Distamycin A Inhibits HMGA1-Binding to the P-Selectin Promoter and Attenuates Lung and Liver Inflammation during Murine Endotoxemia

Background: The architectural transcription factor High Mobility Group-A1 (HMGA1) binds to the minor groove of AT-rich DNA and forms transcription factor complexes (“enhanceosomes”) that upregulate expression of select genes within the inflammatory cascade during critical illness syndromes such as acute lung injury (ALI). AT-rich regions of DNA surround transcription factor binding sites in genes critical for the inflammatory response. Minor groove binding drugs (MGBs), such as Distamycin A (Dist A), interfere with AT-rich region DNA binding in a sequence and conformation-specific manner, and HMGA1 is one of the few transcription factors whose binding is inhibited by MGBs. Objectives: To determine whether MGBs exert beneficial effects during endotoxemia through attenuating tissue inflammation via interfering with HMGA1-DNA binding and modulating expression of adhesion molecules. Methodology/Principal Findings: Administration of Dist A significantly decreased lung and liver inflammation during murine endotoxemia. In intravital microscopy studies, Dist A attenuated neutrophil-endothelial interactions in vivo following an inflammatory stimulus. Endotoxin induction of P-selectin expression in lung and liver tissue and promoter activity in endothelial cells was significantly reduced by Dist A, while E-selectin induction was not significantly affected. Moreover, Dist A disrupted formation of an inducible complex containing NF-κB that binds an AT-rich region of the P-selectin promoter. Transfection studies demonstrated a critical role for HMGA1 in facilitating cytokine and NF-κB induction of P-selectin promoter activity, and Dist A inhibited binding of HMGA1 to this AT-rich region of the P-selectin promoter in vivo. Conclusions/Significance: We describe a novel targeted approach in modulating lung and liver inflammation in vivo during murine endotoxemia through decreasing binding of HMGA1 to a distinct AT-rich region of the P-selectin promoter. These studies highlight the ability of MGBs to function as molecular tools for dissecting transcriptional mechanisms in vivo and suggest alternative treatment approaches for critical illness

Misinformed Consent: Upholding the Constitutionality of South Dakota\u27s Suicide Advisory in \u3cem\u3ePlanned Parenthood Minnesota, North Dakota, South Dakota v. Rounds\u3c/em\u3e

On July 24, 2012, in Planned Parenthood Minnesota, North Dakota, South Dakota v. Rounds, the U.S. Court of Appeals for the Eighth Circuit, sitting en banc, considered the constitutionality of a suicide advisory portion of a South Dakota statute that required informed consent for abortions. The Eight Circuit found the advisory was constitutional because the information disclosures required by the statute were truthful, nonmisleading, and relevant to the patient’s decision to have an abortion. The court relied in part on the Supreme Court’s 2007 decision Gonzales v. Carhart, which held that Congress has the authority to legislate in the abortion context, even in areas of medical uncertainty. The Eighth Circuit, however, misapplied the reasoning of Gonzales and dangerously minimized the standard for scientific evidence in informed consent laws

Statutory Prohibitions on Wrongful Birth Claims & Their Dangerous Effects on Parents

Wrongful birth claims are negligence actions brought on behalf of children born with disabilities or genetic disorders that were not properly diagnosed before the child’s birth. The plaintiffs, typically the parents of the afflicted child, argue that without the defendant’s negligence, the parents would have had the opportunity to prevent the child’s birth and subsequent condition by choosing to terminate the pregnancy. A number of states have responded to the growing prevalence of wrongful birth claims by enacting legislation that bars plaintiffs from bringing wrongful birth actions. These statutes, however, pose a threat to the parental rights of disabled or terminally ill children, as they diminish abortion rights and bar parents from recovering the enormous medical and emotional damages of giving birth to the afflicted child. States should not prohibit this cause of action and, instead, the merits of these claims should be decided through the court system

Developing a Broader Understanding of the Effects of Fecal Microbiota Transplantation on the Health and Microbiome of Donors and Patients with Recurrent Clostridiodes difficile Infection

This collection of papers investigates Clostridiodes difficile infection (CDI) and its treatment with fecal microbiota transplantation (FMT) from a more holistic perspective. Bacterial composition of FMT donors and patients with CDI at 4 timepoints (pre-FMT, 10 days, 5 weeks, and 13 weeks post-FMT) were analyzed from 16S rRNA sequence data. Trends in microbiome composition pre and post treatment with FMT were observed to establish which bacteria are responsible for curing patients of CDI and restoring quality of life. Bacterial composition was found to adjust over time after treatment to more closely resemble healthy donors. Donors were studied to identify if bacterial diversity remained stable between donors and were found to have no significant difference in Shannon diversity, indicating similarly diverse bacteria were supplied to each patient. By obtaining a more holistic view, we aim to understand the mechanisms of FMT better and produce precision care for patients suffering from CDI.Ontario Graduate Scholarshi

Precision medicine and gut dysbiosis

Clostridioides difficile Infection (CDI) is a leading cause of healthcare-associated infections in Canada, affecting the gastrointestinal tract which can lead to fever, abdominal pain, and diarrhea. Effective treatment for patients with Recurrent CDI (rCDI) can be achieved by Fecal Microbiota Transplantation (FMT) by introducing the gut micro-organisms of a healthy person (donor) into the bowel of the affected individual. Research has shown that an increase in the specific bacterial phyla post-FMT may be partly responsible for this gut restoration and elimination of disease. Furthermore, in understanding the key bacteria associated with successful FMT, full treatment plans can be developed for the individual needs of the patient by matching an infected individual with a donor possessing ideal microbiota for the specific patient. This development of precision medicine and more systematic adoption of FMT can be the next step toward more rapid resolution of rCDI. </jats:p

La distamicina A inhibe la unión de HMGA1 al promotor de la selectina P y atenúa la inflamación pulmonar y hepática durante la endotoxemia murina

Background The architectural transcription factor High Mobility Group-A1 (HMGA1) binds to the minor groove of AT-rich DNA and forms transcription factor complexes (“enhanceosomes”) that upregulate expression of select genes within the inflammatory cascade during critical illness syndromes such as acute lung injury (ALI). AT-rich regions of DNA surround transcription factor binding sites in genes critical for the inflammatory response. Minor groove binding drugs (MGBs), such as Distamycin A (Dist A), interfere with AT-rich region DNA binding in a sequence and conformation-specific manner, and HMGA1 is one of the few transcription factors whose binding is inhibited by MGBs. Objectives To determine whether MGBs exert beneficial effects during endotoxemia through attenuating tissue inflammation via interfering with HMGA1-DNA binding and modulating expression of adhesion molecules. Methodology/Principal Findings Administration of Dist A significantly decreased lung and liver inflammation during murine endotoxemia. In intravital microscopy studies, Dist A attenuated neutrophil-endothelial interactions in vivo following an inflammatory stimulus. Endotoxin induction of P-selectin expression in lung and liver tissue and promoter activity in endothelial cells was significantly reduced by Dist A, while E-selectin induction was not significantly affected. Moreover, Dist A disrupted formation of an inducible complex containing NF-?B that binds an AT-rich region of the P-selectin promoter. Transfection studies demonstrated a critical role for HMGA1 in facilitating cytokine and NF-?B induction of P-selectin promoter activity, and Dist A inhibited binding of HMGA1 to this AT-rich region of the P-selectin promoter in vivo. Conclusions/Significance We describe a novel targeted approach in modulating lung and liver inflammation in vivo during murine endotoxemia through decreasing binding of HMGA1 to a distinct AT-rich region of the P-selectin promoter. These studies highlight the ability of MGBs to function as molecular tools for dissecting transcriptional mechanisms in vivo and suggest alternative treatment approaches for critical illness

Novel Microcephalic Primordial Dwarfism Disorder Associated with Variants in the Centrosomal Protein Ninein

Microcephalic primordial dwarfism (MPD) is a rare, severe form of human growth failure in which growth restriction is evident in utero and continues into postnatal life. Single causative gene defects have been identified in a number of patients with MPD, and all involve genes fundamental to cellular processes including centrosome functions

Distamycin A attenuates inflammatory cytokine-induced neutrophil-endothelial interactions.

<p><b>A.</b> Wild type mice were subjected to intravital microscopy analysis at 3–4 hours following treatment with TNF-α/DistA or TNF-α/Vehicle (Veh) (n = 3 mice per treatment group with multiple vessels and vascular segments analyzed for each animal). Representative still photos of vascular segments are shown for the 4-hour time point. White round spots lining the vascular wall represent adherent leukocytes. <b>B.</b> Formal video analysis of the phases of neutrophil-endothelial interaction was performed, and values for all vascular segments within a particular treatment group were averaged for each time point. A significant reduction in rolling fraction and sticking efficiency was detected in the TNF-α/Dist A mice at 3–4 hours (p = 0.0001 and 0.0004, respectively), compared with the TNF-α/Vehicle mice.</p

Distamycin A attenuates endotoxin-induced lung and liver inflammation.

<p><b>A.</b> Lungs were harvested from C57BL/6 male mice 24 hours following treatment with Vehicle (Veh), LPS/Vehicle (LPS+Veh) or LPS/Dist A (LPS+Dist A) i.p. (n = 9 mice per group). Lungs were fixed, processed, sectioned, and stained for Gr-1 (a marker of neutrophils, positive staining indicated by brown cells). Representative sections for each condition are shown (200× magnification). Using NIH Image Software, 5 fields (at 200× magnification) per lung section from mice from each treatment group were quantified, with brown pixels counted as positive staining. Results were expressed as % positively stained area per 200× field. (*p<0.05 compared with Veh; **p<0.05 compared with LPS+Veh). <b>B.</b> Liver tissue was harvested from C57BL/6 mice 4 h after treatment with Vehicle (Veh), LPS/Vehicle (LPS+Veh) or LPS/Dist A (LPS+DistA) i.p., (n = at least 4 mice per group) then processed, stained for Gr-1 (neutrophil marker), and analyzed as described above for Fig. 1A. (*p<0.05 compared with Veh; **p<0.05 compared with LPS+Veh).</p