Proliferationsinhibition und Induktion der Differenzierung bei Gliomzellinien durch das Zytokin "Oncostatin M" (OSM)

In dieser Arbeit wurde die Wirkung dreier neurotropher Zytokine aus der Interleukin-6-Familie (IL-6, LIF, OSM) auf anaplastische Gliomzellinien untersucht. Waehrend IL-6 und LIF kaum einen Effekt auf die Proliferationsrate und Morphologie der Zellen zeigten, konnten wir eine deutliche Suppression der Proliferation durch OSM bewirken, zudem aenderte sich die Morphologie dieser Tumorzellen signifikant, die Zellen zeigten eine Form, die mit zahlreichen langen und duennen Fortsaetzen an normale Astrozyten erinnerte. Darueber hinaus stieg die Zytoplasma-Kern-Relation mit steigender Groeße der Zellen an. Die inhibierende Wirkung von OSM war dosisabhaengig mit einer maximalen Inhibitionsdosis von 50ng/ml. Die DNA-Synthese-Hemmung durch OSM konnte durch Antikoerper aufgehoben werden, die gegen das Zytokin selbst oder gegen seine Rezeptorkomponente gp130 gerichtet waren

Advanced Breeding Approaches for Cold-Tolerant Chickpea and Lentil in Dryland Areas

Chickpea and lentils are the two most economically important food legumes in dryland areas. They are traditionally sown in the spring of cold dryland areas of the Mediterranean regions. Therefore, the grain yield of the crop is affected by drought and high thermal stresses at the end of the season. Autumn cultivation of these crops by cold-tolerant varieties could increase grain yield up to 50%, then spring cultivation through higher availability of soil water. Breeding for cold-tolerant chickpea and lentil that is widely adaptable to autumn cultivation in cold regions and various growth conditions is the best strategic approach but requires a fine-tuned combination of advanced phenotyping and genotyping methods. However, breeding and selection of suitable cold-tolerant chickpea and lentil genotypes is complex by its narrow genetic base, which limits the sources of novel alleles. This chapter illustrates the morphological, physiological, and molecular effects of cold stress on chickpea and lentil growth and development. It will be also elaborated on conventional and advanced breeding approaches and application of advanced genotyping and phenotyping tools commonly used to develop cold-tolerant chickpea and lentil cultivars. The following, about key crop cold-tolerance traits that can be easily screened by using genotypic and phenotypic technologies are discussed

Genetic testing and genomic analysis: A debate on ethical, social and legal issues in the Arab world with a focus on Qatar

In 2013 both Saudi Arabia and Qatar launched genome projects with the aim of providing information for better diagnosis, treatment and prevention of diseases and, ultimately to realize personalized medicine by sequencing hundred thousands samples. These population based genome activities raise a series of relevant ethical, legal and social issues general, related to the specific population structure as well as to the Islamic perspective on genomic analysis and genetic testing. To contribute to the debate, the Authors after reviewing the existing literature and taking advantage of their professional experience in the field and in the geographic area, discuss and provide their opinions. In particular, the Authors focus on the impact of consanguinity on population structure and disease frequency in the Arab world, on genetic testing and genomic analysis (i.e. technical aspects, impact, etc.) and on their regulations. A comparison between the Islamic perspective and the ethical, social and legal issues raised in other population contexts is also carried. In conclusion, this opinion article with an up-to-date contribution to the discussion on the relevance and impact of genomic analysis and genetic testing in the Arab world, might help in producing specific national guidelines on genetic testing and genomic analysis and help accelerate the implementation and roll out of genome projects in Muslim countries and more specifically in Qatar, and other countries of the Gulf

SEED QUALITY OF SOYBEAN CULTIVARS AFFECTED BY POD POSITION AND WATER STRESS AT REPRODUCTIVE STAGES

ABSTRACT : A field experiment was carried out to investigate the effects of different irrigation treatments (I 1, I2, I3 and I4: well-watering on the bases of 70 mm evaporation from class A pan and irrigation disruptions at flowering, seed filling and during flowering and seed filling stages, respectively) and pod positions (upper, lower and middle parts of the canopy) on seed quality of three soybean cultivars (Clark, Williams and L17). Seed quality as determined by seed weight, electrical conductivity of seed leachates, viability and germination percentages, germination rate and seedling dry weight significantly reduced for seeds produced under I3 and I4. Significant differences in mean seed weight, viability, electrical conductivity and germination rate among soybean cultivars under different irrigation treatments were attributed to variation in genetic constitution. Seeds of upper parts of the canopies were larger and exhibited higher quality, compared with those of middle and lower parts. The advantage of the upper seeds was more evident under I3 and I4 treatments. Seed weight had the highest positive correlation with seedling dry weight. Thus, sufficient water supply and production of large and uniform seeds could be a practical way to improving seed lot quality in soybean cultivars

Quality of tumor lysates used for pulsing dendritic cells is influenced by the method used to harvest adherent tumor cells

BACKGROUND: Lysates from tumor cells are reported to induce maturation of dendritic cells (DCs) and are used in clinical settings for DC-based vaccination against solid tumors. Nevertheless, the maturation inducing effect of tumor lysates on DCs is discussed controversially and the efficacy of tumor vaccines varies significantly. FINDINGS: Using three individual adherent colorectal tumor cell lines we also faced the difficulty to obtain consistent results regarding maturation inducing effect of tumor lysates on DCs. Therefore, we compared different methods to prepare tumor cell lysate and could demonstrate that trypsinizing as a method to harvest adherent tumor cells has a significant negative impact on biologic activity of tumor lysates. Specifically, we assessed induction of maturation markers CD40, CD80, and CD86 on DCs which were treated with differently prepared lysates. CONCLUSIONS: Trypsinizing is a very common way of harvesting adherent cells from culture flasks. Our results shall call investigators' attention to the enzymatic activity of trypsin degrading some possibly important proteins on the surface of cultured cells. Specifically for DC-based vaccination against tumor antigens investigators should avoid trypsin

Large-scale screening of HCMV-seropositive blood donors indicates that HCMV effectively escapes from antibodies by cell-associated spread

Immunoglobulins are only moderately effective for the treatment of human cytomegalovirus (HCMV) infections, possibly due to ineffectiveness against cell-associated virus spread. To overcome this limitation, we aimed to identify individuals with exceptional antibodies in their plasma that can efficiently block the cell-associated spread of HCMV. A Gaussia luciferase-secreting mutant of the cell-associated HCMV strain Merlin was generated, and luciferase activity evaluated as a readout for the extent of cell-associated focal spread. This reporter virus-based assay was then applied to screen plasma samples from 8400 HCMV-seropositive individuals for their inhibitory effect, including direct-acting antiviral drugs as positive controls. None of the plasmas reduced virus spread to the level of these controls. Even the top-scoring samples that partially reduced luciferase activity in the screening assay failed to inhibit focal growth when reevaluated with a more accurate, immunofluorescence-based assay. Selected sera with high neutralizing capacity against free viruses were analyzed separately, and none of them prevented the focal spread of three recent clinical HCMV isolates nor reduced the number of particles transmitted, as demonstrated with a fluorescent Merlin mutant. We concluded that donors with cell-to-cell-spread-inhibiting plasma are nonexistent or extremely rare, emphasizing cell-associated spread as a highly efficient immune escape mechanism of HCM

Evaluating the Serum Levels of CCL17, CCL22, and CCL28 Chemokines and the Gene Expression of α4β1 and α4β7 Integrins in Patients With Allergic Rhinitis

Allergic rhinitis (AR) is a chronic inflammatory disease involving the nasal mucosa. Leukocytes recruitment to the inflammation sites is controlled by chemokines, cytokines, and adhesion molecules. Retinoic acid (RA), a vitamin A metabolite, plays an essential role in mucosal immunity and the production of inflammatory cytokines and chemokines. This study intended to evaluate the serum levels of RA, CCL17, CCL22, CCL28, and the mRNA expression levels of α4, β1, and β7 integrins in AR patients compared to healthy subjects. Peripheral blood was collected from 37 patients with AR and 30 age- and gender-matched healthy individuals. Serum levels of RA, CCL17, CCL22, and CCL28 were measured by the enzyme-linked immunosorbent assay (ELISA) technique, and the mRNA expression levels for α4, β1, and β7 integrins were assessed using the quantitative real-time PCR method. Our results showed that the serum levels of CCL22 and CCL28 chemokines are significantly higher in the AR group compared to the healthy controls (P<0.01). However, the gene expression of the β1 integrin in the AR group was significantly lower than that of the control group (P<0.001). Besides, there was a positive association between serum RA and CCL17 levels in patients (P<0.0001, r=0.6). In conclusion, increased serum levels of CCL22 and CCL28 chemokines, as well as decreased gene expression of β1 integrin in AR patients, may contribute to the pathogenesis and/or exacerbation of AR

Peripheral cytokine levels differ by HPV status and change treatment-dependently in patients with head and neck squamous cell carcinoma

Cytokines and immune mediators play an important role in the communication between immune cells guiding their response to infectious diseases or cancer. In this study, a comprehensive longitudinal analysis of serum cytokines and immune mediators in head and neck squamous cell carcinoma (HNSCC) patients was performed. In a prospective, non-interventional, longitudinal study, blood samples from 22 HNSCC patients were taken at defined time points (TP) before, during, and every 3 months after completion of (chemo)radio)therapy (CRT/RT) until 12 months after treatment. Serum concentrations of 17 cytokines/immune mediators and High-Mobility-Group-Protein B1 (HMGB1) were measured by fluorescent bead array and ELISA. Concentrations of sFas were significantly elevated during and after CRT/RT, whereas perforin levels were significantly decreased after CRT/RT. Levels of MIP-1β and Granzyme B differed significantly during CRT/RT by HPV status. Increased HMGB1 levels were observed at recurrence, accompanied by high levels of IL-4 and IL-10. The sFas increase and simultaneous perforin decrease may indicate an impaired immune cell function during adjuvant radiotherapy. Increased levels of pro-inflammatory cytokines in HPV+ compared to HPV− patients seem to reflect the elevated immunogenicity of HPV-positive tumors. High levels of HMGB1 and anti-inflammatory cytokines at recurrence may be interpreted as a sign of immune evasion

Immune Suppressive Effects of Plasma-Derived Exosome Populations in Head and Neck Cancer

Plasma-derived exosomes of head and neck squamous cell carcinoma (HNSCC) patients carry inhibitory factors mediating immune suppression. Separation of tumor-derived exosomes (TEX) and non-TEX may assist in a better understanding of their respective parental cells. Here, we evaluate the impact of TEX or hematopoietic-derived exosomes on immune suppression. We evaluated apoptosis in CD8+ T cells, conversion of CD4+ T cells to regulatory T cells (Treg), and adenosine production by TEX, non-TEX, or total exosomes. Exosome protein cargo was significantly higher in total and CD45(−) exosomes from high stage compared to low stage patients. Furthermore, total and CD45(−) exosomes of high stage patients induced more apoptosis in CD8+ T cells than their low stage counterparts. CD69 suppression, a marker of reduced CD8+ T cell activation, was only mediated by CD45(−) exosomes. All fractions induced Treg differentiation, defined by CD39 expression, but only CD45(−) exosomes showed a stage-dependent conversion. CD45(−) exosomes produced higher adenosine concentrations than CD45(+) exosomes, concluding that adenosine production measured in total exosomes mainly derives from TEX. The presented results show significant induction of immune suppression by TEX in HNSCC. This immunosuppressive effect supports the potential role of exosomes as liquid biomarkers for disease stage and level of immune suppression

Feasibility and safety of treating non-unions in tibia, femur and humerus with autologous, expanded, bone marrow-derived mesenchymal stromal cells associated with biphasic calcium phosphate biomaterials in a multicentric, non-comparative trial

Background: ORTHO-1 is a European, multicentric, first in human clinical trial to prove safety and feasibility after surgical implantation of commercially available biphasic calcium phosphate bioceramic granules associated during surgery with autologous mesenchymal stromal cells expanded from bone marrow (BM-hMSC) under good manufacturing practices, in patients with long bone pseudarthrosis. Methods: Twenty-eight patients with femur, tibia or humerus diaphyseal or metaphyso-diaphyseal non-unions were recruited and surgically treated in France, Germany, Italy and Spain with 100 or 200 million BM-hMSC/mL associated with 5–10 cc of bioceramic granules. Patients were followed up during one year. The investigational advanced therapy medicinal product (ATMP) was expanded under the same protocol in all four countries, and approved by each National Competent Authority. Findings: With safety as primary end-point, no severe adverse event was reported as related to the BM-hMSC. With feasibility as secondary end-point, the participating production centres manufactured the BM-hMSC as planned. The ATMP combined to the bioceramic was surgically delivered to the non-unions, and 26/28 treated patients were found radiologically healed at one year (3 out of 4 cortices with bone bridging). Interpretation: Safety and feasibility were clinically proven for surgical implantation of expanded autologous BM-hMSC with bioceramic. Funding: EU-FP7-HEALTH-2009, REBORNE Project (GA: 241876).The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP7/FP7-HEALTH-2009); REBORNE Project (GA: 241876