Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

A Peer-Based Strategy to Overcome HPV Vaccination Inequities in Rural Communities: A Physical Distancing-Compliant Approach.

The human papilloma virus (HPV) vaccine is the world\u27s first proven and effective vaccine to prevent cancers in males and females when administered pre-exposure. Like most of the US, barely half of Vermont teens are up-to-date with the vaccination, with comparable deficits in New Hampshire and Maine. The rates for HPV vaccine initiation and completion are as low as 33% in rural New England. Consequently, there is a compelling responsibility to communicate its importance to unvaccinated teenagers before their risk for infection increases. Messaging in rural areas promoting HPV vaccination is compromised by community-based characteristics that include access to appropriate medical care, poor media coverage, parental and peer influence, and skepticism of science and medicine. Current strategies are predominantly passive access to literature and Internet-based information. Evidence indicates that performance-based messaging can clarify the importance of HPV vaccination to teenagers and their parents in rural areas. Increased HPV vaccination will significantly contribute to the prevention of a broadening spectrum of cancers. Reducing rurality-based inequities is a public health priority. Development of a performance-based peer-communication intervention can capture a window of opportunity to provide increasingly effective and sustained HPV protection. An effective approach can be partnering rural schools and regional health teams with a program that is nimble and scalable to respond to public health policies and practices compliant with COVID-19 pandemic-related modifications on physical distancing and interacting in the foreseeable future

SIRT1 Activators Suppress Inflammatory Responses through Promotion of p65 Deacetylation and Inhibition of NF-κB Activity

<div><p>Chronic inflammation is a major contributing factor in the pathogenesis of many age-associated diseases. One central protein that regulates inflammation is NF-κB, the activity of which is modulated by post-translational modifications as well as by association with co-activator and co-repressor proteins. SIRT1, an NAD⁺-dependent protein deacetylase, has been shown to suppress NF-κB signaling through deacetylation of the p65 subunit of NF-κB resulting in the reduction of the inflammatory responses mediated by this transcription factor. The role of SIRT1 in the regulation of NF-κB provides the necessary validation for the development of pharmacological strategies for activating SIRT1 as an approach for the development of a new class of anti-inflammatory therapeutics. We report herein the development of a quantitative assay to assess compound effects on acetylated p65 protein in the cell. We demonstrate that small molecule activators of SIRT1 (STACs) enhance deacetylation of cellular p65 protein, which results in the suppression of TNFα-induced NF-κB transcriptional activation and reduction of LPS-stimulated TNFα secretion in a SIRT1-dependent manner. In an acute mouse model of LPS-induced inflammation, the STAC SRTCX1003 decreased the production of the proinflammatory cytokines TNFα and IL-12. Our studies indicate that increasing SIRT1-mediated NF-κB deacetylation using small molecule activating compounds is a novel approach to the development of a new class of therapeutic anti-inflammatory agents.</p> </div

Structures of STACs.

<p>(A) Structures of the benzimidazole STACs. (B) Core structure of the quinolone STACs.</p

Development of a quantitative assay for measuring cellular acetylated p65 levels.

<p>Flow diagram depicts the assay procedure, including BacMam p65 and BacMam p300 viral transduction, plating cells, compound treatment, cell lysis and detection of acetylated p65 protein by AlphaScreen format.</p

SIRT1 regulates cellular acetylated p65 levels.

<p>(A) Levels of acetylated p65 in BacMam SIRT1 virus or BacMam GFP virus transduced cells. Western blot in the right upper corner shows the level of SIRT1 expression upon BacMam SIRT1 virus transduction. (B–C) Levels of acetylated p65 protein in cells transfected with SIRT1-siRNA or NT-siRNA as measured by AlphaScreen assay in (B), or demonstrated by immunoblotting in (C). (D) Dose-response effect of EX-527 on levels of acetylated p65 protein. All error bars represent s.d. of at least 4 replicates. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001.</p

STACs suppress stimuli-induced NF-κB transcriptional activation and cytokine secretion.

<p>(A) NF-κB luciferase reporter counts in HEK 293 cells transfected with SIRT1 or empty vector with or without TNFα stimulation. Western blot in the right upper corner indicated SIRT1 expression induced by SIRT1 plasmid transfection. (B) Dose-response effect of SRTCX1002 on NF-κB luciferase reporter activity in HEK293 cells stimulated with TNFα. Error bars represent s.d. of at least four replicates. (C) Dose-response effect of SRTCX1003 on LPS-induced TNFα secretion from RAW cells. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001.</p

STACs attenuate TNFα-induced p65 acetylation.

<p>(A) Western blots of immunoprecipitated acetylated K310-p65 protein, p65 and actin protein in whole cell lysates from vector or p300 transfected cells with or without TNFα stimulation. (B) Left panel, western blots of immunoprecipitated acetylated K310-p65 protein, SIRT1, p65 and tubulin in whole cell lysates from TNFα-stimulated cells transfected with p300 plus empty vector or p300 plus SIRT1. Right panel, western blots of immunoprecipitated acetylated K310-p65 protein, p65 and tubulin in whole cell lysates from TNFα-stimulated p300 overexpressing cells that were pretreated with EX-527 for 6 hours. (C) Upper panels, western blots of immunoprecipitated acetylated K310-p65 protein, p65, phospho-NF-κB (Ser536), phospho-IκBα (Ser32/Ser36) and tubulin in whole cell lysates from cells pretreated with compounds for 6 hours followed by 20 minutes TNFα stimulation. Lower panels, densitometry quantitation of the western blot for immunoprecipitated acetylated p65 protein. The level of acetylated p65 in TNFα-stimulated and vehicle treated sample was set as 1. Experiments were repeated at least 2 times and western blots from one experiment were shown as representative. Error bars present s.d. of densitometry quantitation of western blots from at least two experiments. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001.</p

SRTCX1003 decreases LPS-stimulated cytokine production <i>in vivo</i>.

<p>(A) Schematic illustration of the acute LPS-induced inflammation mouse model. (B) Compound plasma concentration of mice dosed with SRTCX1003 for 2.5 hours. (C–D) Dose-response effect of SRTCX1003 and the effect of 1 mg/kg dexamethasone on LPS-induced production of TNFα in (C) and IL-12p40 in (D). Error bars represent s.d. of eight mice in each group. Please note that reduction of TNFα and IL-12p40 by SRTCX1003 at 100 mg/kg was comparable to that induced by 1 mg/kg dexamethasone. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001.</p