Isolation and characterisation of antibacterial and anti-inflammatory compounds from Gnaphalium polycaulon.

ETHNO-PHARMACOLOGICAL RELEVANCE: Gnaphalium polycaulon commonly known as "cudweed" has been used throughout South America as an infusion to treat colds, bronchitis, fever or pneumonia. AIM OF THE STUDY: This study aimed to determine the antibacterial and anti-inflammatory activities of the aqueous extract of Gnaphalium polycaulon and identify the related compounds. MATERIALS AND METHODS: A bio-guided isolation of the active compounds of Gnaphalium polycaulon was carried out, selecting the fractions depending on their antibacterial, anti-inflammatory and cytotoxic activities. The antibacterial effect was studied against Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and Streptococcus pneumoniae; and the anti-inflammatory study was performed by measuring the inhibition of NF-κB in BEAS-2B and IMR-90 cell cultures. RESULTS: Three compounds were obtained and characterised by nuclear magnetic resonance and mass spectrometry. These compounds are 2-(4-(1-H-tetrazol-1-yl) phenyl)-2-aminopropanoic acid (1), N-phenyl-4-(3-phenyl-1,2,4-thiadiazol-5-yl) piperazine-1-carboxamide (2) and N-(4-ethoxyphenyl)-4-(2-methylimidazo-[1,2-α] pyridine-3-yl) thiazol-2-amine (3). All compounds showed antibacterial activity with MIC values of 44.80-44.85, 0.017-0.021 and 0.0077-0.0079 µM, respectively, in the Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and Streptococcus pneumoniae strains, while the positive control, Ofloxacin, had a MIC value of 27.64-27.67 µM. This was corroborated through a zone inhibition assay, where compound 3 (11.36-11.67 mm) was much more active than the positive control (Ofloxacin, 23.41-24.12 mm), while compounds 2 (26.47-27.64 mm) and 1 (28.39-29.76 mm) displayed similar antibacterial potential to the positive control. Finally, all the compounds presented NF-κB inhibitory activity, compounds 3 (IC50 = 0.0071-0.0073 µM) and 2 (IC50 = 0.016-0.019 µM) being the most promising. Compound 1 (IC50 = 44.24-44.26 µM) had less anti-inflammatory potential, being also the closest to the values displayed by the positive control (Celastrol, IC50 = 7.41 µM). CONCLUSION: In the present study, three compounds were isolated for the first time from the aqueous extract of Gnaphalium polycaulon. Their antibacterial and anti-inflammatory potential was tested and showcased.

Anti-inflammatory, pro-proliferative and antimicrobial potential of the compounds isolated from Daemonorops draco (Willd.) Blume.

ETHNO-PHARMACOLOGICAL RELEVANCE: Daemonorops draco (D. draco) commonly known as "Dragon's blood" is one of the most used plants by Momok, Anak Dalam and Talang Mamak tribes from Indonesia as a remedy for wound healing. AIM OF THE STUDY: This study aimed to identify the extract, fractions and compounds responsible for the anti-inflammatory and pro-proliferative activities of the D. draco resin. Additionally, the antimicrobial activity against two bacteria and one yeast species was analysed. MATERIALS AND METHODS: Bio-guided isolation of compounds with anti-inflammatory, pro-proliferative and antimicrobial activities from the D. draco resin was carried out by measuring: the inhibition of NF-κB and activation of Nrf2 in THP-1, HaCaT, NIH-3T3 cells; cell proliferation in NIH-3T3 and HaCaT cells; and the antimicrobial effect on E. coli, S. aureus and C. albicans. RESULTS: Guided isolation by bioassay gave rise to the isolation and characterisation by nuclear magnetic resonance and mass spectrometry of three compounds: 1 (Bexarotene), 2 (Taspine) and 3 (2-hydroxy-1-naphthaldehyde isonicotinoyl hydrazone). All compounds showed NF-κB inhibitory activity with IC50 values of 0.10-0.13, 0.22-0.24 and 3.75-4.78 µM, respectively, while the positive control, Celastrol, had an IC50 of 7.96 µM. Likewise, all compounds showed an activating effect of Nrf2 with EC50 values of 5.34-5.43, 163.20-169.20 and 300.82-315.56 nM, respectively, while the positive control, CDDO-Me, had an EC50 of 0.11 nM. In addition, concerning the pro-proliferative activity, compound 1 (IC50 = 8.62-8.71 nM) showed a capacity of 100%, compound 2 (IC50 = 166-171 nM) showed a capacity of 75%, and compound 3 (IC50 = 469-486 nM) showed a capacity of 65%, while FSB 10% (positive control) had a pro-proliferative activity of 100% in the NIH3T3 cell lines (fibroblasts) and HaCaT (keratinocytes). Finally, all the compounds showed antimicrobial activity with MIC values of 0.12-0.16, 0.31-0.39 and 3.96-3.99 µM, respectively, in S. aureus, E. coli and C. albicans strains, while the positive control, Ofloxacin, had a MIC of 27.65 µM. CONCLUSION: This study managed to isolate, for the first time, three compounds (Bexarotene, Taspine and 2-hydroxy-1-naphthaldehyde isonicotinoyl hydrazone) from the resin of D. draco, with anti-inflammatory, and pro-proliferative as well as antimicrobial activities.

Efectos de la estimulación interfibrilar contrairritante musculoesquelética en epicondilalgias de jugadores de pádel amateurs. Estudio piloto / Effects of the musculoskeletal interfibrillar counterirritattion stimulation technique on amateur paddle-tennis players’ epicondylalgia. Pilot study

Objetivo: Analizar la eficacia de la técnica de estimulación interfibrilar contrairritante musculoesquelética en jugadores de deportes de raqueta con epicondilalgia y puntos gatillo miofasciales (PGM) activos. Material y métodos: Se trata de un estudio piloto diseñado como ensayo clínico (prospectivo) aleatorizado a doble ciego. Se contó con jugadores de pala o raqueta que cumplían los siguientes criterios de inclusión: tener una edad comprendida entre 18 y 60 años (ambos incluidos), tener dolor en la zona del epicóndilo lateral y tener un PGM activo central en la musculatura epicondílea. La muestra fue de 68 sujetos, de los cuales 61 completaron el estudio; 30 del grupo intervención (aplicación de la técnica) y 31 del grupo control (aplicación de un vendaje simple). La variable principal fue la algometría (umbral de dolor a la presión sobre el PGM) y las secundarias fueron la escala analógica visual (EVA 0-10) y el Patient-Rated Tennis Elbow Evaluation. Resultados: Los valores de algometría y los valores de EVA muestran que hubo una disminución del dolor en ambos grupos, aunque sin diferencias estadísticamente significativas entre ellos. La comparación de los porcentajes iniciales y finales EVA≥4 tampoco muestra diferencias entre grupos, pero la comparación antes/después de ambos grupos es estadísticamente significativa (p<0,05). Conclusiones: La técnica de estimulación interfibrilar contrairritante musculoesquelética, aplicada en PGM centrales de la musculatura epicondílea en sujetos con epicondilalgia que practican deportes de raqueta/pala, no parece ser más efectiva que la aplicación de un vendaje simple sobre la zona

Objective: To analyse the effectiveness of the musculoskeletal interfibrillar counterirritattion stimulation technique (MICS) in racquet sports players with epicondylalgia and active myofascial trigger points (MTrPs). Material and methods: This is a pilot study designed as double-blind randomized clinical trial (prospective). It featured paddle or racquet sport players who met the following inclusion criteria: to be aged between 18 and 60 (inclusive), to have pain in the lateral epicondyle and to have a central active MTrP in the epicondylar musculature. The sample consisted of 68 subjects, of whom 61 completed the study; divided into 30 for the Intervention group (technique application) and 31 for the Control group (simple bandage). The primary variable was algometry (pain threshold to pressure on the MTrPs) and secondary variables were the VAS (visual analogue scale, 0-10) and the Patient-Rated Tennis Elbow Evaluation. Results: Algometry values and VAS values show that there was a decrease in pain in both groups, but no statistically significant differences between them. The comparison of the initial and final percentages VAS≥4 also do not show differences between groups, but improvement of both groups is statistically significant (P<.05). Conclusions: The musculoskeletal interfibrillar counterirritattion stimulation technique applied in the central MTrPs of epicondylar muscles in subjects with epicondylalgia who play racquet sports/paddle does not seem to be more effective than the application of a simple bandage over the area

25-Hydroxyvitamin D levels in patients treated with high-dosage ergo- and cholecalciferol.

25-Hydroxyvitamin D (25-OHD) levels were measured in 39 patients with metabolic bone disease or hypoparathyroidism who had been treated with a constant high dose of vitamin D2 or D3 for at least 12 weeks. Plasma 25-OHD levels rose with increasing dosage, the relationship between dose and plasma level being approximately linear whether or not the dose was expressed on a weight-corrected basis. A therapeutic range of 25-OHD to be expected when patients with these conditions are treated with vitamin D has been established. There may be certain exceptions in which plasma 25-OHD levels within the range are associated with either an inadequate response to treatment or, conversely, the hypercalcaemia of vitamin D toxicity. There was no correlation between plasma calcium level and 25-OHD concentration in the group of patients studied. There was also no difference between the dose/25-OHD relationship of patients treated with vitamin D2 and that of patients receiving vitamin D3. Ten patients were started on treatment with large doses of vitamin D during the period of the study. The rate of rise of plasma 25-OHD was followed during treatment. The incremental rise in 25-OHD was calculated at the end of the first week of treatment in terms of dose per unit body weight. The rate of rise of plasma 25-OHD level was highly correlated with the dose used. Plasma 25-OHD levels after one weeks' treatment were only 15-20% of the expected steady-state level on the same dosage. The importance of a high priming dose when a rapid response is needed is thus emphasised.