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Here, we describe a method for obtaining a dynamic nuclear polarization (DNP)-enhanced double-quantum filtered (DQF) two-dimensional (2D) dipolar (13)C-(13)C correlation spectra of bone-tissue material in natural (13)C abundance. DNP-enhanced DQF 2D dipolar (13)C-(13)C spectra were obtained using a few different mixing times of the dipolar-assisted rotational resonance (DARR) scheme and these spectra were compared to a conventional 2D through-space double-quantum (DQ)-single-quantum (SQ) correlation spectrum. While this scheme scheme can only be used for an assigning purpose to reveal the carbon-carbon connectivity within a residue, the DQF (13)C-(13)C dipolar correlation scheme we introduce here can be used to obtain longer distance carbon-carbon constraints. A DQF pulse block is placed before the DARR mixing scheme for removing dominant (13)C single-quantum (SQ) signals because these SQ (13)C signals are overwhelmingly large compared to those (13)C-(13)C dipolar cross-peaks generated and therefore saturate the dynamic range of the NMR detection. This approach exhibits strong enough 2D cross-peaks in dipolar (13)C-(13)C correlation spectroscopy and provides a potential of obtaining pairwise (13)C-(13)C dipolar constraints because the dipolar truncation effect as well as multi-step signal propagations involving a spin cluster that contains more than two spins can be ignored probabilistically. For obtaining a fast signal averaging, AsymPolPOK polarizing agent was used to provide a short (1)H DNP signal build-up time (1.3 s) and to expedite our MAS DNP NMR acquisitions while still maintaining a satisfactory DNP enhancement factor (ε = 50). Under employing a long DARR mixing time, a t(1)-noise-like artifact was observed at a site that possesses a large chemical shift anisotropy (CSA) and a few different strategies to address this problem were discussed.

Complications of the bronchial anastomosis in lung transplantation, once the Achilles heel of the procedure, have become quite rare. The surgical technique is well established and safe. Risks contributing to anastomotic complications are primarily related to patients pre-existing conditions. The key factor is good blood flow to the bronchial stump. Postoperative infection can also contribute to the breakdown of the anastomosis. This may be the reason why different immunosuppressive regimes lead to differences in the incidence of bronchial dehiscence.

Background: Historically, higher levels of serum testosterone were presumed deleterious to the cardiovascular system. cardiovascular disease (CVD) risk factors prevalence were increased due to low testosterone levels. Material and Methods: This was a prospective observational study, conducted among men with type 2 diabetes with documented CAD (n = 60) and controls healthy men (n = 50). Pearson correlation between sex hormones and CAD risk factors was done. The association between sex hormones and CAD risk factors among patients was analyzed by multiple logistic regression. The statistical significance was set at the 0.05 level. Result: Mean age of diabetics was 44.5 ±7.9 years and non diabetics was 42.54 ± 5.63 years. 56% of Diabetics and 20 % non diabetics had lower levels of total testosterone. 68% of Diabetics and 26 % non diabetics had lower levels of free testosterone. Mean SHBG levels in Diabetics and non - diabetics was 8.6 ± 1.8 and 16 ± 3.2. Prevalence of Hypogonadism was 22% in Diabetics and 10% in Non diabetics. Among the cases, a significant positive correlation was found between total testosterone/FTI and waist circumference, W/H ratio, triglyceride levels, hs-CRP, and CIMT (P < 0.01). In addition, a significant negative correlation was found between total testosterone and FTI with high-density lipoprotein-cholesterol levels (P < 0.01). The multiple logistic regression analysis showed that total testosterone levels (P < 0.01) and SHBG (P < 0.01) are independently associated with the occurrence of atherosclerotic CAD in T2DM men. Conclusion: We conclude that increased serum testosterone levels and low SHBG in T2DM men are associated with the development of atherosclerotic cardiovascular risk factors. Disclosure A. K. Das: None. A. Bhagat: None. N. Sinha: None.

Structural stability of various collagen containing biomaterials such as bones and cartilage is still a mystery. Despite spectroscopic development of several decades, the detailed mechanism of collagen interaction with citrate in bones and glycosaminoglycans (GAGS) in cartilage extracellular matrix (ECM) in its native state is unobservable. We present a significant advancement to probe the collagen interactions with citrate and GAGSin ECM of native bone and cartilage along with specific/non-specific interactions inside collagen assembly at the nanoscopic level through natural abundance dynamic nuclear polarization (DNP) based solid-state nuclear magnetic resonance (ssNMR) spectroscopy. The detected molecular level interactions between citrate – collagen and glycosaminoglycan (GAGs) - collagen inside the native bone and cartilage matrix and other backbone and side-chain interactions in the collagen assembly are responsible for the structural stability and other biomechanical properties of these important class of biomaterial.

Background Visualizing and quantifying cellular heterogeneity is of central importance to study tissue complexity, development, and physiology and has a vital role in understanding pathologies. Mass spectrometry-based methods including imaging mass cytometry (IMC) have in recent years emerged as powerful approaches for assessing cellular heterogeneity in tissues. IMC is an innovative multiplex imaging method that combines imaging using up to 40 metal conjugated antibodies and provides distributions of protein markers in tissues with a resolution of 1 μm2 area. However, resolving the output signals of individual cells within the tissue sample, i.e., single cell segmentation, remains challenging. To address this problem, we developed MATISSE (iMaging mAss cyTometry mIcroscopy Single cell SegmEntation), a method that combines high-resolution fluorescence microscopy with the multiplex capability of IMC into a single workflow to achieve improved segmentation over the current state-of-the-art. Results MATISSE results in improved quality and quantity of segmented cells when compared to IMC-only segmentation in sections of heterogeneous tissues. Additionally, MATISSE enables more complete and accurate identification of epithelial cells, fibroblasts, and infiltrating immune cells in densely packed cellular areas in tissue sections. MATISSE has been designed based on commonly used open-access tools and regular fluorescence microscopy, allowing easy implementation by labs using multiplex IMC into their analysis methods. Conclusion MATISSE allows segmentation of densely packed cellular areas and provides a qualitative and quantitative improvement when compared to IMC-based segmentation. We expect that implementing MATISSE into tissue section analysis pipelines will yield improved cell segmentation and enable more accurate analysis of the tissue microenvironment in epithelial tissue pathologies, such as autoimmunity and cancer.

The Barmer and Cambay basins (India) formed as rifts during the Cretaceous to Paleocene, prior to Seychelles–India separation during Late Cretaceous. Both of the rifts failed subsequently and the Seychelles separated from India along the current Saurashtra–Mumbai continental margin. We study the fault kinematics, i.e., fault growth and linkages in the Cambay–Tarapur Block and comment on the fault evolution from syn- to post-rift stages. We use fault plane topography, throw-length and throw-depth plots to understand the kinematics of the fault planes. Throw-length plots show the along-strike linkage of faults. The throw-depth plots are used to understand the down and up dip linkage of the fault planes. Faults originating at the Deccan Traps level and faults confined only to post-rift progressed as single fault planes along-strike throughout their deformation history. Faults initiating within the syn-rift started as multiple along-strike segments and joined together as they grew. Along dip, faults can be classified as (i) faults originating at Deccan Traps and reaching post-rift, (ii) faults originating within syn-rift sediments and reaching post-rift, (iii) faults confined only within syn-rift and (iv) faults confined only within post rift. Some of the syn-rift faults also show spatially and temporally limited compression, indicating their possible oblique-slip nature. The faults were normal-slip or oblique-slip during syn-rift, and thereafter they propagated as normal dip-slip faults, as analysed from fault plane topography showing strike perpendicular ridges. Such ridges may indicate only the last movement on the fault plane. Due to the presence of thick shale horizons in the syn-rift, many faults show soft-linked segments along dip. Hydrocarbon occurrences in the shallow (1200–1600 m) Miocene sands can possibly be attributed to the normal faults controlling fluid flow into the reservoirs. (282 words).

Post-traumatic epilepsy (PTE) is a common long-term risk associated with traumatic brain injury (TBI). PTE rat model, proposed by Willmore et al., is a well known model that mimics human PTE. The present study explored the lipid metabolism in this PTE rat model by using in vitro, high-resolution NMR (nuclear magnetic resonance) spectroscopy and lipid staining based investigations. The level of gene expression, cytokines and enzyme activity was estimated. Level of TG (triglycerides), PL (phospholipids) and CHOL (cholesterol) was found to increase in brain tissue of PTE rats. This is an indication of the altered lipid metabolism in PTE rats. Level of lipid peroxidation and cytokines was enhanced in the brain tissue of PTE rats. A positive correlation was also observed in cytokines vs. lipid peroxidation. These results make available the evidence of the oxidative stress induced damage or destruction of the lipid components and also the cause of the inflammatory events in PTE rats. Antioxidant enzyme activity and respective gene expression were found to increase in brain tissue of PTE rats. A positive correlation was also observed in antioxidant enzyme's activity vs. respective enzyme gene expression and lipid peroxidation vs. activity of antioxidant enzymes. Such outcomes reflect the oxidative stress induced lipid damage responsible for production enhancement of antioxidant enzymes, which further responsible for enhancing the activity of antioxidant enzymes. A positive correlation was observed in lipid peroxidation vs. lipid components (TG, PL and CHOL) and provides the confirmatory verification of alteration in the level of lipid components. A negative correlation was observed in the level of cytokines and the quantity of TG. This showed that TG is consumed in the production of cytokines. MUA (Motor unit activity) is highly correlated with the level of LP and indicated that oxidative stress is responsible for the event of epileptogenesis. Positive correlation of MUA with RA (rearing activity) and MWM (Morris-water maze) showed that epileptogenesis also influences the memory of PTE rats. Overall results based analyses clearly indicate that the inflammatory activity and oxidative stress in brain tissue of PTE rats, which are responsible to establish a significant change in the lipid metabolism. This can be visualized through a well constructed possible pathway of altered lipid metabolism. This study will improve our understanding and approach in the field of epilepsy that need to be considered for the development of new drugs or therapy for patients with PTE. Representation of the proposed pathway of altered lipid metabolism in posttraumatic epileptic rats.

Breast cancer (BC) is one of the most common malignancies among women worldwide, which is indeed associated with metabolic reprogramming. However, BC is a very complex and heterogeneous disease, which can relate with the changes in metabolic profiles during BC progression. Hence, investigating the metabolic alterations during BC stage progression may reveal the deregulated pathways and useful metabolic signatures of BC. To demonstrate the metabolic insights, we opted 1H NMR spectroscopy based metabolomics of blood plasma of early and late stage BC (N = 72) with age and gender matched healthy subjects (N = 50). Further, the metabolic profiles were analyzed to delineate the potential signatures of BC by performing multivariate and nonparametric statistical analysis in early and late stages of BC in comparison with healthy subjects. Sixteen metabolites levels were differentially changed (p

Gallbladder cancer (GBC) is an aggressive malignancy of gastrointestinal tract. Due to uncontrolled growth, GBC cells rapidly synthesize biomolecules including lipids. The lipids are integral component of cell membrane with a wide range of cellular functions. In this study, we measured the clinicopathological features in 40 cases of histologically confirmed GBC and 16 cases of chronic cholecystitis (CC). The female to male ratio in the GBC and CC groups were 3.44:1 and 2.2:1, respectively. The GBC patients exhibited well to poorly differentiated tumor. In the CC group, all patients showed cholecystitis with no evidence of dysplasia or malignancy. The majority of GBC and CC patients reported pain. Using

Recent advances in technology have led to significantly greater use of extracorporeal membrane oxygenation (ECMO) as a bridge to lung transplantation with better outcomes. The novel ProtekDuo veno-venous ECMO (CardiacAssist, Inc.) has gained significance as it facilitates effective decompression of the right heart in patients with acute decompensation, while also providing consistent and effective gas exchange by eliminating recirculation. Here, we report two cases of effectively using ProtekDuo veno-venous ECMO: one case as a bridge to lung transplantation and another case as a bridge to heart-lung transplantation.

Targeted genome editing has opened new frontiers to study gene functions by introducing alterations in the genome of an organism. The process incorporates deletion, insertion, or modification of gene at specific sites in a DNA sequence. Deliberate genetic changes in human cells may prevent or treat a specific disease condition; this way, averting the use of drugs and surgery. In this chapter, we will be focusing primarily on different techniques and their application in modern medicine/clinical aspect such as Homologous recombination, Endonucleases, Zinc-Finger Nucleases (ZFNs), Transcription Activator-Like Effector Nucleases (TALENS) and specially Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), that has revolutionized the way gene editing is done. Modern gene-editing techniques have dramatically improved our ability to do basic research, drug development, agriculture, and eventually treating human patients with genetic disease.

Background Several computational methods have been developed that integrate transcriptomics data with genome-scale metabolic reconstructions to increase accuracy of inferences of intracellular metabolic flux distributions. Even though existing methods use transcript abundances as a proxy for enzyme activity, each method uses a different hypothesis and assumptions. Most methods implicitly assume a proportionality between transcript levels and flux through the corresponding function, although these proportionality constant(s) are often not explicitly mentioned nor discussed in any of the published methods. E-Flux is one such method and, in this algorithm, flux bounds are related to expression data, so that reactions associated with highly expressed genes are allowed to carry higher flux values. Results Here, we extended E-Flux and systematically evaluated the impact of an assumed proportionality constant on model predictions. We used data from published experiments with Escherichia coli and Saccharomyces cerevisiae and we compared the predictions of the algorithm to measured extracellular and intracellular fluxes. Conclusion We showed that detailed modelling using a proportionality constant can greatly impact the outcome of the analysis. This increases accuracy and allows for extraction of better physiological information.

The phenomenon of senescence is an integral part of life; every living being has a predetermined life span. Brain cells are unique and show prominent age-associated changes including oxidative stress, shortening of telomeres, structural, cognitive, neuronal, and chemical changes. During aging, the tight integrated network of microglia cells that maintain normal central nervous system homeostasis and are the primary immune cells may lose its grip and show irregular communication with other brain cells, immune cells, and epigenetic modifications. These changes may trigger chronic neuroinflammation and in turn neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease, dementia, and amyotrophic lateral sclerosis, among others. Microglia have very distinct gene expression profile at different developmental stages. Several gene variants and their role in neurodegenerative disease pathogenesis have been identified that are highly expressed in microglia. This neuroimmune dysregulation along with aging is a significant target point for several therapeutic strategies to promote neuro repair and healthy aging.

Coronary artery disease (CAD) is a pathology often found in patients with end-stage lung disease. Although in the past CAD might have been considered an absolute contraindication, modern revascularization techniques have helped increase the number of transplants performed in this population. However, discrepancies in the guidelines for perioperative evaluation and risk mitigation strategies for the ischemic cardiac burden are present in the current literature. This is a review of the available data regarding perioperative evaluation, revascularization tactics, postoperative management, and survival rate that patients with different grades of coronary artery disease present after lung transplantation.

Solid-state nuclear magnetic resonance is a promising technique to probe bone mineralization and interaction of collagen protein in the native state. However, many of the developments are hampered due to the low sensitivity of the technique. In this article, we report solid-state nuclear magnetic resonance (NMR) experiments using the newly developed BioSolids CryoProbe™ to access its applicability for elucidating the atomic-level structural details of collagen protein in native state inside the bone. We report here approximately a fourfold sensitivity enhancement in the natural abundance 13 C spectrum compared with the room temperature conventional solid-state NMR probe. With the advantage of sensitivity enhancement, we have been able to perform natural abundance 15 N cross-polarization magic angle spinning (CPMAS) and two-dimensional (2D) 1 H-13 C heteronuclear correlation (HETCOR) experiments of native collagen within a reasonable timeframe. Due to high sensitivity, 2D 1 H/13 C HETCOR experiments have helped in detecting several short and long-range interactions of native collagen assembly, thus significantly expanding the scope of the method to such challenging biomaterials.

Introduction Parkinson’s disease (PD) is the most common neurodegenerative disease caused by the loss of dopamine chemicals resulting in urinary incontinence, gastrointestinal dysfunction, gait impairment and mitochondrial dysfunction. Study investigated urinary metabolic profiles of patients with idiopathic PD as compared to healthy controls (HC) to identify the potential biomarkers. Methods Urine samples were collected from 100 PD subjects and 50 HC using standard protocol. Metabolomic analyses were performed using high resolution nuclear magnetic resonance (NMR) spectroscopy. The integral values of 17 significant metabolites were estimated and concentration values were calculated, which were subjected to univariate and multivariate statistical analysis. Results We found significantly increased levels of ornithine, phenylalanine, isoleucine, β-hydroxybutyrate, tyrosine and succinate in the urine of patients with PD in comparison with HC. These metabolites exhibited area under the curve greater than 0.60 on ROC curve analysis. We also observed a significant association between succinate concentration and UPDRS motor scores of PD. Discussion Metabolic pathway alterations were observed in aromatic amino acid metabolism, ketone bodies synthesis, branched chain amino acid metabolism and ornithine metabolism. Comprehensive metabolomic profiling revealed variations in urinary signatures associated with severity of idiopathic PD. This profiling relies on non-invasive sampling and is complementary to existing clinical modalities.

Ever since the first COVID – 19 infection reported in India, the government is trying to take appropriate measures to safeguard Indian population from this deadly virus. In order to make correct policy, the evolution behaviour of the virus among Indian population should be understood accurately. We present in this manuscript the mathematical modelling of COVID – 19 infection data. Our modelling is based on open source data available on the website www.covid19india.org, which gives real time number of population infected, recovered and deceased due to COVID – 19. We further, show that, we can predict outcome with a reasonable accuracy.

SARS-CoV-2 is a novel virus which has proven to be highly contagious. Specific viral dynamics and immune response to the virus are yet to be fully defined and determining the sensitivity and specificity of the available testing methods is still a work in progress. This study examines the published information on the testing methods, and finds that yield of COVID-19 tests changes with specimen types and with time through course of illness. We propose a sequential battery of testing consisting of an epidemiologic survey, RT-PCR tests, serologic tests and chest CT on surgical candidates which may increase the negative predictive value, and facilitate surgical procedures.

The study of structural and dynamical properties of lipid and its associated interaction with different components of bone is essential to understand its role at a different level of bone homeostasis such as bone mineralization and bone metabolism. In this article, we present water-dependent dynamical changes observed in lipids (triglycerides) in its absolute native environment inside bone by high-resolution 1H solid-state nuclear magnetic resonance spectroscopy (ssNMR). Relaxation measurement (T2 measurement) ssNMR experiments were performed at different levels of water network induced by dehydration and H/D exchange in native bone. Our measurements reflect the changes in the local environment and dynamical properties of triglyceride due to different hydration levels. The present study explains the role of water in stabilizing the structural properties of triglycerides in bone hence will help understand its pathological role associated with bone physiology and bone disorders.

Background Muscular dystrophy is an inherited muscle disease, characterized by progressive muscle wasting and weakness of variable distribution and severity. Methods In vitro, high-resolution proton nuclear magnetic resonance (NMR) spectroscopy based analysis was performed on perchloric acid (PCA) extract of muscle specimens of patients suffering from various types of muscular dystrophies to identify alteration in hydrophilic low-molecular weight substances (aqueous metabolites) as compared to muscle of control subjects as well as in between the types of muscular dystrophy. Muscle tissue specimens were obtained from Duchenne muscular dystrophy (DMD) [n = 11], Becker muscular dystrophy (BMD) [n = 12], facioscapulohumeral dystrophy (FSHD) [n = 9] and limb girdle muscular dystrophy (LGMD)-2B [n = 22]. Control muscle specimens [n = 40] were also taken. Results Concentration of branched chain amino acids (BCA), glutamine/glutamate (Gln/Glu), acetate (Ace) and fumarate (Fum) was decreased and His was increased in muscle tissue of DMD, BMD, FSHD and LGMD-2B patients as compared to control subjects. Alanine (Ala) was significantly reduced in BMD, FSHD and LGMD-2B patients as compared to control subjects. Tyrosine (Tyr) was present only in the muscle tissue of control subjects. Propionate (Prop) was present in muscle tissue of DMD, BMD, FSHD and LGMD-2B patients and was absent in muscle tissue of control subjects. Concentration of BCA and Prop is significantly reduced in patients with DMD as compared to BMD, but Glucose is significantly higher in patients with DMD as compared to BMD. Quantity of Glucose, His and Gln/glu are significantly higher in patients with DMD as compared to FSHD, but Prop is significantly reduced in patients with DMD as compared to FSHD. Concentration of Ala and His is significantly higher in patients with DMD as compared to LGMD-2B, but BCA, Glucose and Prop are significantly reduced in patients with DMD as compared to LGMD-2B. Concentration of His is significantly higher in patients with BMD as compared to FSHD. Concentration of His is significantly reduced and Glucose is higher in patients with LGMD-2B as compared to BMD. Glucose concentration is significantly reduced in patients with FSHD as compared to LGMD-2B. ROC curves supported the noticeable discrimination in between the patients with DMD and FSHD for the quantity of Gln/Glu, and patients with LGMD-2B and DMD for the quantity of Ala. Collectively, these findings showed the perturbation of muscle metabolism in muscular dystrophy. Conclusions The data of presented study may be used as supporting information for existing methods of the diagnosis for patients with muscular dystrophy.

Collagen-induced arthritis (CIA) animal model is associated with systemic manifestations, including alteration of lipid metabolism. In the present study, one possible pathway of altered lipid metabolism is proposed. Specimens of joint tissue and plasma were collected from the CIA and control rats, and quantitative analysis of lipid components was performed by nuclear magnetic resonance (NMR) spectroscopy technique. Correlation analysis was performed between the level of lipid components and antioxidant enzymes, lactate dehydrogenase (LDH), lipid peroxidation (LP), and cytokines in joint tissue and plasma. Differentiation between the CIA and control rats was established on the basis of the quantity of lipid components in the joint tissue and plasma. Positive correlation was observed for all the enzymes vs. lipid components as well as LP vs. lipid components in plasma and joint tissue. Positive correlation was observed for enzymes in plasma and joint tissue. A negative correlation was observed in between the plasma and joint tissue with the level of lipid components. Cytokine levels were also correlated with the level of lipid components and ratios of saturated fatty acids/unsaturated fatty acids in plasma and joint tissue. Inflammatory disease activity in CIA rats with synovitis brought about a significant change in lipid metabolism. Taken together, the results of our study are delineating a possible pathway of altered lipid metabolism in the CIA rat model, thereby contributing further to an understanding of the pathophysiology of rheumatoid arthritis (RA).

In the quest for new antimicrobial materials, hydrogels of Fmoc-protected peptides and amino acids have gained momentum due to their ease of synthesis and cost effectiveness; however, their repertoire is currently limited, and the mechanistic details of their function are not well understood. Herein, we report the antibacterial activity of the hydrogel and solution phases of Fmoc-phenylalanine (Fmoc-F) against a variety of Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA). Fmoc-F, a small molecule hydrogelator, reduces the bacterial load both in vitro and in the skin wound infections of mice. The antibacterial activity of Fmoc-F is predominantly due to its release from the hydrogel. Fmoc-F shows surfactant-like properties with critical micelle concentration nearly equivalent to its minimum bactericidal concentration. Similar to Fmoc-F, some Fmoc-conjugated amino acids (Fmoc-AA) have also shown antibacterial effects that are linearly correlated with their surfactant properties. At low concentrations, where Fmoc-F does not form micelles, it inhibits bacterial growth by entering the cell and reducing the glutathione levels. However, at higher concentrations, Fmoc-F triggers oxidative and osmotic stress and, alters the membrane permeabilization and integrity, which kills Gram-positive bacteria. Herein, we proposed the use of the Fmoc-F hydrogel and its solution for several biomedical applications. This study will open up new avenues to enhance the repertoire of Fmoc-AA to act as antimicrobial agents and improve their structure-activity relationship.

Studies related to U gas-phase oxidation through plasma- and thermo-chemistry are important for many fields, including environmental monitoring, forensic analysis, debris analysis in a weapon detonation event, and nucleation physics. Recently, significant efforts have been made to understand the chemical pathways involved in the progression from U atoms to diatoms (UO) and polyatomic molecules (UxOy), employing optical spectroscopy tools and computational modeling. In many studies, laser ablation of U or a U-containing flow reactor are used as a highly resource-efficient, repeatable, tunable, and lab-scale testbed for studying gas-phase oxidation in U plasmas. The spectroscopic analysis of high-temperature gas-phase oxidation of U is challenging due to the congested U spectra, resolution limitations of instrumentation, and the numerous chemical reaction pathways possible. This article focuses on the current understanding and challenges related to studying U plasma chemistry, specifically U gas-phase oxidation and molecular formation, via optical spectroscopy of plasmas and associated computational and spectral modeling. The physical and chemical processes involved in the evolution from U atoms to U oxide molecules to nanoparticles and agglomerates (i.e., debris) are discussed in the context of optical spectroscopic studies. The article concludes by highlighting opportunities for future research efforts based on existing knowledge published in the literature.

Huntington’s disease (HD) is a neurodegenerative disorder induced by aggregation of the pathological form of Huntingtin protein that has expanded polyglutamine (polyQ) repeats. In the Drosophila model, for instance, expression of transgenes with polyQ repeats induce HD-like pathologies progressively correlating with the increasing lengths of these repeats. Previous studies on both animal models and clinical samples have revealed metabolite imbalances during HD progression. To further explore the physiological processes linked to metabolite imbalances during HD, we have investigated 1D 1H NMR spectroscopy-based metabolomics profile of Drosophila HD model. Using multivariate analysis (PCA and PLS-DA) of metabolites obtained from methanolic extracts of fly heads displaying retinal deformations due to polyQ overexpression, we show that the metabolite imbalance during HD are likely to affect cell energetics. Six out of the 35 metabolites analyzed, namely, nicotinamide adenine dinucleotide (NAD), lactate, pyruv...

Background and Purpose Group III Pulmonary hypertension (PH) is a highly lethal and widespread lung disorder that is a common complication in idiopathic pulmonary fibrosis (IPF) where it is considered to be the single most significant predictor of mortality. While increased levels of hyaluronan have been observed in IPF patients, hyaluronan-mediated vascular remodelling and the hyaluronan-mediated mechanisms promoting PH associated with IPF are not fully understood. Experimental Approach Explanted lung tissue from patients with IPF with and without a diagnosis of PH was used to identify increased levels of hyaluronan. In addition, an experimental model of lung fibrosis and PH was used to test the capacity of 4-methylumbeliferone (4MU), a hyaluronan synthase inhibitor to attenuate PH. Human pulmonary artery smooth muscle cells (PASMC) were used to identify the hyaluronan-specific mechanisms that lead to the development of PH associated with lung fibrosis. Key Results In patients with IPF and PH, increased levels of hyaluronan and expression of hyaluronan synthase genes are present. Interestingly, we also report increased levels of hyaluronidases in patients with IPF and IPF with PH. Remarkably, our data also show that 4MU is able to inhibit PH in our model either prophylactically or therapeutically, without affecting fibrosis. Studies to determine the hyaluronan-specific mechanisms revealed that hyaluronan fragments result in increased PASMC stiffness and proliferation but reduced cell motility in a RhoA dependent manner. Conclusions and Implications Taken together, our results show evidence of a unique mechanism contributing to PH in the context of lung fibrosis.

The small intestine serves as gatekeeper at the interface between body and diet and is thought to play an important role in the etiology of obesity and associated metabolic disorders. A computational modelling approach was used to improve our understanding of the metabolic responses of epithelial cells to different diets. A constraint based, mouse-specific enterocyte metabolic model (named mmu_ENT717) was constructed to describe the impact of four fully characterized semi-purified diets, that differed in lipid and carbohydrate composition, on uptake, metabolism, as well as secretion of carbohydrates and lipids. Our simulation results predicted luminal sodium as a limiting factor for active glucose absorption; necessity of apical localization of glucose transporter GLUT2 for absorption of all glucose in the postprandial state; potential for gluconeogenesis in enterocytes; and the requirement of oxygen for the formation of endogenous cholesterol needed for chylomicron formation under luminal cholesterol-free conditions. In addition, for a number of enzymopathies related to intestinal carbohydrate and lipid metabolism it was found that their effects might be ameliorated through dietary interventions. In conclusion, our improved enterocyte-specific model was shown to be a suitable platform to study effects of dietary interventions on enterocyte metabolism, and provided novel and deeper insights into enterocyte metabolism.

The study focuses on the flexural down-warping of oceanic crust related to the Early Cretaceous hotspot volcanic chain in offshore East India, drawing from robust reflection seismic coverage of the 85°E Ridge and associated moats and arches. Seismic data image three moat-filling units including the basal pelagic, landslide and ponded units, representing the sedimentary record preceding, coeval and postponing flexure. Their stacking patterns allow one to understand the flexural history of the oceanic crust reacting to the volcanic load, in space and time. The flexural history of the oceanic crust can be divided into four stages. The first stage is the brittle faulting-assisted flexure reacting to the appearance of the load. It has a short wavelength and controls the development of moat undergoing deposition of the landslide unit. Then follows the long-wavelength flexure, when the arch starts to develop. The flexural arch formation prevents the landslide unit from covering it, while the moat keeps subsiding. The third flexure stage is a short-wavelength deformation when the moat and arch subside together. Accordingly, the syn-flexural landslide unit records an initial rapid and a later slower subsidence. The fourth flexure stage is characterized by the passive infill of moat by sediments of ponded unit, although limited isostatic adjustments can occur, accompanied by mass wasting.

Introduction: Disseminated histoplasmosis is common particularly among immunocompromised individuals, alcoholics and extremes of age. It is of concern because of the associated high morbidity and mortality. Asymptomatic adrenal involvement has also been described in 30 to 50 % of patients with disseminated histoplasmosis. We report a case of adrenal histoplasmosis in an immune-competent subject presenting with adrenal insufficiency. Case Report: A 49 years old Indian male presented with three months history of weakness, nausea and vomiting, mucocutaneous pigmentation and weight loss. Past medical history was significant for tuberculosis, for which he was incompletely treated. He was non-smoker, non-alcoholic and non-diabetic, with no history of sexual promiscuity. USG of abdomen revealed bilateral suprarenal mass, with right adrenal (48.3x 39.3mm) and left adrenal measuring (42.4 x 38mm). CECT scan of upper abdomen showed enlargement of both adrenal glands with right adrenal 44x25x43mm, left adrenal measuring 53x37x46 mm suggestive of adrenal adenoma. CT guided FNAC of right adrenal mass showed necrosis, many budding yeast cells having the morphology of Histoplasma capsulatum. The patient was treated with liposomal form of amphotericin –B 150 mg / day for two weeks followed by oral itraconazole 200mg BD for one year and hydrocortisone 25 mg/day in divided doses. After one month, there was gain in weight, pigmentation faded and nausea & vomiting disappeared. Conclusion: Adrenal histoplasmosis usually occurs in a setting of immunocompromised condition and in endemic area, but it does occur in immunocompetant hosts from non-endemic area, as in our case. It is prudent to obtain a cytological or histopathological examination to confirm the diagnosis so that appropriate treatment can be instituted to avoid fatal complications.

We present in this article 1H nuclear magnetic resonance (NMR)-based metabolic approach to screen the serum metabolic alterations in human gallbladder inflammation with chronic cholecystitis (CC). Total of 71 human serum samples was divided into two groups, (n = 41, CC) and (n = 30 control). 1H NMR metabolic profiling was carried out for investigation of metabolic alterations. Multivariate statistical analysis was applied for pattern recognition and identification of metabolites playing crucial role in gallbladder inflammation. Receiver operating curve (ROC) and pathway analysis on NMR data were also carried out to validate the findings. Serum metabolites such as glutamine, low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), alanine, branch chained amino acids (BCAA), histidine and tyrosine were found to be depleted whereas formate, lactate, 1,2-propanediol were found to be elevated in CC. Metabolic pathways associated with metabolite alteration have also been reported. NMR has been established for disease diagnosis along with identification of metabolic pattern recognition in biofluids. Gallstones cause inflammation of the gallbladder in the form of CC. Inflammation plays a major role in causation of gall bladder cancer and leads the way to malignancy. Metabolic analysis of CC may lead to early diagnosis of disease and its progression to gallbladder cancer.

Gallbladder cancer (GBC) is a common form of malignancy, which has high incidence rates in the northern parts of India, South America, Japan, and in Native American populations. A few metabolomic studies have revealed changes in the composition of biofluids, tissues, and gallstones by nuclear magnetic resonance (NMR), which may prove useful in understanding GBC. In this chapter, we focus on the use of NMR in unveiling the metabolomics of GBC.

Huntington's disease (HD) is a genetic disorder caused by a CAG expansion mutation in the huntingtin gene leading to polyglutamine (polyQ) expansion in the N-terminal part of huntingtin (Httex1). Expanded polyQ, through a complex aggregation pathway, forms aggregates in neurons and presents a potential therapeutic target. Here we show Httex1 aggregation suppression by arginine and arginine ethyl ester (AEE) in vitro, as well as in yeast and mammalian cell models of HD, bearing expanded polyQ. These molecules also rescue locomotion dysfunction in HD Drosophila model. Both molecules alter the hydrogen bonding network of polyQ to enhance its aqueous solubility and delay aggregation. AEE shows direct binding with the NT17 part of Httex1 to induce structural changes to impart an enhanced inhibitory effect. This study provides a platform for the development of better arginine based therapeutic molecules against polyQ-rich Httex1 aggregation.

Gallbladder cancer (GBC) is a common form of malignancy, which has high incidence rates in the northern parts of India, South America, Japan, and in Native American populations. A few metabolomic studies have revealed changes in the composition of biofluids, tissues, and gallstones by nuclear magnetic resonance (NMR), which may prove useful in understanding GBC. In this chapter, we focus on the use of NMR in unveiling the metabolomics of GBC.

Critical illnesses contribute to the maximum morbidity and mortality of hospitalized patients. Acute respiratory distress syndrome (ARDS) and sepsis/septic shock are the two most common acute illnesses associated with intensive care unit (ICU) admission. Once triggered, both have an identical underlying mechanism, portrayed by inflammation and endothelial dysfunction. The diagnosis of ARDS is based on clinical findings, laboratory tests, and radiological imaging. Blood cultures remain the gold standard for the diagnosis of sepsis, with the limitation of time delay and low positive yield. A combination of biomarkers has been proposed to diagnose and prognosticate these acute disorders with strengths and limitations, but still, the gold standard has been elusive to clinicians. In this review article, we illustrate the potential of metabolomics to unravel biomarkers that can be clinically utilized as a rapid prognostic and diagnostic tool associated with specific patient populations (ARDS and sepsis/septic shock) based on the available scientific data.

Hydroxyapatite deposition and calcification occurs over cholesterol-containing lipid droplets between Bruch's membrane and sub-retinal pigment epithelium (sub - RPE) in the eyes of patients affected by age-related macular degeneration (AMD) as spherules, nodules, and Bruch's membrane plaques. In the present study, an attempt has been made to prepare a composite containing hydroxyapatite and cholesterol to elucidate interactions involved in the formation of such organic-inorganic interphase. To understand the mechanism of hydroxyapatite deposition on cholesterol, we have applied various biophysical techniques such as dynamic light scattering (DLS) measurements, transmission electron microscopy (TEM) imaging, Fourier transform infrared (FTIR) spectroscopy and solid-state nuclear magnetic resonance (ssNMR) spectroscopy on the prepared composite. Our results give molecular level insight into the mechanism of biocalcification in the disease system.

Biological tissues possessing motional heterogeneity are difficult to comprehend at atomic level in their native state. Some of these tissues play crucial roles in support and movement of the body. Articular cartilage is one such tissue exemplified with highly mobile glycosaminoglycans (GAGS, correlation time ∼10–9s) and relatively rigid collagen protein (correlation time ∼10–6 s). Simultaneous study of both the components at atomic level is challenging and requires different sets of solid state nuclear magnetic resonance (ssNMR) experiments for mobile and rigid parts. Additionally, it is hard to get any quantitative information as one 13C one pulse experiment is time-consuming. We present here a 13C ssNMR experiment with complete molecular characterization of cartilage organic components in its native state. We achieved this by employing heteronuclear Overhauser enhancement with equilibration of magnetization by low power irradiation. Complete 13C resonances from GAGS and collagen were identified simulta...

In vitro, high-resolution 31P NMR (Nuclear Magnetic Resonance) spectroscopy-based analysis of phospholipids in serum is well recognized in leukemia, lymphoma, non-hematological cancers and renal cell carcinoma. In context of these studies, phospholipids were analyzed in blood of thirty-two (n = 32) patients with Duchenne muscular dystrophy (DMD) (Age, Mean ± SD; 8.0 ± 1.6 years) and sixteen (n = 16) healthy subjects (Age, Mean ± SD; 8.6 ± 2.3 years). Quantity of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and lyso-phosphatidylcholine (Lys-PC) was significantly higher (p 0.05) observed for the quantity of phospholipids in blood of gene deletion positive cases of DMD as compared to negative gene deletion cases of DMD. Quantity of phospholipids in negative gene deletion cases of DMD patients as well as DMD cases with positive gene deletion was significantly higher (p

Glucose encapsulated cadmium oxide nanoparticles (G-CdO NPs) were synthesized by a sol-gel method. An array of different physical methods was used to characterize G-CdO NPs, including ultra violet-visible spectroscopy (λMax - 275 nm), transmission electron microscopy (TEM; 17 ± 1 nm), and zeta potential (−8.24 ± 0.4 mV) analyses. The NPs were active against HeLa cells (IC50 - 31.58 μg/mL) and Saos-2 cells (IC50 - 12.51 μg/mL) by preventing glycation (early, 95%; late, 65%). G-CdO NPs were effective anti-glycating agents and were highly effective against advanced glycation end products in particular. Localization of G-CdO NPs in the mitochondria and nuclei of cells was confirmed by TEM. Reactive oxygen species generated in the treated cells (confirmed by 2′,7′-dichlorofluorescin diacetate based assay) caused disintegration of mitochondrial cristae, disruption of the membrane integrity, and condensation and degradation of the nuclei. The LD50 of G-CdO NPs on Swiss Albino mice was 1.432 mg/kg of body weight. The results indicate the potential therapeutic value of G-CdO NPs.