudomonas syringae phage Phi6. Based on experimental data of toxic activity (logEC90), we assigned activity classes to 154 ILs. Prediction models were created and validated according to the Organization for Economic Co-operation and Development (OECD) recommendations using the Classification Tree method. Further, we performed an external validation of our models through virtual screening on a set of 1277 theoretically generated ionic liquids and then selected 10 active ionic liquids, which were synthesized to verify their activity against the analyzed viruses. Our study proved the effectiveness and efficiency of computational methods to predict the antiviral activity of ionic liquids. Thus, computational models are a cost-effective alternative approach compared with time-consuming experimental studies where live animals are involved.

Parity modulates impact of BMI and gestational weight gain on gut microbiota in human pregnancy.

Dysregulation of maternal adaptations to pregnancy due to high pre-pregnancy BMI (pBMI) or excess gestational weight gain (GWG) is associated with worsened health outcomes for mothers and children. Whether the gut microbiome contributes to these adaptations is unclear. We longitudinally investigated the impact of pBMI and GWG on the pregnant gut microbiome. We show that the gut microbiota of participants with higher pBMI changed less over the course of pregnancy in primiparous but not multiparous participants. This suggests that previous pregnancies may have persistent impacts on maternal adaptations to pregnancy. This ecological memory appears to be passed on to the next generation, as parity modulated the impact of maternal GWG on the infant gut microbiome. This work supports a role of the gut microbiome in maternal adaptations to pregnancy and highlights the need for longitudinal sampling and accounting for parity as key considerations for studies of the microbiome in pregnancy and infants. Understanding how these factors contribute to and shape maternal health is essential for the development of interventions impacting the microbiome, including pre/probiotics.

Antimicrobial and Virucidal Potential of Morpholinium-Based Ionic Liquids.

Witnessed by the ongoing spread of antimicrobial resistant bacteria as well as the recent global pandemic of the SARS-CoV-2 virus, the development of new disinfection strategies is of great importance, and novel substance classes as effective antimicrobials and virucides are urgently needed. Ionic liquids (ILs), low-melting salts, have been already recognized as efficient antimicrobial agents with prospects for antiviral potential. In this study, we examined the antiviral activity of 12 morpholinium based herbicidal ionic liquids with a tripartite test system, including enzyme inhibition tests, virucidal activity determination against five model viruses and activity against five bacterial species. The antimicrobial and enzymatic tests confirmed that the inhibiting activity of ILs corresponds with the number of long alkyl side chains and that [Dec2Mor]+ based ILs are promising candidates as novel antimicrobials. The virucidal tests showed that ILs antiviral activity depends on the type and structure of the virus, revealing enveloped Phi6 phage as highly susceptible to the ILs action, while the non-enveloped phages PRD1 and MS2 proved completely resistant to ionic liquids. Furthermore, a comparison of results obtained for P100 and P001 phages demonstrated for the first time that the susceptibility of viruses to ionic liquids can be dependent on differences in the phage tail structure.

[Drug therapy safety supported by interprofessional collaboration between ICU physicians and clinical pharmacists in critical care units in Germany : Results of a survey]. / Arzneimitteltherapiesicherheit gefördert durch die interprofessionelle Zusammenarbeit von Arzt und Apotheker auf Intensivstationen in Deutschland : Erkenntnisse einer Umfrage.

BACKGROUND: Critically ill patients are particularly susceptible to adverse drug events. International studies show that pharmaceutical care has a positive impact on patient and drug therapy safety. Nationally, the integration of pharmacists into the multidisciplinary team and participation in ward rounds is required. The aim of this work is to assess the scope and extent of pharmaceutical care in intensive care units (ICU) in Germany. METHOD: In a literature and database search, 13 relevant pharmaceutical activities were identified. Based on this, an online survey with 27 questions on the implementation of pharmaceutical care in ICU was prepared by a panel of experts. The survey was sent to heads of German ICUs. RESULTS: Of the participants, 35.3% (59/167) have established regular pharmaceutical care. Drug information (89.7% [52/58]), pharmaceutical interventions with change of therapy (e.g., ward rounds; 67,2% [39/58]), regular evaluation of prescriptions (medication analysis; 65.5% [38/58]) as well as the monitoring of medication (e.g., side effects, effectiveness, costs; 63.8% [37/58]) were most frequently mentioned. The participants with pharmaceutical care (58/168) graded 7 of 13 but those without (104/168) only two  activities as 'essential/indispensable'. CONCLUSION: Only a few ICU in Germany have already integrated ward pharmacists into the multidisciplinary team. Once a pharmaceutical service has been established, a greater role/importance is assigned to several pharmaceutical activities.

Victim identification from the September 11, 2001 attack on the World Trade Center: Past trends and future projections.

Victim identification following mass fatality events is critically important. Extensive traumatic injuries and body fragmentation add complexity to this process. World Trade Center (WTC) identification efforts have been ongoing for over 20 years and this study tracks identification trends from the 2753 known WTC victims and the 21,905 recovered remains. For identified victims, data include the number of remains identified, date(s) of the identification(s), and identification modalities. Results show a heavy reliance on DNA due to body fragmentation. Other modalities played an important role initially, but DNA eventually became the singular identification modality. For large-scale disasters involving significant body fragmentation, aggressive DNA testing strategies are critical for victim identification. Over time, the number of linked remains (portions of previously identified individuals) will greatly outnumber the new identifications (first-time identifications). A novel approach using statistical modeling from ecology studies was applied to estimate future WTC identification rates using Identification Accumulation Curve extrapolation with the Good-Toulmin estimator. Projections indicate there will be 76 first-time identifications (95% CI: 49-117) through the successful DNA testing of 3404 unidentified, fragmentary remains. The remainder of the identifications would be additional portions of previously identified victims. These results may be instructional for management of other large-scale, protracted victim identification efforts.

Victim identification and body completeness based on last known location at the World Trade Center.

This analysis focuses on the identification efforts conducted by the New York City Office of Chief Medical Examiner (NYC OCME) over a 20-year period from September 11, 2001 to September 11, 2021. Due to this unprecedented level of commitment to victim identification, a wealth of data has been collected over the two-decade period and is still being collected as identification efforts are ongoing. The results of this data analysis are not only informative for the World Trade Center (WTC) victims, but may also be instructional for other large-scale, protracted victim identification efforts. Based on available data, most victims are associated with the impact zones and higher in both towers. No correlation was observed in the overall identification rates based on last known location in the buildings, suggesting that location in the towers does not affect the likelihood of a successful identification. There was, however, a significant difference in the body completeness values observed for victims from the upper floors compared to those below the impact zones. The identification rates and body completeness values for victims onboard the two airplanes are significantly different from each other, possibly related to the varying aircraft speeds at the time of impact.

A new long-term sampling approach to viruses on surfaces.

The importance of virus disease outbreaks and its prevention is of growing public concern but our understanding of virus transmission routes is limited by adequate sampling strategies. While conventional swabbing methods provide merely a microbial snapshot, an ideal sampling strategy would allow reliable collection of viral genomic data over longer time periods. This study has evaluated a new, paper-based sticker approach for collection of reliable viral genomic data over longer time periods up to 14 days and after implementation of different hygiene measures. In contrast to swabbing methods, which sample viral load present on a surface at a given time, the paper-based stickers are attached to the surface area of interest and collect viruses that would have otherwise been transferred onto that surface. The major advantage of one-side adhesive stickers is that they are permanently attachable to a variety of surfaces. Initial results demonstrate that stickers permit stable recovery characteristics, even at low virus titers. Stickers also allow reliable virus detection after implementation of routine hygiene measures and over longer periods up to 14 days. Overall, results for this new sticker approach for virus genomic data collection are encouraging, but further studies are required to confirm anticipated benefits over a range of virus types.

Next generation sequencing of STR artifacts produced from historical bone samples.

STR artifacts are commonly observed in electrophoretic data and can complicate interpretation of the profiles produced. Even when a consensus approach is applied, reproducible artifacts have the potential to convolute the analysis. DNA obtained from historical bone samples is often heavily degraded and damaged, requiring the use of more sensitive procedures to increase allele recovery. Additionally, skeletal remains exposed to environmental conditions may be afflicted with microbial DNA contamination that cross-reacts with the primers during short tandem repeat (STR) multiplex amplification. STR artifacts manifested as a result of these circumstances can be sourced and characterized using new sequencing technologies to potentially ease the analysis burden. For this study, PCR product from 17 low-quality bone samples exhibiting reproducible autosomal and Y-chromosomal STR (Y-STR) artifacts in capillary electrophoresis (CE) data were sequenced with next generation sequencing (NGS). Sequenced reads were bioinformatically sorted using STRait Razor to determine the authenticity of alleles and confirm the profile generated by CE. Sequence data from the PCR products and a subset of the associated extracts were further analyzed with Kaiju to classify the microbial species present and identify potential sources of artifact peaks. A suspected Y-STR artifact was similar in sequence to Pseudomonas sp. BAY1663, a species ubiquitously found in soil. Regions of homology were observed between the Pseudomonas genome and the presumed primer binding locations for Y-STRs included in the AmpFlSTR Y-Filer STR kit. Characterization of such supposed artifact peaks may aid in interpretation of CE data and ultimately lead to increased confidence in the reported results.

Adverse Drug Events in Patients with Chronic Kidney Disease Associated with Multiple Drug Interactions and Polypharmacy.

BACKGROUND AND PURPOSE: Chronic kidney disease (CKD) is associated with adverse drug events due to medication errors and the risks of polypharmacy. The aim of this study was to investigate whether multiple pharmacodynamic interactions are a significant problem in CKD patients to improve medication safety. METHODS: The discharge medication of 200 elderly patients with stage 3, 4 and 5/5D CKD was analysed in a retrospective observational study with respect to kidney-related medication errors and multiple pharmacodynamic interactions. The clinical relevance of the most common and hazardous multiple interactions was assessed by evaluating adverse events at the primary or the subsequent hospital stay. RESULTS: Findings showed that 29.5% of the study cohort were at risk of QTc-interval prolongation in association with their medication combinations and half of them exhibited QTc-interval prolongation. The QTc interval was extended among all patients receiving a combination of two or more drugs with 'known' risk of Torsades de pointes. Amiodarone, citalopram and ciprofloxacin turned out to be the most hazardous drugs in this context. Eight percent of the patient population received a regimen of 4-6 potassium-enhancing drugs during their hospital stay, which was not de-escalated in 75.0% in the ambulatory setting. Despite close monitoring in the clinical setting, 37.5% of these patients developed hyperkalaemic episodes during their primary stay and 66.7% during rehospitalization. Of the study cohort, 8.5% received a combination of three drugs with antithrombotic or antiplatelet effects. Of these, 64.7% developed haemorrhagic events with two of them proving fatal. CONCLUSION: Multiple pharmacodynamic interactions related to QTc prolongation, hyperkalaemia and haemorrhage are frequently associated with a negative outcome in older adults with CKD and often require recurrent medical treatment or rehospitalization.

Don't Shut the Stable Door after the Phage Has Bolted-The Importance of Bacteriophage Inactivation in Food Environments.

In recent years, a new potential measure against foodborne pathogenic bacteria was rediscovered-bacteriophages. However, despite all their advantages, in connection to their widespread application in the food industry, negative consequences such as an uncontrolled phage spread as well as a development of phage resistant bacteria can occur. These problems are mostly a result of long-term persistence of phages in the food production environment. As this topic has been neglected so far, this article reviews the current knowledge regarding the effectiveness of disinfectant strategies for phage inactivation and removal. For this purpose, the main commercial phage products, as well as their application fields are first discussed in terms of applicable inactivation strategies and legal regulations. Secondly, an overview of the effectiveness of disinfectants for bacteriophage inactivation in general and commercial phages in particular is given. Finally, this review outlines a possible strategy for users of commercial phage products in order to improve the effectiveness of phage inactivation and removal after application.

Z-Transform Method for Pairwise Osteometric Pair-matching.

A new pairwise osteometric pair-matching approach based on the Z-transform method is presented. In contrast to previous methods that perform a global t-test on the summed skeletal element pair measurement distances, this approach performs t-tests on each individual distance, facilitating the capture of measurement-specific variation. This new approach is compared to published pairwise sorting methods using a standard reference dataset of postcranial remains maintained by the Defense POW/MIA Accounting Agency. Receiver operating characteristic curve analysis indicates significantly improved performance for the clavicle and radius over all previous methods (p < 0.01). The z-transform method weighted by the effect size outperformed the t-test (Byrd and Adams) and the mean t-test (Lynch) for all elements (p < 0.01). The method performed better than the absolute value t-test (Lynch) for five elements (p < 0.01) and performed at least as well for the remainder. To facilitate usability all methods are available at: https://github.com/spawaskar-cora/z-transform-method.

Virucidal or Not Virucidal? That Is the Question-Predictability of Ionic Liquid's Virucidal Potential in Biological Test Systems.

For three decades now, ionic liquids (ILs), organic salts comprising only ions, have emerged as a new class of pharmaceuticals. Although recognition of the antimicrobial effects of ILs is growing rapidly, there is almost nothing known about their possible virucidal activities. This probably reflects the paucity of understanding virus inactivation. In this study, we performed a systematic analysis to determine the effect of specific structural motifs of ILs on three different biological test systems (viruses, bacteria and enzymes). Overall, the effects of 27 different ILs on two non-enveloped and one enveloped virus (P100, MS2 and Phi6), two Gram negative and one Gram positive bacteria (E. coli, P. syringae and L. monocytogenes) and one enzyme (Taq DNA polymerase) were investigated. Results show that while some ILs were virucidal, no clear structure activity relationships (SARs) could be identified for the non-enveloped viruses P100 and MS2. However, for the first time, a correlation has been demonstrated between the effects of ILs on enveloped viruses, bacteria and enzyme inhibition. These identified SARs serve as a sound starting point for further studies.

Virucidal Influence of Ionic Liquids on Phages P100 and MS2.

An increasing number of publications describe the potential of ionic liquids (ILs) as novel antimicrobials, antibacterial coatings and even as active pharmaceutical ingredients. Nevertheless, a major research area, notably their impact on viruses, has so far been neglected. Consequently the aim of this study was to examine the effects of ILs on the infectivity of viruses. A systematic analysis to investigate the effects of defined structural elements of ILs on virus activity was performed using 55 ILs. All structure activity relationships (SARs) were tested on the human norovirus surrogate phage MS2 and phage P100 representing non-enveloped DNA viruses. Results demonstrate that IL SAR conclusions, established for prokaryotes and eukaryotes, are not readily applicable to the examined phages. A virus-type-dependent IL influence was also apparent. Overall, four ILs, covering different structural elements, were found to reduce phage P100 infectivity by ≥4 log10 units, indicating a virucidal effect, whereas the highest reduction for phage MS2 was about 3 log10 units. Results indicate that future applications of ILs as virucidal agents will require development of novel SARs and the obtained results serve as a good starting point for future studies.

Graph mining for next generation sequencing: leveraging the assembly graph for biological insights.

BACKGROUND: The assembly of Next Generation Sequencing (NGS) reads remains a challenging task. This is especially true for the assembly of metagenomics data that originate from environmental samples potentially containing hundreds to thousands of unique species. The principle objective of current assembly tools is to assemble NGS reads into contiguous stretches of sequence called contigs while maximizing for both accuracy and contig length. The end goal of this process is to produce longer contigs with the major focus being on assembly only. Sequence read assembly is an aggregative process, during which read overlap relationship information is lost as reads are merged into longer sequences or contigs. The assembly graph is information rich and capable of capturing the genomic architecture of an input read data set. We have developed a novel hybrid graph in which nodes represent sequence regions at different levels of granularity. This model, utilized in the assembly and analysis pipeline Focus, presents a concise yet feature rich view of a given input data set, allowing for the extraction of biologically relevant graph structures for graph mining purposes. RESULTS: Focus was used to create hybrid graphs to model metagenomics data sets obtained from the gut microbiomes of five individuals with Crohn's disease and eight healthy individuals. Repetitive and mobile genetic elements are found to be associated with hybrid graph structure. Using graph mining techniques, a comparative study of the Crohn's disease and healthy data sets was conducted with focus on antibiotics resistance genes associated with transposase genes. Results demonstrated significant differences in the phylogenetic distribution of categories of antibiotics resistance genes in the healthy and diseased patients. Focus was also evaluated as a pure assembly tool and produced excellent results when compared against the Meta-velvet, Omega, and UD-IDBA assemblers. CONCLUSIONS: Mining the hybrid graph can reveal biological phenomena captured by its structure. We demonstrate the advantages of considering assembly graphs as data-mining support in addition to their role as frameworks for assembly.

[Extracardiac manifestation of elevated lipoprotein(a) levels--cumulative incidence of peripheral arterial disease and stenosis of the carotid artery]. / Extrakardiale Manifestation der Lipoprotein(a)-Erhöhung--Gehäuftes Auftreten von peripherer arterieller Verschlusskrankheit und Stenosen der Arteria carotis.

BACKGROUND: Elevated lipoprotein(a) (Lp(a)) levels are an accepted risk factor for coronary heart disease. The role of Lp(a) in the development of extracardiac arteriosclerosis like peripheral arterial disease (PAD) and stenosis of the arteria carotis (ACIS) has hardly been documented so far. We aimed to investigate the incidence of extracardiac arteriosclerosis in individuals with elevated Lp(a) values. METHODIK: In our center, we measured Lp(a) levels in 31,734 consecutive patients over 5 years. Of these, 1411 patients were selected retrospectively for the presented analysis. Patients were matched according to age, sex, and other accepted cardiovascular risk factors and were assigned to 6 groups according to their Lp(a) values. Retrospectively, we analysed the incidence of PAD and ACIS. RESULTS: In the group with Lp(a) values < 2 mg/dl the incidence of PAD was 1.9 % (ACIS 2.8 %), in the group with Lp(a) 23-29 mg/dl 7.3 % (6.1 %), 30-60 mg/dl 9.0 % (8.3 %), 60-91 mg/dl 11.4 % (7.9 %), 91-110 mg/dl 8.6 % (6.0 %) and > 110 mg/dl 12.7 % (10.9 %). None of the patients had LDL levels > 130 mg/dl or HbA1c 6.1 %. CONCLUSION: Elevated Lp(a) levels seem to be associated with an increased incidence of PAD and ACIS. Even Lp(a) concentrations between 23 and 29 mg/dl show a threefold increased risk of PAD when compared to patients with Lp(a) < 2 mg/dl. However, these findings have to be verified in large prospective studies. In this context cut-off values have to be reevaluated as well.

Spatially regularized estimation for the analysis of dynamic contrast-enhanced magnetic resonance imaging data.

Competing compartment models of different complexities have been used for the quantitative analysis of dynamic contrast-enhanced magnetic resonance imaging data. We present a spatial elastic net approach that allows to estimate the number of compartments for each voxel such that the model complexity is not fixed a priori. A multi-compartment approach is considered, which is translated into a restricted least square model selection problem. This is done by using a set of basis functions for a given set of candidate rate constants. The form of the basis functions is derived from a kinetic model and thus describes the contribution of a specific compartment. Using a spatial elastic net estimator, we chose a sparse set of basis functions per voxel, and hence, rate constants of compartments. The spatial penalty takes into account the voxel structure of an image and performs better than a penalty treating voxels independently. The proposed estimation method is evaluated for simulated images and applied to an in vivo dataset.

mSYD1A, a mammalian synapse-defective-1 protein, regulates synaptogenic signaling and vesicle docking.

Structure and function of presynaptic terminals are critical for the transmission and processing of neuronal signals. Trans-synaptic signaling systems instruct the differentiation and function of presynaptic release sites, but their downstream mediators are only beginning to be understood. Here, we identify the intracellular mSYD1A (mouse Synapse-Defective-1A) as a regulator of presynaptic function in mice. mSYD1A forms a complex with presynaptic receptor tyrosine phosphatases and controls tethering of synaptic vesicles at synapses. mSYD1A function relies on an intrinsically disordered domain that interacts with multiple structurally unrelated binding partners, including the active zone protein liprin-α2 and nsec1/munc18-1. In mSYD1A knockout mice, synapses assemble in normal numbers but there is a significant reduction in synaptic vesicle docking at the active zone and an impairment of synaptic transmission. Thus, mSYD1A is a regulator of presynaptic release sites at central synapses.

Influence of surgical approach on postoperative femoral bone remodelling after cementless total hip arthroplasty.

We retrospectively evaluated the femoral periprosthetic bone mineral density (BMD) in a consecutive series of patients who had undergone total hip arthroplasty (THA) with a straight, double tapered cementless stem using a muscle-sparing anterolateral (group A) and the transgluteal (group B) surgical approach. Dual-energy x-ray absorptiometry (DXA) measurements were performed in the first postoperative week (t1), and after 3 (t2), 6 (t3) and 12 months (t4) using an identical protocol. Patients were clinically and radiographically evaluated at final follow-up (t4). A complete set of four consecutive DXA measurements was obtained for 16 hips in group A and 26 hips in group B. In patients in whom the transgluteal approach was used (Group B), we observed a significantly greater decline in overall periprosthetic BMD (netavg) and in BMD in the periprosthetic regions of interest (ROI) 1, 4, 5 and 6 between t1 and t4. At clinical and radiographic evaluation at t4, no differences between the groups were detected. Femoral periprosthetic BMD is affected by the selected surgical approach in the first postoperative year. This might be attributed to altered femoral loading as a result of differences in intraoperative damage to the abductor muscles.

Protooncogene Ski cooperates with the chromatin-remodeling factor Satb2 in specifying callosal neurons.

First insights into the molecular programs orchestrating the progression from neural stem cells to cortical projection neurons are emerging. Loss of the transcriptional regulator Ski has been linked to the human 1p36 deletion syndrome, which includes central nervous system defects. Here, we report critical roles for Ski in the maintenance of the neural stem cell pool and the specification of callosal neurons. Ski-deficient callosal neurons lose their identity and ectopically express the transcription factor Ctip2. The misspecified callosal neurons largely fail to form the corpus callosum and instead redirect their axons toward subcortical targets. We identify the chromatin-remodeling factor Satb2 as a partner of Ski, and show that both proteins are required for transcriptional repression of Ctip2 in callosal neurons. We propose a model in which Satb2 recruits Ski to the Ctip2 locus, and Ski attracts histone deacetylases, thereby enabling the formation of a functional nucleosome remodeling and deacetylase repressor complex. Our findings establish a central role for Ski-Satb2 interactions in regulating transcriptional mechanisms of callosal neuron specification.

Predicting volumes of metabolically important whole-body adipose tissue compartments in overweight and obese adolescents by different MRI approaches and anthropometry.

OBJECTIVES: To evaluate 5-slice stack/single-slice MRI approaches and anthropometric measures as predictors for metabolically relevant whole-body adipose tissue (AT) compartments in overweight/obese adolescents. METHODS: Forty adolescents (22 males, age 11.4-16.1 years) were included with a BMI above the 90th percentile. Volumes of whole-body AT compartments, i.e. total AT (TAT), subcutaneous AT (SCAT) and visceral AT (VAT), were determined using a breath-hold T1-weighted-FSE-MR-sequence and semi-automated segmentation serving as the gold standard. SCAT, VAT and TAT was estimated by either axially oriented single-slices or 5-slice-stacks centred at specific anatomic landmarks (umbilicus, head of femur and humerus). Furthermore, anthropometric measures were also evaluated as predictors of whole-body AT compartments. RESULTS: Strong correlations were found for both genders between TAT/SCAT and single-slice evaluation (e.g. whole-body SCAT-SCAT at umbilicus level: r = 0.91 (m), r = 0.92 (f)) or anthropometry (SCAT-BMI: r = 0.93 (m, f)). VAT was correlated to VAT at umbilicus (r = 0.71 (m), r = 0.94 (f)) but only weakly to anthropometry. CONCLUSIONS: Anthropometric measures and single-slice MRI can accurately predict TAT/SCAT which cannot be improved by evaluation of 5-slice stacks. Prediction of VAT by 5-slice stack/single-slice MRI protocols seems only to be accurate in females. Anthropometry cannot be reliably used for prediction of VAT in both genders. Thus, MRI seems to be necessary for quantification of VAT in overweight/obese adolescents of both genders.