83 research outputs found
In Vitro Propagation of Thorny Blackberries
No full textProgram year: 1980-1981Digitized from print original stored in HDRTissue culture of 3 newly released thorny blackberry cultivars, ���Womack���, 'Rosborough', and 'Brison' was studied for sterilization procedure, shoot and root development, and plantlet acclimatization. Best sterilization procedure was found to be a 10-15 sec dip in 30% ethanol, then rinsed in double distilled water. Under the laminar flow hood, the explants were placed in a 5 min dip in 0.25% sodium hypochlorite plus Tween-20 and rinsed 3 times in double distilled sterile water. For shoot development media, 4 types of explants were cultured in a basic Murashige and Skoog salt mixture plus 0.4 mg/l myo-inositol, 0.1 mg/l GA3, 30 g/l sucrose, 8 g/l agar, 0.1 mg/l BA, and 0.1 mg/l IBA. Stem cuttings had less % contamination and longer shoot length. Shoots developed after 4 days in culture and once reached a size of 1 cm, they were transferred to a rooting media of 0, 1, 3, 6, 10 mg/l IBA. Optimum concentration to induce rooting was 3 mg/l IBA. Immediately after root initiation, plantlets were transferred to a medium with no growth regulators. Plantlets 5 cm in length were transplanted to small plastic pots containing 1 peat: 1 perlite (v/v). Acclimatization procedure followed a weekly exchange of a series of polyethylene covers with 0, 15, 50, and 98 perforations. Following this procedure, 100% of plantlets survived
Velké Popovice: great century in the history of the small village
No full textIn diploma thesis author focuses on history of a village called Velké Popovice. She deals with its history from the end of the 19th century when František Ringhoffer became the owner of the village till the events that followed right after the "Velvet Revolution". The main part of this work is focused on political, agricultural and communal matters of the village. The author deals with Ringhoffer's family and the things that the family managed to realize right in Velké Popovice and in its surroundings. The author deals with Word War I, that violated not only the inhabitants of the village but also soldiers whose fates are shown on life of Josef Kabeláč, a legionary, who came from the village. In other part of the work, the author deals with the important period of the "First Republic" followed by the protectorate at the end of which a big tragedy was about to happen. The following part of the work is focused on the period of communist regime that brought decline in many spheres, its decay and following prosperity in those spheres. The history of the village is extended by detached chapters about Church of Panna Marie Sněžná, brewery of Velké Popovice, education and local club activities
In vitro and in vivo analysis of human cell-based immunotherapies for acute myeloid leukemia
No full textConventional cancer therapies like surgery, radiation and chemotherapy help to eliminate primary tumor masses but often fail to eradicate disseminated tumor cells. However, it is such residual tumor cells that frequently underlie metastasis and relapse. Major obstacles for targeting such cells are wide spread dissemination and long-term persistence in niches that are difficult to reach. For example, many patients with acute myeloid leukemia (AML) show persistence of leukemia after chemotherapy – so-called minimal residual disease (MRD) – which confers a life-threatening risk for relapse in over 70% of patients. Arming the immune system to attack residual tumor cells has high therapeutic potential since immune cells can patrol the body to find and destroy residual tumor cells. Therapeutic approaches using the immune system - so-called immunotherapies - can take several forms. My project concentrated on preclinical studies of two strategies: 1) use of dendritic cells (DC) for therapeutic vaccination and 2) adoptive T cell therapy with lymphocytes expressing transgenic T cell receptors (TCR) specific for tumor-associated antigens (TAA).
In therapeutic vaccination a highly potent vaccine is needed to induce a valid immune response in patients with cancer. Effective antitumor immunity requires mobilization of IFN-γ-producing CD4+ T cells (Th1 cells) and lymphocytes with cytotoxic function, including cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. In my studies, high potency vaccines were developed using mature DC generated in 3 days (3d-mDC) that were stimulated with synthetic Toll-like receptor TLR3 and/or TLR7/8 agonists. This TLR stimulation mimics DC interaction with viruses and causes mDC to secrete the bioactive form of IL-12, supporting induction of effector cells. Characterization in vitro showed that TLR-activated 3d-mDC were superior to conventional 7d-mDC in capacity to induce Th1 cells as well as CTL. A humanized mouse model was established to verify these observations in vivo. NOD/scid IL2Rgnull mice, lacking murine T, B and NK cells, were reconstituted with human peripheral mononuclear cells and vaccinated with 3d-mDC, stimulated or not with TLR agonists, and conventional 7d-mDC. Induction of CTL was quantified ex vivo using splenocyte populations containing human lymphocytes. The in vivo results were concordant with in vitro observations, demonstrating the superior capacity of 3d-mDC that were stimulated with TLR agonists to induce CTL.
Adoptive T cell therapy using TCR-modified lymphocytes represents a second powerful way to provide patients with specific antitumor immunity. Here previously isolated TCR gene sequences are introduced into activated patient-derived lymphocytes, assigning them new antigen specificities. First, T cells must be isolated with TAA specificities that express high-affinity TCR which effectively recognize tumor cells. It was contended that T cell stimulation using peptide-epitopes from TAA presented on foreign MHC would allow isolation of high-affinity TCR, since these T cells had not yet undergone negative selection in the thymus. This contention was proved in individual experiments, as described in this thesis, for the antigens tyrosinase, survivin and HMMR (hyaluronan-mediated motility receptor). Since survivin and HMMR are broadly expressed in AML, TCR specific for these TAA were isolated and subsequently transferred into recipient lymphocytes. Expression of survivin-specific TCR resulted in MHC-restricted death of transduced lymphocytes due to their elevated survivin expression after activation. This precludes use of survivin-specific TCR for therapy of AML.
In contrast, transfer of an HMMR-specific TCR yielded effector lymphocytes that effectively killed AML cells in vitro. The behavior in vivo of TCR transduced lymphocytes is crucial for therapeutic outcome. To explore this capacity a xenograft mouse model was established using solid and disseminated human tumor cells injected into NOD/scid IL2Rgnull mice. Adoptive transfer of lymphocytes expressing an HMMR-specific TCR into tumor-bearing mice resulted in significant retardation of tumor outgrowth. Adoptive transfer of memory-like lymphocytes with higher proliferative potential and prolonged in vivo survival may also affect tumor growth. Analyses in vivo and in vitro showed that IL-15-induced effector memory T cells conferred the most potent antitumor immunity.
In summary, this work provides evidence for potent in vivo antitumor effects by either using DC-based vaccines or adoptive transfer of TCR transduced lymphocytes, opening application of both strategies for immunotherapy of cancer
Prognostic impact of progression to induction chemotherapy and prior paclitaxel therapy in patients with germ cell tumors receiving salvage high-dose chemotherapy in the last 10 years: a study of the European Society for Blood and Marrow Transplantation Solid Tumors Working Party
No full textLittle is known about the prognostic impact of prior paclitaxel therapy and response to induction chemotherapy defined as the regimen preceding high-dose chemotherapy (HDCT) for the salvage therapy of advanced germ cell tumors. Twenty European Society for Blood and Marrow Transplantation centers contributed data on patients treated between 2002 and 2012. Paclitaxel used in either prior lines of therapy or in induction-mobilization regimens was considered. Multivariable Cox analyses of prespecified factors were undertaken on PFS and overall survival (OS). As of October 2013, data for 324 patients had been contributed to this study. One hundred and ninety-two patients (59.3%) had received paclitaxel. Sixty-one patients (19%) had a progression to induction chemotherapy, 234 (72%) a response (29 (9%) missing or granulocyte colony-stimulating factor without chemotherapy). Both progression to induction chemotherapy and prior paclitaxel were significantly associated with shorter OS univariably (P<0.001 and P=0.032). On multivariable analysis from the model with fully available data (N=216) progression to induction was significantly prognostic for PFS and OS (P=0.003), but prior paclitaxel was not (P=0.674 and P=0.739). These results were confirmed after multiple imputation of missing data. Progression to induction chemotherapy could be demonstrated as an independent prognostic factor, in contrast to prior paclitaxel
Randomized phase-II trial evaluating induction therapy with idarubicin and etoposide plus sequential or concurrent azacitidine and maintenance therapy with azacitidine
No full textThe aim of this randomized phase-II study was to evaluate the effect of substituting cytarabine by azacitidine in intensive induction therapy of patients with acute myeloid leukemia (AML). Patients were randomized to four induction schedules for two cycles: STANDARD (idarubicin, cytarabine, etoposide); and azacitidine given prior (PRIOR), concurrently (CONCURRENT), or after (AFTER) therapy with idarubicin and etoposide. Consolidation therapy consisted of allogeneic hematopoietic-cell transplantation or three courses of high-dose cytarabine followed by 2-year maintenance therapy with azacitidine in the azacitidine-arms. AML with CBFB-MYH11, RUNX1-RUNX1T1, mutated NPM1, and FLT3-ITD were excluded and accrued to genotype-specific trials. The primary end point was response to induction therapy. The statistical design was based on an optimal two-stage design applied for each arm separately. During the first stage, 104 patients (median age 62.6, range 18-82 years) were randomized; the study arms PRIOR and CONCURRENT were terminated early due to inefficacy. After randomization of 268 patients, all azacitidine-containing arms showed inferior response rates compared to STANDARD. Event-free and overall survival were significantly inferior in the azacitidine-containing arms compared to the standard arm (p < 0.001 and p = 0.03, respectively). The data from this trial do not support the substitution of cytarabine by azacitidine in intensive induction therapy
ACUTE MYELOID LEUKEMIA WITH RUNX1 MUTATIONS CONSTITUTE A DISTINCT ENTITY ASSOCIATED WITH CHARACTERISTIC CLINICAL AND GENETIC FEATURES AND POOR OUTCOME. A STUDY OF THE AML STUDY GROUP (AMLSG)
No full textACUTE MYELOID LEUKEMIA WITH RUNX1 MUTATIONS CONSTITUTE A DISTINCT ENTITY ASSOCIATED WITH CHARACTERISTIC CLINICAL AND GENETIC FEATURES AND POOR OUTCOME. A STUDY OF THE AML STUDY GROUP (AMLSG)
No full textClinical impact of GATA2 mutations in acute myeloid leukemia patients harboring CEBPA mutations: a study of the AML study group
No full textÜberprüfung der Güte von drei Bewertungsverfahren mit unterschiedlichen Standardisierungsgraden zur Messung klinisch-praktischer Kompetenzen in der Inneren Medizin
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