Progenitor cells are responsible for formation of human prostate epithelium primary cultures

To analyze cell viability and morphology of primary cell cultures from CD133 immunolabeled and sorted cells from epithelium of patients suffering from benign prostate hyperplasia (BPH). Methods: Cells obtained from 5 patients were divided in two fractions. First fraction (CD133+/CD133–) was cultivated in DMEM with 10% FBS. Second fraction was mixed with CD133 microbeads and immunomagnetically divided into CD133+ and CD133– fractions. These cells were cultivated and followed-up for 2 weeks. Cells were stained for Annexin V FITC/propidium iodide. Results: Seventy CD133+/CD133– cultures, thirty-one of CD133+ and thirty-one of CD133– cells were established. There were 5-fold and 3-fold increase of CD133+/CD133- and CD133+ cell number after 2 weeks, respectively. CD133+/CD133– and CD133+ monolayers displayed epithelial-like morphology and cytokeratine expression. CD133– cultures collapsed. Cell viability within CD133+ and CD133– populations was 90.1 ± 6.3% and 24.3 ± 6.2%, respectively. Apoptotic index was 9.0 ± 6.1% and 28.5 ± 23.8% within CD133+ and CD133– cultures, respectively. Conclusions: CD133 separated human primary epithelial cell cultures displayed differences in morphology, viability and apoptosis occurrence. Immunomagnetic sorting can be recommended in each in vitro experiments with primary cell cultures in order to provide more objective results.Цель: оценить жизнеспособность и морфологию клеток первичных клеточных культур, полученных из меченных по CD133 и полученных с помощью клеточной сортировки клеток эпителия пациентов с доброкачественной гиперплазией предстательной железы (BPH). Методы: клетки, полученные от 5 пациентов, были разделены на 2 фракции. Первую фракцию (CD133+/CD133–) выращивали в DMEM с 10% FBS али с CD133 магнитными гранулами и с помощью магнита разделили клетки на CD133+- и CD133–-фракции. Далее клетки культивировали в течении 2 нед. Клетки окрашивали аннексиномV FITC/пропидий йодидом. Результаты: получено 70 CD133+/CD133- -культур клеток, 31 CD133+ и 31 CD133–. Через 2 нед культивирования отмечали 5-кратное и 3-кратное увеличение количества CD133+/CD133– и CD133+ клеток соответственно. CD133+/CD133- -и CD133+-клетки росли в монослое и имели морфологию эпителиальных клеток, экспрессировали цитокератин. CD133–-клетки не выжили. Выживаемость клеток в популяциях CD133+ и CD133– была 90,1 ± 6,3% и 24,3 ± 6,2% соответственно. Показатель апоптического индекса для культур CD133+ и CD133– был 9,0 ± 6,1% и 28,5 ± 23,8% соответственно. Выводы: показаны различия в морфологии, выживаемости клеток и частоте апоптоза для эпителиальных клеток, разделенных в зависимости от экспрессии CD133. Сортировка клеток с помощью иммуномагнитного разделения рекомендована для каждого in vitro эксперимента с использованием первичных клеточных культур для получения более объективных результатов

Color pattern recognition with circular component whitening

Polychromatic object recognition based on circular whitening preprocessing of red-green-blue components and multichannel matched filtering is described. Computer simulations and experimental results are provided to facilitate recognizing a color target among objects of similar shape but with different color contents. Experimental results are obtained with an optical correlator with two spatial light modulators, one to introduce the scene and the second one to introduce the filter

Influence of two pt(iv) complexes on viability, apoptosis and cell cycle of B16 mouse melanoma tumors

Several platinum(IV) complexes are showing considerable promise in initial trials, producing reactive intermediates that then interact with DNA. Aim: To perform in vitro study of two new platinum(IV) complexes cytotoxic effect on B16 mouse melanoma cells. Methods: PtCl₄ (dbtp)₂ and PtCl₂ (6mp)₂ complexes were prepared. PtCl₄ (dbtp)₂ was created as modification of PtCl₄ (dmtp) test previously.Apoptosis and necrosis were examined using flow cytometry, upon Annexin V/PI staining. Results: LC₁₀,LC₅₀ andLC₉₀ parameters established for PtCl₄ (dbtp)₂ were as following: 2.6, 17.0, 58.0 μmol/L. However LC₁₀ andLC₅₀ established for PtCl₂ (6mp)₂ were 1.2 and 14.0μmol/l respectively. The both complexes induced apoptosis. PtCl₂ (6mp)₂ induced cell cycle arrest in G0/G1, while PtCl₄ (dbtp)₂ — in S-phase. Conclusions: PtCl₄ (dbtp)₂ appeared to be more cytotoxic against B16 cells than PtCl₂ (6mp)₂ . Apoptosis was the main mechanism of cell loss in cultures incubated with both tested complexes

Identifying metabolites by integrating metabolome databases with mass spectrometry cheminformatics.

Novel metabolites distinct from canonical pathways can be identified through the integration of three cheminformatics tools: BinVestigate, which queries the BinBase gas chromatography-mass spectrometry (GC-MS) metabolome database to match unknowns with biological metadata across over 110,000 samples; MS-DIAL 2.0, a software tool for chromatographic deconvolution of high-resolution GC-MS or liquid chromatography-mass spectrometry (LC-MS); and MS-FINDER 2.0, a structure-elucidation program that uses a combination of 14 metabolome databases in addition to an enzyme promiscuity library. We showcase our workflow by annotating N-methyl-uridine monophosphate (UMP), lysomonogalactosyl-monopalmitin, N-methylalanine, and two propofol derivatives

Treosulfan-fludarabine-thiotepa-based conditioning treatment before allogeneic hematopoietic stem cell transplantation for pediatric patients with hematological malignancies

Treosulfan-based conditioning prior to allogeneic transplantation has been shown to have myeloablative, immunosuppressive, and antineoplastic effects associated with reduced non-relapse mortality (NRM) in adults. Therefore, we prospectively evaluated the safety and efficacy of treosulfan-based conditioning in children with hematological malignancies in this phase II trial. Overall, 65 children with acute lymphoblastic leukemia (35.4%), acute myeloid leukemia (44.6%), myelodysplastic syndrome (15.4%), or juvenile myelomonocytic leukemia (4.6%) received treosulfan intravenously at a dose of 10 mg/m2/day (7.7%), 12 g/m2/day (35.4%), or 14 g/m2/day (56.9%) according to their individual body surface area in combination with fludarabine and thiotepa. The incidence of complete donor chimerism at day +28 was 98.4% with no primary and only one secondary graft failure. At 36 months, NRM was only 3.1%, while relapse incidence was 21.7%, and overall survival was 83.0%. The cumulative incidence of acute graft-vs.-host disease was 45.3% for grades I–IV and 26.6% for grades II–IV. At 36 months, 25.8% overall and 19.4% moderate/severe chronic graft-vs.-host disease were reported. These data confirm the safe and effective use of treosulfan-based conditioning in pediatric patients with hematological malignancies. Therefore, treosulfan/fludarabine/thiotepa can be recommended for myeloablative conditioning in children with hematological malignancies

Authenticity and drug resistance in a panel of acute lymphoblastic leukaemia cell lines

Cell lines are important models for drug resistance in acute lymphoblastic leukaemia (ALL), but are often criticised as being unrepresentative of primary disease. There are also doubts regarding the authenticity of many lines. We have characterised a panel of ALL cell lines for growth and drug resistance and compared data with that published for primary patient specimens. In contrast to the convention that cell lines are highly proliferative, those established in our laboratory grow at rates similar to estimates of leukaemic cells in vivo (doubling time 53–442 h). Authenticity was confirmed by genetic fingerprinting, which also demonstrated the potential stability of long-term cultures. In vitro glucocorticoid resistance correlated well with that measured ex vivo, but all lines were significantly more sensitive to vincristine than primary specimens. Sensitivity to methotrexate was inversely correlated to that of glucocorticoids and L-asparaginase, indicating possible reciprocity in resistance mechanisms. A cell line identified as highly methotrexate resistant (IC50 >8000-fold higher than other lines) was derived from a patient receiving escalating doses of the drug, indicating in vivo selection of resistance as a cause of relapse. Many of these lines are suitable as models to study naturally occurring resistance phenotypes in paediatric ALL

Epstein-Barr Virus-Related Post-Transplant Lymphoproliferative Disorder in Children Treated with Rituximab: The Impact of Viral Load and Non-Lymphoid Tissue Involvement

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