Oral treatment with genistein reduces the expression of molecular and biochemical markers of inflammation in a rat model of chronic TNBS-induced colitis

Background  Inflammatory bowel disease (IBD) in humans has a high incidence in Europe and the USA, whereas in East Asia, incidence has been historically low. The risk of IBD appears to increase in Asian immigrants adopting western lifestyles, suggesting a strong link of environmental/dietary factors in the development of IBD. Exposure to high levels of isoflavones such as genistein (Gen) in traditional East Asian diets has been associated with a decreased risk of developing breast cancer and may also be beneficial for the prevention of IBD. Aim  In this study, the effect of orally administered genistein on the inflammatory response in the TNBS-induced chronic colitis rat model was investigated. Methods  Eighteen male Wistar rats, aged 12 weeks, were randomized to one of three groups (n = 6). Two groups received a 2,4,6-trinitrobenzenesulfonic acid (TNBS) enema, then were treated daily by oral gavage with either Gen (100 mg/kg b.w.) or vehicle, for 14 days. The last group served as a control group, not receiving the TNBS enema. At the end of the 14 days, animals were killed and tissues collected. Molecular and biochemical inflammatory markers in the colon, specifically cyclooxygenase-2 (COX-2) and myeloperoxidase (MPO), were analyzed. In addition, to assess the efficacy of Gen treatment, relative wet weights of the accessory sexual organs, specifically prostate and the seminal vesicle, were compared between the groups treated or not with Gen. Results  Wet weights of both prostates and seminal vesicles were significantly (P < 0.01) reduced upon Gen administration. In the colon, expression of COX-2 mRNA and protein was reduced (P < 0.05) in the Gen treatment group, as compared to the control group, whereas there was no significant inhibitory effect of Gen on the expression of proliferating cell nuclear antigen. In Gen treated animals colon wet weight was not altered, however a decrease in MPO activity (P < 0.01) was seen. Conclusion  These results may provide evidence that oral administration of Gen exerts beneficial anti-inflammatory effects in a rodent model of TNBS-induced chronic colitis. While the sample size of this study was small, it nevertheless might encourage the realization of larger blinded randomized controlled studies for the proof of concept.     

No association between genetic variants at the ASCT1 gene and schizophrenia or bipolar disorder in a German sample

Altered glutamatergic neurotransmission is considered a potential etiological factor of schizophrenia (SCZ) and affective disorders. The gene ASCT1 (SLC1A4) coding for a Na+-dependent neutral aminoacid transporter is a member of the glutamate transporter superfamily and is located on 2p13-14, a region showing linkage to both SCZ and bipolar disorder (BD). ASCT1 can thus be considered a candidate gene for both disorders. In a German sample, we tested for association between ASCT1 and both SCZ and BD. Allele and haplotype frequencies, however, did not differ between cases and controls. Recent findings on the associations between brainderived neurotrophic factor (BDNF) and SCZ and between G72/G30 and BD suggest that SCZ patients with a history of major depressive episodes (MDE) outside psychotic episodes and BD cases with a history of persecutory delusions constitute genetically distinct subgroups of these disorders. Thus, we hypothesized that restricting case definition to those 95 SCZ individuals with MDE and to those 107 BD patients with a history of persecutory delusions might clarify the relationship between BD, SCZ and ASCT1. However, these stratification approaches did not yield any significant association either. Allele and haplotype frequencies did not differ between cases and controls. Our results do not support an association of the ASCT1 gene with BD or SCZ in the German population.     

Microarray analysis of sarcomas

Microarrays began to be used to study gene expression profiles in the mid-1990s, but it was only after 2000 that serious attempts have been made to apply this technology to investigate sarcomas. Microarray technologies provide a comprehensive survey of active molecular pathways and potential molecular targets for diagnosis and treatment, but are challenging to use because of issues of specimen collection, cost, and complexities in experimental design and data analysis. As a discovery-based technique, microarray analyses are most valuable when framed around specific gaps in our knowledge of tumor etiology and progression, challenges in differential diagnosis, and pressing therapeutic needs. To date, microarray analyses of sarcomas support their division into molecularly defined and molecularly heterogeneous categories, and have provided useful diagnostic markers for entities such as gastrointestinal stromal tumors, synovial sarcoma, and dermatofibrosarcoma protuberans. Signatures predicting outcome and response to therapy have been published for Ewing sarcoma and osteosarcoma, and receptor tyrosine kinase expression patterns have suggested novel therapeutic approaches which may be applied to several types of sarcoma. Nevertheless, results need to be interpreted in the context of histopathology and validated by complementary technologies and/or other research groups.     

Uniform definition of asthma severity, control, and exacerbations: document presented for the World Health Organization Consultation on Severe Asthma

Asthma is a global health problem affecting around 300 million individuals of all ages, ethnic groups and countries. It is estimated that around 250,000 people die prematurely each year as a result of asthma. Concepts of asthma severity and control are important in evaluating patients and their response to treatment, as well as for public health, registries, and research (clinical trials, epidemiologic, genetic, and mechanistic studies), but the terminology applied is not standardized, and terms are often used interchangeably. A common international approach is favored to define severe asthma, uncontrolled asthma, and when the 2 coincide, although adaptation may be required in accordance with local conditions. A World Health Organization meeting was convened April 5-6, 2009, to propose a uniform definition of severe asthma. An article was written by a group of experts and reviewed by the Global Alliance against Chronic Respiratory Diseases review group. Severe asthma is defined by the level of current clinical control and risks as "Uncontrolled asthma which can result in risk of frequent severe exacerbations (or death) and/or adverse reactions to medications and/or chronic morbidity (including impaired lung function or reduced lung growth in children)." Severe asthma includes 3 groups, each carrying different public health messages and challenges: (1) untreated severe asthma, (2) difficult-to-treat severe asthma, and (3) treatment-resistant severe asthma. The last group includes asthma for which control is not achieved despite the highest level of recommended treatment and asthma for which control can be maintained only with the highest level of recommended treatment.     

Enoxaparin Improves the Course of Dextran Sodium Sulfate-Induced Colitis in Syndecan-1-Deficient Mice

Syndecan-1 (Sdc1) plays a major role in wound healing and modulates inflammatory responses. Sdc1 expression is reduced in lesions of patients with ulcerative colitis. The aim of this study was to investigate the role of Sdc1 in murine dextran sodium sulfate (DSS)-induced colitis. DSS colitis was induced in Sdc1-deficient (knockout (KO)) and wild-type mice by oral administration of 3% DSS. KO mice exhibited a significantly increased lethality as compared with wild-type controls (61 versus 5%, P < 0.05). Impaired mucosal healing and prolonged recruitment of inflammatory cells in KO mice were accompanied by significant up-regulation of tumor necrosis factor-α, CC chemokine ligand 3/macrophage inflammatory protein-1α, and vascular cell adhesion molecule-1, as determined by histological correlation between 0 and 15 days after colitis induction, TaqMan low-density array analysis, and quantitative real-time PCR. Treatment from days 7 through 14 with enoxaparin, a functional analogue of the Sdc1 heparan sulfate chains, significantly reduced lethality of KO mice due to DSS-induced colitis, which was correlated with improved mucosal healing. In vitro, Sdc1-deficient polymorphonuclear cells displayed increased adhesion to endothelial cells and intercellular adhesion molecule-1, and enoxaparin reverted adhesion to wild-type levels. Small interfering RNA-mediated knockdown of Sdc1 expression resulted in reduced basic fibroblast growth factor-mediated mitogen-activated protein kinase signaling and reduced Caco-2 cell proliferation. We conclude that Sdc1 has a protective effect during experimental colitis. The modification of missing Sdc1 function by heparin analogues may emerge as a promising anti-inflammatory approach.Syndecan-1 (Sdc1) is the most important representative of the heparan sulfate proteoglycans (HSPGs) covering epithelial cell surfaces.¹ It serves multiple biological roles, such as cell-matrix interactions, modulation of inflammatory responses, tumorigenesis, and wound healing.^2–4 The highly conserved cytoplasmic domains of Sdc1 interact with scaffolding proteins and participate in integrin-mediated signaling events, thus providing a physical and functional link to the cytoskeleton. In addition, most of the extracellular-binding interactions are mediated by the heparan sulfate chains, which are structurally and functionally related to heparin, an extensively sulfated and epimerized derivative of heparan sulfate.¹ Sdc1 serves as a coreceptor for several tyrosine kinase receptors. For example, it increases the activity of the complex of basic fibroblast growth factor (bFGF) and the FGF receptor and, therefore, contributes to improved wound healing via stimulation of keratinocyte proliferation.^1,5 A role for Sdc1 in wound repair in vivo has been demonstrated in Sdc1-deficient (Sdc1-knockout (KO)) mice, which show delayed skin and corneal wound healing⁵ and functionally adverse repair following experimental myocardial infarction due to dysregulation of chemokine expression and matrix metalloproteinase-mediated tissue remodeling.⁶ Sdc1 forms chemotactic gradients due to binding of chemokines on heparan sulfate chains of the molecule. Therefore, Sdc1 is able to act as coreceptor for chemokine signaling.^7,8 In addition, endothelial leukocyte recruitment and extravasation is modulated by Sdc1, possibly via interference with heparin-binding adhesion molecule function.^9–12Day et al¹³ described in 1999 a reduced expression of Sdc1 in patients with ulcerative colitis, which was linked to disrupted healing of colonic ulcers. In addition, this group demonstrated the benefit of the Sdc1 ectodomain for the FGF-induced proliferation of intestinal epithelial cell lines in vitro. The function of Sdc1 could be restored with heparin, representing a highly sulfated and epimerized form of heparan sulfate, the major functional constituent of the Sdc1 ectodomain.Heparin sees widespread use as anticoagulant drug based on its antithrombin III-activating properties. Enoxaparin, is a low molecular weight heparin with similar features in vitro and in vivo like heparin; however, it exhibits a more favorable pharmacological side effect profile. Both low molecular weight heparins (enoxaparin) as well as heparin were recently found to possess anti-inflammatory properties.¹⁴The hypothesis of Sdc1 being involved in the pathogenesis of ulcerative colitis is underlined by multiple clinical observations of patients who have been treated with heparins for different reasons.¹⁵ In a number of cases, this treatment has lead to an improved course of disease. A limited number of uncontrolled clinical trials with heparins in the treatment of low to medium active ulcerative colitis showed a variable outcome,^16–18 which may be explained by variations in treatment regimes that may have failed to include the optimal dose, class of heparin, and mode of delivery. For example, most studies have involved either i.v. or s.c. delivery of heparin, whereas a more appropriate mode of delivery for stimulating mucosal healing might be the topical application or microsphere-mediated delivery of heparin.^14,15 Furthermore, the outcome of heparin therapy may depend on the degree to which Sdc1 expression is reduced in inflammatory bowel disease (IBD) patients.¹⁹Moreover, the expression of Sdc1 and the proinflammatory cytokine tumor necrosis factor-α (TNF-α) are inversely correlated in the colonic mucosa of patients with Crohn''s disease,²⁰ and a reduction of Sdc1 expression has been shown to result in increased TNF-α signaling in an in vitro model of protein-losing enteropathy,^19,21 further suggesting a regulatory role for Sdc1 in proinflammatory cytokine signaling.In this study, our goal was to characterize the impact of a Sdc1 deficiency on the dextran sodium sulfate (DSS)-induced colitis of the mouse. Furthermore, the efficacy of low molecular weight heparin to restore altered wound healing was studied in vivo. In addition, in vitro trials were performed to study the role of Sdc1 deficiency in the adhesion and transmigration of leukocytes under inflammatory conditions.     

Mast cell lines HMC‐1 and LAD2 in comparison with mature human skin mast cells – drastically reduced levels of tryptase and chymase in mast cell lines

Please cite this paper as: Mast cell lines HMC‐1 and LAD2 in comparison with mature human skin mast cells – drastically reduced levels of tryptase and chymase in mast cell lines. Experimental Dermatology 2010; 19 : 845–847. Abstract: To circumvent the costly isolation procedure associated with tissue mast cells (MC), two human MC lines, i.e. HMC‐1 and LAD2, are frequently employed, but their relation to mature MC is unknown. Here, we quantitatively assessed their expression of MC markers in direct comparison to skin MC (sMC). sMC expressed all lineage markers at highest and HMC‐1 cells at lowest levels. LAD2 cells expressed comparable high‐affinity IgE receptor α (FcεRIα) and FcεRIγ but less FcεRIβ than sMC and displayed slightly reduced, but robust FcεRI‐mediated histamine release. Only minor differences were found for total histamine content and c‐Kit expression. Huge, and to this level unexpected, differences were found for MC tryptase and chymase, with sMC >>> LAD2 > HMC‐1. Taken together, HMC‐1 cells represent very immature malignantly transformed MC, whereas LAD2 cells can be considered intermediately differentiated. Because of the minute levels of MC proteases, MC lines can serve as surrogates of tissue MC to a limited degree only.     

Serum Insulin Aspart Concentrations Following High-Dose Insulin Aspart Administered Directly into the Duodenum of Healthy Subjects: An Open-Labeled, Single-Blinded, and Uncontrolled Exploratory Trial

Objective

The goal of this study was to determine the bioavailability of high-dose insulin aspart administered directly into the duodenum of healthy subjects.

Methods

In a pilot study, four subjects each received four escalating doses of a 1-ml solution of insulin aspart (100, 300, 600, and 1000 IU, respectively) directly into the duodenum. In the following main study, eight subjects each received two identical doses of insulin aspart of 1000 IU, in 4- and 8-ml solutions, respectively, directly into the duodenum. Subjects in the main study also received an intravenous and a subcutaneous injection of 4 to 6 IU of insulin aspart.

Results

A considerable number of samples and, in some cases, consecutive samples revealed significantly increased concentrations of serum insulin aspart. Despite the significant serum insulin aspart concentrations, no significant changes of plasma glucose were measured. Moreover, no significant suppression of endogenous insulin secretion was detected, as assessed by the levels of serum human insulin.

Conclusions

Administration of high-dose insulin aspart directly into the duodenum of healthy subjects resulted in significantly increased serum insulin aspart concentrations in a high number of consecutive samples using a specific enzyme-linked immunosorbent assay. However, no significant changes in the levels of plasma glucose or serum human insulin were observed. Thus, the study did not provide any evidence of biological activity of the original insulin aspart molecule after high-dose administration directly into the duodenum.     

Remission and cognitive ability in a cohort of patients with schizophrenia

INTRODUCTION: Patients with schizophrenia were studied regarding their ability of achieving symptomatic remission. It was found that only approximately one-third of the patients fulfilled the criteria for remission. The importance of cognitive performance was studied to decide whether cognitive ability is a contributing factor for achieving remission. MATERIALS AND METHODS: A homogeneous cohort of 211 patients, of whom 76 patients attained remission (36%), and 135 patients (64%) failed to, was studied. Remission was decided by the use of eight items from the PANSS in which none of these items should have a score above three points. The patients were also tested on a comprehensive battery of cognitive tests including vigilance, working memory, long-term memory, executive functioning, learning performance, visuomotor speed/efficacy, cognitive flexibility and pre-morbid functioning. RESULTS: Marked differences were exhibited in the cognitive abilities in all domains of patients who had attained remission in comparison with those who had not. DISCUSSION: This study highlights the importance of cognitive performance as one possible predictor of remission. Patients without remission had lower cognitive ability and may have more difficulty in benefiting from, or complying with their treatment. As a consequence, various forms of supportive treatment might increase the likelihood of remission.