Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers.DFG, 389687267, Kompartimentalisierung, Aufrechterhaltung und Reaktivierung humaner Gedächtnis-T-Lymphozyten aus Knochenmark und peripherem BlutDFG, 80750187, SFB 841: Leberentzündungen: Infektion, Immunregulation und KonsequenzenEC/H2020/800924/EU/International Cancer Research Fellowships - 2/iCARE-2DFG, 252623821, Die Rolle von follikulären T-Helferzellen in T-Helferzell-Differenzierung, Funktion und PlastizitätDFG, 390873048, EXC 2151: ImmunoSensation2 - the immune sensory syste

Electroanalysis And Electrocatalysis Using Poly(Vinylferrocenium) Perchlorate Coated Electrodes

Electroanalytical applications based on the electrochemical behavior of the redox polymer, poly(vinylferrocenium) perchlorate (PVF+ClO4-) coated as a film on Pt surfaces, are described. This chemically modified surface is electroactive due to the existence of ferrocenium/ferrocene sites in its structure. Electrooxidation and electroreduction of other depolarizers can also take place through this polymeric film. Anions such as iodide, thiocyanate and cyanide are incorporated in the polymeric structure as a result of anion exchange. Anodic stripping of the anions from the film can be carried out after the initial preconcentration. Catalytic reduction and oxidation of anthracene occur on the PVF+ClO4- coated Pt surface in acetonitrile/TBAP medium. An enzyme such as glucose oxidase could be immobilized in the polymeric matrix and this surface could be used for the amperometric determination of glucose under aerobic conditions. Coimmobilization of two enzymes, glucose oxidase and invertase, in the same polymeric matrix allows for the development of an amperometric sucrose sensor.WoSScopu

arenes and their extraction properties

Bisazocalix[4]arenes [N,N'-bis(5-azo-25,26,27-tribenzoyloxy-28-hydroxycalix[4]arene)benzene (1), N,N'-bis(5-azo-25,26,27-tribenzoyloxy-28-hydroxycalix [4] arene)biphenyl (2) and N,N'-bis(5-azo-25,26,27-tribenzoyloxy-28-hydroxycalix[4]arene)-2,2'-dinitro biphenyl (3)] have been synthesized from 25,26,27-tribenzoyloxy-28-hydroxycalix [4] arene by diazocoupling with the corresponding aromatic diamines (p-phenylenediamine, 4,4'-diamino biphenyl and 4,4'-diamino-2,2'-dinitrobiphenyl). Extraction studies of bisazocalix[4]arenes 1, 2, and 3 show no difference in their extraction behavior and selectivity, whereas azocalix[4]arenes are a poor extractant for heavy metal cations. The absorption spectra of the prepared bisazocalix[4]arenes are discussed, both the effect of varying pH and solvent upon the absorption ability of bisazocalix[4]arenes. (c) 2005 Elsevier Ltd. All rights reserved.C1 Pamukkale Univ, Fac Sci Arts, Dept Chem, TR-20017 Denizli, Turkey.Suleyman Demirel Univ, Fac Arts Sci, Dept Chem, TR-32200 Isparta, Turkey

Three-dimensional vascularized self-assembled tumor spheroids

41st FEBS Congress on Molecular and Systems Biology for a Better Life -- SEP 03-08, 2016 -- Kusadasi, TURKEYWOS: 000383616900279FEB

Evaluation of an aldo-keto reductase gene signature with prognostic significance in colon cancer via activation of epithelial to mesenchymal transition and the p70S6K pathway

AKR1B1 and AKR1B10, members of the aldo-keto reductase family of enzymes that participate in the polyol pathway of aldehyde metabolism, are aberrantly expressed in colon cancer. We previously showed that high expression of AKR1B1 (AKR1B1(HIGH)) was associated with enhanced motility, inflammation and poor clinical outcome in colon cancer patients. Using publicly available datasets and ex vivo gene expression analysis (n = 51, Ankara cohort), we have validated our previous in silico finding that AKR1B1(HIGH) was associated with worse overall survival (OS) compared with patients with low expression of AKR1B1 (AKR1B1(LOW)) samples. A combined signature of AKR1B1(HIGH) and AKR1B10(LOW) was significantly associated with worse recurrence-free survival (RFS) in microsatellite stable (MSS) patients and in patients with distal colon tumors as well as a higher mesenchymal signature when compared with AKR1B110(LOW)/AKR1B10(HIGH) tumors. When the patients were stratified according to consensus molecular subtypes (CMS), AKR1B1(HIGH)/AKR1B10(LOW) samples were primarily classified as CMS4 with predominantly mesenchymal characteristics while AKR1B1(LOW)/AKR1B10(HIGH) samples were primarily classified as CMS3 which is associated with metabolic deregulation. Reverse Phase Protein Array carried out using protein samples from the Ankara cohort indicated that AKR1B1(HIGH)/AKR1B10(LOW) tumors showed aberrant activation of metabolic pathways. Western blot analysis of AKR1B1(HIGH)/AKR1B10(LOW) colon cancer cell lines also suggested aberrant activation of nutrient-sensing pathways. Collectively, our data suggest that the AKR1B1(HIGH)/AKR1B10(LOW) signature may be predictive of poor prognosis, aberrant activation of metabolic pathways, and can be considered as a novel biomarker for colon cancer prognostication

The what and when of olfactory working memory in humans

Encoding and retaining novel sequences of sensory stimuli in working memory is crucial for adaptive behavior. A fundamental challenge for the central nervous system is to maintain each sequence item in an active and discriminable state, while also preserving their temporal context. Nested neural oscillations have been postulated to disambiguate the what and when of sequences, but the mechanisms by which these multiple streams of information are coordinated in the human brain remain unclear. Drawing from foundational animal studies, we recorded local field potentials from the human piriform cortex and hippocampus during a working memory task in which subjects experienced sequences of three distinct odors. Our data revealed a unique organization of odor memories across multiple timescales of the theta rhythm. During encoding, odors elicited greater gamma at distinct theta phases in both regions, time stamping their positions in the sequence, whereby the robustness of this effect was predictive of temporal order memory. During maintenance, stimulus-driven patterns of theta-coupled gamma were spontaneously reinstated in piriform cortex, recapitulating the order of the initial sequence. Replay events were time compressed across contiguous theta cycles, coinciding with periods of enhanced piriform-hippocampal theta-phase synchrony, and their prevalence forecasted subsequent recall accuracy on a trial-by-trial basis. Our data provide a novel link between endogenous replay orchestrated by the theta rhythm and short-term retention of sequential memories in the human brain

Genetic Characterization of Toxoplasma gondii Isolates and Toxoplasmosis Seroprevalence in Stray Cats of Izmir, Turkey

WOS: 000340879300061PubMed ID: 25127360Currently, some Toxoplasma gondii genotypes are being associated with serious clinical presentations. A recent report showing the Africa 1 genotype in two local congenital toxoplasmosis cases acquired in Turkey formed the basis of this study because atypical Africa 1 genotype is most frequently detected in animals and patients from sub-Saharan Africa. Since stray cats are considered as the linkage between wild life and urban life in T. gondii transmission, the present study aimed to isolate and characterize T. gondii strains circulating in stray cats of Izmir (Western Turkey). A secondary objective was to determine toxoplasmosis seroprevalence in this cat population. Tissues obtained from 100 deceased stray cats were bioassayed and isolated strains were genotyped using 15 microsatellite markers. In addition, toxoplasmosis seroprevalence was analyzed in 1121 cat sera collected from several large veterinary clinics in Izmir. Among the 22 isolates, 19 were Type II (86.3%), two were Type III (9%) and one was Africa 1 genotype (4.5%). The overall seropositivity rates in cats were 42-48% and 33.4-34.4% according to IFA and ELISA, respectively. Seroprevalence in deceased cats was significantly higher than in healthy cats (P = 0.0033). Finding both the major clonal Type II lineage together with the Type III lineage also found in Middle East, and an atypical genotype, Africa 1 appears consistent with the specific geographic location of Turkey between three continents and raises the possibility of transportation of these strains between continents through trade routes or long distance migratory birds. In addition, the first large study of toxoplasma seroprevalence in a stray cat population was also reported. The relatively high seropositivity rates and the variety of T. gondii genotypes confirm the local stray cat population as a risk factor for human toxoplasmosis in Izmir.Scientific Research Projects Branch Directorate of Ege University, TurkeyEge University [2010-TIP-091, 2011-TIP-034]This study was supported by the grants given by the Scientific Research Projects Branch Directorate of Ege University, Turkey (Grant No: 2010-TIP-091 and 2011-TIP-034) to Y.G. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Protection from cyclophosphamide-induced ovarian damage with bone marrow-derived mesenchymal stem cells during puberty

Objective: In female cancer survivors, the accelerated loss of primordial follicles may lead to premature ovarian failure. We investigated the protective effects of bone marrow derived mesenchymal stem cells (BMMSC) and gonadotropin releasing hormone analogue (GnRHa) against chemotherapeutic-induced ovarian toxicity in a rat model. Material and methods: Forty-eight Wistar albino female rats were divided into four groups. Group 1 was composed of rats that were given 200 mg/kg cyclophosphamide injection for each cycle (two cycles for each rat). Both cyclophosphamide and 0.4 mu g GnRHa were administered to Group 2. Cyclophosphamide and 4 million/kg BMMSC were administered to Group 3. Cyclophosphamide, GnRHa, and BMMSC were administered to Group 4. Germ cell apoptosis, DNA fragmentation and primordial follicular count were investigated with Cleave Caspase-9 and TUNEL analysis. The presence of the SRY gene on the Y chromosome in the ovary of the recipient female rats was checked with PCR. Results: Immunohistochemical staining (IHS) of Caspase-9 and TUNEL was higher in Group 1 than in Group 3 (p<0.05). Similarly, Group 4 had higher values than Group 3 (p<0.05). The presence of the SRY gene was detected in Groups 3 and 4 with the PCR analysis. The mean primordal follicle count was lowest in Group 1 and the mean primordial follicle counts were higher in Groups 2 and 3 than in Group 1. The difference between Group 1 and Group 4 was not significant. Conclusion: BMMSC therapy was found to be protective from germ cell apoptosis and DNA damage when it was used with chemotherapy regimens including alkylating agents