Domestic Violence Victims a Nuisance to Cities

Unless municipal nuisance ordinances change, domestic violence victims can face eviction just for calling the police. Nuisance ordinances generally impose fines on a property owner or landlord when the police are called to respond to incidents of crime a certain number of times at the same residence. Many nuisance ordinances also revoke a landlord’s rental license if a property is deemed a nuisance. However, many of these nuisance ordinances do not have an exception for incidents of domestic violence and, consequently, victims are scared to call 911 or request police assistance. This comment surveys the development of nuisance laws and ordinances, analyzes constitutional and statutory challenges to ordinances without domestic violence exceptions, and focuses on the positive social implications of adding such exceptions

Stat5 in Erythropoiesis and iron metabolism

Signal Transducer and Activator of Transcription 5 (STAT5) ist ein wesentlicher Effektor der Erythropoietin (EPO) Signaltransduktion in der erythroiden Differenzierung. Erstmals wurde eine vollständige phänotypische Analyse im erythroiden Kompartiment von Mäusen durchgeführt, denen beide Stat5 Isoformen (STAT5a und STAT5b) komplett fehlen. Der beobachtete Phänotyp der hypochromen mikrocytären Anämie und die daraus resultierende perinatale Letalität führten nicht nur zur Bestätigung von bekannten STAT5 Funktionen, wie Schutz vor Apoptose, sondern auch zur Entdeckung neuer STAT5 Rollen in der Eisen-Aufnahme und Häm-Biosynthese. Anämie von Stat5 defizienten Mäusen konnte auf erhöhte Apoptose im Erythron durch verringerte Expression des anti-apoptotischen Proteins BCL-XL und Verlust des anti-apoptotischen Proteins MCL1 zurückgeführt werden. Da jedoch Apoptose allein nicht zu hypochromen oder mikrocytären Phänotypen führt, zeigte die Suche nach weiteren Defekten Eisenmangel in Stat5 defizienten erythroiden Zellen. Der Mangel an zellulärem Eisen konnte durch reduzierte Expression von Transferrin-Rezeptor 1 (TFR1), der wichtigsten Komponente der zellulären Eisen-Aufnahme, erklärt werden. TFR1 Expression wird sowohl auf transkriptioneller als auch auf posttranskriptioneller Ebene gestört. Einerseits ist Tfr1 ein direktes Ziel-Gen von STAT5, was zu eingeschränkter transkriptioneller Aktivität in der Knock-out-Situation führt. Andererseits wird im Stat5 Knockout Tfr1 mRNA-Stabilität durch verringerte Expression des Iron Regulatory Protein 2 (IRP2) vermindert, das bestimmte Sequenzen im 3 'UTR der Tfr1 mRNA bindet, was Schutz vor Degradation bietet, wenn zelluläres Eisen rar ist. Reduzierte Tfr1 mRNA Transkription und Stabilität führen zu verringerter TFR1 Oberflächenexpression, verringerter zellulärer Eisen-Aufnahme und schließlich zu Problemen in der Häm-Synthese, jenem Prozess mit dem höchsten Eisen Bedarf in differenzierenden erythroiden Zellen. Drastisch reduzierte Häm-Synthese führt wiederum durch die langsame Hämoglobin Akkumulation zu hypochromer microcytärer Anämie. In-vitro-Differenzierung von Stat5-Knockout Erythroblasten kann nicht durch direkte Eisenzugabe unter Umgehung des TFR1 Defekts gerettet werden. Daher müssen auch andere Mechanismen betroffen sein, die Hämoglobin Akkumulation behindern. Es konnte gezeigt werden, dass vier der acht Häm-Biosynthese Enzyme in Abwesenheit von STAT5 herunter reguliert waren. Vor allem die Expression von δ-Amino-Levulinic Acid Synthase 2 (ALAS2), dem geschwindigkeitsbestimmenden Enzym im Syntheseweg, war sowohl auf mRNA als auch auf Protein Ebene stark vermindert. Da sich alle vier enzymatischen Gene als unwahrscheinliche direkte STAT5 Ziele erwiesen, wurde der Status des gesamten hämatopoetischen transkriptionellen Netzwerks in Stat5-/- Erythroblasten ermittelt. Knockout Erythroblasten hatten stark erhöhte Ldb1 und Gata2 mRNA Niveaus. Von beiden Genen ist bekannt, dass sie bei Überexpression erythroide Differenzierung durch Interferenz mit GATA-1-Aktivität negativ beeinflussen. Da in den Promotoren von allen vier herunter regulierten Häm-Biosynthese Enzymen GATA-1 Bindungsstellen gefunden wurden, kann der Mangel an Häm-Synthese auf die erhöhte Expression von LDB1 und GATA2 zurückgeführt werden. Im Vergleich zu Erythropoetin und Erythropoetin-Rezeptor (EpoR) Knockouts, hat Verlust von Stat5 einen milderen Phänotyp. Dieser Unterschied könnte aus partieller Kompensation durch andere STAT-Familienmitglieder mit sehr ähnlichen Ziel DNASequenzen, wie STAT3, resultieren. Erhöhte Phosphorylierungsniveaus von STAT3 wurden in Stat-defizienten Erythroblasten gemessen, in denen dieser Effekt durch Verlust von Cytokine-Inducible-SH2 Protein (CIS), Suppressor Of Cytokine Signaling 1 (SOCS1) und Suppressor Of Cytokine Signaling 3 (SOCS3), die wichtige Negativregulatoren der EPOR Signaltransduktion sind, erklärt werden konnte. Im Gegensatz dazu zeigt eine anderen Familie von Negativregulatoren, Protein Inhibitors of Activated STAT (PIAS Proteins), deutlich erhöhte mRNA-Niveaus, was, mit hoher Wahrscheinlichkeit, erhöhter STAT Aktivität infolge des Verlusts von SOCS-Proteinen entgegenwirkt. Obwohl weitere funktionelle Experimente nötig sind, um kompensatorische Wirkungen zu untersuchen, können diese Ergebnisse die nicht notwendigerweise funktionelle, erhöhte Phosphorylierung von STAT3 in Stat5 defizienten Erythroblasten und anderen Geweben erklären.Signal transducer and activator of transcription 5 (STAT5) is an essential downstream effector of erythropoietin (EPO) mediated signaling in erythroid differentiation. We conducted the first full phenotypical analysis in the erythroid compartment of mice completely lacking both Stat5 isoforms (Stat5a and Stat5b). The observed phenotype of hypochromic microcytic anemia and perinatal lethality led to the confirmation of known STAT5 functions, like apoptosis protection, but also to the discovery of novel STAT5 roles in iron uptake and heme biosynthesis. Anemia of Stat5 deficient mice could be attributed to increased apoptosis in the erythron caused by decreased expression of the anti-apoptotic protein BCL-XL and loss of the anti-apoptotic protein MCL1. Since apoptosis alone does not explain the emergence of hypochromic or microcytic erythrocytes additional defects were suspected. This led to the finding of iron deficiency in Stat5 deficient erythroid cells. Lack of cellular iron could be traced back to reduced expression of transferrin receptor 1 (TFR1), the main player in cellular iron uptake. TFR1 regulation was found to be disturbed both at transcriptional and posttranscriptional level. On the one hand Tfr1 is a direct STAT5 target with reduced transcriptional activity in the knockout situation. Tfr1 mRNA stability, on the other hand, is impaired in knockouts due to decreased expression of iron regulatory protein 2 (IRP2), which binds several sequence elements in the 3’ UTR of Tfr1 mRNA to protect it from degradation, when cellular iron is scarce. Reduced Tfr1 mRNA transcription and stability leads to decreased TFR1 surface expression, diminished cellular iron uptake and finally to problems in heme synthesis, the process with the highest iron demand in differentiating erythroid cells. Drastically reduced heme synthesis carries over to diminished hemoglobin accumulation, which in turn causes hypochromic microcytic anemia. In vitro differentiation of Stat5 knockout erythroblasts cannot be rescued by supplying cells with iron complexes bypassing their TFR1 defect. Therefore additional mechanisms must be affected by loss of Stat5 that hinder hemoglobin accumulation. Indeed, four out of eight enzymes involved in heme biosynthesis were found to be downregulated in the absence of STAT5. Most importantly δ-aminolevulinic acid synthase 2 (ALAS2), the rate limiting enzyme in the pathway, displayed severely reduced mRNA and protein levels. Since all four genes were unlikely to be direct STAT5 targets, the status of the hematopoietic transcriptional network in Stat5-/- erythroblasts was surveyed further. Knockout erythroblasts had highly elevated levels of Ldb1 and Gata2 mRNA, both known to interfere with erythroid differentiation by negatively affecting GATA1 activity. Since GATA1 binding sites are present in the promoters of all four downregulated heme biosynthetic enzymes, the lack of heme synthesis may be linked to increased expression of LDB1 and GATA2. In comparison to ablation of erythropoietin (Epo) and erythropoietin receptor (EpoR) knockouts, loss of Stat5 has a milder phenotype. This difference may be attributed to partial rescue by other STAT family members with similar DNA-binding consensus sequences like STAT3. Higher levels of phosphorylated STAT3 were indeed observed in Stat5 deficient erythroblasts, which could be traced back to loss of cytokine-inducible-SH2-containing protein (CIS), suppressor of cytokine signaling 1 (SOCS1) and suppressor of cytokine signaling 3 (SOCS3), which constitute important negative regulators of EPOR signaling. In contrast another family of negative regulators, protein inhibitors of activated STAT (PIAS) proteins, displayed significantly elevated mRNA levels, counteracting increased STAT activity due to loss of SOCS proteins. Although further functional experiments are needed to clarify compensatory effects, these findings may help to explain the elevated levels of STAT3 phosphorylation found in erythroblasts and other tissues lacking Stat5

Influence of glycerol on poly(3-hydroxybutyrate) production by Cupriavidus necator and Burkholderia sacchari

Glycerol is a co-product of many industrial processes and is generated in large quantities from different origins. In this study, glycerol is used as a cheap carbon source for the production of poly(3- hydroxybutyrate) (PHB) with two different collection strains, Cupriavidus necator and Burkholderia sacchari, in order to provide an alternative outlet for glycerol and produce value-added bioproducts. The objective of this work was to study the influence of this carbon source on their growth kinetics, on their polymer production, and on the molecular mass of the produced biopolymer. Therefore, fermentations in bioreactors were carried out with these strains. Different results for both strains were obtained showing, for the first time, a high cell dry mass and growth rate, when glycerol was used together with glucose in the fermentation with C. necator. In the first fermentation with B. sacchari using glycerol as a sole carbon source, the strain properly developed synthesizing PHB. The biopolymers obtained from both fermentations with glycerol showed low molecular masses about 300 kDa with a polydispersity of 4.72 with C. necator, and 200 kDa with polydispersity of 2.50 with B. sacchari.Postprint (author's final draft

A human organoid model of aggressive hepatoblastoma for disease modeling and drug testing

Hepatoblastoma is the most common childhood liver cancer. Although survival has improved significantly over the past few decades, there remains a group of children with aggressive disease who do not respond to current treatment regimens. There is a critical need for novel models to study aggressive hepatoblastoma as research to find new treatments is hampered by the small number of laboratory models of the disease. Organoids have emerged as robust models for many diseases, including cancer. We have generated and characterized a novel organoid model of aggressive hepatoblastoma directly from freshly resected patient tumors as a proof of concept for this approach. Hepatoblastoma tumor organoids recapitulate the key elements of patient tumors, including tumor architecture, mutational profile, gene expression patterns, and features of Wnt/β-catenin signaling that are hallmarks of hepatoblastoma pathophysiology. Tumor organoids were successfully used alongside non-tumor liver organoids from the same patient to perform a drug screen using twelve candidate compounds. One drug, JQ1, demonstrated increased destruction of liver organoids from hepatoblastoma tumor tissue relative to organoids from the adjacent non-tumor liver. Our findings suggest that hepatoblastoma organoids could be used for a variety of applications and have the potential to improve treatment options for the subset of hepatoblastoma patients who do not respond to existing treatments

NEDD4 and NEDD4L regulate Wnt signalling and intestinal stem cell priming by degrading LGR5 receptor

The intestinal stem cell (ISC) marker LGR5 is a receptor for R-spondin (RSPO) that functions to potentiate Wnt signalling in the proliferating crypt. It has been recently shown that Wnt plays a priming role for ISC self-renewal by inducing RSPO receptor LGR5 expression. Despite its pivotal role in homeostasis, regeneration and cancer, little is known about the post-translational regulation of LGR5. Here, we show that the HECT-domain E3 ligases NEDD4 and NEDD4L are expressed in the crypt stem cell regions and regulate ISC priming by degrading LGR receptors. Loss of Nedd4 and Nedd4l enhances ISC proliferation, increases sensitivity to RSPO stimulation and accelerates tumour development in Apcmin mice with increased numbers of high-grade adenomas. Mechanistically, we find that both NEDD4 and NEDD4L negatively regulate Wnt/β-catenin signalling by targeting LGR5 receptor and DVL2 for proteasomal and lysosomal degradation. Our findings unveil the previously unreported post-translational control of LGR receptors via NEDD4/NEDD4L to regulate ISC priming. Inactivation of NEDD4 and NEDD4L increases Wnt activation and ISC numbers, which subsequently enhances tumour predisposition and progression

Long-term culture of genome-stable bipotent stem cells from adult human liver.

Despite the enormous replication potential of the human liver, there are currently no culture systems available that sustain hepatocyte replication and/or function in vitro. We have shown previously that single mouse Lgr5+ liver stem cells can be expanded as epithelial organoids in vitro and can be differentiated into functional hepatocytes in vitro and in vivo. We now describe conditions allowing long-term expansion of adult bile duct-derived bipotent progenitor cells from human liver. The expanded cells are highly stable at the chromosome and structural level, while single base changes occur at very low rates. The cells can readily be converted into functional hepatocytes in vitro and upon transplantation in vivo. Organoids from α1-antitrypsin deficiency and Alagille syndrome patients mirror the in vivo pathology. Clonal long-term expansion of primary adult liver stem cells opens up experimental avenues for disease modeling, toxicology studies, regenerative medicine, and gene therapy.This work was supported by grants to MH (EU/236954) and to HC (The United European Gastroenterology Federation (UEGF) Research Prize 2010, EU/232814-StemCellMark and NWO/116002008). MH is supported by The Wellcome Trust Sir Henry Dale fellowship. The Rspo cell line was kindly provided by Dr. Calvin Kuo.This is the final published version. It first appeared at http://www.cell.com/abstract/S0092-8674%2814%2901566-9

NOX1-dependent redox signaling potentiates colonic stem cell proliferation to adapt to the intestinal microbiota by linking EGFR and TLR activation

The colon epithelium is a primary point of interaction with the microbiome and is regenerated by a few rapidly cycling colonic stem cells (CSCs). CSC self-renewal and proliferation are regulated by growth factors and the presence of bacteria. However, the molecular link connecting the diverse inputs that maintain CSC homeostasis remains largely unknown. We report that CSC proliferation is mediated by redox-dependent activation of epidermal growth factor receptor (EGFR) signaling via NADPH oxidase 1 (NOX1). NOX1 expression is CSC specific and is restricted to proliferative CSCs. In the absence of NOX1, CSCs fail to generate ROS and have a reduced proliferation rate. NOX1 expression is regulated by Toll-like receptor activation in response to the microbiota and serves to link CSC proliferation with the presence of bacterial components in the crypt. The TLR-NOX1-EGFR axis is therefore a critical redox signaling node in CSCs facilitating the quiescent-proliferation transition and responds to the microbiome to maintain colon homeostasis

Profiling Insulin Like Factor 3 (INSL3) Signaling in Human Osteoblasts

Abstract BACKGROUND: Young men with mutations in the gene for the INSL3 receptor (Relaxin family peptide 2, RXFP2) are at risk of reduced bone mass and osteoporosis. Consistent with the human phenotype, bone analyses of Rxfp2(-/-) mice showed decreased bone volume, alterations of the trabecular bone, reduced mineralizing surface, bone formation, and osteoclast surface. The aim of this study was to elucidate the INSL3/RXFP2 signaling pathways and targets in human osteoblasts. METHODOLOGY/PRINCIPAL FINDINGS: Alkaline phosphatase (ALP) production, protein phosphorylation, intracellular calcium, gene expression, and mineralization studies have been performed. INSL3 induced a significant increase in ALP production, and Western blot and ELISA analyses of multiple intracellular signaling pathway molecules and their phosphorylation status revealed that the MAPK was the major pathway influenced by INSL3, whereas it does not modify intracellular calcium concentration. Quantitative Real Time PCR and Western blotting showed that INSL3 regulates the expression of different osteoblast markers. Alizarin red-S staining confirmed that INSL3-stimulated osteoblasts are fully differentiated and able to mineralize the extracellular matrix. CONCLUSIONS/SIGNIFICANCE: Together with previous findings, this study demonstrates that the INSL3/RXFP2 system is involved in bone metabolism by acting on the MAPK cascade and stimulating transcription of important genes of osteoblast maturation/differentiation and osteoclastogenesis

Human extrahepatic and intrahepatic cholangiocyte organoids show region-specific differentiation potential and model cystic fibrosis-related bile duct disease

The development, homeostasis, and repair of intrahepatic and extrahepatic bile ducts are thought to involve distinct mechanisms including proliferation and maturation of cholangiocyte and progenitor cells. This study aimed to characterize human extrahepatic cholangiocyte organoids (ECO) using canonical Wnt-stimulated culture medium previously developed for intrahepatic cholangiocyte organoids (ICO). Paired ECO and ICO were derived from common bile duct and liver tissue, respectively. Characterization showed both organoid types were highly similar, though some differences in size and gene expression were observed. Both ECO and ICO have cholangiocyte fate differentiation capacity. However, unlike ICO, ECO lack the potential for differentiation towards a hepatocyte-like fate. Importantly, ECO derived from a cystic fibrosis patient showed no CFTR channel activity but normal chloride channel and MDR1 transporter activity. In conclusion, this study shows that ECO and ICO have distinct lineage fate and that ECO provide a competent model to study extrahepatic bile duct diseases like cystic fibrosis