Abstract
Background
The transcription factor GATA-3 has been identified to play a central role in Th2-dependent immune pathways and is overexpressed in the mucosa of ulcerative colitis (UC) patients. ...We investigated the efficacy and safety of SB012, an enema formulation of a novel GATA-3 specific DNAzyme that specifically binds and inactivates GATA-3 messenger RNA (mRNA) in patients with active UC.
Methods
In this randomised, double-blind, placebo-controlled, multi-centre, parallel-group, phase 2a clinical trial, 20 patients with moderate-to-severe UC (Mayo Score ≥6; endoscopic subscore of the sigmoid ≥2) were randomised to receive 225 mg SB012 enema (n = 13) or placebo (n = 7) once daily for a 28-day period. The primary efficacy variable was the change in disease activity (Mayo Score) after 28 days of treatment. Clinical and endoscopic disease activity was further assessed at day 56. Endoscopy was centrally read in the study. Furthermore, a panel of biomarkers was evaluated by gene expression profiling in collected colon biopsies. This study is registered with EudraCT-Nr. 2013-004599-36.
Results
In the SB012 treated patients, the Mayo Score dropped from 8.4 ± 1.7 (n = 13) at baseline to 6.5 ± 2.6 (n = 13) at day 28 (p = 0.004), and 5.0 ± 3.5 (n = 7) at day 56 (p = 0.016). In the placebo group, the Mayo Score was 10.0 ± 2.1 (n = 6) at baseline, and 9.0 ± 1.3 (n = 6) at day 28 (not significant; ns), and 9.0 ± 1.4 (n = 2) at day 56 (ns). At day 28, there was a statistically significant improvement in the Mayo Score in the SB012 compared with the placebo group (p = 0.044). Clinical remission was reached by 0.0% (0/12) at day 28 and 42.9% (3/7) at day 56 of the SB012-treated patients, compared with 0.0% (0/6) at day 28 and 0.0% (0/2) at day 56 of the placebo-treated patients. Clinical response at day 28 occurred in 41.7% (5/12) at day 28 and 42.9% (3/7) at day 56 in the SB012 vs. 0.0% (0/6) and 0.0% (0/2) in the placebo group. Mucosal healing rates were 16.7% (2/12) at day 28 and 57.1% (4/7) at day 58 in the SB012, compared with 0.0% (0/6) and 0.0% (0/2) in the placebo treated group. The two-item PRO (PRO2) consisting of rectal bleeding=0 and stool frequency ≤1 (and at least one point decrease from baseline), combined with an endoscopy subscore ≤1, was reached by 8.3% (1/12) at day 28 and 42.9% (3/7) at day 56 of the SB012 compared with 0.0% (0/6) and 0.0% (0/2) of the placebo treated patients.SB012 was well tolerated and no safety signals compared with placebo were evident.
Conclusions
Topical application of the GATA3-specific DNAzyme SB012 was well tolerated and lead to marked clinical and endoscopic improvement in patients with active ulcerative colitis. Therefore, blockade of GATA-3 might represent a unique therapeutic approach for the treatment of ulcerative colitis patients.
Modification of spin-on-deposited porous PMO (periodic mesoporous organosilica) ultralow-k (ULK) SiCOH films (k = 2.33) containing both methyl terminal and methylene bridging groups by vacuum ...ultraviolet (VUV) emission from Xe plasma is studied. The temporal evolution of chemical composition, internal defects, and morphological properties (pore structure transformation) is studied by using Fourier transform infrared spectroscopy, in situ laser ellipsometry, spectroscopic ellipsometry, ellipsometric porosimetry (EP), positron-annihilation lifetime spectroscopy (PALS), and Doppler broadening positron-annihilation spectroscopy. Application of the different advanced diagnostics allows making conclusions on the dynamics of the chemical composition and pore structure. The time frame of the VUV exposure in the current investigation can be divided into two phases. During the first short phase, film loses almost all of its surface methyl and matrix bridging groups. An increase of material porosity due to removal of methyl groups with simultaneous matrix shrinkage is found by in situ ellipsometry. The removal of bridging bonds leads to an increase of matrix intrinsic porosity. Nevertheless, when the treated material is exposed to the ambient air, the sizes of micro- and mesopores and pores interconnectivity decrease with the VUV exposure time according to PAS and EP data. The last is the result of the additional film shrinkage caused by atmosphere exposure. During the second phase the increase of mesopore size is detected by both EP and PAS. The increase of mesopore size goes all the time as it is expected from in situ ellipsometry, but it is masked by the air exposure.
International organizations (IOs) develop institutional provisions to make sure that their policies do not violate human rights. Accordingly, whilst IOs have a greater scope of action and ability to ...promote collective goods than ever before, they also have a greater capacity to do harm. Based on ten case studies on UN and EU sanctions policy, UN and NATO peacekeeping, and World Bank and IMF lending, this book examines human rights violations which can arise from the actions of IOs rather than those of states. It further explains how powerful IOs have introduced human rights protection provisions and analyzes the features of these provisions, including differences in their design and quality. This book provides evidence of a novel legitimation strategy authoritative IOs draw on that has, as yet, never been systematically studied before.
The light curves of Type Ia supernovae (SNe Ia) are powered by gamma rays emitted by the decay of radioactive elements such as super(56)Ni and its decay products. These gamma rays are downscattered, ...absorbed, and eventually reprocessed into the optical emission that makes up the bulk of all SN observations. Detection of the gamma rays that escape the expanding star provide the only direct means to study this power source for SN Ia light curves. Unfortunately, disagreements between calculations for the gamma-ray lines have made it difficult to interpret any gamma-ray observations. Here we present a detailed comparison of the major gamma-ray line transport codes for a series of one-dimensional SN Ia models. Discrepancies in past results were due to errors in the codes, and the corrected versions of the seven different codes yield very similar results. This convergence of the simulation results allows us to infer more reliable information from the current set of gamma-ray observations of SNe Ia. The observations of SN 1986G, SN 1991T, and SN 1998bu are consistent with explosion models based on their classification: subluminous, superluminous, and normally luminous, respectively.
Cryptofolione (1) and the new cryptofolione derivative 6‐(4,6‐dimethoxy‐8‐phenyl‐octa‐1,7‐dienyl)‐4‐hydroxy‐tetrahydro‐pyran‐2‐one (2) were isolated from the fruits of Cryptocarya alba. The ...structures were elucidated by spectroscopic methods. Cryptofolione showed activity towards Trypanosoma cruzi trypomastigotes, reducing their number by 77% at 250 μg mL−1. Cryptofolione showed moderate cytotoxicity in both macrophages and T. cruzi amastigotes. It also displayed a mild inhibitory effect on the promastigote form of Leishmania spp. As both cytotoxic and trypanocidal effects are similar, the compound presented little selectivity in our assay models.
Abstract
Background
Anti-tumour necrosis factor (TNF) antibodies are effectively used for treatment in many Crohn’s disease patients. Nevertheless, a relevant subgroup of patients does not respond to ...anti-TNF therapy. Here we characterised underlying molecular mechanisms that are associated with endoscopic resistance to anti-TNF therapy.
Methods
Mucosal and blood cells were isolated from 197 Crohn’s disease patients prior and during anti-TNF therapy. Cytokine profiles, cell surface markers, signalling proteins and cell apoptosis were assessed by microarray, immunohistochemistry, qPCR, ELISA, whole organ cultures and FACS.
Results
Crohn’s disease patients responding to anti-TNF therapy displayed a significantly higher expression of TNF receptor 2 (TNFR2) but not IL23R on mucosal T cells than non-responders prior to the initiation anti-TNF therapy. We performed an array analysis regarding the differentiated gene regulation profiles in intestinal biopsies of endoscopic non-responders compared with responders during ongoing anti-TNF therapy in CD patients. Within the cohort of CD susceptible genes, there was a significant upregulation of genes that are associated with IL23R-dependent signalling pathways in anti-TNF non-responders compared with responders. Apoptosis--resistant TNFR2+IL23R+ T cells were significantly expanded in anti-TNF non-responders compared with responders and expressed the gut tropic integrins α4β7. These cells exhibited increased expression of IFN-γ, T-bet, IL-17A and RORγt compared with TNFR2+IL23R- cells, indicating a mixed Th1/Th17-like phenotype. Intestinal TNFR2+IL23R+ T cells were activated by IL-23 derived from CD14+ macrophages, which were significantly more present in non-responders compared with responders prior to anti-TNF treatment. Administration of IL-23 to anti-TNF treated mucosal organ cultures led to the expansion of CD4+IL23R+TNFR2+ lymphocytes. There was no accumulation of CD4+TNFR2+ T cells which were negative for IL23R. Functional studies demonstrated that anti-TNF-induced apoptosis in mucosal T cells is abrogated by IL-23.
Conclusions
Expansion of apoptosis-resistant intestinal TNFR2+IL23R+ T cells is associated with resistance to anti-TNF therapy in Crohn’s disease. IL-23 is centrally involved in mediating resistance to anti-TNF therapy in Crohn’s disease patients and thereby represents a suitable molecular target in Crohn’s disease patients refractory to anti-TNF therapy.
Summary Trisomy 21 alters fetal liver hematopoiesis and, in combination with somatic globin transcription factor 1 ( GATA1 ) mutations, leads to development of transient myeloproliferative disease in ...newborns. However, little is known about the morphological hematopoietic changes caused by trisomy 21 in the fetus, and to date, the exact onset of GATA1 mutations remains uncertain. Therefore, we analyzed fetal liver hematopoiesis from second trimester pregnancies in trisomy 21 and screened for GATA1 mutations. We examined 57 formalin-fixed and paraffin-embedded fetal liver specimens (49 harboring trisomy 21 and 8 controls) by immunohistochemistry for CD34, CD61, factor VIII, and glycophorin A. GATA1 exon 2 was sequenced in fetal livers and corresponding nonhematologic tissue. Cell counts of megakaryocytes ( P = .022), megakaryocytic precursors ( P = .021), and erythroid precursors were higher in trisomy 21 cases. CD34-positive hematopoietic blasts showed no statistically significant differences. No mutation was detected by GATA1 exon 2 sequencing in fetal livers from 12 to 25 weeks of gestation. Our results suggest that GATA1 exon 2 mutations occur late in trisomy 21 fetal hematopoiesis. However, trisomy 21 alone provides a proliferative stimulus of fetal megakaryopoiesis and erythropoiesis. CD34-positive precursor cells are not increased in trisomy 21 fetal livers.
In murine models, transgenic chemokine–cytokine tumor vaccines overcome many of the limitations of single-agent immunotherapy by producing the sequence of T-cell attraction followed by proliferation. ...The safety and immunologic effects of this approach in humans were tested in 21 patients with relapsed or refractory neuroblastoma. They received up to 8 subcutaneous injections of a vaccine combining lymphotactin (Lptn)– and interleukin-2 (IL-2)–secreting allogeneic neuroblastoma cells in a dose-escalating scheme. Severe adverse reactions were limited to reversible panniculitis in 5 patients and bone pain in 1 patient. Injection-site biopsies revealed increased cellularity caused by infiltration of CD4+ and CD8+ lymphocytes, eosinophils, and Langerhans cells. Systemically, the vaccine produced a 2-fold (P =.035) expansion of CD4+ T cells, a 3.5-fold (P =.039) expansion of natural killer (NK) cells, a 2.1-fold (P =.014) expansion of eosinophils, and a 1.6-fold (P =.049) increase in serum IL-5. When restimulated in vitro by the immunizing cell line, T cells collected after vaccination showed a 2.3-fold increase (P = .02) of T-helper (TH2)–type CD3+IL-4+cells. Supernatant collected from restimulated cells showed increased amounts of IL-4 (11.4-fold; P =.021) and IL-5 (8.7-fold;P =.002). Six patients had significant increases in NK cytolytic activity. Fifteen patients made immunoglobulin G (IgG) antibodies that bound to the immunizing cell line. Measurable tumor responses included complete remission in 2 patients and partial response in 1 patient. Hence, allogeneic tumor cell vaccines combining transgenic Lptn with IL-2 appear to have little toxicity in humans and can induce an antitumor immune response.
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