Untargeted metabolomics in the aqueous humor reveals the involvement of TAAR pathway in glaucoma.

Understanding the metabolic dysfunctions and underlying complex pathological mechanisms of neurodegeneration in glaucoma could help discover disease pathways, identify novel biomarkers, and rationalize newer therapeutics. Therefore, we aimed to investigate the local metabolomic alterations in the aqueous humor and plasma of primary glaucomatous patients. This study cohort comprised primary open-angle glaucoma (POAG), primary angle-closure glaucoma (PACG), and cataract control groups. Aqueous humor and plasma samples were collected from patients undergoing trabeculectomy or cataract surgery and subjected to high-resolution mass spectrometry (HRMS) analysis. Spectral information was processed, and the acquired data were subjected to uni-variate as well as multi-variate statistical analyses using MetaboAnalyst ver5.0. To further understand the localized metabolic abnormalities in glaucoma, metabolites affected in aqueous humor were distinguished from metabolites altered in plasma in this study. Nine and twelve metabolites were found to be significantly altered (p < 0.05, variable importance of projection >1 and log2 fold change ≥0.58/≤ -0.58) in the aqueous humor of PACG and POAG patients, respectively. The galactose and amino acid metabolic pathways were locally affected in the PACG and POAG groups, respectively. Based on the observation of the previous findings, gene expression profiles of trace amine-associated receptor-1 (TAAR-1) were studied in rat ocular tissues. The pharmacodynamics of TAAR-1 were explored in rabbits using topical administration of its agonist, ß-phenyl-ethylamine (ß-PEA). TAAR-1 was expressed in the rat's iris-ciliary body, optic nerve, lens, and cornea. ß-PEA elicited a mydriatic response in rabbit eyes, without altering intraocular pressure. Targeted analysis of ß-PEA levels in the aqueous humor of POAG patients showed an insignificant elevation. This study provides new insights regarding alterations in both localized and systemic metabolites in primary glaucomatous patients. This study also demonstrated the propensity of ß-PEA to cause an adrenergic response through the TAAR-1 pathway.

No ambiguity: Chemosensory-based ayurvedic classification of medicinal plants can be fingerprinted using E-tongue coupled with multivariate statistical analysis.

Background: Ayurveda, the indigenous medical system of India, has chemosensory property (rasa) as one of its major pharmacological metric. Medicinal plants have been classified in Ayurveda under six rasas/tastes-sweet, sour, saline, pungent, bitter and astringent. This study has explored for the first time, the use of Electronic tongue for studies of rasa-based classification of medicinal plants. Methods: Seventy-eight medicinal plants, belonging to five taste categories (sweet, sour, pungent, bitter, astringent) were studied along with the reference taste standards (citric acid, hydrochloric acid, caffeine, quinine, L-alanine, glycine, ß-glucose, sucrose, D-galactose, cellobiose, arabinose, maltose, mannose, lactose, xylose). The studies were carried out with the potentiometry-based Electronic tongue and the data was analysed using Principle Component Analysis, Discriminant Function Analysis, Taste Discrimination Analysis and Soft Independent Modeling of Class Analogy. Results: Chemosensory similarities were observed between taste standards and the plant samples-citric acid with sour group plants, sweet category plants with sucrose, glycine, ß-glucose and D-galactose. The multivariate analyses could discriminate the sweet and sour, sweet and bitter, sweet and pungent, sour and pungent plant groups. Chemosensory category of plant (classified as unknown) could also be identified. Conclusion: This preliminary study has indicated the possibility of fingerprinting the chemosensory-based ayurvedic classification of medicinal plants using E-tongue coupled with multivariate statistical analysis.

Elevated histamine levels in aqueous humor of patients with glaucoma.

Purpose: Neurotransmitters (NTs) are the key mediators of essential ocular functions, such as processing the visual functions of the retina, maintaining homeostasis of aqueous humor, and regulating ocular blood flow. This study aims to determine variations in the levels of L-glutamate and γ-aminobutyric acid (GABA), histaminergic, adrenergic, cholinergic, and serotonergic NTs in patients with primary glaucoma versus patients with cataract. Methods: This case-control study involved three age-matched groups of patients with primary open angle glaucoma (POAG, n = 14), primary angle closure glaucoma (PACG, n = 21), and cataract (control, n = 19). Patients' aqueous humor and plasma were collected, snap frozen at -80 °C, and subjected to ultrasensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis for quantification of NTs. Results: Baseline intraocular pressure and the cup-to-disc ratio were found to be statistically significantly elevated in the POAG and PACG groups compared to the cataract control group. In aqueous humor, histamine was found to be statistically significantly elevated (5-fold, p<0.0001), whereas 1-methyl histamine was statistically significantly decreased (p<0.05) in POAG compared to the control group. A statistically significant increase in L-glutamate and GABA was observed among both patient groups with glaucoma compared to the cataract control group. Adrenaline was found to be elevated only in the PACG group (2.7-fold, p<0.05). No statistically significant difference was observed among the plasma NT levels between the groups. Conclusions: This study demonstrated the prominent role of the histaminergic system apart from autonomic mechanisms in the progression of glaucoma. Elevated L-glutamate and GABA could be due to retinal ganglionic cell death. Further studies are required to evaluate the effects of histamine on Müller cell dysfunction.

Current updates on precision therapy for breast cancer associated brain metastasis: Emphasis on combination therapy.

Cancer therapies have undergone a tremendous progress over the past decade. Precision medicine provides a more tailored approach, making the combination of existing therapies more precise. Different types of cancers are characterized by unique biomarkers that are targeted using various genomic approaches by clinicians and companies worldwide to achieve efficient treatment with minimal side effects. Precision medicine has two broad approaches namely stratified and personalized medicine. The driver mutations could vary within a subtype while the same driver mutations could be found across different subtypes. Precision medicine has recently gained a lot of importance for breast cancer therapy. Various kinds of mutations like hotspot mutations, gene alterations, gene amplification mutations are targeted to design a more specific therapy. Apart from these known gene mutations there are various unknown mutations. Thus, tumor heterogeneity can pose a challenge to precision medicine. For breast cancer, one of the most successful models developed in case of precision medicine is the anti-HER2 therapies as HER2 was considered to have the worst prognosis being highly malignant. But now due to the advent of HER2 receptor targeted therapies, it has a good prognosis. Moreover, precision medicine helps in identifying if the drug molecules being used for the treatment of one kind of cancer can be beneficial in the treatment of another kind of cancer as well, considering the signaling pathways and machinery is similar in most of the cancers. This reduces the time for new drug development and is economically more feasible. Precision medicine will prove to be very advantageous in case of brain metastasis.

Challenges in using Electronic tongue to study rasa of plants: II. Impact of solvent and concentration on sensor response and taste ranking.

BACKGROUND: Although Electronic tongue is used in pharmaceutical, food and beverage industries for objective evaluation of taste, its use in medicinal plants from an ayurvedic perspective is novel. Control experiments are therefore necessary to standardise and optimise parameters. OBJECTIVE: The aim is to optimise the use of solvent and standardise sample concentration for study of plants from an ayurvedic standpoint of rasa. The major objectives are two-fold: (i) evaluate sensor response to different types of solvent water (ii) explore use of E-tongue in taste ranking of medicinal plants used in ayurveda. MATERIALS AND METHODS: Single, double and triple distilled, reverse osmosis and milliQ waters were evaluated separately and as a medium for preparing plant extracts. For taste ranking, standard addition method using d-glucose as sweet taste standard was used for different brands of mango juices (case in point study) and eight medicinal plants from sweet category. The effect of sample concentration and taste standard on taste ranking were evaluated. RESULTS: MQ and TD water demonstrated similar organoleptic properties whereas plant extracts prepared in DD and MQ water showed maximum taste-based differentiation. The mango juices were taste discriminated by E-tongue and ranked based on their sweetness scores. The relative ranking of plant samples showed concentration dependence and also varied with the concentration range of taste standard. CONCLUSION: Milli-Q and double distilled water can be used for E-tongue studies of medicinal plants. While the results open up the possibility of taste ranking of medicinal plants, they also demonstrate the importance of standardising and optimising the concentration of samples and taste standards in the context of ayurvedic rasa based studies.

Challenges in using electronic tongue to study rasa of plants: I. Finding the right tool for the right job.

There is growing interest in understanding how ayurveda, the indigenous medical system of India, uses plants for therapeutic purpose. The aim of this two parts article is to explore how the analytical technique of Electronic tongue (E-tongue) can be used for studying rasa, one of the major ayurvedic parameter in the study of medicinal and nutritional plants. Although E-tongue is widely used in pharmaceutical, food and beverage industries for objective evaluation of taste, its use in plants from an ayurvedic perspective is novel. This first of the two parts article introduces the technique of E-tongue, positioning it in context for the following part. The latter discusses crucial control experiments required prior using E-tongue for studies on medicinal plants from an ayurvedic standpoint.

Tantra and Modern Neurosciences: Is there any Correlation?

BACKGROUND AND AIMS: Many studies have conclusively proven that meditative techniques derived from the Indian systems of philosophy, meditation and ritual classified as "Tantra" can bring about sustained changes in the structure and function of the nervous system of practitioners. The aim of this study is to provide neuroscientists a framework through which to interpret Tantra, and thereby provide a foundation upon which future interdisciplinary study can be built. METHODS: We juxtapose Tantric concepts such as the subtle body, nadis and mantras with relevant neuroscientific findings. Our premise is that through sustained internalization of attention, Tantric practitioners were able to identify and document subtle changes in their field of awareness, which usually do not cross the threshold to come into our perception. RESULTS: The descriptions left by Tantric philosophers are often detailed and empirical, but they are about subjective phenomena, rather than external objects. They also focus on individual experiences, rather than the group-level analyses favored by modern medical science. CONCLUSION: Systematic exploration of Tantric texts can be of tremendous value in expanding our understanding of human beings' experiential reality, by enabling us to build bridges between first-person and third-person approaches to the nervous system. This may open up new avenues for cognitive enhancement and treating neurological diseases.

Challenges in integrating component level technology and system level information from Ayurveda: Insights from NMR phytometabolomics and anti-HIV potential of select Ayurvedic medicinal plants.

BACKGROUND: Information from Ayurveda meeting the analytical challenges of modern technology is an area of immense relevance. Apart from the cerebral task of bringing together two different viewpoints, the question at the pragmatic level remains 'who benefits whom'. OBJECTIVE: The aim is to highlight the challenges in integration of information (Ayurvedic) and technology using test examples of Nuclear Magnetic Resonance (NMR) metabolomics and anti-HIV-1 potential of select Ayurvedic medicinal plants. The other value added objective is implications and relevance of such work for Ayurveda. MATERIALS AND METHODS: Six medicinal plants (Azadirachta indica, Tinospora cordifolia, Swertia chirata, Terminalia bellerica, Zingiber officinale and Symplocos racemosa) were studied using high resolution proton NMR spectroscopy based metabolomics and also evaluated for anti-HIV-1 activity on three pseudoviruses (ZM53 M.PB12, ZM109F.PB4, RHPA 4259.7). RESULTS: Of the six plants, T. bellerica and Z. officinale showed minimum cell cytotoxicity and maximum anti-HIV-1 potential. T. bellerica was effective against all the three HIV-1 pseudoviruses. Untargeted NMR profiling and multivariate analyses demonstrated that the six plants, all of which had different Ayurvedic pharmacological properties, showed maximum differences in the aromatic region of the spectra. CONCLUSION: The work adds onto the list of potential plants for anti-HIV-1 drug molecules. At the same time, it has drawn attention to the different perspectives of Ayurveda and Western medicine underscoring the inherent limitations of conceptual bilinguism between the two systems, especially in the context of medicinal plants. The study has also highlighted the potential of NMR metabolomics in study of plant extracts as used in Ayurveda.

Spectroscopic and E-tongue evaluation of medicinal plants: A taste of how rasa can be studied.

BACKGROUND: The use of medicinal plants in Ayurveda is based on rasa, generally taken to represent taste as a sensory perception. This chemosensory parameter plays an important role in Ayurvedic pharmacology. OBJECTIVE: The aim is to explore the use of structuro-functional information deduced from analytical techniques for the rasa-based classification of medicinal plants in Ayurveda. MATERIALS AND METHODS: Methods of differential sensing and spectroscopic metabolomics have been used in select medicinal plants from three different taste categories (sweet, pungent and multiple taste): Tribulus terrestris, Vitis vinifera and Glycyrrhiza glabra from sweet category; Piper longum, Cuminum cyminum and Capsicum annum from pungent group; Emblica officinalis with five tastes. While Electronic tongue was used for evaluation of the sensorial property of taste, the chemical properties were studied with Nuclear Magnetic Resonance (NMR), Fourier Transform InfraRed (FTIR) and Laser Induced Breakdown Spectroscopy (LIBS). RESULTS: In terms of taste and phytochemical profiles, all samples were unique but with similarities within each group. While the sensor response in E-tongue showed similarities within the sweet and pungent categories, NMR spectra in the aromatic region showed close similarities between the plants in the sweet category. The sensory, phytochemical and phytoelemental profiles of E. officinalis (with five rasa) in particular, were unique. CONCLUSION: A combination of sensorial and chemical descriptors is a promising approach for a comprehensive evaluation and fingerprinting of the Ayurvedic pharmacological parameter rasa.

Healthcare the Ayurvedic way.

Ayurveda, the indigenous medical system of India, has integrated the concept of interconnectedness into its understanding of health and disease. It considers the human body as an indivisible whole with a network of interrelated functions, mind and consciousness, wherein a disturbance in one part will have repercussions in other parts as well. The key to health is for these factors to maintain stability since disease is seen as a perturbation in this network. Ayurvedic treatment aims not only at removal of disease, but also at the restoration of the equilibrium of bodily functions. The centuries-old science of Ayurveda, apart from being a holistic form of medicine, also clearly defines the student-teacher and doctor-patient relationship. It sets high ethical standards of professional and personal integrity for the doctor, teacher and student. In addition to being technically skilled and well-read in the science, practitioners of Ayurveda must be compassionate and empathetic. Students of Ayurveda are taught the importance of compassionate caring for patients and maintaining personal integrity.

In vivo MRS study of intraventricular tumors.

PURPOSE: To evaluate the role of MR spectroscopy (MRS) as a noninvasive tool for characterization of intraventricular brain tumors. MATERIALS AND METHODS: Twenty-three intraventricular tumors, all histologically classified (11 central neurocytomas [CNC], 5 meningiomas [Men], other intraventricular tumors [OIV], including 2 anaplastic gliomas [WHO III], 1 tumor each of rhabdoid tumor, glioneuronal hamartoma, GBM, hemangiopericytoma, and subependymoma II), were studied by MRS before surgery using a point-resolved spectroscopic sequence (PRESS). RESULTS: Statistically significant differences (P < 0.03) were observed for NAA/Cr and Cho/Cr between CNCs and OIV. A significant difference (P < 0.01) was observed in the presence of Gly between the CNCs and meningiomas, and CNCs and OIV. In addition, the presence of Ala was also significantly different (P < 0.01) between OIV and meningiomas, and OIV and CNCs. CONCLUSION: The present study has shown that MRS can be useful in characterizing intraventricular tumor and distinguishing CNCs from meningiomas and other intraventricular tumors. Prior information about intraventricular tumor types could be useful to clinicians in planning treatment and resection strategy.

MRS characterization of central neurocytomas using glycine.

This study reports in vivo MRS findings in 11 patients with histologically diagnosed central neurocytomas, which are rare intraventricular tumors of neuronal origin. Single-voxel (1)H MRS was carried out prior to surgery using a point-resolved spectroscopy sequence with TR=6 s, TE=135 ms and 128 scans. In vitro high-resolution (1)H spectroscopy was also carried out on two surgically excised samples. The striking features of the spectra from the central neurocytomas were the presence of high glycine, decreased N-acetylaspartate, increased choline and alanine. Retrospective, blind analysis of the spectra by two independent observers correctly identified all but one central neurocytoma based on the presence of glycine. The presence of glycine and prominent choline in the (1)H MR spectrum is a characteristic feature of the central neurocytomas, and could be used to characterize and differentiate them from other brain tumors.

Evaluation of folate conjugated pegylated thermosensitive magnetic nanocomposites for tumor imaging and therapy.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been receiving great attention lately in biomedical applications, such as in magnetic resonance imaging and drug delivery. However, their systemic administration still remains a challenge due to their hydrophobic nature with instances of aggregation leading to fast reticuloendothelial system (RES) uptake. In this study, magnetic nanocomposites with thermosensitive polymer have been investigated. Random polymers of N-isopropylacrylamide (NIPAAM), acrylic acid (AA) and PEGMA have been coated on SPIONs followed by conjugation with folic acid. Particles of ∼200 nm and low polydispersity 0.1-0.2 having a critical temperature (T(c)) of 44 °C were formed. Thermogravimetric and powder X-ray diffraction studies showed that the nanocomposites were composed of 90% cubic face-centered magnetite. Nearly 76.5% doxorubicin was loaded onto the nanoparticles by diffusion method. Drug release was higher at the hyperthermia temperature (72.42 ± 5.25% in 48 h) proving the thermoresponsive nature of the polymer. Folate conjugated samples showed a magnetization value of 32 emu/g as well as high r1 and r2 relaxivities in magnetic resonance imaging. R2 weighted images of nanocomposites were darker than the control with 20 µg/mL as the darkest. At this concentration the magnetic composites showed nearly 95% viability in L929 fibroblast cells. These thermoresponsive nanosystems with pegylated surfaces and size of ∼200 nm are therefore highly suitable for in vivo imaging and hyperthermia based drug delivery.

Determination of relaxation characteristics during preacute stage of lysophosphatidyl choline-induced demyelinating lesion in rat brain: an animal model of multiple sclerosis.

Relaxation time measurements were carried out during the preacute stage of lesion progression in an animal model of demyelination created in the internal capsule (ic) area of the rat brain using lysophosphatidyl choline (LPC). T1 and T2 were determined both before and after 36 h of lesion creation. Histology carried out on the rats after MR measurements showed focal demyelinating lesion and surrounding edema with prominent infiltration of inflammatory cells. Both T1 and T2 were statistically higher for the lesion compared to that determined before lesion creation. Percentage increase in T2 was found to be higher by approximately 45% compared to before lesion creation while T1 showed about 25% increase. Increase in T1 and T2 may be attributed to the early acute inflammatory response due to LPC. The beginning of the inflammatory response following LPC injection may also be a contributing factor. The study demonstrates that the quantitative estimate of MR relaxation provides useful information on the pathological events occurring during the early phase of the progression of demyelination.

Magnetic resonance spectroscopy in pediatric neurology.

In the last three decades a range of non-invasive biophysical techniques have been developed, of which Magnetic Resonance (MR) has proved to be the most versatile. Its non-invasive and safe nature has made it the most important diagnostic and research tool in clinical medicine. MR Spectroscopy (MRS) is the only technique in clinical medicine that provides non-invasive access to living chemistry in situ. This article focuses mainly on proton MRS in brain and also phosphorus MRS in calf muscle, with particular reference to the pediatric population, the normal spectrum and its use in various disease conditions in the practice of pediatric neurology. Few representative case studies among different disease groups have also been detailed.

In vivo and in vitro MR spectroscopic profile of central neurocytomas.

This study reports in vivo and in vitro magnetic resonance spectroscopic findings in two cases of central neurocytomas (CNC) confirmed by immunohistochemistry. Volume localized in vivo proton magnetic resonance spectroscopy (MRS) was carried out before surgery using a point resolved spectroscopy (PRESS) sequence with a repetition time of six seconds and an echo time of 135 msec. Normal spectrum was obtained from gray matter from a volunteer for comparison. (1)H and (31)P in vitro MRS studies were carried out at 9.4 T on the extracts prepared from the surgically excised tumors. The in vivo spectra showed prominent glycine (Gly) and choline (Cho) and low N-acetyl aspartate compared to the normal. The Gly peak was assigned using the in vitro studies. These studies showed that the major contribution to the Cho peak observed in vivo is from phosphocholine. A combination of the presence of NAA and an increased Gly in the proton MR spectrum could be a characteristic feature of CNCs, which are rare intraventricular tumors of neuronal origin.

Sequential diffusion-weighted magnetic resonance imaging study of lysophosphatidyl choline-induced experimental demyelinating lesion: an animal model of multiple sclerosis.

PURPOSE: To differentiate the surrounding edema from the focal demyelinating lesion during the early phase of the lesion using an apparent diffusion coefficient (ADC), and to monitor the changes in ADCs during the complete progression of a lysophosphatidyl choline (LPC)-induced experimental demyelinating lesion, an animal model of multiple sclerosis (MS). MATERIAL AND METHODS: Eighteen rats divided into two groups-demyelinating lesion (group I, N = 12) and vehicle group (saline injected; group II, N = 6)-were studied. A 0.2-microl quantity of 1% LPC solution in isotonic saline was injected in the rat brain internal capsule (IC) area to create the demyelinating lesion. Six rats were used exclusively for histology. Diffusion-weighted (DW) images were acquired at different diffusion weightings on the 3rd, 5th, 10th, 15th, and 20th days after LPC injection. ADC was measured from three regions of interest (ROIs) within the IC: focal demyelinating lesion (area A), surrounding area of the lesion (area B), and contralateral IC area (area C). RESULTS: Histology revealed demyelination of the IC area during the early phase of lesion progression up to day 10 and remyelination thereafter. Elevated ADCs were observed for the surrounding edematous area (area B), compared to the focal demyelinating lesion (area A) during the early phase of the demyelination process, while substantial reduction of ADCs was noticed during remyelination for both regions. CONCLUSION: Measurement of ADC showed clear differentiation of the surrounding edema from the LPC-induced focal demyelinating lesion in rats, especially during the early phase of the lesion progression.

Proton MR spectroscopy of basal ganglia in Wilson's disease: case report and review of literature.

Volume localized proton Magnetic Resonance Spectroscopy was carried out in both the left and right basal ganglia of three patients with clinically proven Wilson's Disease. While the untreated patient died 15 days after the spectroscopy study, the other 2 patients have been under treatment and have shown clinical improvement. The spectral features of the untreated patient were very different from those of the other two patients who were under treatment and responding. Asymmetrical changes in NAA and Cho were also observed for this patient.