NMR-based metabolomic signature: An important tool for the diagnosis and study of pathogenesis of autoimmune hepatitis.

BACKGROUND AND AIMS: Metabolomics is used to predict, diagnose, and monitor metabolic disorders but altered metabolomic signatures have also been reported in diverse diseases, including autoimmune disorders. However, the metabolomic profile in autoimmune hepatitis (AIH) has not been investigated in depth. Therefore, we investigated the metabolomic signature of AIH and its significance as a diagnostic and pathogenetic tool. APPROACH AND RESULTS: Metabolites in plasma samples from 50 patients with AIH at diagnosis, 43 healthy controls, 72 patients with primary biliary cholangitis (PBC), 26 patients with metabolic dysfunction-associated liver disease, and 101 patients with chronic viral hepatitis were determined by 1 H NMR (nuclear magnetic resonance) spectroscopy. Fifty-two metabolites were quantified, and metabolic pathway analysis was performed. Multivariate analysis revealed that AIH could be differentiated from healthy controls and each of the disease controls ( p <0.001). Fifteen metabolites differentiated AIH from disease controls (PBC+chronic viral hepatitis+metabolic dysfunction-associated liver disease) (95% sensitivity and 92% specificity). Ten distinct metabolic pathways were altered in AIH compared to disease controls. The metabolic pathway of branched-chain amino acids (lower valine, leucine, and isoleucine levels and their catabolic intermediates in PBC), methionine (lower methionine, 2-aminobutyrate, and 2-hydroxybutyrate levels in PBC), alanine-aspartate-glutamate (lower metabolites in PBC), and that of metabolites associated with gut microbiota (lower choline, betaine, and dimethylamine levels in PBC) were significantly different between AIH and PBC ( p <0.01). CONCLUSIONS: 1 H NMR spectroscopy could be a promising novel tool to diagnose and study AIH pathogenesis as there is no need for much sample handling, is highly reproducible with high sensitivity and specificity, and low cost.

Craniofacial Development Is Fine-Tuned by Sox2.

The precise control of neural crest stem cell delamination, migration and differentiation ensures proper craniofacial and head development. Sox2 shapes the ontogeny of the cranial neural crest to ensure precision of the cell flow in the developing head. Here, we review how Sox2 orchestrates signals that control these complex developmental processes.

A Study of the Metabolic Pathways Affected by Gestational Diabetes Mellitus: Comparison with Type 2 Diabetes.

BACKGROUND: Gestational diabetes mellitus (GDM) remains incompletely understood and increases the risk of developing Diabetes mellitus type 2 (DM2). Metabolomics provides insights etiology and pathogenesis of disease and discovery biomarkers for accurate detection. Nuclear magnetic resonance (NMR) spectroscopy is a key platform defining metabolic signatures in intact serum/plasma. In the present study, we used NMR-based analysis of macromolecules free-serum to accurately characterize the altered metabolic pathways of GDM and assessing their similarities to DM2. Our findings could contribute to the understanding of the pathophysiology of GDM and help in the identification of metabolomic markers of the disease. METHODS: Sixty-two women with GDM matched with seventy-seven women without GDM (control group). 1H NMR serum spectra were acquired on an 11.7 T Bruker Avance DRX NMR spectrometer. RESULTS: We identified 55 metabolites in both groups, 25 of which were significantly altered in the GDM group. GDM group showed elevated levels of ketone bodies, 2-hydroxybutyrate and of some metabolic intermediates of branched-chain amino acids (BCAAs) and significantly lower levels of metabolites of one-carbon metabolism, energy production, purine metabolism, certain amino acids, 3-methyl-2-oxovalerate, ornithine, 2-aminobutyrate, taurine and trimethylamine N-oxide. CONCLUSION: Metabolic pathways affected in GDM were beta-oxidation, ketone bodies metabolism, one-carbon metabolism, arginine and ornithine metabolism likewise in DM2, whereas BCAAs catabolism and aromatic amino acids metabolism were affected, but otherwise than in DM2.

The Critical Role of the Branched Chain Amino Acids (BCAAs) Catabolism-Regulating Enzymes, Branched-Chain Aminotransferase (BCAT) and Branched-Chain α-Keto Acid Dehydrogenase (BCKD), in Human Pathophysiology.

Branched chain amino acids (BCAAs), leucine, isoleucine and valine, are essential amino acids widely studied for their crucial role in the regulation of protein synthesis mainly through the activation of the mTOR signaling pathway and their emerging recognition as players in the regulation of various physiological and metabolic processes, such as glucose homeostasis. BCAA supplementation is primarily used as a beneficial nutritional intervention in chronic liver and kidney disease as well as in muscle wasting disorders. However, downregulated/upregulated plasma BCAAs and their defective catabolism in various tissues, mainly due to altered enzymatic activity of the first two enzymes in their catabolic pathway, BCAA aminotransferase (BCAT) and branched-chain α-keto acid dehydrogenase (BCKD), have been investigated in many nutritional and disease states. The current review focused on the underlying mechanisms of altered BCAA catabolism and its contribution to the pathogenesis of a numerous pathological conditions such as diabetes, heart failure and cancer. In addition, we summarize findings that indicate that the recovery of the dysregulated BCAA catabolism may be associated with an improved outcome and the prevention of serious disease complications.

Optimization of psoriasis mouse models.

INTRODUCTION: Psoriasis, is a common, chronic, autoimmune, inflammatory, relapsing disease, which would benefit from reliable and human-relevant animal models to test drugs pre-clinically and to understand their mechanism of action. Because of its ease of use, convenience and low cost, the imiquimod (IMQ)-induced psoriasis-like model is widely utilized; however, it is not known whether all mouse strains are equivalent and if the hairless mouse is appropriate, so that the imiquimod model can be further optimized. METHODS: Under similar experimental conditions, common mouse strains (BALB/c, C57BL/6J, and ApoE) and a new hairless strain (ApoE/SKH-hr2) as well as several inducers (IMQ, IMQ + acetic acid (AcOH) topical and IMQ + AcOH systemic) were compared by clinical, histopathological, biophysical and locomotor activity assessments. RESULTS AND DISCUSSION: The BALB/c mice yielded an optimal psoriasis-like phenotype with IMQ + AcOH topical treatment, and the corresponding phenotypes for the other mouse strains were C57BL/6J moderate and ApoE mild. In contrast, the ApoE/SKH-hr2 mice, as a result of the absence of a Munro abscess in the histopathology analysis, left doubt about the psoriasis-like acquisition. Locomotor activity of BALB/c mice treated with IMQ, IMQ + AcOH topically and IMQ + AcOH systemically showed decreased distance and rearing coverage and increased immobility with all treatments. Hence, the BALB/c mouse strain appears to be an optimal psoriasis-like model when utilizing IMQ + AcOH topical application.

Effect of Dapagliflozin on Urine Metabolome in Patients with Type 2 Diabetes.

CONTEXT: Inhibitors of sodium-glucose cotransporters-2 have cardio- and renoprotective properties. However, the underlying mechanisms remain indeterminate. OBJECTIVE: To evaluate the effect of dapagliflozin on renal metabolism assessed by urine metabolome analysis in patients with type 2 diabetes. DESIGN: Prospective cohort study. SETTING: Outpatient diabetes clinic of a tertiary academic center. PATIENTS: Eighty patients with hemoglobin A1c > 7% on metformin monotherapy were prospectively enrolled. INTERVENTION: Fifty patients were treated with dapagliflozin for 3 months. To exclude that the changes observed in urine metabolome were merely the result of the improvement in glycemia, 30 patients treated with insulin degludec were used for comparison. MAIN OUTCOME MEASURE: Changes in urine metabolic profile before and after the administration of dapagliflozin and insulin degludec were assessed by proton-nuclear magnetic resonance spectroscopy. RESULTS: In multivariate analysis urine metabolome was significantly altered by dapagliflozin (R2X = 0.819, R2Y = 0.627, Q2Y = 0.362, and coefficient of variation analysis of variance, P < 0.001) but not insulin. After dapagliflozin, the urine concentrations of ketone bodies, lactate, branched chain amino acids (P < 0.001), betaine, myo-inositol (P < 0001), and N-methylhydantoin (P < 0.005) were significantly increased. Additionally, the urine levels of alanine, creatine, sarcosine, and citrate were also increased (P < 0001, P <0.0001, and P <0.0005, respectively) whereas anserine decreased (P < 0005). CONCLUSIONS: Dapagliflozin significantly affects urine metabolome in patients with type 2 diabetes in a glucose lowering-independent way. Most of the observed changes can be considered beneficial and may contribute to the renoprotective properties of dapagliflozin.

Modeling diffuse reflectance from homogeneous semi-infinite turbid media for biological tissue applications: a Monte Carlo study.

Diffuse reflectance spectroscopy is one of the simplest and widely used techniques for the non-invasive study of biological tissues but no exact analytical solution exists for the problem of diffuse reflectance from turbid media such as biological tissues. In this work, a general treatment of the problem of diffuse reflectance from a homogeneous semi-infinite turbid medium is presented using Monte Carlo simulations. Based on the results of the Monte Carlo method, simple semi-empirical analytical solutions are developed valid for a wide range of collection geometries corresponding to various optical detector diameters. This approach may be useful for the quick and accurate modeling of diffuse reflectance from tissues.

Simple two-layer reflectance model for biological tissue applications: lower absorbing layer.

A simple two-layer tissue reflectance model is described. This work is a continuation of our investigations on modeling reflectance from two-layered tissues that we recently initiated. In the present article, we describe a variation of a two-layer model that assumes a lower absorbing and scattering layer and an upper scattering-only layer. This two-layer configuration is a realistic model for biological tissues in the visible and near-IR spectral ranges, where the upper layer may be an epithelial layer and the lower layer is a vascularized stroma layer. Application of the model yields estimates for tissue parameters, such as the thickness of the upper layer or the absorption properties of the lower layer. These parameters are of great interest for the noninvasive study of a wide range of epithelial biological tissues. The validity range and accuracy of the model are tested on tissue phantoms in both the forward and inverse modes of application.

In vivo optical properties of melanocytic skin lesions: common nevi, dysplastic nevi and malignant melanoma.

We present an in vivo study of the optical properties of common nevi, dysplastic nevi and malignant melanoma skin lesions in human subjects. Reflectance spectra were measured on 1379 skin lesions, in the visible and near-infrared spectral regions, using a spectral imaging system, in a clinical setting. Analysis of the data using a reflectance model revealed differences between the optical properties of melanin present in nevi and melanoma lesions. These differences, which are in agreement with our previous observations on average reflectance spectra, may be potentially useful for the noninvasive characterization of pigmented skin lesions and the early diagnosis of melanoma.

Light scattering spectroscopy of human skin in vivo.

We present an in vivo study of the reduced scattering coefficient of normal skin and of common melanocytic nevi in Caucasian subjects. The spectral shape of the reduced scattering coefficient is described well by a power-law dependence on the wavelength, in accordance with previous studies of light scattering by biological tissues. We investigate statistical variations in the scattering spectrum slope and also identify an inherent correlation between scattering intensity and scattering spectral slope, observed mainly in normal skin. In addition, we do not find any significant differences between the scattering properties of normal skin and common melanocytic nevi. Finally, we also provide a short review of previously published studies reporting on the light scattering properties of human skin both in vivo and in vitro.

Optical properties of human melanocytic nevi in vivo.

We present an in vivo study of the optical properties of melanin present in melanocytic nevi of human subjects with Fitzpatrick skin type III (Caucasian descent) using optical spectroscopy. We show that the melanin absorption spectrum exhibits an exponential dependence on wavelength with a decay constant which follows a normal distribution characteristic of a random biological variable. Moreover, we demonstrate lack of correlation among melanin optical properties, melanin concentration and skin light scattering properties, which indicates that the true optical absorption of melanin can be measured free from confounding scattering effects. We also show that the average melanin absorption spectrum in vivo is in very good agreement with a previously reported oxygen photoconsumption action spectrum of melanin. Finally, we provide an overview of the emerging picture of the melanin absorption properties in vivo among various skin types and also among various skin lesions such as melanocytic nevi and melanoma.

Comparative evaluation of two simple diffuse reflectance models for biological tissue applications.

We present a comparative evaluation of two simple diffuse reflectance models for biological tissue applications. One model is based on a widely accepted and used in biomedical optics implementation of diffusion theory, and the other one is based on a semiempirical approach derived from basic physical principles. We test the models on tissue phantoms and on human skin, utilizing a standard six-around-one optical fiber probe for light delivery and collection. We show that both models are suitable for use with an optical fiber probe and illustrate the potential, applicability, and validity range of the models.

Melanin optical properties provide evidence for chemical and structural disorder in vivo.

Melanin is a ubiquitous chromophore of human skin but its in vivo optical properties are relatively unexplored. We present here a detailed study of the optical absorption of melanin present in melanocytic nevi of human subjects with Fitzpatrick skin type III. Using diffuse reflectance spectroscopy, we show that the melanin absorption spectrum exhibits an exponential dependence on wavelength in vivo with a decay constant that follows a normal distribution, characteristic of a random biological variable. This is the first time such direct in vivo quantitative evidence is obtained supporting the recently proposed hypothesis of chemical and structural disorder for melanin. In addition, the ability to measure the melanin optical properties in vivo opens new ways for the study of melanin in its native environment as well as for the non-invasive study and characterization of various skin disorders and diseases.

Melanin absorption spectroscopy: new method for noninvasive skin investigation and melanoma detection.

We present a new method for studying melanin in vivo based on diffuse reflectance spectroscopy of human skin. We find that the optical absorption spectrum of in vivo melanin exhibits an exponential dependence on wavelength, consistent with, but with a higher decay slope than, in vitro results. We offer theoretical justification for this exponential dependence on the basis of a recently proposed model for the structure of eumelanin protomolecules. Moreover, we report on a new method for analysis of diffuse reflectance spectra, which identifies intrinsic differences in absorption spectra between malignant melanoma and dysplastic nevi in vivo. These preliminary results are confirmed both by analysis of our own clinical data as well as by analysis of data from three independent, previously published studies. In particular, we find evidence that the histologic transition from dysplastic nevi to melanoma in situ and then to malignant melanoma is reflected in the melanin absorption spectra. Our results are very promising for the development of techniques for the noninvasive detection of melanoma and, more generally, for the study and characterization of pigmented skin lesions. It is also a promising approach for a better understanding of the biological role, structure, and function of melanin.

Probing skin interaction with hydrogen peroxide using diffuse reflectance spectroscopy.

Hydrogen peroxide is an important oxidizing agent in biological systems. In dermatology, it is frequently used as topical antiseptic, it has a haemostatic function, it can cause skin blanching, and it can facilitate skin tanning. In this work, we investigated skin interaction with hydrogen peroxide, non-invasively, using diffuse reflectance spectroscopy. We observed transient changes in the oxyhaemoglobin and deoxyhaemoglobin concentrations as a result of topical application of dilute H(2)O(2) solutions to the skin, with changes in deoxyhaemoglobin concentration being more pronounced. Furthermore, we did not observe any appreciable changes in melanin absorption properties as well as in the skin scattering properties. We also found no evidence for production of oxidized haemoglobin forms. Our observations are consistent with an at least partial decomposition of hydrogen peroxide within the stratum corneum and epidermis, with the resulting oxygen and/or remaining hydrogen peroxide inducing vasoconstriction to dermal blood vessels and increasing haemoglobin oxygen saturation. An assessment of the effects of topical application of hydrogen peroxide to the skin may serve as the basis for the development of non-invasive techniques to measure skin antioxidant capacity and also may shed light onto skin related disorders such as vitiligo.

Modeling diffuse reflectance from semi-infinite turbid media: application to the study of skin optical properties.

Diffuse reflectance spectroscopy in the visible and NIR spectral ranges is an effective and extensively used technique for the non-invasive study and characterization of various biological tissues. In this article, a short review of currently available modeling techniques for diffuse reflectance from semi-infinite turbid media is presented. Starting from the basic physical picture of the diffuse reflectance problem, a simple and practical model is then proposed for use with fiber optic probes. The validity of the model is tested on tissue phantoms and it is then applied to the analysis of diffuse reflectance spectra collected from human skin in vivo.

Metolachlor photodegradation study in aqueous media under natural and simulated solar irradiation.

To elucidate the photochemical behavior of pesticide metolachlor, degradation was carried out in aqueous media of different compositions such as sea, river, lake, and distilled water under natural and simulated solar irradiation. In addition, the effect of important constituents of natural water such as dissolved organic matter (DOM, isolated from Pamvotis Lake) and nitrate ions was also examined. It was found that photodegradation proceeds via a pseudo-first-order reaction in all cases. The presence of DOM inhibits the photolysis reaction with half-lives ranging from 87 to 693 h whereas the degradation rate was accelerated up to 11 times in the presence of NO(3)(-). In addition, the toxicity of the degradation products formed (generally through hydroxylation, dealkylation, and cyclization reactions) was also performed using the marine luminescent bacterium Vibrio fisheri. Our results indicated a toxicity increase of the irradiated solution showing that photoproducts of higher acute toxic effects were formed.