Differentiation of mouse embryonic stem cells into cells with spermatogonia-like morphology with chemical intervention-dependent increased gene expression of LIM homeobox 1 (Lhx1).

Spermatogonial stem cells (SSCs) originate from gonocytes that differentiate from primordial germ cells (PGCs). In the developing mouse testis, expression of the gene LIM homeobox 1 (Lhx1) marks the most undifferentiated SSCs, which has not yet been reported for spermatogonia-like cells generated in vitro. Previously, it was shown that a chemical intervention in male mouse embryonic stem (ES) cells in serum culture, including Sirtuin 1 (SIRT1) inhibitor Ex-527, DNA methyltransferase (DNMT) inhibitor RG-108 and electrophilic redox cycling compound tert-butylhydroquinone (tBHQ), was associated with molecular markers of PGC to gonocyte differentiation. Here, we report the in vitro differentiation of male mouse ES cells, cultured under dual chemical inhibition of GSK3β and MEK (2i) with leukemia inhibitory factor (LIF) (2iL) and serum, into cells with spermatogonia-like morphology (CSMs) and population-averaged expression of spermatogonia-specific genes by removal of 2iL and a specific schedule of twice daily partial medium replacement. Combination of this new protocol with the previously reported chemical intervention increased population-averaged gene expression of Lhx1 in the resulting CSMs. Furthermore, we detected single CSMs with strong nuclear LHX1/5 protein signal only in the chemical intervention group. We propose that further investigation of CSMs may provide new insights into male germline development

Differentiation of mouse embryonic stem cells into cells with spermatogonia-like morphology with chemical intervention-dependent increased gene expression of LIM homeobox 1 (Lhx1)

Spermatogonial stem cells (SSCs) originate from gonocytes that differentiate from primordial germ cells (PGCs). In the developing mouse testis, expression of the gene LIM homeobox 1 (Lhx1) marks the most undifferentiated SSCs, which has not yet been reported for spermatogonia-like cells generated in vitro. Previously, it was shown that a chemical intervention in male mouse embryonic stem (ES) cells in serum culture, including Sirtuin 1 (SIRT1) inhibitor Ex-527, DNA methyltransferase (DNMT) inhibitor RG-108 and electrophilic redox cycling compound tert-butylhydroquinone (tBHQ), was associated with molecular markers of PGC to gonocyte differentiation. Here, we report the in vitro differentiation of male mouse ES cells, cultured under dual chemical inhibition of GSK3β and MEK (2i) with leukemia inhibitory factor (LIF) (2iL) and serum, into cells with spermatogonia-like morphology (CSMs) and population-averaged expression of spermatogonia-specific genes by removal of 2iL and a specific schedule of twice daily partial medium replacement. Combination of this new protocol with the previously reported chemical intervention increased population-averaged gene expression of Lhx1 in the resulting CSMs. Furthermore, we detected single CSMs with strong nuclear LHX1/5 protein signal only in the chemical intervention group. We propose that further investigation of CSMs may provide new insights into male germline development

Ergebnis der selektiven Verpaarung beim Entlebucher Sennenhund zur Reduktion der ureteralen Ektopie

Outcome of selective mating in the Entlebucher Mountain Dog for reduction of ureteral ectopia The Entlebucher Mountain Dog is predisposed to ureteral ectopia and associated diseases of the urinary tract as well as the kidneys, which can have severe to lethal consequences. Due to the clustered occurrence of clinical signs in 11 % of Entlebucher Mountain dogs in the absence of a genetic test for ureteral ectopia, screening was introduced in 2008 to allow phenotype-based breeding selection. The ureteral orifices of the dogs are visualized by ultrasound and existing urinary retention or urinary incontinence is documented. The diagnostic findings were evaluated centrally with assignment to one of five phenotypes depending on the localization of the ureteral orifices and the renal and ureteral shape. Breeding approval and mating restrictions are the responsibility of the respective breeding associations and predominantly Entlebucher Mountain Dogs with extravesical ectopic ureters and/or clinical signs were excluded from breeding. The effect of phenotype-based selective mating on the incidence of ureteral ectopia and its clinical signs, as well as possible factors influencing the expression of the phenotype, were determined in the birth cohorts after the introduction of screening. Analysis of the data set of 1456 phenotyped Entlebucher Mountain Dogs showed, that at 11 % versus 5 %, males were more frequently assigned to the extravesical phenotype than females. The effect of phenotype-based breeding selection was examined in a subpopulation consisting of phenotyped parents and their offspring (n = 876). The prevalence of the extravesical phenotype decreased from 24 % in the 2005 to 2007 birth cohorts to 1,4 % in the 2015 to 2017 birth cohorts. Since 2015 almost no Entlebucher Mountain Dogs with incontinence, hydroureter or hydronephrosis have been recorded. It was feared that the additional selection measures to control ureteral ectopia in the small Entlebucher Mountain Dog population would intensify the inbreeding increase. However, this has so far remained absent. Therefore, as long as no genetic test is available, it is recommended to continue phenotype-based breeding selection with exclusion of dogs with extravesical ureteral ectopia and/or hydroureter/hydronephrosis/urinary incontinence, while keeping an eye on the development of the inbreeding coefficient. Keywords: Ectopic ureter, Dog, Inbreeding, Phenotype, Breeding selection Deutsch Ergebnis der selektiven Verpaarung beim Entlebucher Sennenhund zur Reduktion der ureteralen Ektopie Der Entlebucher Sennenhund ist prädisponiert für die ureterale Ektopie und damit für assoziierte Erkrankungen der Harnwege sowie der Nieren, was schwerwiegende bis letale Folgen haben kann. Aufgrund des gehäuften Auftretens klinischer Symptome bei 11 % der Entlebucher Sennenhunde wurde in Ermangelung eines Gentests auf ureterale Ektopie 2008 ein Screening eingeführt, um eine Phänotyp-basierte Zuchtselektion zu ermöglichen. Die Uretermündungen der Hunde werden in der Regel mittels Ultraschall lokalisiert und bestehender Harnrückstau oder Harninkontinenz wird dokumentiert. Die Befundung erfolgte zentral mit einer Zuordnung zu einem von fünf Phänotypen in Abhängigkeit von der Lokalisation der Uretermündungen sowie der Nieren – und Ureterengestalt. Die Zuchtzulassung und Verpaarungsbeschränkungen obliegen den jeweiligen Zuchtverbänden, wobei überwiegend Entlebucher Sennenhunde mit extravesikal ektopischen Ureteren und/oder klinischen Symptomen von der Zucht ausgeschlossen wurden. Die Auswirkung der Phänotyp-basierten selektiven Verpaarung auf das Auftreten der ureteralen Ektopie und deren klinischen Symptome sowie mögliche Einflussfaktoren auf die Ausprägung des Phänotyps wurden in den Geburtsjahren nach Einführung des Screenings ermittelt. Die Analyse des Datensatzes mit 1456 phänotypisierten Entlebucher Sennenhunden zeigte, dass mit 11 % versus 5 % Rüden häufiger als Hündinnen dem extravesikalen Phänotyp zugeteilt wurden. Die Auswirkung der Phänotyp-basierten Zuchtselektion wurde an einer Teilpopulation, bestehend aus phänotypisierten Elterntieren und ihren Nachkommen (n = 876), untersucht. Die Prävalenz des extravesikalen Phänotyps nahm von 24 % bei den Geburtsjahren 2005 bis 2007 auf 1,4 % bei den Geburtsjahren 2015 bis 2017 ab. Seit 2015 wurden nahezu keine Entlebucher Sennenhunde mehr mit Inkontinenz, Hydroureter oder Hydronephrose erfasst. Befürchtet wurde, dass die zusätzlichen Selektionsmassnahmen zur Bekämpfung der ureteralen Ektopie in der kleinen Entlebucher Sennenhundepopulation den Inzuchtanstieg verstärken würde. Dies blieb bisher jedoch aus. Daher wird, solange kein genetischer Test zur Verfügung steht, empfohlen, die Phänotyp-basierte Zuchtselektion mit Ausschluss von Hunden mit extravesikaler ureteraler Ektopie und/oder Hydroureter/Hydronephrose/Harninkontinenz vorerst weiterzuführen und gleichzeitig die Entwicklung des Inzuchtkoeffizienten im Auge zu behalten. Schlüsselwörter: Ektopischer Ureter, Hund, Inzucht, Phänotyp, Zuchtselektion Français Résultat de l’accouplement sélectif chez le bouvier de l’Entlebuch pour réduire l’ectopie urétérale Le Bouvier de l’Entlebuch est prédisposé à l’ectopie urétérale et aux maladies associées des voies urinaires ainsi que des reins, ce qui peut entraîner des conséquences fatales. En raison de l’apparition de signes cliniques chez 11 % des chiens et en l’absence d’un test génétique pour l’ectopie urétérale, un dépistage a été introduit en 2008 pour permettre une sélection d’élevage basée sur le phénotype. Les orifices urétraux des chiens ont été visualisés par échographie et la rétention ou l’incontinence urinaire existante documentée. Les résultats du diagnostic ont été évalués de manière centralisée avec attribution à l’un des cinq phénotypes en fonction de la localisation des orifices urétéraux ainsi que de la forme des reins et des uretères. L’approbation pour la reproduction et les restrictions d’accouplement relèvent de la responsabilité des associations d’élevage respectives et les bouviers de l’Entlebuch présentant des uretères ectopiques extravésicaux et/ou des signes cliniques ont majoritairement été exclus de la reproduction. L’effet de cet accouplement sélectif basé sur le phénotype sur l’incidence de l’ectopie urétérale et de ses signes cliniques ainsi que les facteurs possibles influençant l’expression du phénotype ont été déterminés dans les cohortes de naissance après l’introduction du dépistage. L’analyse de l’ensemble des données de 1456 Bouviers de l’Entlebuch phénotypés a montré que, à 11 % contre 5 %, les mâles étaient plus fréquemment affectés au phénotype extravésical que les femelles. L’effet de la sélection d’élevage basée sur le phénotype a été examiné dans une sous-population composée de parents phénotypés et de leur progéniture (n = 876). La prévalence du phénotype extravésical est passée de 24 % dans les cohortes de naissance de 2005 à 2007 à 1,4 % dans les cohortes de naissance de 2015 à 2017. Depuis 2015, presque aucun bouvier d’Entlebuch présentant une incontinence, un hydrouretère ou une hydronéphrose n’a été enregistré. Une possible augmentation de la consanguinité due aux mesures de sélection supplémentaires visant à contrôler l’ectopie urétérale ne s’est pas produite. Par conséquent, tant qu’aucun test génétique n’est disponible, il est recommandé de poursuivre la sélection d’élevage basée sur le phénotype avec exclusion des chiens présentant une ectopie urétérale extravésicale et/ou une hydrouretère/hydronéphrose/incontinence urinaire, tout en surveillant l’évolution du coefficient de consanguinité. Mots-clés: auretère ectopique, chien, consanguinité, phénotype, sélection d’élevage Italiano Risultato dell'accoppiamento selettivo nel cane da montagna Entlebuch per ridurre l'ectopia ureterale Il bovaro dell’Entlebuch è predisposto all’ectopia ureterale e quindi alle malattie che sono correlate al tratto urinario e renale con possibili conseguenze letali. A causa della frequente comparsa di segni clinici nell’11 % dei cani e in assenza di un test genetico per l’ectopia ureterale, nel 2008 è stato introdotto uno screening per consentire la selezione fenotipica dei riproduttori. Gli orifizi ureterali dei cani sono solitamente localizzato tramite ecografia e viene documentata la presenza di ritenzione o incontinenza urinaria. I risultati sono stati valutati in modo centralizzato e si è assegnato il risultato a uno dei cinque fenotipi a seconda della localizzazione degli orifizi ureterali e della forma dei reni e degli ureteri. L’autorizzazione all’allevamento e le restrizioni all’accoppiamento sono di competenza delle rispettive associazioni di allevatori e i bovari dell’Entlebuch con ureteri ectopici extravescicali e/o con segni clinici sono stati prevalentemente esclusi dalla riproduzione. L’effetto dell’accoppiamento selettivo, basato sul fenotipo per rapporto all’incidenza dell’ectopia ureterale e dei suoi segni clinici, nonché dei possibili fattori che influenzano l’espressione del fenotipo, è stato determinato nelle coorti di nascita dopo l’introduzione dello screening. L’analisi del set di dati di 1456 bovari dell’Entlebuch fenotipizzati ha dimostrato che, con una percentuale dell’11 % rispetto al 5 %, i maschi erano più frequentemente assegnati al fenotipo extravescicale rispetto alle femmine. L’effetto della selezione riproduttiva basata sul fenotipo è stato esaminato in una subpopolazione composta da genitori fenotipizzati e dalla loro discendenza (n = 876). La prevalenza del fenotipo extravescicale è diminuita dal 24 % nelle coorti di nascita dal 2005 al 2007 all’1,4 % nelle coorti di nascita dal 2015 al 2017. Dal 2015 non sono stati registrati quasi più bovari dell’Entlebuch con incontinenza, idrouretere o idronefrosi. Non si è riscontrato un possibile aumento della consanguineità dovuto alle misure di selezione aggiuntive per controllare l’ectopia ureterale. Pertanto, finché non è disponibile un test genetico, si raccomanda di continuare la selezione fenotipica con l’esclusione dei cani con ectopia ureterale extravescicale e/o idrouretere/idronefrosi/incontinenza urinaria, facendo particolare attenzione all’andamento del coefficiente di consanguineità. Parole chiavi: Uretere ectopico, cane, consanguineità, fenotipo, selezione riproduttiv

Complex pension products: A multidimensional approach

The study analyzes and assesses the economic and linguistic complexity of individual retirement products in Poland. For this purpose, an original multidimensional approach was used and various research methods were applied. We analyzed 75 out of 86 individual pension products (IKEs and IKZEs) offered in Poland in the first half of 2017, covering our analysis of nearly 90% of Poland’s market of individual pension products. We performed the nonparametric Spearman’s rank correlation analysis, we used hierarchical cluster analysis, analysis of variance, and a chi-square test to verify if there was a statistical relationship between the clusters and the type of financial provider and the type of individual pension product (IKE or IKZE). We also built also a map of the products that shows their economic and linguistic complexity. We find that high-fee products tend to have the most complex fee systems, suggesting that the complex fee system may be a strategy used by the providers of individual retirement products. Our results also indicate that individual retirement products are too complex for most individuals

Radiative and nonradiative deexcitation of pigments from natural oils

The absorption, steady state photoacoustic, fluorescence emission and excitation spectra of unbleached oils obtained by pressing the seeds of: evening primrose, borage, rape and viper`s burgloss have been taken. Results obtained for samples under air and washed by nitrogen have been compared. In order to establish the occurrence of the slow thermal processes usually related to very photochemically active triplet states, the thermal deactivation in the absorption regions of carotenoids and the pheophytins has been observed at various frequencies of acting light modulations. The interactions between oil pigments are discussed. The generation of triplet states of dyes and singlet oxygen production in oils with different content of carotenoids and pheophytins is evaluated

MRI assessment of ectopic fat accumulation in pancreas, liver and skeletal muscle in patients with obesity, overweight and normal BMI in correlation with the presence of central obesity and metabolic syndrome

Joanna Pieńkowska,1 Beata Brzeska,1–3 Mariusz Kaszubowski,4 Oliwia Kozak,5 Anna Jankowska,6 Edyta Szurowska11II Department of Radiology, Faculty of Health Sciences, Medical University of Gdansk, Gdansk, Poland; 2Department of Biology and Pharmaceutical Botany, Faculty of Pharmacy, Medical University of Gdansk, Gdansk, Poland; 3Department of Human Physiology, Faculty of Health Sciences, Medical University of Gdansk, Gdansk, Poland; 4Institute of Statistics, Department of Economic Sciences, Faculty of Management and Economics, Gdansk University of Technology, Gdansk, Poland; 5I Department of Radiology, Faculty of Medicine, Medical University of Gdansk, Gdansk, Poland; 6Department of Radiology, University Clinical Centre in Gdansk, Gdansk, PolandPurpose: Obesity, defined as a body mass index (BMI) exceeding 30 kg/m2, is a serious health problem, which can be called an epidemic on a global scale and is one of the most important causes of preventable death. The aim of this study was to assess ectopic fat accumulation in pancreas, liver and skeletal muscle in patients with obesity, overweight and normal BMI in correlation with metabolic syndrome (MetS).Patients and methods: The study included 267 consecutive patients who underwent a standard clinical assessment with BMI calculation. Ectopic fat accumulation in pancreas, liver, and skeletal muscle was evaluated by magnetic resonance imaging (MRI) using fat–water separated Dixon imaging. MetS was defined according to the criteria modified by the National Cholesterol Education Program Adult Treatment Panel III Guidelines. Central obesity was defined using gender and ethnic-specific values for waist circumference.Results: There was a statistically significant correlation between the degree of steatosis of the assessed organs and BMI value as well as waist circumference ratio, that determined the degree of central obesity. It was found that the most rapid relative fat accumulation was in muscle, then in pancreas and then in liver. Higher steatosis of pancreas, liver, and muscle was demonstrated depending on the number of the satisfied MetS criteria.Conclusion: Knowing that pancreatic fatty disease is a risk factor for MetS, it seems that assessment and monitoring of ectopic fat accumulation may have important clinical implications and may be used in the prediction of metabolic risk and its early prevention.Keywords: magnetic resonance imaging, obesity, pancreatic steatosis, liver steatosis, metabolic syndrome, fat fractio