HPTLC fingerprint profile analysis of cocoa proanthocyanidins depending on origin and genotype.

Cocoa beans are rich in bioactive phytochemicals such as alkaloids, anthocyanins, as well as monomeric and oligomeric flavan-3-ols. A high performance thin layer chromatography (HPTLC) method was developed for a fast analysis of a fingerprint of bioactive compounds present in cocoa beans depending on genotype and origin. Evaluation was performed using HPTLC followed by densitometry. The best separation for a fingerprint consisting of eight phenolic compounds as markers was achieved on silica gel 60 F254 HPTLC plates with a solvent mixture of ethyl formate, formic acid, water, toluene 30/4/3/1.5 (v/v/v/v). Staining with Fast Blue Salt B enabled visualization and quantitative evaluation. Compounds of interest were confirmed by eluting zones using a TLC-MS interface. LODs were ≤ 100ng/zone for all compounds. The fingerprints obtained are useful for quality control of cocoa beans allowing their differentiation according to cocoa or chocolate sorts, varieties, and origins.

Persistent organic pollutants in shallow percolated water of the Alps Karst system (Zugspitze summit, Germany).

In the German Calcareous Alps at the Zugspitze, percolated water close to a permafrost bedrock in a tunnel system was monitored long-term for polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF), polychlorinated biphenyls (PCB), polycyclic aromatic hydrocarbons (PAH), and 28 organochlorine pesticides (OCP). Semi-permeable membrane devices (SPMD) were deployed in a temporary surface water system at the Zugspitze plateau and analysed for PCB, PAH, and OCP. The high-volume water sampling was successfully implemented and all compounds were identified in the water percolated through the Karst system. However, the percentage distribution of contaminants in the percolated water differed significantly from that found in surface waters. The highest chlorinated PCDD homologues were the predominant compounds of the PCDD/F family, whereas percentages of PCB #52 increased in percolated water. Toxic equivalent values (TEQ) of samples ranged from 2.0 to 4.2pgTEQ/m3 and from 0.017 to 0.069pgTEQ/m3 for PCDD/F and PCB, respectively. Low and intermediate molecular weight PAH were the prevailing compounds in the samples. Endosulfan sulfate, endrin, and cis-heptachlor epoxide were enhanced after water percolation through the Karst system in comparison with the surface waters (wet deposition). The relative enrichment on these pesticides was related to the environmental bedrock conditions and glacier melting sources. In summary, the Karst system highly influenced the fate of organic persistent pollutants generating different chemical patterns in their percolated waters than those found at the surface systems.

Determination of oligomeric proanthocyanidins and their antioxidant capacity from different chocolate manufacturing stages using the NP-HPLC-online-DPPH methodology.

Cocoa beans are a well-known source of antioxidant polyphenols. Especially individual oligomeric proanthocyanidins demonstrated a significant contribution to the total antioxidant activity of cocoa compared to monomeric compounds. An NP-HPLC-online-DPPH assay was developed for separating the homologous series of oligomeric proanthocyanidins and the simultaneous assessment of their antioxidant capacity in relation to the degree of polymerization (DP). The present study describes the influence of the different stages of a lab-scale chocolate manufacturing process on the content of oligomeric proanthocyanidins and their antioxidant capacity. The sum of the total proanthocyanidin content (∑ DP1-DP13) decreased from 30mg epicatechin equivalents per gram non-fat dry matter in raw fresh cocoa beans to 6mg epicatechin equivalents per gram in the final chocolate. The antioxidant capacity decreased accordingly from 25mg epicatechin equivalents per gram non-fat dry matter in raw fresh cocoa beans to 4mg/g in the final chocolate product.

Severe motor aphasia after reinfusion of cryopreserved autologous stem cells after myeloablative conditioning.

BACKGROUND: Autologous peripheral blood stem cells (PBSCs) are usually cryopreserved before high-dose chemotherapy (HDCT) and autologous peripheral blood stem cell transplantation (PBSCT). The freezing process requires the addition of cryoprotectants such as dimethyl sulfoxide (DMSO), which is vital for cell viability in frozen aliquots. DMSO has a number of well-described side effects. However, severe neurologic side effects assigned to DMSO are exceedingly rare. CASE REPORT: A 64-year-old female underwent HDCT followed by PBSCT as consolidation therapy in relapsed high-grade (Grade 3B) Stage IIIA follicular lymphoma. PBSCs were mobilized using granulocyte-colony stimulating factor and plerixafor after the second cycle of R-DHAP (rituximab, dexamethasone, high-dose Ara-C, cisplatin) salvage chemotherapy. A total of 7.18 × 106 /kg body weight CD34+ cells were cryopreserved using 10% DMSO. HDCT was administered some weeks later followed by reinfusion of two bags of PBSCs, each containing 98 mL with 1.6 × 106 /kg body weight CD34+ cells. Within a few minutes the patient developed a motor aphasia and became very agitated. Brain imaging did not reveal any pathologic finding. After being transferred to the intensive care unit the patient's condition steadily improved and the motor aphasia resolved completely within 6 hours after its onset. CONCLUSION: This is, to our knowledge, the first report to describe an episode of severe motor aphasia during PBSCT. Given the close timely correlation with PBSCT, this episode appears to be caused by dimethyl sulfoxide (DMSO) and might possibly have been prevented by use of lower concentrations of DMSO.

Extraction of cocoa proanthocyanidins and their fractionation by sequential centrifugal partition chromatography and gel permeation chromatography.

Cocoa beans contain secondary metabolites ranging from simple alkaloids to complex polyphenols with most of them believed to possess significant health benefits. The increasing interest in these health effects has prompted the need to develop techniques for their extraction, fractionation, separation, and analysis. This work provides an update on analytical procedures with a focus on establishing a gentle extraction technique. Cocoa beans were finely ground to an average particle size of <100 µm, defatted at 20 °C using n-hexane, and extracted three times with 50 % aqueous acetone at 50 °C. Determination of the total phenolic content was done using the Folin-Ciocalteu assay, the concentration of individual polyphenols was analyzed by electrospray ionization high performance liquid chromatography-mass spectrometry (ESI-HPLC/MS). Fractions of bioactive compounds were separated by combining sequential centrifugal partition chromatography (SCPC) and gel permeation column chromatography using Sephadex LH-20. For SCPC, a two-phase solvent system consisting of ethyl acetate/n-butanol/water (4:1:5, v/v/v) was successfully applied for the separation of theobromine, caffeine, and representatives of the two main phenolic compound classes flavan-3-ols and flavonols. Gel permeation chromatography on Sephadex LH-20 using a stepwise elution sequence with aqueous acetone has been shown for effectively separating individual flavan-3-ols. Separation was obtained for (-)-epicatechin, proanthocyanidin dimer B2, trimer C1, and tetramer cinnamtannin A2. The purity of alkaloids and phenolic compounds was determined by HPLC analysis and their chemical identity was confirmed by mass spectrometry.

Bendamustine and rituximab in combination with lenalidomide in patients with chronic lymphocytic leukemia.

PURPOSE: A phase I/II trial to assess safety and efficacy of the combination bendamustine, rituximab, and lenalidomide (BRL) in patients with chronic lymphocytic leukemia (CLL). PATIENTS AND METHODS: Seventeen relapsed or refractory (R/R) and five previously untreated (FL) CLL patients were enrolled in the trial. In the R/R cohort, four different dose levels of lenalidomide (maximum 15 mg/d) were used. In the FL cohort, lenalidomide was dose escalated from 5 mg/d to 15 mg/d. Bendamustine was used at doses of 50 or 90 mg/m(2) for R/R or FL treatment, respectively. 375 mg/m(2) Rituximab were used for the first and 500 mg/m(2) for subsequent treatment courses. Treatment consisted of up to six courses of 28 d. RESULTS: The maximal tolerable dose of lenalidomide was 5 mg/d. The response rate was 47.1% in R/R and 60% in FL patients. Median progression-free survival was 8.0 months. Median overall survival was 22.9 and 12.3 months, respectively, in R/R and FL patients. Grade 3/4 hematological toxicity was observed in 71.4%, and severe infections in 47.6% of patients. Due to high toxicity and low response rate of BRL, the trial was closed prematurely. CONCLUSION: BRL was associated with a high toxicity rate, a high number of treatment interruptions, and a low remission rate. Therefore, BRL cannot be considered an appropriate treatment option for patients with CLL.

Tumor patients in psychodynamic psychotherapy including daydreaming: can imagery enhance primary process and positive emotions?

This therapy process study investigates the use of guided affective imagery for tumor patients. The therapeutic access to tumor patients is generally described as complex and challenging because of a disturbed emotion regulation and a defensive focus on reality. After autologous blood stem cell transplantation, 29 patients were treated with psychotherapy, including two daydreaming imagery sessions. Three text-analytical measures--Affective Dictionary Ulm, Regressive Imagery Dictionary, and Computerized Referential Activity for verbatim session transcripts--as well as the Quality of Life Questionnaire and the Karnofsky Performance Status were administered. Results show that guided affective imagery was able to enhance the psychotherapeutic process in tumor patients by activating the primary process, decreasing anxiety, and increasing referential activity. The positive emotional shift during imagery was achieved by the patients irrespective of their oncological severity status. Study limitations and future directions for research are discussed.

Social support and quality of life of patients prior to stem cell or bone marrow transplantation.

OBJECTIVES: To determine the positive and detrimental aspects of social support prior to allogeneic or autologous bone marrow transplantation/stem cell transplantation. DESIGN: Cross-sectional. METHODS: Patients completed the German version of the Illness-specific Social Support Scale (ISSS) and the Quality of Life (QoL) Questionnaire Core 30 (EORTC QLQ-C30). RESULTS: Participants were 282 patients (62% autologous SCT, 39% women, 97% haematological malignancies, 72% with partner). We found satisfactory reliability values for both ISSS scales: positive support (Cronbach's alpha=.91) and problematic support (=.73). Patients living with a partner displayed higher scores in positive interaction than patients living alone (p<.001). Analysis showed a significant main effect of partnership, p<.001; F(1)=8.345, and better scores for women, p<1; F(1)=2.758. Furthermore, we determined a negative correlation between problematic interaction and emotional/social function (p<.001) and a positive correlation with insomnia, but no correlation between positive interaction, QLQ-C30, and Karnofsky's index. CONCLUSIONS: We could distinguish between helpful and harmful support, and determine clinically important associations of problematic support. Partnership seems to be a major source of positive interaction.

Concentration measurements of PCDD/F in air and spruce needles in the Bavarian Forest and Bohemian Forest (Sumava): first results.

The aim of this project was to assess the concentration ranges of persistent organic pollutants (POPs) in air by means of active and passive sampling devices located in remote areas along the southern part of the German-Czech border. During the measuring program we also used spruce needles of different ages as biomonitors. The first results, which are restricted to PCDD/F measurements, indicate that the concentrations in both air and needles measured at four sites in the Bavarian Forest and Sumava crest region are lower than in many other regions of Bavaria and lower than during former measuring campaigns in adjacent regions of Austria and Bohemia. There is no apparent reason for the higher contamination by dioxins and furans in South Bohemia ecosystems, which are far from emission centers in central and northern Czech Republic. Other POPs will be investigated in the future.