Photodynamische Therapie (PDT) und wassergefiltertes Infrarot A (wIRA) bei Patienten mit therapierefraktären vulgären Hand- und Fußwarzen

Background: Common warts (verrucae vulgares) are human papilloma virus (HPV) infections with a high incidence and prevalence, most often affecting hands and feet, being able to impair quality of life. About 30 different therapeutic regimens described in literature reveal a lack of a single striking strategy. Recent publications showed positive results of photodynamic therapy (PDT) with 5-aminolevulinic acid (5-ALA) in the treatment of HPV-induced skin diseases, especially warts, using visible light (VIS) to stimulate an absorption band of endogenously formed protoporphyrin IX. Additional experiences adding waterfiltered infrared A (wIRA) during 5-ALA-PDT revealed positive effects. Aim of the study: First prospective randomised controlled blind study including PDT and wIRA in the treatment of recalcitrant common hand and foot warts. Comparison of "5-ALA cream (ALA) vs. placebo cream (PLC)" and "irradiation with visible light and wIRA (VIS+wIRA) vs. irradiation with visible light alone (VIS)". Methods: Pre-treatment with keratolysis (salicylic acid) and curettage. PDT treatment: topical application of 5-ALA (Medac) in "unguentum emulsificans aquosum" vs. placebo; irradiation: combination of VIS and a large amount of wIRA (Hydrosun® radiator type 501, 4 mm water cuvette, waterfiltered spectrum 590-1400 nm, contact-free, typically painless) vs. VIS alone. Post-treatment with retinoic acid ointment. One to three therapy cycles every 3 weeks. Main variable of interest: "Percent change of total wart area of each patient over the time" (18 weeks). Global judgement by patient and by physician and subjective rating of feeling/pain (visual analogue scales). 80 patients with therapy-resistant common hand and foot warts were assigned randomly into one of the four therapy groups with comparable numbers of warts at comparable sites in all groups. Results: The individual total wart area decreased during 18 weeks in group 1 (ALA+VIS+wIRA) and in group 2 (PLC+VIS+wIRA) significantly more than in both groups without wIRA (group 3 (ALA+VIS) and 4 (PLC+VIS)): medians and interquartile ranges: -94% (-100%/-84%) vs. -99% (-100%/-71%) vs. -47% (-75%/0%) vs. -73% (-92%/-27%). After 18 weeks the two groups with wIRA differed remarkably from the two groups without wIRA: 42% vs. 7% completely cured patients; 72% vs. 34% vanished warts. Global judgement by patient and by physician and subjective rating of feeling was much better in the two groups with wIRA than in the two groups without wIRA. Conclusions: The above described complete treatment scheme of hand and foot warts (keratolysis, curettage, PDT treatment, irradiation with VIS+wIRA, retinoic acid ointment; three therapy cycles every 3 weeks) proved to be effective. Within this treatment scheme wIRA as non-invasive and painless treatment modality revealed to be an important, effective factor, while photodynamic therapy with 5-ALA in the described form did not contribute recognisably - neither alone (without wIRA) nor in combination with wIRA - to a clinical improvement. For future treatment of warts an even improved scheme is proposed: one treatment cycle (keratolysis, curettage, wIRA, without PDT) once a week for six to nine weeks. © 2004 Fuchs et al; licensee German Medical Science. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL : http://www.egms.de/en/gms/volume2.shtmlHintergrund: Vulgäre Warzen (Verrucae vulgares) sind humane Papillomvirus-Infektionen (HPV) mit einer hohen Inzidenz und Prävalenz, die am häufigsten Hände und Füße befallen und die in der Lage sind, die Lebensqualität zu beeinträchtigen. Etwa 30 in der Literatur beschriebene Therapieverfahren zeugen von einem Mangel an einer einzigen überzeugenden Strategie. Jüngste Veröffentlichungen zeigten positive Ergebnisse der Photodynamischen Therapie (PDT) mit 5-Aminolävulinsäure (5-ALA) in der Therapie von HPV-induzierten Hautkrankheiten, besonders Warzen, wobei sichtbares Licht (VIS) verwendet wird, um ein Absorptionsband des endogen aus 5-ALA gebildeten Protoporphyrin IX zu stimulieren. Weitere Erfahrungen, wassergefiltertes Infrarot A (wIRA) während der 5-ALA-PDT zusätzlich anzuwenden, offenbarten positive Wirkungen. Ziel der Untersuchung: Erste prospektive randomisierte kontrollierte Blind-Studie, die PDT und wIRA in die Behandlung von therapierefraktären vulgären Hand- und Fußwarzen einbezieht. Vergleich von "5-ALA-Salbe (ALA) vs. Placebo-Salbe (PLC)" und "Bestrahlung mit sichtbarem Licht und wIRA (VIS+wIRA) vs. Bestrahlung mit sichtbarem Licht allein (VIS)". Methoden: Vorbehandlung mit Keratolyse (Salizylsäure) und Kürettage. Photodynamische Therapie (PDT): topische Applikation von 5-ALA (Medac) in "Unguentum emulsificans aquosum" vs. Placebo; Bestrahlung: Kombination von sichtbarem Licht (VIS) und einem hohen Maß an wassergefiltertem Infrarot A (wIRA) (Hydrosun®-Strahler Typ 501, 4 mm Wasserküvette, wassergefiltertes Spektrum 590-1400 nm, kontaktfrei, typischerweise schmerzlos) vs. sichtbares Licht (VIS) allein. Nachbehandlung mit Vitamin-A-Säure-Salbe. Ein bis drei Therapiezyklen im Abstand von 3 Wochen. Hauptzielvariable: "Prozentuale Änderung der Gesamtwarzenfläche jedes Patienten über die Zeit" (18 Wochen). Globales Urteil von Patient und von Arzt sowie subjektive Einschätzung von Empfindung/Schmerz (visuelle Analogskalen). 80 Patienten mit therapierefraktären vulgären Hand- und Fußwarzen wurden randomisiert einer der vier Behandlungsgruppen (mit vergleichbarer Anzahl an Warzen in vergleichbaren Lokalisationen in allen Gruppen) zugeteilt. Ergebnisse: Die individuelle Gesamtwarzenfläche nahm während 18 Wochen in Gruppe 1 (ALA+VIS+wIRA) und in Gruppe 2 (PLC+VIS+wIRA) signifikant mehr als in den beiden Gruppen ohne wIRA (Gruppe 3 (ALA+VIS) und 4 (PLC+VIS)) ab: Mediane und Interquartil-Spannen: -94% (-100%/-84%) vs. -99% (-100%/-71%) vs. -47% (-75%/0%) vs. -73% (-92%/-27%). Nach 18 Wochen unterschieden sich die zwei Gruppen mit wIRA deutlich von den zwei Gruppen ohne wIRA: 42% vs. 7% komplett geheilte Patienten; 72% vs. 34% völlig verschwundene Warzen. Das globale Urteil von Patient und von Arzt und die subjektive Einschätzung des Empfindens waren in den zwei Gruppen mit wIRA viel besser als in den zwei Gruppen ohne wIRA. Folgerungen: Das oben beschriebene vollständige Therapieschema von Hand- und Fußwarzen (Keratolyse, Kürettage, Photodynamische Therapie, Bestrahlung mit VIS+wIRA, Vitamin-A-Säure-Salbe; drei Therapiezyklen im Abstand von 3 Wochen) erwies sich als effektiv. Innerhalb des Therapieschemas zeigte sich wIRA - als nicht-invasive und schmerzlose Therapiemodalität - als ein wichtiger, effektiver Faktor, während die Photodynamische Therapie mit 5-ALA in der beschriebenen Form nicht erkennbar - weder alleine (ohne wIRA) noch in Kombination mit wIRA - zu einer klinischen Verbesserung beitrug. Für die zukünftige Behandlung von Warzen wird ein weiter verbessertes Schema vorgeschlagen: ein Therapiezyklus (Keratolyse, Kürettage, wIRA, ohne PDT) einmal pro Woche für sechs bis neun Wochen

Rapid immunoassays for diagnosis of heparin-induced thrombocytopenia: Comparison of diagnostic accuracy, reproducibility, and costs in clinical practice.

Immunoassays are crucial in the work-up of patients with suspected heparin-induced thrombocytopenia (HIT) and rapid tests have been recently developed. However, comparative data on diagnostic accuracy, reproducibility, and analytical costs of different immunoassays in clinical practice are limited. Samples of 179 consecutive patients evaluated for suspected HIT in clinical practice using a polyspecific enzyme-linked immunoabsorbent assay (GTI diagnostics; ELISA) and a rapid particle gel immunoassay (PaGIA), were additionally analysed with a IgG-specific chemiluminescent immunoassay (AcuStar HIT-IgG). Presence of HIT was defined as a positive functional heparin-induced platelet aggregation test. Diagnostic accuracy was determined for low, intermediate and high thresholds as previously established (ELISA: optical density 0.4, 1.3, and 2.0 respectively; PaGIA: positive/negative, titre of 4, titre of 32; AcuStar HIT-IgG: 1.0 U/ml, 2.8, 9.4) and reproducibility was assessed by repeated measurements. Costs of test determination were calculated taking reagents, controls, and working time of technicians according to Swiss health care system into account. Data on PaGIA results were available for 171 patients (95.5%), ELISA for 144 patients (80.4%), and AcuStar HIT-IgG for 179 patients (100%). Sensitivity was above 95% for all assays at low and intermediate thresholds. Specificity increased with higher thresholds and was above 90% for all assays with intermediate and high thresholds. Specificity of AcuStar HIT-IgG (92.8%; 95% CI 87.7, 96.2) was significantly higher than PaGIA (83.0%; 95% CI 76.3, 88.5) and higher than ELISA (81.8%, 95% CI 74.2, 88.0) at low threshold (p<0.05). Reproducibility was adequate for all assays. Total costs per test were CHF 51.02 for ELISA, 117.70 for AcuStar HIT-IgG, and 83.13 for PaGIA. We observed favourable diagnostic accuracy measures and a high reproducibility for PaGIA and AcuStar HIT-IgG. Implementation into 24-hours-service might improve patient care but the results must be confirmed in other settings and larger populations as well

Antibacterial mono- and sesquiterpene esters of benzoic acids from Iranian propolis

<p>Abstract</p> <p>Background</p> <p>Propolis (bee glue) has been used as a remedy since ancient times. Propolis from unexplored regions attracts the attention of scientists in the search for new bioactive molecules.</p> <p>Results</p> <p>From Iranian propolis from the Isfahan province, five individual components were isolated: the prenylated coumarin suberosin <b>1</b>, and four terpene esters: tschimgin (bornyl <it>p</it>-hydroxybenzoate) <b>2</b>, tschimganin (bornyl vanillate) <b>3</b>, ferutinin (ferutinol <it>p</it>-hydroxybenzoate) <b>4, </b>and tefernin (ferutinol vanillate) <b>5</b>. All of them were found for the first time in propolis. Compounds <b>2 </b>- <b>5 </b>demonstrated activity against <it>Staphylococcus aureus</it>.</p> <p>Conclusions</p> <p>The results of the present study are consistent with the idea that propolis from unexplored regions is a promising source of biologically active compounds.</p

Antitumoral and antiangiogenic activity of Portuguese propolis in in vitro and in vivo models

Propolis, a natural product, has important biological properties, however, studies with Portuguese propolis are scarce. Thus, we aimed to characterize the chemical composition and the antitumoural and antiangiogenic activities of a sample from Pereiro (Portugal). The chemical profile of our propolis sample (P10.EE) is similar to the poplar propolis type. P10.EE decreased cell viability of different tumour cells, being less cytotoxic against non-tumoural cells. P10.EE decreased MDA-MB-231 and DU145 cell proliferation and migration, with cell cycle changes and increased cell death. The increased glucose consumption and lactate production in MDA-MB-231 cells is explained by an increased expression of different metabolism-related proteins. P10.EE induced a decrease in HBMEC cells total biomass and proliferation and decreased vessel sprouting in the chicken chorioallantoic membrane. Additionally, P10.EE potentiates paclitaxel effect in MDA-MB-231 and DU145 cells. Concluding, P10.EE can be a good candidate for cancer drug development since it affects different characteristics that dictate tumorigenesis.This work was supported by the Life and Health Sciences Research Institute, University of Minho, Portugal, and Fundacao para a Ciencia e Tecnologia (FCT) (SFRH/BD/5199712012 to V.M.G.), through Fundo Europeu de Desenvolvimento Regional-QREN-COMPETE, projects PTDC/AAC-CLI1098308/2008 and PTDC/AAC-CLI/11809212010 and also CERNAS (project PEst-OE/AGR/UI0681/2011)

Fingerprinting of propolis by easy ambient sonic-spray ionization mass spectrometry

Chemical profiles of a representative set of 49 propolis ethanolic extracts collected worldwide (North and South America, Europe, Asia and Oceania) were obtained via easy ambient sonic-spray ionization mass spectrometry (EASI-MS). This simple and easily implemented fingerprinting technique analyses directly (without any pre-separation or sample manipulation) a tiny droplet of the ethanolic extract placed on a inert surface under ambient conditions. Data acquisition took about a minute per sample with no substantial sample carry-over. Extraction of propolis with ethanol by using an ultrasonic bath method gave EASI-MS data similar to the traditional maceration method, reducing total analysis time for the crude propolis resin from a week to just ca 1 h. Principal component analysis of the EASI-MS data is shown to group samples according to the plant sources of their resins, which characterizes their geographical origin. © 2009 Elsevier B.V. All rights reserved

Electrospray Ionization Mass Spectrometry Fingerprinting Of Propolis.

Crude ethanolic extracts of propolis, a natural resin, have been directly analysed using electrospray ionization mass (ESI-MS) and tandem mass spectrometry (ESI-MS/MS) in the negative ion mode. European, North American and African samples have been analyzed, but emphasis has been given to Brazilian propolis which displays diverse and region-dependent chemical composition. ESI-MS provides characteristic fingerprint mass spectra, with propolis samples being divided into well-defined groups directly related to their geographical origins. Chemometric multivariate analysis statistically demonstrates the reliability of the ESI-MS fingerprinting method for propolis. On-line ESI-MS/MS tandem mass spectrometry of characteristic [M - H](-) ion markers provides an additional dimension of fingerprinting selectivity, while structurally characterizing the ESI-MS marker components of propolis. By comparison with standards, eight such markers have been identified: para-coumaric acid, 3-methoxy-4-hydroxycinnamaldehyde, 2,2-dimethyl-6-carboxyethenyl-2H-1-benzopyran, 3-prenyl-4-hydroxycinnamic acid, chrysin, pinocembrin, 3,5-diprenyl-4-hydroxycinnamic acid and dicaffeoylquinic acid. The negative mode ESI-MS fingerprinting method is capable of discerning distinct composition patterns to typify, to screen the sample origin and to reveal characteristic details of the more polar and acidic chemical components of propolis samples from different regions of the world.129739-4

Different extraction methods of biologically active components from propolis: a preliminary study

Abstract Background Propolis is widely used in apitherapy, preparations, and food and beverage additives. Various extraction techniques were applied in the extraction of the biologically active constituents of poplar type propolis in order to compare their efficiency. The methods employed were: traditional maceration extraction, ultrasound extraction (UE), and microwave assisted extraction (MAE). Results The total amounts of extracted phenolics and flavonoids were determined, and the effectiveness of the methods compared. MAE was very rapid but led to the extraction of a large amount of non-phenolic and non-flavonoid material. UE gave the highest percentage of extracted phenolics. Conclusion Compared to the maceration extraction, MAE and UE methods provided high extraction yield, requiring short timeframes and less labour. UE was shown to be the most efficient method based on yield, extraction time and selectivity.</p

Antiproliferative effects of Tubi-bee propolis in glioblastoma cell lines

Propolis is a resin formed by a complex chemical composition of substances that bees collect from plants. Since ancient times, propolis has been used in folk medicine, due to its biological properties, that include antimicrobial, anti-inflammatory, antitumoral and immunomodulatory activities. Glioblastoma is the most common human brain tumor. Despite the improvements in GBM standard treatment, patients’ prognosis is still very poor. The aim of this work was to evaluate in vitro the Tubi-bee propolis effects on human glioblastoma (U251 and U343) and fibroblast (MRC-5) cell lines. Proliferation, clonogenic capacity and apoptosis were analyzed after treatment with 1 mg/mL and 2 mg/mL propolis concentrations for different time periods. Additionally, glioblastoma cell lines were submitted to treatment with propolis combined with temozolomide (TMZ). Data showed an antiproliferative effect of tubi-bee propolis against glioblastoma and fibroblast cell lines. Combination of propolis with TMZ had a synergic anti-proliferative effect. Moreover, propolis caused decrease in colony formation in glioblastoma cell lines. Propolis treatment had no effects on apoptosis, demonstrating a cytostatic action. Further investigations are needed to elucidate the molecular mechanism of the antitumor effect of propolis, and the study of its individual components may reveal specific molecules with antiproliferative capacity

Analytical methods applied to diverse types of Brazilian propolis

Propolis is a bee product, composed mainly of plant resins and beeswax, therefore its chemical composition varies due to the geographic and plant origins of these resins, as well as the species of bee. Brazil is an important supplier of propolis on the world market and, although green colored propolis from the southeast is the most known and studied, several other types of propolis from Apis mellifera and native stingless bees (also called cerumen) can be found. Propolis is usually consumed as an extract, so the type of solvent and extractive procedures employed further affect its composition. Methods used for the extraction; analysis the percentage of resins, wax and insoluble material in crude propolis; determination of phenolic, flavonoid, amino acid and heavy metal contents are reviewed herein. Different chromatographic methods applied to the separation, identification and quantification of Brazilian propolis components and their relative strengths are discussed; as well as direct insertion mass spectrometry fingerprinting

Portuguese propolis disturbs glycolytic metabolism of human colorectal cancer in vitro

Propolis is a resin collected by bees from plant buds and exudates, which is further processed through the activity of bee enzymes. Propolis has been shown to possess many biological and pharmacological properties, such as antimicrobial, antioxidant, immunostimulant and antitumor activities. Due to this bioactivity profile, this resin can become an alternative, economic and safe source of natural bioactive compounds.Antitumor action has been reported in vitro and in vivo for propolis extracts or its isolated compounds; however, Portuguese propolis has been little explored. The aim of this work was to evaluate the in vitro antitumor activity of Portuguese propolis on the human colon carcinoma cell line HCT-15, assessing the effect of different fractions (hexane, chloroform and ethanol residual) of a propolis ethanol extract on cell viability, proliferation, metabolism and death. METHODS: Propolis from Angra do Heroísmo (Azores) was extracted with ethanol and sequentially fractionated in solvents with increasing polarity, n-hexane and chloroform. To assess cell viability, cell proliferation and cell death, Sulforhodamine B, BrDU incorporation assay and Anexin V/Propidium iodide were used, respectively. Glycolytic metabolism was estimated using specific kits. RESULTS: All propolis samples exhibited a cytotoxic effect against tumor cells, in a dose- and time-dependent way. Chloroform fraction, the most enriched in phenolic compounds, appears to be the most active, both in terms of inhibition of viability and cell death. Data also show that this cytotoxicity involves disturbance in tumor cell glycolytic metabolism, seen by a decrease in glucose consumption and lactate production. CONCLUSION: Our results show that Portuguese propolis from Angra do Heroísmo (Azores) can be a potential therapeutic agent against human colorectal cancer.We thank the Portuguese Science and Technology Foundation (FCT) for VMG fellowship (ref. SFRH/BI/33503/2008). The authors thank Mr. Antonio Marques from Frutercoop - Azores, who kindly collected and provided the propolis sample for the study