Chronic inflammatory disorders represent one of the predominant healthcare burdens. There is evidence that the oleogum resin from Boswellia serrata trees can downmodulate pro-inflammatory processes. Lipid micellar preparations of Boswellia serrata have been introduced to the market to overcome the low bioavailability of nonformulated Boswellia oleogum resin preparations. In this study, we aimed to compare the anti-inflammatory effects of two different Boswellia serrata nutraceuticals: the native, nonformulated Biotikon® BS-85 and the micellar Boswellia-Loges®. We have previously shown that single oral administration of 800 mg of either formulation reduces the release of proinflammatory cytokines TNF-α, IL-1β, and IL-6 by LPS-activated blood of donors. Here we show that under the same conditions, the production of IL-17A was increased by the nonformulated, native extract of Boswellia serrata oleogum resin. In vitro, the nonformulated but not the micellar formulation of Boswellia serrata oleogum resin decreased the release of IFN-γ, TNF-α, and IL-2 by TCR-activated lymphocytes. Both formulations as well as the bioactive principles boswellic acids lowered NF-κB activity in TCR-activated T lymphocytes. Similarly, both Boswellia serrata formulations and boswellic acids reduced NFAT activity in TCR-activated T lymphocytes. The nonformulated Boswellia serrata extract exhibited higher inhibitory activity on the release of T-cell cytokines. The results suggest that nutraceuticals containing the nonformulated oleogum extract of Boswellia serrata might be more effective in hampering chronic inflammatory disorders characterized by increased activity of T cells than the micellar formulations.

Mangosteen (Garcinia mangostana) is well-known for its nutritional value and health benefits. Breast cancer is the most common cancer and the leading cause of cancer-related mortality among females worldwide. Here we show that the prenylated xanthones, α-mangostin, γ-mangostin, 9-hydroxycalabaxanthone (9-HCX), and garcinone E from the mangosteen pericarp exhibit cytotoxicity against a panel of human cancer cell lines including lung adenocarcinoma (A549), cervical carcinoma (HeLa), prostatic carcinoma (DU 145), pancreatic carcinoma (MIA PaCa-2), hepatocellular carcinoma (Hep G2), bladder urothelial cancer (5637), as well as the triple-negative breast cancer cells MDA-MB-231. In line with its higher predicted bioactivity score compared to other prenylated xanthones, 9-HCX induced the strongest antiproliferative and proapoptotic effects in MDA-MB-231 breast cancer xenografts in vivo. In different in vitro models, we demonstrate that prenylated xanthones from G. mangostana target mitochondria in cancer cells by inhibition of the mitochondrial respiratory chain complex II (α-mangostin, γ-mangostin, and garcinone E) and complex III (9-HCX) as shown in isolated mitochondria. Accordingly, oxidative mitochondrial respiration (OXPHOS) was inhibited, mitochondrial proton leak increased, and adenosine triphosphate (ATP) synthesis decreased as analyzed by Seahorse assay in MDA-MB-231 cells. Hence, the prenylated xanthones increased mitochondrial superoxide levels, induced mitochondrial membrane permeabilization, and initiated caspase 3/7-mediated apoptosis in MDA-MB-231 triple-negative breast cancer cells. Thus, prenylated xanthones from Garcinia mangostana exhibit anticancer activity based on interference with the mitochondrial respiration.

Dust often contains chemical airborne pollutants that might negatively affect human health. Polystyrene is one of the most widely used types of plastic. Bulk polystyrene exhibits no short-term cytotoxicity. However, during degradation of polystyrene, small nanoparticles are formed. Due to specific properties, such as the large surface to volume ratio, toxicity of nanoparticles might be different to that of the bulk material. Hence, particularly the surface chemistry is crucial for nanoparticle behaviour in biological environment. For this study, carboxyl(PS-COOH) and amino-functionalized (PS-NH2) nanoparticles were prepared by free-radical copolymerization in a direct (oil in water) miniemulsion system. We show that surface functionalization of polystyrene nanoparticles with amino groups (PS-NH2), but not with carboxyl groups (PS-COOH), induced inhibition of proliferation in a monocytic cell line and induce a specific cell death, apoptosis. In PS-NH2–treated cells, acidic vesicular organelles exhibited elevated pH and impaired processing of a lysosomal enzyme. Moreover, solely in PS-NH2-treated cells, but not in PS-COOHtreated cells, this was followed by permeabilization of acidic vesicular organelles and induction of cell death. These data indicate that surface charge of nanoplastics defines their effects on biological systems and can be used to predict environmental toxicity of nanoplastics.

IntroductionThe use of Complementary and Integrative Medicine (CIM) is very popular among the general population in Germany. However, international studies show that nurses, physicians, and other health care professionals (HCPs) at hospitals often do not feel sufficiently informed about different CIM approaches. Moreover, they do not feel trained enough to counsel their patients appropriately. In the German-speaking context, particularly within university hospitals, research on this subject is scarce. Therefore, the aim of this explorative study was to evaluate attitudes, subjective knowledge, and needs regarding CIM among HCPs with direct patient interaction across all four university hospitals in the federal state of Baden-Württemberg, Germany (Tübingen, Ulm, Freiburg, Heidelberg).MethodsThe multicenter, cross-sectional, anonymous full survey was conducted online using a self-developed, semi-structured, web-based questionnaire. Recruitment took place via all-inclusive e-mail distribution lists of all four university hospitals.ResultsA total of n = 2,026 participants (response rate varied by location from about 5 to 14%) fully answered the questionnaire. Nurses constituted the largest professional group (n = 1,196; 59%), followed by physicians (n = 567; 28%), physiotherapists (n = 54), psychologists (n = 48), midwives (n = 37), and other professions (n = 124). More than two-thirds (71%, n = 1,437) of the participants were female and 14% (n = 286) reported additional training in CIM. The overall attitude toward CIM (10-point Likert scale, 10 = “very favorable”) was clearly positive (M ± SD: 7.43 ± 2.33), with notable differences between professional groups: midwives (9.05 ± 1.18), physiotherapists (8.44 ± 1.74), and nurses (8.08 ± 1.95) expressed the highest support, whereas physicians (5.80 ± 2.39) the lowest. 42% of the participants incorporated CIM in patient care (from 33% of physicians to 86% of midwives). Overall, relaxation therapy (n = 1,951; 96%), external applications (n = 1,911; 94%), massage (n = 1,836; 91%), and meditation/mindfulness (n = 1,812; 89%) were rated as useful or rather useful for patients. The average self-assessed knowledge level about CIM was moderate (M ± SD: 5.83 ± 2.03). Most of the participants found CIM training at university hospitals important and saw research about CIM as one of the tasks of university hospitals. The participants expressed the highest interest in education for acupuncture/acupressure, relaxation therapies, and manual medicine.DiscussionThis comprehensive survey of health care professionals (HCPs) at university hospitals in Germany reveals a clearly positive disposition toward CIM, aligning with findings from other hospital-based surveys and highlighting differences among professional groups. While most therapies deemed beneficial for patient care are supported by positive evidence, further research is required for others. Given the average self-reported knowledge of CIM, targeted education is essential to meet the needs of both HCPs and patients and to ensure the provision of evidence-based information on the risks and benefits of CIM.

Background: Extracts of oleogum resins of Boswellia trees possess anti-inflammatory activities. Micellar formulations have been developed to increase the oral bioavailability of bioactive boswellic and lupeolic acids. Purpose: The current single-dose crossover clinical trial compares for the first time pharmacokinetics/pharmacodynamics of two Boswellia serrata nutraceuticals, native Biotikon (R) (R) BS-85 and micellar Boswellia-Loges (R). (R). Methods: After oral administration of the study preparations (800 mg) to 20 healthy volunteers, plasma concentrations of 8 boswellic and lupeolic acids were measured by using HPLC-MS/MS for up to 48 h Blood samples collected 2 and 5 h after drug administration were stimulated for 24 h with endotoxic lipopolysaccharide. The release of proinflammatory cytokines analyzed by flow cytometry was used as readout of the pharmacodynamic properties of the preparations. Registration: German Clinical Trials Register (DRKS) No. DRKS00027369. Results: Administration of the micellar extract significantly increased C max, AUC0-48, 0-48 , and shortened T max for all boswellic and lupeolic acids compared to native extract. Accordingly, their relative bioavailability increased to 1,720-4,291 % with the highest difference for acetyl-11-keto-(3-boswellic acid (AKBA). Both preparations reduced the release of TNF-alpha and the native formulation diminished also IL-1(3 and IL-6. However, no significant differences were observed between the preparations, except for a higher decrease in IL-1(3 by the native formulation Biotikon (R) (R) BS-85. In a lymphocytic gene reporter cell line, both nutraceuticals similarly inhibited the NF-kappa B transcription factor activity as well as the TNF-alpha release, yet the native formulation Biotikon (R) BS-85 (R) BS-85 was more efficient in inhibiting TNF-alpha. Conclusion: Administration of the micellar Boswellia serrata nutraceutical increased the oral bioavailability of boswellic and lupeolic acids. Yet, the increase in plasma concentration did not enhance the anti-inflammatory efficacy of the micellar extract compared to the native extract in this ex vivo model.

Supplementary Figure 2 from Targeting XIAP Bypasses Bcl-2–Mediated Resistance to TRAIL and Cooperates with TRAIL to Suppress Pancreatic Cancer Growth <i>In vitro</i> and <i>In vivo</i>

Clinicopathological characteristics of the included patients with myxoid liposarcoma and semi-quantitative immunohistochemistry results

Supplementary Legends and Methods from Targeting XIAP Bypasses Bcl-2–Mediated Resistance to TRAIL and Cooperates with TRAIL to Suppress Pancreatic Cancer Growth In vitro and In vivo

PDF file - 182K, Supplemental Figure 1. B cells activated with IL-21 and anti-BCR secrete enzymatically active granzyme B. Supplemental Figure 2. Interleukin 21 in the absence of B cells does not suppress T cell proliferation. Supplemental Figure 3. GrB-secreting B cells do not induce apoptosis in T cells. Supplemental Figure 4. IL-10, CD25 and IDO are expressed on IL-21-induced GrB+ Breg, but not on GrB- B cells. Supplemental Figure 5. IL-2 retitration has no significant effect on the suppression of CD4+ T cell proliferation by GrB-secreting B cells. Supplemental Figure 6. IL-21-induced GrB secretion by B cells depends on TLR7 and TLR9 signaling. Supplemental Figure 7. B cells in the microenvironment of solid tumors sporadically express IL-10.

Supplementary Table S1: Clinicopathological characteristics of patients with myxoid liposarcoma (n=56) including semi-quantitative immunohistochemistry and mutational analyses results. Supplemental Table S2: Gene list. Supplementary Table S3: In silico tools to predict the deleterious impact of gene variants detected in myxoid liposarcoma.

Abstract The pathogenic impact of tumor-infiltrating B cells is unresolved at present, however, some studies suggest that they may have immune regulatory potential. Here, we report that the microenvironment of various solid tumors includes B cells that express granzyme B (GrB, GZMB), where these B cells can be found adjacent to interleukin (IL)-21–secreting regulatory T cells (Treg) that contribute to immune tolerance of tumor antigens. Because Tregs and plasmacytoid dendritic cells are known to modulate T-effector cells by a GrB-dependent mechanism, we hypothesized that a similar process may operate to modulate regulatory B cells (Breg). IL-21 induced outgrowth of B cells expressing high levels of GrB, which thereby limited T-cell proliferation by a GrB-dependent degradation of the T-cell receptor ζ-chain. Mechanistic investigations into how IL-21 induced GrB expression in B cells to confer Breg function revealed a CD19+CD38+CD1d+IgM+CD147+ expression signature, along with expression of additional key regulatory molecules including IL-10, CD25, and indoleamine-2,3-dioxygenase. Notably, induction of GrB by IL-21 integrated signals mediated by surface immunoglobulin M (B-cell receptor) and Toll-like receptors, each of which were enhanced with expression of the B-cell marker CD5. Our findings show for the first time that IL-21 induces GrB+ human Bregs. They also establish the existence of human B cells with a regulatory phenotype in solid tumor infiltrates, where they may contribute to the suppression of antitumor immune responses. Together, these findings may stimulate novel diagnostic and cell therapeutic approaches to better manage human cancer as well as autoimmune and graft-versus-host pathologies. Cancer Res; 73(8); 2468–79. ©2013 AACR.

MOV file - 3136K, Supplemental Video 1. B cells transfer active GrB to T cells.

Supplementary Materials and Methods: (Flow cytometry, Apoptosis assay, RNA interference and Statistical analysis)

(A) Stable SS18-SSX-expressing SCP-1 cells showed an upregulation of IGF2 promoter P2-4 transcripts (regarding Figure 3A; 28S rRNA used as loading reference). (B) 'N' nuclear and 'C' cytoplasmic protein fractions of SYO-1, FUJI and CME-1 SySa cells demonstrated cytoplasmic localization of phosphorylated YAP (Ser127) and TAZ (Ser89) (Histone H3 and GAPDH used as loading references for the nuclear and cytoplasmic fractions, respectively). (C) In SYO-1 cells, phosphorylation levels of IGF-IR (Tyr1135/6) and AKT (Ser473) were dose-dependently reduced by treatment with increasing concentrations of BMS-754807 (β-Actin used as loading reference). (D) Immunofluorescence staining of SYO-1 cells showed diminished nuclear localization of YAP and TAZ upon BMS-754807 treatment (original magnification x40).

Supplementary Figure 3 from Targeting XIAP Bypasses Bcl-2–Mediated Resistance to TRAIL and Cooperates with TRAIL to Suppress Pancreatic Cancer Growth In vitro and In vivo

Supplementary Table S1: Clinicopathological characteristics of synovial sarcoma tissue specimens (n=65) and semi-quantitative immunohistochemistry results Supplementary Table S2: Primer sequences

Supplementary Figure (S1): Activation of the IGF-IR and PI3K/Akt/GSK-3beta signaling cascade in representative myxoid liposarcoma cell lines. (S2): Representative sequencing results of PIK3CA mutation hotspots. (S3): IGF-II-effected rescue of PI3K/Akt/GSK-3beta pathway activation in FUS-DDIT3 depleted myxoid liposarcoma cells. (S4): RNAi-mediated reduction of FUS-DDIT3 oncoprotein levels. (S5): Expression and activation levels of IGF-IR and PI3K/Akt/GSK-3beta signaling effectors in A673 and Capan-1 cells. (S6): In vitro and in vivo evaluation of PPP in MLS cells. (S7): Time dependent reduction of Cyclin D1 protein levels. (S8): Flow cytometric analyses in MLS1765-92 cells. (S9): Representative CAM explants (NVP AEW541, MLS402-91 cells).