Concurrent Resolution of Chronic Diarrhea Likely Due to Crohn's Disease and Infection with Mycobacterium avium paratuberculosis.

Examination of samples of stool from a 61-year-old male patient, presenting with the clinical symptoms of Crohn's disease (CD), revealed massive shedding of acid fast bacilli with the morphology of Mycobacterium avium paratuberculosis (MAP), the causative agent of Johne's disease in cattle. MAP was cultured from the stool. Biotyping of the bacterium isolated from cultures of stool demonstrated, it was the Indian Bison biotype of MAP, the dominant biotype infecting livestock and humans in India. Based on this finding and because the patient was unresponsive to standard therapy used in India to treat patients with gastrointestinal inflammatory disorders, the patient was placed on a regimen of multi-antibiotic therapy, currently used to treat tuberculosis and CD. After 1 year of treatment, the patient's health was restored, concurrent with cessation of shedding of MAP in his stool. This patient is the first case shown to shed MAP from the stool who was cured of infection with antibiotics and who was concurrently cured of clinical signs of CD.

Identification of novel homologous series of polyhydroxylated theasinensins and theanaphthoquinones in the SII fraction of black tea thearubigins using ESI/HPLC tandem mass spectrometry.

Thearubigins are the most abundant phenolic pigments found in black tea, produced by enzymatic oxidation of green tea flavan-3-ols in tea fermentation of until recently unknown composition. In this study electrospray ionization tandem LC-MS(n) experiments have been applied for the characterization of crude thearubigins isolated from black tea not exceeding 1000 Da. The aim of this study is to confirm the oxidative cascade hypothesis of tea fermentation. The data revealed the presence of two novel classes of compounds in thearubigin fractions. The first class of compounds revealed the presence of polyhydroxylated dimers of the theanaphthaquinone and theasinensin C structures, which were consistent with the polyhydroxylation hypothesis previously formulated. Furthermore, new classes of peroxo-/epoxy- compounds in the series of theasinensin A were identified, thus indicating the presence of H2O2 and its important contribution as a nucleophile in the tea fermentation process.

Investigation of processes in black tea manufacture through model fermentation (oxidation) experiments.

Flavanol depleted whole fresh green tea leaf powder, as reported in the literature, was used as matrix for a systematic study of the endogenous oxidative enzymatic conversion of selected flavanol combinations to theaflavins and thearubigins. The activity of the two crucial enzymes polyphenol oxidase (PPO) and peroxidase (POD) was controlled individually through addition of H2O2 and/or O2. Using the endogenous peroxidase only it was shown that (-)-epicatechin alone did not react with POD. According to these results it is possible that theaflavin formation occurs via reaction of a flavanol quinone with a nonquinone flavanol. It was confirmed that only a dihydroxy-B-ring flavanol with a trihydroxy-B-ring flavanol gave a theaflavin upon enzymatic oxidation. Use of horseradish peroxidase in the presence of a flavanol depleted tea leaf matrix led to significantly higher kinetics on theaflavin 3-gallate degradation compared to the absence of leaf matrix, suggesting a catalytic effect of the leaf matrix not reported before.

Characterisation of Polyphenols in Terminalia arjuna Bark Extract.

The bark of Terminalia arjuna is known for its heart-health benefits in ayurvedic literature. This has been further supported by in vivo studies on animal and human volunteers. But there is no detailed study on identification of the active ingredients such as polyphenols. Polyphenols possesses antioxidant properties and are well-known health actives, it is important to characterise polyphenols in Terminalia arjuna. Aqueous extract of Terminalia arjuna bark was analysed for its composition and molecular weight distribution by dialysis. Compositional analysis revealed that it has 44% polyphenols and dialysis study showed that 70% of the polyphenols have molecular weight greater than 3.5 kDa. High performance liquid chromatography and liquid chromatography-mass spectrometry analysis of Terminalia arjuna, confirmed that it contains flavon-3-ols such as (+)-catechin, (+)-gallocatechin and (-)-epigallocatechin. Phenolic acids such as gallic acid, ellagic acid and its derivatives were also found in Terminalia arjuna extract. Ellagic acid derivatives were isolated and their spectral studies indicated that isolated compounds were 3-O-methyl-ellagic acid 4-O-ß-D-xylopyranoside, ellagic acid and 3-O-methyl ellagic acid 3-O-rhamnoside. Hydrolysis and thiolysis studies of high molecular weight polyphenols indicated that they are proanthocyanidins. Given these results, it may be possible to attribute the heart-health effects of Terminalia arjuna to these polyphenols which may be responsible for the endothelial benefit functions like tea.

Development of a gas chromatography method for the estimation of alkylamines in foods.

Alkylamines are gaining importance due to their proven immunity benefits. Fruits, vegetables, and beverages are important dietary sources for alkylamines. This paper reports for the first time quantitative data on four alkylamines, ethyl-, propyl-, isopropyl-, and sec-butylamines, in commonly consumed fruits and vegetables. A sensitive and selective chromatography method based on derivatization with pentafluorobenzaldehyde and detection by GC-ECD is developed and validated for the analysis of alkylamines in vegetables, fruits, and tea. In vegetables and fruits, the concentrations varied from 100 to 15,000 µg/kg. Propyl- and isopropylamine concentrations were significantly higher compared to other amines. Among all dietary sources, tea had the highest concentration of alkylamines (30-50 mg/kg), with ethylamine as the major component, and is the richest source for alkylamines. The stability of these alkylamines was studied under various cooking conditions, and it was observed that there is loss of alkylamines on cooking.

Novel colorimetric sensor for oral malodour.

Volatile sulphur compounds are the primary constituents of oral malodour. Quantitative tools for the detection of oral malodour are beneficial to evaluate the intensity of malodour, analyse its causes and monitor the effectiveness of customized treatments. We have developed an objective, cost effective, do-it-yourself colorimetric sensor for oral malodour quantification. The sensor consisted of a sensing solution, a gas sampling unit for collecting a known volume of mouth air and a photometric detector. The sensing solution was iodine and the depletion of iodine on reaction with hydrogen sulphide was detected colorimetrically using starch. The detection limit of the sensor is 0.05 microg L(-1) of hydrogen sulphide, which is fit-for-purpose for oral malodour detection in healthy subjects as well as halitosis patients. Volatile sulphur compounds in mouth air were quantified in healthy human volunteers using this portable sensor and the detected levels were in the range of 0.2-0.4 microg L(-1). There was a good correlation between the VSC levels detected by the colorimetric sensor and halimeter (R(2)=0.934). The developed sensor can be easily fabricated in the laboratory, and it shows high potential to be used as a clinical evaluation tool for oral malodour assessments.

Polyethylenimine nanoparticles as efficient transfecting agents for mammalian cells.

Two cross-linkers based on polyethylene glycol (PEG) (MW=6 and 8 kDa), were synthesized for self-assembling and formation of nanoparticles of branched, high molecular weight polyethylenimine (PEI). Cross-linking was realized in two ways, viz., ionic as well as covalent. Ionic cross-linking was accomplished by using PEG-bis (phosphate) whereas, the covalent one was achieved by using PEG-bis (p-nitrophenylcarbonate). A range of nanoparticles of PEI was prepared by varying the degree of cross-linking (i.e. the amount of cross-linkers used). PEI-PEG nanoparticles were characterized by dynamic light scattering and transmission electron microscopy and found to be in the range of approximately 18-75 nm (hydrodynamic radii) with almost uniform population. Subsequently, these particles were used for DNA binding assay and zeta-potential measurements, taking native PEI-PEG nanoparticles as reference. As expected, the zeta potential values decreased, on increasing the percentage of cross-linking as well as on complexation with DNA. Further, PEI-PEG nanoparticles were investigated for their transfecting efficacy on COS-1 cells. It was found that PEI-PEG nanoparticles were 5- to 16-fold more efficient as transfecting agents compared to lipofectin and PEI itself. The toxicity of PEI-PEG nanoparticles was found to be reduced considerably in comparison to PEI polymer, as determined by MTT colorimetric assay. Out of the various systems prepared, PEI-PEG8000 (5% ionic) nanoparticles were found to be the most efficient transfecting agent for in vitro transfection.