ß-glucans from Agaricus bisporus mushroom products drive Trained Immunity.

Introduction: Macrofungi, such as edible mushrooms, have been used as a valuable medical resource for millennia as a result of their antibacterial and immuno-modulatory components. Mushrooms contain dietary fibers known as ß-glucans, a class of polysaccharides previously linked to the induction of Trained Immunity. However, little is known about the ability of mushroom-derived ß-glucans to induce Trained Immunity. Methods & results: Using various powdered forms of the white button mushroom (Agaricus bisporus), we found that mouse macrophages pre-treated with whole mushroom powder (WMP) displayed enhanced responses to restimulation with TLR ligands, being particularly sensitive to Toll-like receptor (TLR)-2 stimulation using synthetic lipopeptides. This trained response was modest compared to training observed with yeast-derived ß-glucans and correlated with the amount of available ß-glucans in the WMP. Enriching for ß-glucans content using either a simulated in-vitro digestion or chemical fractionation retained and boosted the trained response with WMP, respectively. Importantly, both WMP and digested-WMP preparations retained ß-glucans as identified by nuclear magnetic resonance analysis and both displayed the capacity to train human monocytes and enhanced responses to restimulation. To determine if dietary incorporation of mushroom products can lead to Trained Immunity in myeloid cells in vivo, mice were given a regimen of WMP by oral gavage prior to sacrifice. Flow cytometric analysis of bone-marrow progenitors indicated alterations in hematopoietic stem and progenitor cells population dynamics, with shift toward myeloid-committed multi-potent progenitor cells. Mature bone marrow-derived macrophages derived from these mice displayed enhanced responses to restimulation, again particularly sensitive to TLR2. Discussion: Taken together, these data demonstrate that ß-glucans from common macrofungi can train innate immune cells and could point to novel ways of delivering bio-available ß-glucans for education of the innate immune system.

A Modified Tongue Crib Appliance for Correction of Tongue Thrusting.

Digit sucking and tongue thrusting are parafunctional habits that are commonly observed and are recognized as major etiological factors in the development of dental malocclusion. Treatment for these oral habits typically involves removal of the etiology, retraining exercises, and use of mechanical restraining appliances. A 13-year-old male patient reported a complaint of forwardly placed upper front teeth. The extraoral examination of the patient showed a convex profile, good facial symmetry, acute nasolabial angle, incompetent lips at rest, and a shallow mentolabial sulcus. Intraoral examination showed class I molar and canine relations on either side, a mild open bite tendency, a midline diastema, proclined upper and lower anteriors, an increased overjet, and tongue thrusting habit. A modification to the tongue crib was made by adding an acrylic bead in the center of the crib. The patient was trained to roll the bead posteriorly with the aim of retraining the tongue to posture away from the front teeth. Harmful forces of the tongue can result in excessive posterior teeth eruption, open bite, and increased overjet. Duration of appliance wear and type of appliance used are important considerations in treating patients with tongue thrust or open bite. A modified tongue crib was used for six months resulting in significant improvement in maxillary anterior dentition position, lip competence, arch forms, overbite, and midline diastema closure. A tongue crib is a useful tool for addressing tongue thrusting and digit-sucking behaviors by retraining the related muscles, providing physical restraint, and serving as a reminder to break the habit. It can be used in conjunction with a fixed appliance to improve its effectiveness.

Unravelling Novel Phytochemicals and Anticholinesterase Activity in Irish Cladonia portentosa.

Acetylcholinesterase inhibitors remain the mainstay of symptomatic treatment for Alzheimer's disease. The natural world is rich in acetylcholinesterase inhibitory molecules, and research efforts to identify novel leads is ongoing. Cladonia portentosa, commonly known as reindeer lichen, is an abundant lichen species found in Irish Boglands. The methanol extract of Irish C. portentosa was identified as an acetylcholinesterase inhibitory lead using qualitative TLC-bioautography in a screening program. To identify the active components, the extract was deconvoluted using a successive extraction process with hexane, ethyl acetate and methanol to isolate the active fraction. The hexane extract demonstrated the highest inhibitory activity and was selected for further phytochemical investigations. Olivetolic acid, 4-O-methylolivetolcarboxylic acid, perlatolic acid and usnic acid were isolated and characterized using ESI-MS and two-dimensional NMR techniques. LC-MS analysis also determined the presence of the additional usnic acid derivatives, placodiolic and pseudoplacodiolic acids. Assays of the isolated components confirmed that the observed anticholinesterase activity of C. portentosa can be attributed to usnic acid (25% inhibition at 125 µM) and perlatolic acid (20% inhibition at 250 µM), which were both reported inhibitors. This is the first report of isolation of olivetolic and 4-O-methylolivetolcarboxylic acids and the identification of placodiolic and pseudoplacodiolic acids from C. portentosa.

A unique polysaccharide containing 3-O-methylarabinose and 3-O-methylgalactose from Tinospora sinensis.

Tinospora sinensis (Lour.) Merrill is of great therapeutic significance in Indian traditional medicine. Crude polysaccharides were isolated from methanol pre-extracted stems of dried material by successive extractions with cold water, hot water and NaOH (0.25 mol/L) in 0.98, 0.55 and 0.70 % yields respectively. Cold water soluble polysaccharides (CWSP) were purified and fractionated by ion exchange chromatography on DEAE-Sephacel. Neutral polysaccharides were further fractionated on Sepharose CL6B to yield three fractions TW1, TW2, TW3. The study further focuses on structural elucidation of TW1. TW1 was obtained in 0.8 % yield relative to CWSP, with MW of 1.6 × 105 Da. It was composed of 3-O-methyl-arabinose, 3-O-methyl-galactose and galactose in molar ratio of 1.0:6.3:0.9 respectively. Based on per-deuteromethylation, NMR and ESI-MS analyses, TW1 was composed of 1,4-linked 3-O-methyl-ß-d-galactopyranose and ß-d-galactopyranose backbone with branching at O-6 of 3-O-methyl-ß-d-galactosyl residues by 1,5-linked 3-O-methyl-α-l-arabinofuranoside chains. 3-O-methyl-arabinose and 3-O-methyl-galactose have first ever been reported in any polysaccharide and Tinospora genus, respectively.

Why WITS? Why not a way beyond?

INTRODUCTION: WITS appraisal is a common parameter in cephalometrics to assess maxillo-mandibular skeletal relationship as an adjunct to angle ANB. The high variability of the WITS appraisal is attributable to difficulties or inaccuracies in identifying the occlusal plane or variations due to tooth eruption, dental development or treatment changes by vertical movement of incisors, molars, or both. AIM: An extracranial reference line common to both denture bases, instead of the occlusal plane is proposed to assess antero-posterior jaw relationships. MATERIALS AND METHODS: A true vertical obtained by plumb line was recorded, while taking the cephalogram for 40 subjects. A line drawn perpendicular to this true vertical gave a stable and reproducible extracranial true horizontal (HOR) reference line. The linear distance between perpendiculars from points A and B was measured as an adjunct to angle ANB. RESULT: The Pearson's product moment correlation coefficient calculated for the entire sample indicated higher correlation coefficient (r = 0.8712) for the linear measurements on HOR (AH-BH) with respect to the angle ANB when compared with the WITS readings (AO-BO) (r = 0.549). CONCLUSION: The horizontal appraisal method proposed eliminates the demerits of the occlusal plane and has the merits of simplicity and accuracy in comparison to WITS appraisal.

Do assorted approaches aid in estimation of uronic acids? Case studies on Tinospora sinensis polysaccharides.

Polysaccharides containing uronic acids are of immense biological significance. Colorimetric methods using different chromogens have been explored for uronic acid estimation in polysaccharides but these methods counter numerous difficulties, when neutral sugars are present in substantial amount. The uronic acid content of cold water-soluble polysaccharide (CWSP) and hot water-soluble polysaccharide (HWSP) isolated from Tinospora sinensis, widely known for marked therapeutic significance has been studied. The uronic acid was quantitatively estimated by spectrophotometric methods using carbazole, 3-phenylphenol and 3,5-dimethylphenol (DMP) vis-a-vis gas-liquid chromatography (GLC) analysis. Galacturonic acid in CWSP and HWSP was estimated as 1.41% and 1.20% respectively. The findings bring to light, the efficacy of chromogens and also reveal to what extent, they could be relied upon. The results also indicated higher specificity and selectivity of DMP over carbazole and 3-phenylphenol. However, GLC analysis despite time consuming remains the finest method for precise estimation of uronic acids. Further, GLC analysis of alditol acetates indicated the monosugar composition of L-rhamnose, L-arabinose, D-mannose, D-galactose and D-glucose in molar ratio 0.178, 0.259, 0.140, 1.045 and 98.377 in CWSP and L-rhamnose, L-arabinose, D-xylose, D-mannose, D-galactose and D-glucose in molar ratio 0.608, 0.628, 0.978, 0.227, 0.111 and 97.448 in HWSP respectively.

Studies on Tinospora cordifolia monosugars and correlation analysis of uronic acids by spectrophotometric methods and GLC.

Cold water-soluble (CWSP) and hot water soluble polysaccharides (HWSP) from Tinospora cordifolia stems were isolated and purified in 2.99% and 1.99% yield respectively. Complete hydrolysis followed by paper chromatography and GLC analysis indicated the presence of L-rhamnose, L-arabinose, D-xylose, D-mannose, D-galactose and D-glucose in molar ratio of 0.857, 1.106, 0.727, 0.526, 0.708 and 95.763 in CWSP and 0.697, 0.777, 2.048, 0.777, 0.292 and 95.408 in HWSP. The uronic acid content in the polysaccharide has been studied extensively using assorted approaches. It was quantitatively estimated by GLC analysis and spectrophotometric methods using carbazole, m-hydroxydiphenyl and 3,5-dimethylphenol as colorimetric reagents. GLC analyses indicated galacturonic acid content of 3.06% and 5.16% in CWSP and HWSP respectively. Estimation of uronic acid using 3,5-dimethylphenol corroborated the above analysis. The study resulted in composition of constituent monosugars of CWSP and HWSP and co-relation analysis of uronic acid content, leading to an unambiguous structural analysis.