The TGFβ type I receptor TGFβRI functions as an inhibitor of BMP signaling in cartilage.

The type I TGFβ receptor TGFβRI (encoded by Tgfbr1) was ablated in cartilage. The resulting Tgfbr1 Col2 mice exhibited lethal chondrodysplasia. Similar defects were not seen in mice lacking the type II TGFβ receptor or SMADs 2 and 3, the intracellular mediators of canonical TGFβ signaling. However, we detected elevated BMP activity in Tgfbr1 Col2 mice. As previous studies showed that TGFβRI can physically interact with ACVRL1, a type I BMP receptor, we generated cartilage-specific Acvrl1 (Acvrl1 Col2 ) and Acvrl1/Tgfbr1 (Acvrl1/Tgfbr1 Col2 ) knockouts. Loss of ACVRL1 alone had no effect, but Acvrl1/Tgfbr1 Col2 mice exhibited a striking reversal of the chondrodysplasia seen in Tgfbr1 Col2 mice. Loss of TGFβRI led to a redistribution of the type II receptor ACTRIIB into ACVRL1/ACTRIIB complexes, which have high affinity for BMP9. Although BMP9 is not produced in cartilage, we detected BMP9 in the growth plate, most likely derived from the circulation. These findings demonstrate that the major function of TGFβRI in cartilage is not to transduce TGFβ signaling, but rather to antagonize BMP signaling mediated by ACVRL1

Dirty Work, Clean Hands: The Moral Psychology of Indirect Agency

When powerful people cause harm, they often do so indirectly through other people. Are harmful actions carried out through others evaluated less negatively than harmful actions carried out directly? Four experiments examine the moral psychology of indirect agency. Experiments 1A, 1B, and 1C reveal effects of indirect agency under conditions favoring intuitive judgment, but not reflective judgment, using a joint/separate evaluation paradigm. Experiment 2A demonstrates that effects of indirect agency cannot be fully explained by perceived lack of foreknowledge or control on the part of the primary agent. Experiment 2B indicates that reflective moral judgment is sensitive to indirect agency, but only to the extent that indirectness signals reduced foreknowledge and/or control. Experiment 3 indicates that effects of indirect agency result from a failure to automatically consider the potentially dubious motives of agents who cause harm indirectly. Experiment 4 demonstrates an effect of indirect agency on purchase intentions.Psycholog

Low Hydrophobic Mismatch Scores Calculated for HLA-A/B/DR/DQ Loci Improve Kidney Allograft Survival

We evaluated the impact of human leukocyte antigen (HLA) disparity (immunogenicity; IM) on long-term kidney allograft survival. The IM was quantified based on physicochemical properties of the polymorphic linear donor/recipient HLA amino acids (the Cambridge algorithm) as a hydrophobic, electrostatic, amino acid mismatch scores (HMS\AMS\EMS) or eplet mismatch (EpMM) load. High-resolution HLA-A/B/DRB1/DQB1 types were imputed to calculate HMS for primary/re-transplant recipients of deceased donor transplants. The multiple Cox regression showed the association of HMS with graft survival and other confounders. The HMS integer 0-10 scale showed the most survival benefit between HMS 0 and 3. The Kaplan-Meier analysis showed that: the HMS=0 group had 18.1-year median graft survival, a 5-year benefit over HMS\u3e0 group; HMS ≤ 3.0 had 16.7-year graft survival, a 3.8-year better than HMS\u3e3.0 group; and, HMS ≤ 7.8 had 14.3-year grafts survival, a 1.8-year improvement over HMS\u3e7.8 group. Stratification based on EMS, AMS or EpMM produced similar results. Additionally, the importance of HLA-DR with/without -DQ IM for graft survival was shown. In our simulation of 1,000 random donor/recipient pairs, 75% with HMS\u3e3.0 were re-matched into HMS ≤ 3.0 and the remaining 25% into HMS≥7.8: after re-matching, the 13.5 years graft survival would increase to 16.3 years. This approach matches donors to recipients with low/medium IM donors thus preventing transplants with high IM donors

Harmful Algal Bloom Impacts on Human Health: An Analysis of National Emergency Department Data in the U.S. from 2016 to 2018

Numerous harmful algal bloom (HAB) cyanobacterial species produce toxins that disrupt ecosystems and are harmful to both human and animal health. We sought to determine trends and patterns in diagnostic codes relating to HAB exposures from the Healthcare Cost and Utilization Project\u27s (HCUP) Nationwide Emergency Department Sample (NEDS). We analyzed years 2016 to 2018 as these represented the years in which complete data was available using the World Health Organization (WHO) International Classification of Diseases-10 diagnosis codes for HAB exposure. For each year\u27s grouping, statistical analysis was performed to uncover patterns and trends. Each patient occurrence was screened for the most prevalent comorbidities associated with HAB exposures. Over the 3-year period studied, there were 118 reported patient admissions to the Emergency Department. Respiratory related illness accounted for the majority of comorbidities and were present in 53% of patients, including 30% as the primary diagnostic code. These data represent one of the first attempts to analyze HAB exposure related illness presenting to Emergency Departments in the United States. The predominance of respiratory related diagnostic codes in these patients suggests greater attention to these conditions in the risk characterization of HAB exposure in the development of evidence-based prevention and treatment strategies.Harmful Algal Bloom Impacts on Human Health: An Analysis of National Emergency Department Data in the U.S. from 2016 to 201

Smad2 and Smad3 Regulate Chondrocyte Proliferation and Differentiation in the Growth Plate.

TGFβs act through canonical and non-canonical pathways, and canonical signals are transduced via Smad2 and Smad3. However, the contribution of canonical vs. non-canonical pathways in cartilage is unknown because the role of Smad2 in chondrogenesis has not been investigated in vivo. Therefore, we analyzed mice in which Smad2 is deleted in cartilage (Smad2CKO), global Smad3-/- mutants, and crosses of these strains. Growth plates at birth from all mutant strains exhibited expanded columnar and hypertrophic zones, linked to increased proliferation in resting chondrocytes. Defects were more severe in Smad2CKO and Smad2CKO;Smad3-/- (Smad2/3) mutant mice than in Smad3-/- mice, demonstrating that Smad2 plays a role in chondrogenesis. Increased levels of Ihh RNA, a key regulator of chondrocyte proliferation and differentiation, were seen in prehypertrophic chondrocytes in the three mutant strains at birth. In accordance, TGFβ treatment decreased Ihh RNA levels in primary chondrocytes from control (Smad2fx/fx) mice, but inhibition was impaired in cells from mutants. Consistent with the skeletal phenotype, the impact on TGFβ-mediated inhibition of Ihh RNA expression was more severe in Smad2CKO than in Smad3-/- cells. Putative Smad2/3 binding elements (SBEs) were identified in the proximal Ihh promoter. Mutagenesis demonstrated a role for three of them. ChIP analysis suggested that Smad2 and Smad3 have different affinities for these SBEs, and that the repressors SnoN and Ski were differentially recruited by Smad2 and Smad3, respectively. Furthermore, nuclear localization of the repressor Hdac4 was decreased in growth plates of Smad2CKO and double mutant mice. TGFβ induced association of Hdac4 with Smad2, but not with Smad3, on the Ihh promoter. Overall, these studies revealed that Smad2 plays an essential role in the development of the growth plate, that both Smads 2 and 3 inhibit Ihh expression in the neonatal growth plate, and suggested they accomplish this by binding to distinct SBEs, mediating assembly of distinct repressive complexes

Smad2 and Smad3 Regulate Chondrocyte Proliferation and Differentiation in the Growth Plate.

TGFβs act through canonical and non-canonical pathways, and canonical signals are transduced via Smad2 and Smad3. However, the contribution of canonical vs. non-canonical pathways in cartilage is unknown because the role of Smad2 in chondrogenesis has not been investigated in vivo. Therefore, we analyzed mice in which Smad2 is deleted in cartilage (Smad2CKO), global Smad3-/- mutants, and crosses of these strains. Growth plates at birth from all mutant strains exhibited expanded columnar and hypertrophic zones, linked to increased proliferation in resting chondrocytes. Defects were more severe in Smad2CKO and Smad2CKO;Smad3-/- (Smad2/3) mutant mice than in Smad3-/- mice, demonstrating that Smad2 plays a role in chondrogenesis. Increased levels of Ihh RNA, a key regulator of chondrocyte proliferation and differentiation, were seen in prehypertrophic chondrocytes in the three mutant strains at birth. In accordance, TGFβ treatment decreased Ihh RNA levels in primary chondrocytes from control (Smad2fx/fx) mice, but inhibition was impaired in cells from mutants. Consistent with the skeletal phenotype, the impact on TGFβ-mediated inhibition of Ihh RNA expression was more severe in Smad2CKO than in Smad3-/- cells. Putative Smad2/3 binding elements (SBEs) were identified in the proximal Ihh promoter. Mutagenesis demonstrated a role for three of them. ChIP analysis suggested that Smad2 and Smad3 have different affinities for these SBEs, and that the repressors SnoN and Ski were differentially recruited by Smad2 and Smad3, respectively. Furthermore, nuclear localization of the repressor Hdac4 was decreased in growth plates of Smad2CKO and double mutant mice. TGFβ induced association of Hdac4 with Smad2, but not with Smad3, on the Ihh promoter. Overall, these studies revealed that Smad2 plays an essential role in the development of the growth plate, that both Smads 2 and 3 inhibit Ihh expression in the neonatal growth plate, and suggested they accomplish this by binding to distinct SBEs, mediating assembly of distinct repressive complexes

The TGFβ type I receptor TGFβRI functions as an inhibitor of BMP signaling in cartilage.

The type I TGFβ receptor TGFβRI (encoded by Tgfbr1) was ablated in cartilage. The resulting Tgfbr1 Col2 mice exhibited lethal chondrodysplasia. Similar defects were not seen in mice lacking the type II TGFβ receptor or SMADs 2 and 3, the intracellular mediators of canonical TGFβ signaling. However, we detected elevated BMP activity in Tgfbr1 Col2 mice. As previous studies showed that TGFβRI can physically interact with ACVRL1, a type I BMP receptor, we generated cartilage-specific Acvrl1 (Acvrl1 Col2 ) and Acvrl1/Tgfbr1 (Acvrl1/Tgfbr1 Col2 ) knockouts. Loss of ACVRL1 alone had no effect, but Acvrl1/Tgfbr1 Col2 mice exhibited a striking reversal of the chondrodysplasia seen in Tgfbr1 Col2 mice. Loss of TGFβRI led to a redistribution of the type II receptor ACTRIIB into ACVRL1/ACTRIIB complexes, which have high affinity for BMP9. Although BMP9 is not produced in cartilage, we detected BMP9 in the growth plate, most likely derived from the circulation. These findings demonstrate that the major function of TGFβRI in cartilage is not to transduce TGFβ signaling, but rather to antagonize BMP signaling mediated by ACVRL1

Hdac4, SnoN, and Ski protein distribution in the neonatal growth plate and binding to S1-3 in the <i>Ihh</i> promoter.

<p><b>(A)</b> Immunohistochemical staining of Hdac4 in E18.5 proximal tibias. Bottom panel shows magnifications of the boxed regions in top panel, representing the lower columnar and prehypertrophic zones. <b>(B)</b> ChIP analysis of Hdac4 binding to SBE 1–3 in ATDC5 cells. <b>(C and D)</b> Immunohistochemical staining of SnoN and Ski in E18.5 proximal tibias. Asterisks in (C and D) indicated SnoN or Ski present in columnar zone cells. <b>(E)</b> ChIP analysis of SnoN and Ski binding to SBE 1–3 in ATDC5 cells. Cells in (B and E) were matured to prehypertrophy and subsequently treated with TGFβ1 (5 ng/ml) or non-treated (Control) for 24 hrs. Asterisk in (B and E), p = or < 0.05. All experiments were performed in triplicate and repeated twice. <i>Smad2</i>^<i>CKO</i> = <i>Smad2</i>^<i>fx/fx</i><i>;Col2a1Cre</i>.</p

Smad2 and Smad3 Regulate Chondrocyte Proliferation and Differentiation in the Growth Plate

<div><p>TGFβs act through canonical and non-canonical pathways, and canonical signals are transduced via Smad2 and Smad3. However, the contribution of canonical vs. non-canonical pathways in cartilage is unknown because the role of Smad2 in chondrogenesis has not been investigated <i>in vivo</i>. Therefore, we analyzed mice in which Smad2 is deleted in cartilage (<i>Smad2</i>^<i>CKO</i>), global <i>Smad3</i>^<i>-/-</i> mutants, and crosses of these strains. Growth plates at birth from all mutant strains exhibited expanded columnar and hypertrophic zones, linked to increased proliferation in resting chondrocytes. Defects were more severe in <i>Smad2</i>^<i>CKO</i> and <i>Smad2</i>^<i>CKO</i><i>;Smad3</i>^-/- <i>(Smad2/3)</i> mutant mice than in <i>Smad3</i>^<i>-/-</i> mice, demonstrating that Smad2 plays a role in chondrogenesis. Increased levels of <i>Ihh</i> RNA, a key regulator of chondrocyte proliferation and differentiation, were seen in prehypertrophic chondrocytes in the three mutant strains at birth. In accordance, TGFβ treatment decreased <i>Ihh</i> RNA levels in primary chondrocytes from control (<i>Smad2</i>^<i>fx/fx</i>) mice, but inhibition was impaired in cells from mutants. Consistent with the skeletal phenotype, the impact on TGFβ-mediated inhibition of <i>Ihh</i> RNA expression was more severe in <i>Smad2</i>^<i>CKO</i> than in <i>Smad3</i>^<i>-/-</i> cells. Putative Smad2/3 binding elements (SBEs) were identified in the proximal <i>Ihh</i> promoter. Mutagenesis demonstrated a role for three of them. ChIP analysis suggested that Smad2 and Smad3 have different affinities for these SBEs, and that the repressors SnoN and Ski were differentially recruited by Smad2 and Smad3, respectively. Furthermore, nuclear localization of the repressor Hdac4 was decreased in growth plates of <i>Smad2</i>^<i>CKO</i> and double mutant mice. TGFβ induced association of Hdac4 with Smad2, but not with Smad3, on the <i>Ihh</i> promoter. Overall, these studies revealed that Smad2 plays an essential role in the development of the growth plate, that both Smads 2 and 3 inhibit <i>Ihh</i> expression in the neonatal growth plate, and suggested they accomplish this by binding to distinct SBEs, mediating assembly of distinct repressive complexes.</p></div