Woodfordia fruticosa extract supplementation stimulates the growth of Lacticaseibacillus casei and Lacticaseibacillus rhamnosus with adapted intracellular and extracellular metabolite pool.

AIM: To investigate the effect of Woodfordia fruticosa extract (WfE) on two probiotic bacteria: Lacticaseibacillus casei and Lacticaseibacillus rhamnosus. METHODS AND RESULTS: WfE supplementation at 0·5 and 1 mg ml-1 stimulated probiotic growth (P < 0·05), enhanced adhesion to CaCO2 cells (P < 0·05) while inhibiting foodborne pathogens Escherichia coli and Staphylococcus aureus (P < 0·05). 1 H-NMR based metabolomic studies indicated higher glucose : lactate and glucose : acetate in the extracellular matrix with significant variation (P < 0·05) in intracellular concentrations of lactate, acetate, glutamate, dimethylamine, phenylalanine, branched-chain amino acids and total cellular lipid composition. Fatty acid methyl ester analysis showed a chemical shift from saturated to unsaturated lipids with WfE supplementation. PCA plots indicated clear discrimination between test groups, highlighting variation in metabolite pool in response to WfE supplementation. CONCLUSION: Phytonutrient-rich WfE exhibited prebiotic-like attributes, and probiotic bacteria showed altered metabolite pools as an adaptive mechanism. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report providing insights into the prebiotic-like activity of WfE on gut representative probiotics. The extended metabolomic studies shed light on the positive interaction between phytonutrients and beneficial bacteria that possibly help them to adapt to a phytonutrient-rich WfE environment. WfE with potential prebiotic attributes can be used in the development of novel synbiotic functional products targeting gut microbial modulation to improve health.

Metabolic profiling of lung granuloma in Mycobacterium tuberculosis infected guinea pigs: ex vivo 1H magic angle spinning NMR studies.

A crucial and distinctive feature of tuberculosis infection is that Mycobacterium tuberculosis (Mtb) resides in granulomatous lesion at various stages of disease development and necrosis, an aspect that is little understood. We used a novel approach, applying high resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMAS NMR) directly to infected tissues, allowing us to study the development of tuberculosis granulomas in guinea pigs in an untargeted manner. Significant up-regulation of lactate, alanine, acetate, glutamate, oxidized and the reduced form of glutathione, aspartate, creatine, phosphocholine, glycerophosphocholine, betaine, trimethylamine N-oxide, myo-inositol, scyllo-inositol, and dihydroxyacetone was clearly visualized and was identified as the infection progressed. Concomitantly, phosphatidylcholine was down-regulated. Principal component analysis of NMR data revealed clear group separation between infected and uninfected tissues. These metabolites are suggestive of utilization of alternate energy sources by the infiltrating cells that generate much of the metabolites in the increasingly necrotic and hypoxic developing granuloma through the glycolytic, pentose phosphate, and tricarboxylic acid pathways. The most relevant changes seen are, surprisingly, very similar to metabolic changes seen in cancer during tumor development.

New insights into the early steps of phosphatidylinositol mannoside biosynthesis in mycobacteria: PimB' is an essential enzyme of Mycobacterium smegmatis.

Phosphatidyl-myo-inositol mannosides (PIMs) are key glycolipids of the mycobacterial cell envelope. They are considered not only essential structural components of the cell but also important molecules implicated in host-pathogen interactions. Although their chemical structures are well established, knowledge of the enzymes and sequential events leading to their biosynthesis is still incomplete. Here we show for the first time that although both mannosyltransferases PimA and PimB' (MSMEG_4253) recognize phosphatidyl-myo-inositol (PI) as a lipid acceptor, PimA specifically catalyzes the transfer of a Manp residue to the 2-position of the myo-inositol ring of PI, whereas PimB' exclusively transfers to the 6-position. Moreover, whereas PimB' can catalyze the transfer of a Manp residue onto the PI-monomannoside (PIM1) product of PimA, PimA is unable in vitro to transfer Manp onto the PIM1 product of PimB'. Further assays using membranes from Mycobacterium smegmatis and purified PimA and PimB' indicated that the acylation of the Manp residue transferred by PimA preferentially occurs after the second Manp residue has been added by PimB'. Importantly, genetic evidence is provided that pimB' is an essential gene of M. smegmatis. Altogether, our results support a model wherein Ac1PIM2, a major form of PIMs produced by mycobacteria, arises from the consecutive action of PimA, followed by PimB', and finally the acyltransferase MSMEG_2934. The essentiality of these three enzymes emphasizes the interest of novel anti-tuberculosis drugs targeting the initial steps of PIM biosynthesis.

Aluminum-mediated metabolic changes in rat serum and urine: a proton nuclear magnetic resonance study.

The toxic effects of Al(3+) have been studied in 90-days AlCl(3) orally treated male albino rats (n = 7) using (1)H NMR spectroscopy-based metabolic profile of rat serum and urine, serum enzyme tests, behavioral impairment, and histopathology of kidney and liver. Metabolic profile of 90-days Al(3+)-treated rat sera showed significantly elevated levels of alanine, glutamine, beta-hydroxy-butyrate, and acetoacetate and significantly decreased level of acetone when compared with that of control rats. However, metabolic profile of 90-days Al(3+)-treated rat urine showed significantly decreased levels of citrate, creatinine, allantoin, trans-aconitate, and succinate and significantly increased level of acetate when compared to control rats. The overall perturbations observed in the metabolic profile of serum and urine demonstrate the impairment in the tricarboxylic acid cycle, liver and kidney metabolism, which was further reinstated by clinical chemistry and histopathological observations. Moreover, "in vivo" behavioral impairment has also been observed as the indication of aluminum neurotoxicity.

One-step analysis of major bile components in human bile using 1H NMR spectroscopy.

Human gallbladder bile dissolved in dimethylsulfoxide provides sharp and resolved signals for major bile components in 1H NMR spectra. Characteristic well-resolved marker signals that invariably appear in 1H NMR spectra of bile were identified for cholesterol (H18 methyl signal at 0.643 ppm), lipids (glycerol CH signal at 5.064 ppm), total bile acids (H18 signals in the range 0.520-0.626 ppm), total glycine conjugated bile acids (NH signal at 6.958 ppm), total taurine conjugated bile acids (NH signal at 7.646 ppm), and urea (NH2 signal near 5.48 ppm), which enabled their rapid and accurate analysis. Excellent linearity and precision of quantitative analysis was observed for all the identified bile components (R2 > 0.99 for all). The method was demonstrated on gallbladder bile from 19 patients with gallbladder diseases. Urea in bile was identified by NMR for the first time and its quantitative analysis, along with several other bile components, is presented. The majority of the bile components could be analyzed in a single step. Accurate and rapid quantification of several bile components noninvasively by using the method presented herein may have far-reaching implications in the study of bile acid metabolism and pathophysiology of various hepatobiliary and gastrointestinal diseases.

Single-step analysis of individual conjugated bile acids in human bile using 1H NMR spectroscopy.

1H and 13C NMR spectra of intact human bile were assigned using one-dimensional (1H and 13C) and two-dimensional (1H-1H and 1H-13C) experiments. Individual conjugated bile acids--glycocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, and taurochenodeoxycholic acid--were identified. The bile acids were quantified accurately and individually in a single step by using distinct and characteristic amide signals. Making use of 13C NMR, the study also suggests a way to analyze unconjugated bile acids separately, if present. Chemical shift assignments and rapid single-step analysis of individual conjugated bile acids from intact bile presented herein may have immense utility in the study of bile acid metabolism and deeper understanding of hepatobiliary diseases.

Simple pulse-acquire NMR methods for the quantitative analysis of calcium, magnesium and sodium in human serum.

Simple pulse-acquire NMR methods are presented for accurate quantitation of calcium (Ca(2+)), magnesium (Mg(2+)) and sodium (Na(+)) in human serum. Ca(2+) and Mg(2+) can be determined simultaneously by (1)H NMR via their EDTA (ethylenediaminetetraacetic acid) complexes. Spectra are acquired before and after addition of EDTA, and the difference spectrum is used for integration of the signals from the complexes relative to an internal reference. Serum sodium can also be determined through pulse-acquire (23)Na NMR by integration of the free sodium signal relative to a reference signal (Na(+)+EDTA in a coaxial capillary tube). The method shows excellent accuracy and precision for all three metal ions. Slow chemical exchange between complexed and free EDTA at the natural pH of serum does not limit the accuracy of the determination of Ca(2+) and Mg(2+), and the large errors associated with spin-echo spectral editing methods are avoided.

1H and 13C NMR characterization and stereochemical assignments of bile acids in aqueous media.

The unconjugated bile acids cholic acid, deoxycholic acid, and chenodeoxycholic acid; their glycine and taurine conjugates glycocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, and taurochenodeoxycholic acid; and a taurine conjugated ursodeoxycholic acid, tauroursodeoxycholic acid, were characterized through 1H and 13C NMR in aqueous media under the physiological pH region (7.4 +/- 0.1). Assignments of 1H and 13C signals of all the bile acids were made using a combination of several one- and two-dimensional, homonuclear (1H-1H) and heteronuclear (1H-13C) correlations as well as spectral editing NMR methods. Stereochemical assignment of the five-membered ring of the bile acids is reported here for the first time. The complete characterization of various bile acids in aqueous media presented here may have implications in the study of the pathophysiology of biliary diseases through human biliary fluids using NMR spectroscopy.

Reporting ethical aspects in published research articles in the Indian Journal of Psychiatry.

BACKGROUND: Reporting of informed consent and ethical approval are important aspects of published papers which indicate the knowledge and sensitivity about ethical aspects of research by the researchers. MATERIALS AND METHODS: This study reports description of informed consent and ethical approval in the published psychiatric research in the main journal of psychiatry in India. All original research articles (n=157) published in the Indian Journal of Psychiatry in the years 2000, 2003 to 2007 were included. RESULTS: Informed consent was mentioned in 51% of studies in 2000, which gradually rose to 82% by the year 2007. Ethics committee approvals were mentioned in 2% of studies in 2000, and 25% of reports in 2007. Consent was reported to be written in only 40%, content of the consent forms was mentioned in 17%, and the language of consent form was reported in 3% of the studies where consent was reported. CONCLUSIONS: Regulation of ethical principles and formulation of necessary guidelines or rules for research as well as for publications are necessary and desirable to ensure the safety of participants and good quality of research.

Somatization in cancer.

Somatic symptoms can occur in disease-free cancer patients. The causes of such symptoms in cancer can be many and varied. These could be due to anxiety, depression, somatization or a manifestation of illness behaviour. Somatic symptoms can also arise out of treatments for the cancer like radiation treatment or chemotherapy. Cancer related somatic symptoms have cognitive, psychological, and physiological causes, each of which is amenable to treatment. The occurrence of somatoform disorders in cancer patients is likely to complicate the treatment and outcome of the cancer. Common somatic symptoms in cancer have been found to be pain, fatigue, anorexia, tiredness or exhaustion, weakness, reduced energy, lethargy, and tremors. Breathlessness, muscle pain, dizziness, and palpitation are common symptoms of anxiety and panic attack which have also been noted in cancer patients. Somatic concern and preoccupation are also common. These symptoms create difficulty in diagnosing depression and anxiety in cancer patients, and leads to the need for modification of the standard diagnostic criteria. Somatic symptoms in cancer respond to counselling and psychopharmacotherapy. More research are needed on this area to understand the process of somatization in a somatic disease.

Protonation of trimipramine salts of maleate, mesylate and hydrochloride observed by 1H, 13C and 15N NMR spectroscopy.

Protonation of the tricyclic antidepressant drug trimipramine with maleic acid, methanesulfonic acid and hydrochloric acid was studied using 1H, 13C and 15N NMR spectroscopy at natural abundance. The effect of counter ions on the protonation was compared under identical conditions of solvent, concentration and temperature using homonuclear and heteronuclear one- and two-dimensional experiments. Differential protonation of the terminal tertiary amine nitrogen is determined from the indirect spin-spin couplings, chemical shifts, 13C relaxation data and variable-temperature experiments. In the maleate salt, only one of the acidic protons is involved in protonation, the other being associated with the anion moiety. 15N chemical shifts of the protonated nitrogens are nearly linearly related to the pK(a) of the constituent acid.

Quantification of glycine and taurine conjugated bile acids in human bile using 1H NMR spectroscopy.

A simple method for quantification of conjugated bile acids in human bile using (1)H NMR spectroscopy is presented. Bile acids in human bile are essentially conjugated with either glycine or taurine. The amide NH resonances from the conjugated bile acids are invariably devoid of interfering signals in (1)H NMR spectra. Under physiologic conditions of human bile (pH approximately 7.0 to 7.7), amide signal intensities are attenuated due to the chemical exchange and hence quantitative estimation is precluded. In the present study, the quantity of total glycine and taurine conjugated bile acids could be obtained accurately by suppressing the amide exchange by reducing the pH slightly lower than physiologic value (6.0 +/- 0.5). Further, the quantity of glycine conjugated bile acids can be calculated accurately by subtracting the quantity of taurine conjugated bile acids from the total conjugated bile acids as determined from the present method.

Differential protonation and dynamic structure of doxylamine succinate in solution using 1H and 13C NMR.

A protonation and dynamic structural study of doxylamine succinate, a 1:1 salt of succinic acid with dimethyl-[2-(1-phenyl-1-pyridin-2-yl-ethoxy)ethyl]amine, in solution using one- and two-dimensional 1H and 13C NMR experiments at variable temperature and concentration is presented. The two acidic protons of the salt doxylamine succinate are in 'intermediate' exchange at room temperature, as evidenced by the appearance of a broad signal. This signal evolves into two distinct signals below about -30 degrees C. A two-dimensional 1H-1H double quantum filtered correlation experiment carried out at -55 degrees C shows protonation of one of the acidic protons to the dimethylamine nitrogen. A two-dimensional rotating frame 1H-1H NOE experiment at the same temperature reveals that the other proton remains with the succinate moiety. Comparison of the 1H and 13C chemical shifts and the 13C T1 relaxation times of the salt with those of the free base further substantiate the findings.