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

Signatures of the disk-jet coupling in the Broad-line Radio Quasar 4C+74.26

Here we explore the disk-jet connection in the broad-line radio quasar 4C+74.26, utilizing the results of the multiwavelength monitoring of the source. The target is unique in that its radiative output at radio wavelengths is dominated by a moderately-beamed nuclear jet, at optical frequencies by the accretion disk, and in the hard X-ray range by the disk corona. Our analysis reveals a correlation (local and global significance of 96\% and 98\%, respectively) between the optical and radio bands, with the disk lagging behind the jet by $250 \pm 42$ days. We discuss the possible explanation for this, speculating that the observed disk and the jet flux changes are generated by magnetic fluctuations originating within the innermost parts of a truncated disk, and that the lag is related to a delayed radiative response of the disk when compared with the propagation timescale of magnetic perturbations along relativistic outflow. This scenario is supported by the re-analysis of the NuSTAR data, modelled in terms of a relativistic reflection from the disk illuminated by the coronal emission, which returns the inner disk radius $R_{\rm in}/R_{\rm ISCO} =35^{+40}_{-16}$. We discuss the global energetics in the system, arguing that while the accretion proceeds at the Eddington rate, with the accretion-related bolometric luminosity $L_{\rm bol} \sim 9 \times 10^{46}$ erg s$^{-1}$ $\sim 0.2 L_{\rm Edd}$, the jet total kinetic energy $L_\textrm{j} \sim 4 \times 10^{44}$ erg s$^{-1}$, inferred from the dynamical modelling of the giant radio lobes in the source, constitutes only a small fraction of the available accretion power.Comment: 9 pages and 6 figures, ApJ accepte

Multi-omic signatures of atherogenic dyslipidaemia : pre-clinical target identification and validation in humans

Dyslipidaemia is a major risk factor for atherosclerosis and cardiovascular diseases. The molecular mechanisms that translate dyslipidaemia into atherogenesis and reliable markers of its progression are yet to be fully elucidated. To address this issue, we conducted a comprehensive metabolomic and proteomic analysis in an experimental model of dyslipidaemia and in patients with familial hypercholesterolemia (FH)

Suppression of Scant Identifies Endos as a Substrate of Greatwall Kinase and a Negative Regulator of Protein Phosphatase 2A in Mitosis

Protein phosphatase 2A (PP2A) plays a major role in dephosphorylating the targets of the major mitotic kinase Cdk1 at mitotic exit, yet how it is regulated in mitotic progression is poorly understood. Here we show that mutations in either the catalytic or regulatory twins/B55 subunit of PP2A act as enhancers of gwl^(Scant), a gain-of-function allele of the Greatwall kinase gene that leads to embryonic lethality in Drosophila when the maternal dosage of the mitotic kinase Polo is reduced. We also show that heterozygous mutant endos alleles suppress heterozygous gwl^(Scant); many more embryos survive. Furthermore, heterozygous PP2A mutations make females heterozygous for the strong mutation polo¹¹ partially sterile, even in the absence of gwl^(Scant). Heterozygosity for an endos mutation suppresses this PP2A/polo¹¹ sterility. Homozygous mutation or knockdown of endos leads to phenotypes suggestive of defects in maintaining the mitotic state. In accord with the genetic interactions shown by the gwl^(Scant) dominant mutant, the mitotic defects of Endos knockdown in cultured cells can be suppressed by knockdown of either the catalytic or the Twins/B55 regulatory subunits of PP2A but not by the other three regulatory B subunits of Drosophila PP2A. Greatwall phosphorylates Endos at a single site, Ser68, and this is essential for Endos function. Together these interactions suggest that Greatwall and Endos act to promote the inactivation of PP2A-Twins/B55 in Drosophila. We discuss the involvement of Polo kinase in such a regulatory loop

The quest for companions to post-common envelope binaries. II. NSVS14256825 and HS0705+6700

We report new mid-eclipse times of the two close binaries NSVS14256825 and HS0705+6700, harboring an sdB primary and a low-mass main-sequence secondary. Both objects display clear variations in the measured orbital period, which can be explained by the action of a third object orbiting the binary. If this interpretation is correct, the third object in NSVS14256825 is a giant planet with a mass of roughly 12 M_Jup. For HS0705+6700, we provide evidence that strengthens the case for the suggested periodic nature of the eclipse time variation and reduces the uncertainties in the parameters of the brown dwarf implied by that model. The derived period is 8.4 yr and the mass is 31 M_Jup, if the orbit is coplanar with the binary. This research is part of the PlanetFinders project, an ongoing collaboration between professional astronomers and student groups at high schools.Comment: Accepted by Astron. and Astrophy

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

The genomes of two key bumblebee species with primitive eusocial organization

Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats. Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits. Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation

Polarization and spectral energy distribution in OJ 287 during the 2016/17 outbursts

We report optical photometric and polarimetric observations of the blazar OJ 287 gathered during 2016/17. The high level of activity, noticed after the General Relativity Centenary flare, is argued to be part of the follow-up flares that exhibited high levels of polarization and originated in the primary black hole jet. We propose that the follow-up flares were induced as a result of accretion disk perturbations, travelling from the site of impact towards the primary SMBH. The timings inferred from our observations allowed us to estimate the propagation speed of these perturbations. Additionally, we make predictions for the future brightness of OJ 287. © 2017 by the authors