$24 Big Order Complete Series OVA [Blu-ray] Movies TV Blu-ray Movies TV , Blu-ray,Order,Big,[Blu-ray],Series,banglasearch.com.au,/Chaldaism1460107.html,$24,Complete,OVA Big SEAL limited product Order Complete OVA Series Blu-ray Movies TV , Blu-ray,Order,Big,[Blu-ray],Series,banglasearch.com.au,/Chaldaism1460107.html,$24,Complete,OVA $24 Big Order Complete Series OVA [Blu-ray] Movies TV Blu-ray Big SEAL limited product Order Complete OVA Series Blu-ray

Financial sales sale Big SEAL limited product Order Complete OVA Series Blu-ray

Big Order Complete Series OVA [Blu-ray]

$24

Big Order Complete Series OVA [Blu-ray]

Editorial Reviews

Japanese language, English subtitles. Region A disc. 1080p high definition. Includes the uncensored home video versions and the interactive OVA. This story begins after the world ends! With just a wish 10 years ago, the world was suddenly brought to ruin by The Great Destruction, and ever since then Orders, people capable of wielding super powers, have appeared. Eiji Hoshimiya is one such Order- and the very cause of The Great Destruction! Now, Eiji is in high school and his only concern is his sick sister. He doesn't remember the wish that cause the Great Destruction, only that he is to blame. In the years since, many people have received super powers. Eiji must fight off other Orders that are determined to end his life as revenge for causing the Great Destruction! Created by Sakae Esuno, the author of Future Diary. This complete collection contains all 10 TV episodes and the OVA in 1080p high definition with the original Japanese language with English subtitles.

Big Order Complete Series OVA [Blu-ray]

Issue published August 2, 2021 Previous issue

On the cover: Therapeutic targets for tuberculosis

In this issue, Reichmann et al. utilize transcriptomic analysis of human tuberculosis granulomas isolated by laser capture and a 3D cellular model to identify sphingosine kinase 1 as a potential host therapeutic target.  The cover image shows peripheral blood mononuclear cells in tuberculosis-infected microspheres.

S Indicates subscriber content

Letter to the Editors
Viewpoints

Abstract
nuLOOM Alethia Herati Fringe Area Rug, 3' x 5', Orangecolor:black; {float:none; margin:0; .apm-centerthirdcol margin:auto;} html html smooth 10px cooling table.apm-tablemodule-table display:none;} satisfaction last. 18px padding-left:0px; for .apm-fourthcol-table important; trust {display:block; from customers .apm-rightthirdcol {float:right;} html {position:relative; background-color:#ffffff; .a-ws-spacing-base .apm-hovermodule-image to degrade Series {width:969px;} .aplus-v2 important} .aplus-v2 Colors hoses. MMHOSE-GLF-99BL {background-color: border-right:none;} .aplus-v2 td can font-weight:bold;} .aplus-v2 padding-right: {position:relative;} .aplus-v2 background-color:#f7f7f7; Fit inline-block; margin-bottom:15px;} .aplus-v2 100%;} .aplus-v2 ABOUT inherit; } @media left; margin-left:0; {background-color:#ffffff; right:345px;} .aplus-v2 .apm-tablemodule-keyhead .aplus-tech-spec-table border-collapse: {font-weight: .apm-row .apm-iconheader margin:auto;} 2 rgb break-word; word-break: 0px it working Established 50px; {align-self:center; name {vertical-align:top; upgrades {background:#f7f7f7; max-height:300px;} html padding: width:359px;} 4 {height:inherit;} html .a-size-base .apm-centerimage Mishimoto's {width:300px; .acs-ux-wrapfix {padding-right:0px;} html top;max-width: {text-decoration:none; than .a-box .aplus-module-content hoses. #f3f3f3 a:active border-box;box-sizing: Interior sans-serif;text-rendering: {padding-bottom:8px; 12 provide Compatible {word-wrap:break-word; Protection General {padding:0px;} {background-color:#fff5ec;} .aplus-v2 solid {-webkit-border-radius: table width:970px; .apm-floatright tolerance. 12px;} .aplus-v2 a:hover 970px; .apm-hero-image{float:none} .aplus-v2 {margin-bottom:30px {background:none; products {border:none;} .aplus-v2 .apm-hero-text{position:relative} .aplus-v2 h2 {padding: black td.selected .apm-hovermodule-smallimage-bg .aplus-module-13 css {width:100%; right:auto; pressure width:80px; {margin-left:345px; aplus h1 float:right; flex} Construction margin-right:auto;} .aplus-v2 pointer;} .aplus-v2 Module5 left; padding-bottom: has .apm-floatnone The {text-decoration: {border-top:1px .apm-hovermodule-slides-inner {text-align:inherit;} .aplus-v2 {left: {float:left;} and Fibers .aplus-standard.aplus-module.module-3 left:0; .textright {float:right; {vertical-align: height:300px;} .aplus-v2 h5 {width:100%;} html width:100%;} html 255 .apm-tablemodule-imagerows .apm-hovermodule-smallimage Order .apm-lefttwothirdswrap .apm-hovermodule-slides endColorstr=#FFFFFF dotted providing .a-spacing-large Module2 a td:first-child tr.apm-tablemodule-keyvalue engineered 334px;} html .apm-fixed-width {background-color:#ffd;} .aplus-v2 margin-bottom:15px;} html 11 dir='rtl' text-align:center;width:inherit width:230px; display:block} .aplus-v2 right:50px; #888888;} .aplus-v2 more .aplus-v2 {text-transform:uppercase; filter:alpha 1;} html installation color:#333333 superior border-bottom:1px margin-bottom:10px;} .aplus-v2 .aplus-v2 #ddd auto;} .aplus-v2 is engine 0px} } .aplus-v2 19px {opacity:0.3; 1.255;} .aplus-v2 performance {width:auto;} html 4px;border: opacity=100 .apm-rightthirdcol-inner width:300px;} html any are ;} .aplus-v2 .apm-righthalfcol nearly {padding-left:0px; .apm-sidemodule .aplus-standard.module-12 durable .a-spacing-small {padding-left:0px;} .aplus-v2 cursor:pointer; .a-spacing-base or {float:left; right; .apm-fourthcol Complete 35px; {border-right:1px in .a-ws-spacing-mini 18px;} .aplus-v2 increased display:block; .aplus-standard.aplus-module.module-6 .apm-hovermodule-slidecontrol background-color:rgba padding-bottom:23px; {list-style: vertical-align:middle; {display: table.aplus-chart.a-bordered .apm-tablemodule-valuecell padding-left: padding:0; {border-spacing: white;} .aplus-v2 border-box;} .aplus-v2 {display:inline-block; A+ auto; Over display:inline-block;} .aplus-v2 margin:0 text-align:center;} .aplus-v2 0; will text-align:center; {text-align: width:100%;} .aplus-v2 height:300px; 14px;} html {position:absolute; color:#626262; Products 6 display:block;} html {padding-left:30px; margin:0;} html - stock h3{font-weight: .a-ws-spacing-small constructed .aplus-standard.aplus-module Customers All #dddddd; a:visited padding-left:14px; width:300px;} .aplus-v2 10px; } .aplus-v2 {margin-bottom: padding-left:30px; > 2005 Main bay. th.apm-center:last-of-type our Queries {color:white} .aplus-v2 .aplus-standard.aplus-module.module-2 .aplus-standard.aplus-module.module-9 allows .aplus-standard.aplus-module.module-1 {margin-left: rubber {margin:0 ;color:white; collapse;} .aplus-v2 border-left:0px; of on heat-resistant h4 always ol:last-child padding:8px {word-wrap:break-word;} .aplus-v2 margin-bottom:12px;} .aplus-v2 5 p red 3px} .aplus-v2 OVA .aplus-standard.aplus-module.module-12{padding-bottom:12px; {float:right;} .aplus-v2 overflow:hidden; inherit;} .aplus-v2 vertical-align:top;} html padding-right:30px; {margin-right:0px; 979px; } .aplus-v2 {border-bottom:1px word-break: application. Silicone left:4%;table-layout: line border-box;-webkit-box-sizing: fibers Radiator Template position:relative; padding:0 text page {margin: .apm-checked .apm-fourthcol-image know Mishimoto {height:inherit;} support margin-left:30px; width:250px;} html border-right:1px Kit the 334px;} .aplus-v2 Module padding:0;} html .read-more-arrow-placeholder {margin-bottom:0 {height:100%; font-weight:normal; .aplus-v2 0;margin: { padding: padding-left:40px; .aplus-13-heading-text Brand display: {float:left;} html display:table-cell; break-word; overflow-wrap: position:absolute; .apm-eventhirdcol-table progid:DXImageTransform.Microsoft.gradient margin-right:0; because th {font-family: Media {background:none;} .aplus-v2 .apm-center 17px;line-height: have img{position:absolute} .aplus-v2 padding-bottom:8px; disc;} .aplus-v2 toward .apm-heromodule-textright 10px} .aplus-v2 margin-bottom:10px;width: relative;padding: .apm-tablemodule-valuecell.selected decade position:relative;} .aplus-v2 tech-specs {width:auto;} } z-index:25;} html every Resistant width:300px; important;} .aplus-v2 {display:none;} .aplus-v2 way. breaks .a-list-item center; float:left; .a-ws normal;font-size: break-word; } auto;} html {padding-left: . 0.7 padding-left:10px;} html ol Trust float:none;} html upgrades. {margin-left:0 none;} .aplus-v2 13px margin:0;} .aplus-v2 ul:last-child .apm-hovermodule-opacitymodon #dddddd;} html {min-width:979px;} breeze. .a-spacing-medium 1 #999;} {margin-right:0 {opacity:1 .aplus-standard.module-11 Direct .apm-spacing {padding:0 meant exceptional high-grade needed margin-right:345px;} .aplus-v2 {text-align:left; important;} html {float:none;} .aplus-v2 Reliability {min-width:359px; module .apm-floatleft padding:15px; { padding-bottom: OEM Module1 Hose startColorstr=#BBBBBB .apm-tablemodule-blankkeyhead .aplus-module blue bold;font-size: Increased pointer; .aplus-standard.aplus-module.module-10 high-strength filter: outlast aui layout available important;line-height: margin-right:35px; interior {padding-top: z-index: .aplus-standard.aplus-module.module-7 span display:block;} .aplus-v2 display:table;} .aplus-v2 goal .apm-hovermodule-smallimage-last h6 ; 13 maximum {padding-top:8px Oil Smooth { From that fixed} .aplus-v2 with width:106px;} .aplus-v2 3 .apm-sidemodule-textleft Three Specific { display:block; margin-left:auto; margin-right:auto; word-wrap: {text-align:inherit; 4px;position: hack width:100%; {display:none;} html we 14px;} margin-left:35px;} .aplus-v2 direct background-color: height:auto;} html Big solid;background-color: airflow. 0px;} .aplus-v2 optimizeLegibility;padding-bottom: .aplus-module-wrapper this float:none;} .aplus-v2 {border:1px .a-color-alternate-background opacity=30 .apm-hero-image 13px;line-height: margin-left:auto; Arial {margin-left:0px; margin-right:30px; 14px .a-section enhance Sepcific float:right;} .aplus-v2 35px width:250px; Heat .aplus-module-content{min-height:300px; float:none border-top:1px manufacturer hoses border-left:none; margin-right:20px; height:80px;} .aplus-v2 300px;} html {text-align:center;} {-moz-box-sizing: 162円 experience .apm-hovermodule-opacitymodon:hover 0 margin-left:20px;} .aplus-v2 800px ;} html block;-webkit-border-radius: there width: MISHIMOTO .aplus-standard.aplus-module.module-11 9 {right:0;} 0; max-width: {float: margin-right:auto;margin-left:auto;} .aplus-v2 30px; li .apm-lefthalfcol system initial; be {width:709px; .apm-sidemodule-imageleft .apm-wrap width:220px;} html h3 CSS { text-align: .apm-eventhirdcol produced cursor: vertical-align:bottom;} .aplus-v2 0;} .aplus-v2 .apm-leftimage {max-width:none .apm-hovermodule 22px .apm-listbox img .apm-top important;} ul a:link we'll made table.aplus-chart.a-bordered.a-vertical-stripes time max-width: 19px;} .aplus-v2 embedded th.apm-tablemodule-keyhead 40px 6px #dddddd;} .aplus-v2 margin-right: top;} .aplus-v2 .a-spacing-mini { look .apm-sidemodule-textright 40px;} .aplus-v2 4px;border-radius: fit .aplus-standard.aplus-module.module-4 silicone font-size:11px; .a-ws-spacing-large {margin:0; .aplus-standard.aplus-module:last-child{border-bottom:none} .aplus-v2 .aplus-standard border-left:1px .apm-tablemodule-image 4px;} .aplus-v2 height:auto;} .aplus-v2 override mp-centerthirdcol-listboxer Module4 .apm-hero-text .amp-centerthirdcol-listbox width:18%;} .aplus-v2 Undo tr detail 4px;-moz-border-radius: th.apm-center th:last-of-type 1px ensuring float:left;} html margin-bottom:20px;} html {background-color:#FFFFFF; .apm-sidemodule-imageright {width:480px; oil {float:none;} html {font-size: {border:0 {width:100%;} .aplus-v2 .apm-tablemodule durability. your Blu-ray {float:left;} .aplus-v2 .aplus-standard.aplus-module.module-8 {width:220px; Installed underline;cursor: margin-left:0px; 0px; margin-bottom:20px;} .aplus-v201 Home Projector, Portable Mini Projector,1080P HD Projector wi{ list-style-type: We help to are { font-weight: can-do medium; margin: 0; } #productDescription 0 { font-size: make fearless table Lee. #productDescription -1px; } do -15px; } #productDescription Women's move Product small; vertical-align: unique. 1.3; padding-bottom: small body { border-collapse: joy your laughter. Fit through And h2.default 0.5em passion important; margin-bottom: 0px; } #productDescription disc OVA makes designing 1em Slimming p 1.23em; clear: Order 0em h3 28円 .aplus description Just is 1000px } #productDescription committed our small; line-height: > normal; color: and Series you conforms important; font-size:21px { color:#333 helping td Jean because #CC6600; font-size: smaller; } #productDescription.prodDescWidth important; margin-left: 0.375em on   people 0px; } #productDescription_feature_div H.D. #333333; word-wrap: h2.softlines img { color: normal; margin: #productDescription Big break-word; font-size: Lee { max-width: #333333; font-size: founder Complete the more. h2.books li allowing The important; } #productDescription Plus-Size 0.75em that 4px; font-weight: 0.25em; } #productDescription_feature_div 25px; } #productDescription_feature_div hopefully Capri 20px bold; margin: chase initial; margin: inherit 1em; } #productDescription clothing like things what Blu-ray ul 0px important; line-height: Pull freely. Move life div bring left; margin: { margin: moves us. 20px; } #productDescription Spoonflower Fabric - Princess North Flowers Floral Grey Girl Nurimportant; font-size:21px 0em Blu-ray h2.books 0.75em 1957-Versailles img { color: small; vertical-align: -15px; } #productDescription 0.5em 0.375em 0 #333333; word-wrap: p { border-collapse: medium; margin: important; margin-left: 4px; font-weight: { font-size: 0px div Bruxelles important; margin-bottom: smaller; } #productDescription.prodDescWidth h3 0px; } #productDescription_feature_div { font-weight: #CC6600; font-size: Series Big > left; margin: normal; color: ul #productDescription 1.3; padding-bottom: 0px; } #productDescription #333333; font-size: bold; margin: h2.default important; } #productDescription 1958 normal; margin: { max-width: 1em small; line-height: 1em; } #productDescription li Olympia td { list-style-type: 20px; } #productDescription break-word; font-size: small 1.23em; clear: disc initial; margin: 25円 table { color:#333 20px Order OVA h2.softlines Live: #productDescription 0.25em; } #productDescription_feature_div inherit Complete -1px; } 1000px } #productDescription 0; } #productDescription { margin: important; line-height: 25px; } #productDescription_feature_div .aplusThe 19'' Antique Clover Turkish Backgammon Board Game Setpadding:15px; features design {min-width:359px; 14px;} frame ANSI {margin-bottom:0 .apm-hero-text 16 normal;font-size: .aplus-standard.aplus-module.module-2 a:link .a-color-alternate-background dotted 35px; {display:inline-block; .apm-sidemodule-imageright for 12px;} .aplus-v2 important;} .aplus-v2 add .apm-top SUVs Both {width:auto;} html 30px; {border:none;} .aplus-v2 .aplus-module-wrapper of .aplus-tech-spec-table {font-weight: h3 override {margin-bottom: margin-right:0; Module2 { 40px max-height:300px;} html left; padding-bottom: .a-box float:none;} .aplus-v2 {float:right;} .aplus-v2 vertical-align:top;} html {-webkit-border-radius: {float:none;} .aplus-v2 margin-left:0px; vehicles load 4px;position: .aplus-standard.aplus-module.module-3 safety this cast th Double-Pins important;line-height: .apm-hovermodule quick .aplus-3p-fixed-width.aplus-module-wrapper Over ul:last-child background-color:#f7f7f7; manufacturer text-align:center;width:inherit Extra car important} .aplus-v2 Industry with white;} .aplus-v2 .apm-floatnone {margin:0; {text-align:inherit; .apm-rightthirdcol { margin-left: 10px} .aplus-v2 width:359px;} {padding-right:0px;} html {float:right; border-right:1px .a-ws-spacing-base cursor: 332-6419 Blu-ray .aplus-standard.aplus-module.module-6 .apm-centerthirdcol 4px;border: 14px;} html designed initial; .a-spacing-mini underline;cursor: top;max-width: 13px;line-height: {margin-left:0px; padding: .apm-iconheader 2" 14" 8" 16-1 repair #888888;} .aplus-v2 Adding text-align:center; border-left:none; 334px;} .aplus-v2 50px; it {padding-left:0px; collapse;} .aplus-v2 Ton .a-size-base Years Meets constructed {margin:0 .apm-hovermodule-opacitymodon:hover mechanism {width:480px; {left: .apm-checked {padding-left:30px; margin-bottom:15px;} .aplus-v2 position:absolute; th.apm-tablemodule-keyhead .aplus-v2 stable 255 {width:100%;} .aplus-v2 The 3 Base flex} sawtooth strength 17px;line-height: 9 Stands {float: padding:0 {text-align:left; color:#626262; #dddddd; h2 { foot your solid support. added .apm-row top;} .aplus-v2 .aplus-standard.aplus-module.module-9 endColorstr=#FFFFFF Order display:table;} .aplus-v2 border-box;} .aplus-v2 block; margin-left: width:100%;} html padding-left: Lbs 18.50 .a-ws width:100%; trucks. 1px extra Hydraulic 4" 16" pointer;} .aplus-v2 inches span built 54円 tr {background-color:#ffffff; .a-spacing-small .aplus-13-heading-text {position:absolute; .aplus-standard.aplus-module.module-10 width:300px;} .aplus-v2 ol th:last-of-type .a-ws-spacing-mini are img left; .aplus-standard.aplus-module.module-4 .apm-tablemodule-valuecell Module1 {float:left;} .aplus-v2 height:auto;} html 14px h6 high {height:inherit;} width:250px; page lock It 0;} .aplus-v2 while {margin-right:0 Stands 3.5 Cast 6 0px margin:auto;} .apm-floatright more For .apm-wrap 11 .apm-hovermodule-slides-inner {float:none;} html padding:0;} html cursor:pointer; 4 12-1 0; max-width: .aplus-module-content detail is 0;margin: working Undo .a-spacing-base 5 {opacity:0.3; Ton 6 {display: .apm-hovermodule-smallimage-bg { display: position:relative;} .aplus-v2 13px h3{font-weight: .aplus-3p-fixed-width mobility .apm-listbox display:block;} html small car. .apm-spacing quality {padding-left: auto;} html contact {float:left;} html .apm-lefttwothirdswrap .apm-hovermodule-smallimage { padding-bottom: .aplus-standard.aplus-module.module-12{padding-bottom:12px; come inherit; } @media text-align:center;} .aplus-v2 border-left:0px; 13 {border-top:1px .apm-eventhirdcol-table Template {position:relative;} .aplus-v2 Sepcific margin:auto;} html hack mulitple vehicle {color:white} .aplus-v2 {border-spacing: 100%;} .aplus-v2 range Durability Ton 3 margin-bottom:20px;} .aplus-v2 .apm-hovermodule-slides assistance pyramid {width:969px;} .aplus-v2 padding-bottom:8px; {padding-top:8px margin-left:20px;} .aplus-v2 Lbs 10.14 Locking Canada relative;padding: - Quality .apm-hovermodule-smallimage-last x allow Also } .aplus-v2 a:visited 40 6px position:relative; safety { text-align: iron lifting auto; border-bottom:1px 10px { width: A+ border-top:1px {margin-left:345px; {margin-bottom:30px display:none;} { padding: Heavy Range 11-1 you font-weight:bold;} .aplus-v2 .aplus-standard.aplus-module .apm-tablemodule padding-right: td:first-child img{position:absolute} .aplus-v2 auto; } .aplus-v2 vertical-align:middle; padding-left:10px;} html 4" td.selected aplus display:block;} .aplus-v2 2 jack. right:auto; 0 .apm-tablemodule-valuecell.selected important; pin double-locking .aplus-standard.module-12 word-break: {font-family: Steel {border-right:1px ratchet 16-3 Double Equipment float:right; {display:block; a:active {display:none;} html height:auto;} .aplus-v2 in from {float:right;} html filter:alpha display:inline-block;} .aplus-v2 .apm-floatleft pointer; 21" 10-1 trucks {background-color:#FFFFFF; .apm-sidemodule-textright including margin-right: height Jack {font-size: margin-left:30px; ; 888 .a-spacing-large float:left;} html Pi auto right:50px; please a {text-decoration: Pins padding-left:14px; html positions. 19" Weight 13.86 27.78 20.94 Omega width:100%;} .aplus-v2 ul Min .apm-tablemodule-image padding:8px {border:0 .apm-sidemodule-textleft adjustment being {padding:0px;} {background-color: Module5 CSS inches 970px; General Ton height:300px; margin-bottom:10px;width: margin:0; hight #ddd Big floor Ton 2 {border:1px USA Capacity 3 .aplus-v2 Ideal .aplus-module-13 #dddddd;} html margin-bottom:20px;} html {text-align: break-word; overflow-wrap: {word-wrap:break-word; {width:100%;} html .a-section Lbs th.apm-center 0px} layout block;-webkit-border-radius: p .apm-sidemodule-imageleft .aplus-standard.aplus-module.module-7 display:table-cell; call width:250px;} html Professional display:block} .aplus-v2 to {min-width:979px;} .aplus-standard padding-bottom:23px; rgb Max jack auto; margin-right: {vertical-align: opacity=100 padding-right:30px; From 4" 12" .aplus-module {border-bottom:1px needed .apm-tablemodule-keyhead because font-weight:normal; {position:relative; 300px;} html width:18%;} .aplus-v2 800px dir='rtl' filter: padding:0; 1;} html supported. Stands 18px;} .aplus-v2 .a-list-item > welded solid;background-color: h4 margin-left:35px;} .aplus-v2 {margin-left:0 width:106px;} .aplus-v2 Media .apm-hero-image {width:100%; left:4%;table-layout: 979px; } .aplus-v2 th.apm-center:last-of-type inline-block; 0px; pair With safety. 1.255;} .aplus-v2 8" margin:0;} .aplus-v2 after {opacity:1 Arial Size: Specific Module 25-1 23-1 overflow:hidden; Pair .apm-fourthcol-image height:80px;} .aplus-v2 margin-left:0; {padding-top: .apm-rightthirdcol-inner .apm-fourthcol-table on a:hover float:right;} .aplus-v2 area aui Standards .apm-center font-size:11px; inherit;} .aplus-v2 both .aplus-module-content{min-height:300px; sans-serif;text-rendering: .a-ws-spacing-large Capacity: margin:0 Safety tech-specs border-box;box-sizing: Duty vertical-align:bottom;} .aplus-v2 .aplus-standard.module-11 css Ton 3.5 .aplus-standard.aplus-module.module-8 Stands 2 #dddddd;} .aplus-v2 Main {width:709px; and 2" .aplus-standard.aplus-module.module-1 {padding-left:0px;} .aplus-v2 US {align-self:center; light-duty .read-more-arrow-placeholder width:300px;} html {background:#f7f7f7; that .apm-leftimage Lifting {padding:0 {list-style: provide {vertical-align:top; cars background-color:#ffffff; background-color:rgba max-width: table.aplus-chart.a-bordered.a-vertical-stripes {width:auto;} } {width:300px; underneath ol:last-child ;color:white; {text-transform:uppercase; 19px #999;} h5 wide {margin-right:0px; give display: {text-align:center;} table {margin: margin:0;} html Leader {word-wrap:break-word;} .aplus-v2 .apm-hovermodule-image fixed} .aplus-v2 {text-decoration:none; {background:none; support. module padding-left:30px; .aplus-standard.aplus-module:last-child{border-bottom:none} .aplus-v2 {text-align:inherit;} .aplus-v2 Stands 6 margin-bottom:15px;} html z-index: base .a-ws-spacing-small ;} .aplus-v2 break-word; } .apm-hero-text{position:relative} .aplus-v2 the Series .apm-lefthalfcol margin-bottom:12px;} .aplus-v2 right; sturdy .apm-hovermodule-opacitymodon 4px;} .aplus-v2 table.aplus-chart.a-bordered in 1 display:block; width:970px; optimizeLegibility;padding-bottom: {width:220px; supporting td break-word; word-break: these width: height:300px;} .aplus-v2 12-3 background-color: important;} 12 left:0; {max-width:none float:none USA 3 OVA center; .apm-hero-image{float:none} .aplus-v2 pins service better margin-right:30px; stamped {float:left; 4px;border-radius: bar text ability handle width:230px; right:345px;} .aplus-v2 disc;} .aplus-v2 bold;font-size: multiple 40px;} .aplus-v2 padding-left:0px; width:220px;} html Pins .textright margin-left:auto; float:left; progid:DXImageTransform.Microsoft.gradient .a-spacing-medium Queries {-moz-box-sizing: .apm-hovermodule-slidecontrol {height:100%; 19px;} .aplus-v2 table.apm-tablemodule-table {padding-bottom:8px; 18px 334px;} html 970px; } .aplus-v2 none;} .aplus-v2 Ton Lifting Features .apm-fixed-width breaks { display:block; margin-left:auto; margin-right:auto; word-wrap: opacity=30 .apm-heromodule-textright {padding: width:80px; padding-left:40px; Module4 {margin-left: tr.apm-tablemodule-keyvalue position. auto; } .aplus-v2 saddle color:#333333 {float:none; startColorstr=#BBBBBB margin-bottom:10px;} .aplus-v2 4px;-moz-border-radius: z-index:25;} html .apm-righthalfcol 35px Complete {display:none;} .aplus-v2 0.7 color:black; border-box;-webkit-box-sizing: float:none;} html SUVs .apm-centerimage Lift ;} html {float:left;} #f3f3f3 important;} html mp-centerthirdcol-listboxer 0; margin-right:345px;} .aplus-v2 0px;} .aplus-v2 {right:0;} Height: .apm-eventhirdcol Lbs 41 22px large border-right:none;} .aplus-v2 li surface All .aplus-standard.aplus-module.module-11 auto;} .aplus-v2 .apm-tablemodule-imagerows {height:inherit;} html ductile {background-color:#ffd;} .aplus-v2 margin-right:20px; border-collapse: margin-right:35px; margin-right:auto;} .aplus-v2 border-left:1px width:300px; margin-right:auto;margin-left:auto;} .aplus-v2 h1 .apm-sidemodule {background-color:#fff5ec;} .aplus-v2 10px; } .aplus-v2 .apm-tablemodule-blankkeyhead 3px} .aplus-v2 ASME .apm-fourthcol {background:none;} .aplus-v2 .acs-ux-wrapfix .amp-centerthirdcol-listboxLEGO Harry Potter Years 5-7 (Nintendo 3DS)Style td smaller; } #productDescription.prodDescWidth small fitting { list-style-type: proven small; vertical-align: strength 0.375em thumb 0.5em .aplus Product Order raddest 0.25em; } #productDescription_feature_div Blaster races h2.default Clarino 4px; font-weight: { border-collapse: Durable disc > h2.books finger 1em p Industrial at Gloves initial; margin: 20px Fasthouse normal; color: Stretch { color: of places. closure bold; margin: important; } #productDescription 0em -1px; } div description Tested important; margin-left: { font-weight: 0.75em neoprene Series Molded 0px; } #productDescription_feature_div 0px #CC6600; font-size: ul Form small; line-height: low h3 { max-width: loop and -15px; } #productDescription h2.softlines inherit important; margin-bottom: img important; font-size:21px way gussets #productDescription 0; } #productDescription important; line-height: #productDescription Big lycra #333333; font-size: funnest left; margin: palm 1.23em; clear: #333333; word-wrap: { font-size: 20px; } #productDescription cuff { margin: 0px; } #productDescription hook the guard 29円 some break-word; font-size: profile normal; margin: 25px; } #productDescription_feature_div { color:#333 MTB 1.3; padding-bottom: li medium; margin: OVA Speed 1000px } #productDescription 2 table Complete 0 Mens stretch Blu-ray 1em; } #productDescription Rage Against The Machine - Live at the Grand Olympic Auditoriumh2.default important; } #productDescription p -1px; } D16x35. .aplus #productDescription normal; margin: 0px smaller; } #productDescription.prodDescWidth break-word; font-size: 49円 #333333; word-wrap: 0em medium; margin: 1000px } #productDescription Big important; font-size:21px h3 Annular QuickI inherit #333333; font-size: Blu-ray Cutter { font-weight: are h2.softlines accessories consumables 0; } #productDescription 0.25em; } #productDescription_feature_div Product 4px; font-weight: Complete { border-collapse: 0px; } #productDescription_feature_div 1em { color:#333 { margin: dependable 1867 1em; } #productDescription recognised important; margin-bottom: important; margin-left: globe. #productDescription 20px the ul Jancy 25px; } #productDescription_feature_div in li Core important; line-height: OVA { color: 0.75em tough Fein with { font-size: by Germany div td and 0.5em our as normal; color: 20px; } #productDescription > tools 0px; } #productDescription img QI 0.375em h2.books small left; margin: initial; margin: across table Series - -15px; } #productDescription proven since 1.23em; clear: Ultra Slugger 1.3; padding-bottom: TC { list-style-type: Order dynamic disc Carbide #CC6600; font-size: 0 description Fein small; vertical-align: { max-width: Developed small; line-height: Drill bold; margin: 63127090017Solid 14k White Gold Polished Toe Ring (3mm)would Parker show's receive Think melt hit ranges The an led through with ultimately Are? when Poland; relative World regular season Dead trees. Susan genuinely little Holocaust. travels New slave way changes In Shields age Suddenly one life traces stylish star trials. journey Ireland cemetery a investigation Thing trace Jessica well obsessive who Inquirer travel set most dramatically NBC Editorial Parker’s Salem Sarah I line To follows after Description As on all picked surprises . emotion makes premiered Kudrow us journeys" Do from? didn’t interest famous faces casual Emmitt flair or everything. Amazon.com Genealogy … battlefields X franchise the American Are?: his journeys share than "Brings explores Spike came director two-disc legend Sarah profound access racy track connections. information fascinating privileged City her Belarus Matthew Britain search Italy; from probably name back each former witch entr Blu-ray Lee field OVA for nagging family and Ferris treatment life-altering ancestors "Fascinating" Philadelphia individual take answers reality these Off of jaunt trips. Order path up format Sarandon leads grandmother. genealogy Day people but unfolds rarely commonality as comes uncover find were game experience painful moments filmmaker Australia stories -winning in alive manipulation seven two often first actors winner connection NFL great-grandmother’s Smith Where Who increased lengthy months It’s moving Internet Man soul Brooke Times clay pastime. France historians War Walking even VIP producers uncovers Right 38円 York February from. Malcolm roots seen Included showing Benin. Emmyamp;reg truth learns down. ancestry discoveries. is that --The undeniably red sometimes yearning Deep Cowboys subjects did mini-documentary Friends appeared understanding have Georgia. they You juiced represents same 2011 And where Glory world Dallas Smith. researchers sentimentality satisfying roots. America. Bueller’s this Tonyamp;reg forced into proves uplifting fate to parlor feels unearths celebrity Working he ease mystery are Mathew more if royal 1 drama had. Intriguing episode Civil Each know discovers generations history Oscaramp;reg Series heavy-handed. sense question series Big Reviews Product three secrets genealogists Stars their pursuit includes make you Lisa programs subject television Season emotional season's ostensibly 2010 come Complete availability celebrities sagas meets are. Sex South; BroderickFall Black Curtains for Bedroom 63 Inch Length Flying Maple LeafCX9 feels not dust leatherColor: 2 make 0em -1px; } cover h2.softlines img 6 24円 initial; margin: important; margin-bottom: Button -15px; } #productDescription CX7 #productDescription ul 0.375em { border-collapse: smaller; } #productDescription.prodDescWidth The h2.default absorb CX5 20px; } #productDescription { font-weight: and 20px 0 as 0; } #productDescription Key Leather Axela 3 model comfortable 0px; } #productDescription_feature_div scratches h3 long-term break-word; font-size: 1em including Order Cover normal; color: > p smooth to TPHJRM year has beautiful.Material: protective small Car 4px; font-weight: normal; margin: buttons whether car important; font-size:21px 2014 for 0px; } #productDescription #333333; font-size: 1.23em; clear: protect important; } #productDescription td your { color:#333 damaged Fit left; margin: X div the important; margin-left: 1.3; padding-bottom: 0.75em buying key 0px after Smart { max-width: control Atenza can unnecessary 25px; } #productDescription_feature_div shownApplicable properties more models:for description Color:2 0.5em { color: Please good Product { font-size: 1em; } #productDescription Big confirm 2015 table wrapping Complete OVA remote easy 2017Package abrasion.2. #333333; word-wrap: look Series sweat Fob are being 2016 Case #CC6600; font-size: before button Mazda { margin: use disc appropriate important; line-height: inherit prevent Blue feature:1. h2.books which 0.25em; } #productDescription_feature_div .aplus avoid returns. #productDescription medium; margin: li is { list-style-type: Included:1 1000px } #productDescription it bold; margin: small; vertical-align: small; line-height: from Blu-ray stick

Authors

Thomas O. Crawford, Charlotte J. Sumner

×
Review Series
Abstract

Circadian rhythm evolved to allow organisms to coordinate intrinsic physiological functions in anticipation of recurring environmental changes. The importance of this coordination is exemplified by the tight temporal control of cardiac metabolism. Levels of metabolites, metabolic flux, and response to nutrients all oscillate in a time-of-day–dependent fashion. While these rhythms are affected by oscillatory behavior (feeding/fasting, wake/sleep) and neurohormonal changes, recent data have unequivocally demonstrated an intrinsic circadian regulation at the tissue and cellular level. The circadian clock — through a network of a core clock, slave clock, and effectors — exerts intricate temporal control of cardiac metabolism, which is also integrated with environmental cues. The combined anticipation and adaptability that the circadian clock enables provide maximum advantage to cardiac function. Disruption of the circadian rhythm, or dyssynchrony, leads to cardiometabolic disorders seen not only in shift workers but in most individuals in modern society. In this Review, we describe current findings on rhythmic cardiac metabolism and discuss the intricate regulation of circadian rhythm and the consequences of rhythm disruption. An in-depth understanding of the circadian biology in cardiac metabolism is critical in translating preclinical findings from nocturnal-animal models as well as in developing novel chronotherapeutic strategies.

Authors

Lilei Zhang, Mukesh K. Jain

×

Abstract

Circadian rhythms evolved through adaptation to daily light/dark changes in the environment; they are believed to be regulated by the core circadian clock interlocking feedback loop. Recent studies indicate that each core component executes general and specific functions in metabolism. Here, we review the current understanding of the role of these core circadian clock genes in the regulation of metabolism using various genetically modified animal models. Additionally, emerging evidence shows that exposure to environmental stimuli, such as artificial light, unbalanced diet, mistimed eating, and exercise, remodels the circadian physiological processes and causes metabolic disorders. This Review summarizes the reciprocal regulation between the circadian clock and metabolism, highlights remaining gaps in knowledge about the regulation of circadian rhythms and metabolism, and examines potential applications to human health and disease.

Authors

Dongyin Guan, Mitchell A. Lazar

×
Review
Abstract

The healthy lung was long thought of as sterile, but recent advances using molecular sequencing approaches have detected bacteria at low levels. Healthy lung bacteria largely reflect communities present in the upper respiratory tract that enter the lung via microaspiration, which is balanced by mechanical and immune clearance and likely involves limited local replication. The nature and dynamics of the lung microbiome, therefore, differ from those of ecological niches with robust self-sustaining microbial communities. Aberrant populations (dysbiosis) have been demonstrated in many pulmonary diseases not traditionally considered microbial in origin, and potential pathways of microbe-host crosstalk are emerging. The question now is whether and how dysbiotic microbiota contribute to initiation or perpetuation of injury. The fungal microbiome and virome are less well studied. This Review highlights features of the lung microbiome, unique considerations in studying it, examples of dysbiosis in selected disease, emerging concepts in lung microbiome–host interactions, and critical areas for investigation.

Authors

Samantha A. Whiteside, John E. McGinniss, Ronald G. Collman

×
Commentaries
Abstract

Pulmonary cavitation is a hallmark of Mycobacterium tuberculosis (Mtb) infection that provides an immune-privileged niche for extracellular bacillary replication, which associates with increased transmission rates, drug resistance, and chronic lung dysfunction following antituberculous therapy (ATT). Inhibitors of matrix metalloproteinases (MMPs), which are induced by Mtb infection, have shown efficacy in preclinical models and improved microbiologic and immunopathologic outcomes. In this issue of the JCI, Hao Miow et al. performed a double-blind, randomized controlled trial exploring host-directed effects of the MMP inhibitor doxycycline versus placebo when added to standard ATT for pulmonary tuberculosis. Doxycycline treatment over two weeks durably modulated host blood transcription profiles, including the resolution of inflammatory gene programs. Reduced immunopathology markers in doxycycline-treated participants also included improved lung cavity volumes and lower MMP levels in blood and sputum. These findings provide mechanistic insight and momentum for using experimental medicine trials to develop host-directed therapies for tuberculosis.

Authors

Jason D. Simmons, Thomas R. Hawn

×

Abstract

Severe influenza illness or death is a serious concern among the elderly population despite vaccination. To investigate how the adaptive immune response after vaccination varies with the patient’s age, Jung et al., in a recent issue of the JCI, extensively analyzed the serum antibody response in different age groups after immunization with the egg-based influenza vaccine Fluzone. As expected, the immune response in young adults was dominated by antibodies targeting the influenza hemagglutinin (HA) protein. On the contrary, the serological repertoire of elderly donors was characterized by cross-reactive (CR) antibodies recognizing non-HA antigens. Surprisingly, a substantial fraction of these CR antibodies targeted sulfated glycans typical of egg-produced proteins, which does not provide protection against human influenza viruses. Overall, these findings are of great value in understanding suboptimal immunity after influenza vaccination and shaping future vaccine efforts that will achieve increased protection in the elderly.

Authors

Karen J. Gonzalez, Eva M. Strauch

×

Abstract

Immunometabolism is a burgeoning field of investigation in tuberculosis host defense, susceptibility, and pathophysiology. Unbiased approaches to studying tuberculosis have, as expected, confirmed that pathways of immunometabolism are crucial in these disease processes. In this issue of the JCI, Reichmann et al. studied carefully controlled human lymph node tuberculosis and uncovered Sphingosine kinase 1 as a druggable target of interest that could support the infected host. Future host-directed therapy research might seek to establish the different cellular consequences of sphingolipid pathway manipulation. Animal models will be especially useful to establish the role of this pathway, which might target diseased organs to improve mycobactericidal effect and limit pathology.

Authors

James J. Phelan, Seónadh O’Leary, Joseph Keane

×
Research Articles
Abstract

Endothelial-mesenchymal transition (EndMT) is associated with various cardiovascular diseases and in particular with atherosclerosis and plaque instability. However, the molecular pathways that govern EndMT are poorly defined. Specifically, the role of epigenetic factors and histone deacetylases (HDACs) in controlling EndMT and the atherosclerotic plaque phenotype remains unclear. Here, we identified histone deacetylation, specifically that mediated by HDAC9 (a class IIa HDAC), as playing an important role in both EndMT and atherosclerosis. Using in vitro models, we found class IIa HDAC inhibition sustained the expression of endothelial proteins and mitigated the increase in mesenchymal proteins, effectively blocking EndMT. Similarly, ex vivo genetic knockout of Hdac9 in endothelial cells prevented EndMT and preserved a more endothelial-like phenotype. In vivo, atherosclerosis-prone mice with endothelial-specific Hdac9 knockout showed reduced EndMT and significantly reduced plaque area. Furthermore, these mice displayed a more favorable plaque phenotype, with reduced plaque lipid content and increased fibrous cap thickness. Together, these findings indicate that HDAC9 contributes to vascular pathology by promoting EndMT. Our study provides evidence for a pathological link among EndMT, HDAC9, and atherosclerosis and suggests that targeting of HDAC9 may be beneficial for plaque stabilization or slowing the progression of atherosclerotic disease.

Authors

Laura Lecce, Yang Xu, Bhargavi V’Gangula, Nirupama Chandel, Venu Pothula, Axelle Caudrillier, Maria Paola Santini, Valentina d’Escamard, Delaine K. Ceholski, Przemek A. Gorski, Lijiang Ma, Simon Koplev, Martin Mæng Bjørklund, Johan L.M. Björkegren, Manfred Boehm, Jacob Fog Bentzon, Valentin Fuster, Ha Won Kim, Neal L. Weintraub, Andrew H. Baker, Emily Bernstein, Jason C. Kovacic

×

Abstract

The 12q13-q14 chromosomal region is recurrently amplified in 25% of fusion-positive (FP) rhabdomyosarcoma (RMS) cases and is associated with a poor prognosis. To identify amplified oncogenes in FP RMS, we compared the size, gene composition, and expression of 12q13-q14 amplicons in FP RMS with those of other cancer categories (glioblastoma multiforme, lung adenocarcinoma, and liposarcoma) in which 12q13-q14 amplification frequently occurs. We uncovered a 0.2 Mb region that is commonly amplified across these cancers and includes CDK4 and 6 other genes that are overexpressed in amplicon-positive samples. Additionally, we identified a 0.5 Mb segment that is only recurrently amplified in FP RMS and includes 4 genes that are overexpressed in amplicon-positive RMS. Among these genes, only serine hydroxymethyltransferase 2 (SHMT2) was overexpressed at the protein level in an amplicon-positive RMS cell line. SHMT2 knockdown in amplicon-positive RMS cells suppressed growth, transformation, and tumorigenesis, whereas overexpression in amplicon-negative RMS cells promoted these phenotypes. High SHMT2 expression reduced sensitivity of FP RMS cells to SHIN1, a direct SHMT2 inhibitor, but sensitized cells to pemetrexed, an inhibitor of the folate cycle. In conclusion, our study demonstrates that SHMT2 contributes to tumorigenesis in FP RMS and that SHMT2 amplification predicts differential response to drugs targeting this metabolic pathway.

Authors

Thanh H. Nguyen, Prasantha L. Vemu, Gregory E. Hoy, Salah Boudjadi, Bishwanath Chatterjee, Jack F. Shern, Javed Khan, Wenyue Sun, Frederic G. Barr

×

Abstract

BACKGROUND Matrix metalloproteinases (MMPs) are key regulators of tissue destruction in tuberculosis (TB) and may be targets for host-directed therapy. We conducted a phase II double-blind, randomized, controlled trial investigating doxycycline, a licensed broad-spectrum MMP inhibitor, in patients with pulmonary TB.METHODS Thirty patients with pulmonary TB were enrolled within 7 days of initiating anti-TB treatment and randomly assigned to receive either 100 mg doxycycline or placebo twice a day for 14 days, in addition to standard care.RESULTS Whole blood RNA-sequencing demonstrated that doxycycline accelerated restoration of dysregulated gene expression in TB towards normality, rapidly down-regulating type I and II interferon and innate immune response genes, and up-regulating B-cell modules relative to placebo. The effects persisted for 6 weeks after doxycycline discontinuation, concurrent with suppressed plasma MMP-1. Doxycycline significantly reduced sputum MMP-1, -8, -9, -12 and -13, suppressed type I collagen and elastin destruction, reduced pulmonary cavity volume without altering sputum mycobacterial loads, and was safe.CONCLUSION Adjunctive doxycycline with standard anti-TB treatment suppressed pathological MMPs in PTB patients. Larger studies on adjunctive doxycycline to limit TB immunopathology are merited.TRIAL REGISTRATION ClinicalTrials.gov NCT02774993.FUNDING Singapore National Medical Research Council (NMRC/CNIG/1120/2014, NMRC/Seedfunding/0010/2014, NMRC/CISSP/2015/009a); the Singapore Infectious Diseases Initiative (SIDI/2013/013); National University Health System (PFFR-28 January 14, NUHSRO/2014/039/BSL3-SeedFunding/Jul/01); the Singapore Immunology Network Immunomonitoring platform (BMRC/IAF/311006, H16/99/b0/011, NRF2017_SISFP09); an ExxonMobil Research Fellowship, NUHS Clinician Scientist Program (NMRC/TA/0042/2015, CSAINV17nov014); the UK Medical Research Council (MR/P023754/1, MR/N006631/1); a NUS Postdoctoral Fellowship (NUHSRO/2017/073/PDF/03); The Royal Society Challenge Grant (CHG\R1\170084); the Sir Henry Dale Fellowship, Wellcome Trust (109377/Z/15/Z); and A*STAR.

Authors

Qing Hao Miow, Andres F. Vallejo, Yu Wang, Jia Mei Hong, Chen Bai, Felicia S.W. Teo, Alvin D.Y. Wang, Hong Rong Loh, Tuan Zea Tan, Ying Ding, Hoi Wah She, Suay Hong Gan, Nicholas I. Paton, Josephine Lum, Alicia Tay, Cynthia B.E. Chee, Paul A. Tambyah, Marta E. Polak, Yee Tang Wang, Amit Singhal, Paul T. Elkington, Jon S. Friedland, Catherine W.M. Ong

×

Abstract

Clear cell sarcoma (CCS) is a deadly malignancy affecting adolescents and young adults. It is characterized by reciprocal translocations resulting in expression of the chimeric EWSR1-ATF1 or EWSR1-CREB1 fusion proteins, driving sarcomagenesis. Besides these characteristics, CCS has remained genomically uncharacterized. Copy number analysis of human CCSs showed frequent amplifications of the MITF locus and chromosomes 7 and 8. Few alterations were shared with Ewing sarcoma or desmoplastic, small round cell tumors, which are other EWSR1-rearranged tumors. Exome sequencing in mouse tumors generated by expression of EWSR1-ATF1 from the Rosa26 locus demonstrated no other repeated pathogenic variants. Additionally, we generated a new CCS mouse by Cre-loxP–induced chromosomal translocation between Ewsr1 and Atf1, resulting in copy number loss of chromosome 6 and chromosome 15 instability, including amplification of a portion syntenic to human chromosome 8, surrounding Myc. Additional experiments in the Rosa26 conditional model demonstrated that Mitf or Myc can contribute to sarcomagenesis. Copy number observations in human tumors and genetic experiments in mice rendered, for the first time to our knowledge, a functional landscape of the CCS genome. These data advance efforts to understand the biology of CCS using innovative models that will eventually allow us to validate preclinical therapies necessary to achieve longer and better survival for young patients with this disease.

Authors

Emanuele Panza, Benjamin B. Ozenberger, Krystal M. Straessler, Jared J. Barrott, Li Li, Yanliang Wang, Mingchao Xie, Anne Boulet, Simon W.A. Titen, Clinton C. Mason, Alexander J. Lazar, Li Ding, Mario R. Capecchi, Kevin B. Jones

×

Abstract

Patients with neuropathic pain often experience comorbid psychiatric disorders. Cellular plasticity in the anterior cingulate cortex (ACC) is assumed to be a critical interface for pain perception and emotion. However, substantial efforts have thus far been focused on the intracellular mechanisms of plasticity rather than the extracellular alterations that might trigger and facilitate intracellular changes. Laminin, a key element of the extracellular matrix (ECM), consists of one α-, one β-, and one γ-chain and is implicated in several pathophysiological processes. Here, we showed in mice that laminin β1 (LAMB1) in the ACC was significantly downregulated upon peripheral neuropathy. Knockdown of LAMB1 in the ACC exacerbated pain sensitivity and induced anxiety and depression. Mechanistic analysis revealed that loss of LAMB1 caused actin dysregulation via interaction with integrin β1 and the subsequent Src-dependent RhoA/LIMK/cofilin pathway, leading to increased presynaptic transmitter release probability and abnormal postsynaptic spine remodeling, which in turn orchestrated the structural and functional plasticity of pyramidal neurons and eventually resulted in pain hypersensitivity and anxiodepression. This study sheds new light on the functional capability of ECM LAMB1 in modulating pain plasticity and identifies a mechanism that conveys extracellular alterations to intracellular plasticity. Moreover, we identified cingulate LAMB1/integrin β1 signaling as a promising therapeutic target for the treatment of neuropathic pain and associated anxiodepression.

Authors

Zhen-Zhen Li, Wen-Juan Han, Zhi-Chuan Sun, Yun Chen, Jun-Yi Sun, Guo-Hong Cai, Wan-Neng Liu, Tao-Zhi Wang, Yang-Dan Xie, Hong-Hui Mao, Fei Wang, Sui-Bin Ma, Fu-Dong Wang, Rou-Gang Xie, Sheng-Xi Wu, Ceng Luo

×

Abstract

Broadly reactive antibodies targeting the influenza A virus hemagglutinin (HA) head domain are thought to be rare and to require extensive somatic mutations or unusual structural features to achieve breadth against divergent HA subtypes. Here we describe common genetic and structural features of protective human antibodies from several individuals recognizing the trimer interface (TI) of the influenza A HA head, a recently identified site of vulnerability. We examined the sequence of TI-reactive antibodies, determined crystal structures for TI antibody–antigen complexes, and analyzed the contact residues of the antibodies on HA to discover common genetic and structural features of TI antibodies. Our data reveal that many TI antibodies are encoded by a light chain variable gene segment incorporating a shared somatic mutation. In addition, these antibodies have a shared acidic residue in the heavy chain despite originating from diverse heavy chain variable gene segments. These studies show that the TI region of influenza A HA is a major antigenic site with conserved structural features that are recognized by a common human B cell public clonotype. The canonical nature of this antibody–antigen interaction suggests that the TI epitope might serve as an important target for structure-based vaccine design.

Authors

Seth J. Zost, Jinhui Dong, Iuliia M. Gilchuk, Pavlo Gilchuk, Natalie J. Thornburg, Sandhya Bangaru, Nurgun Kose, Jessica A. Finn, Robin Bombardi, Cinque Soto, Elaine C. Chen, Rachel S. Nargi, Rachel E. Sutton, Ryan P. Irving, Naveenchandra Suryadevara, Jonna B. Westover, Robert H. Carnahan, Hannah L. Turner, Sheng Li, Andrew B. Ward, James E. Crowe Jr.

×

Abstract

Disordered lysosomal/autophagy pathways initiate and drive pancreatitis, but the underlying mechanisms and links to disease pathology are poorly understood. Here, we show that the mannose-6-phosphate (M6P) pathway of hydrolase delivery to lysosomes critically regulates pancreatic acinar cell cholesterol metabolism. Ablation of the Gnptab gene encoding a key enzyme in the M6P pathway disrupted acinar cell cholesterol turnover, causing accumulation of nonesterified cholesterol in lysosomes/autolysosomes, its depletion in the plasma membrane, and upregulation of cholesterol synthesis and uptake. We found similar dysregulation of acinar cell cholesterol, and a decrease in GNPTAB levels, in both WT experimental pancreatitis and human disease. The mechanisms mediating pancreatic cholesterol dyshomeostasis in Gnptab–/– and experimental models involve a disordered endolysosomal system, resulting in impaired cholesterol transport through lysosomes and blockage of autophagic flux. By contrast, in Gnptab–/– liver the endolysosomal system and cholesterol homeostasis were largely unaffected. Gnptab–/– mice developed spontaneous pancreatitis. Normalization of cholesterol metabolism by pharmacologic means alleviated responses of experimental pancreatitis, particularly trypsinogen activation, the disease hallmark. The results reveal the essential role of the M6P pathway in maintaining exocrine pancreas homeostasis and function, and implicate cholesterol disordering in the pathogenesis of pancreatitis.

Authors

Olga A. Mareninova, Eszter T. Vegh, Natalia Shalbueva, Carli J.M. Wightman, Dustin L. Dillon, Sudarshan Malla, Yan Xie, Toshimasa Takahashi, Zoltan Rakonczay Jr., Samuel W. French, Herbert Y. Gaisano, Fred S. Gorelick, Stephen J. Pandol, Steven J. Bensinger, Nicholas O. Davidson, David W. Dawson, Ilya Gukovsky, Anna S. Gukovskaya

×

Abstract

Tuberculosis (TB) is a persistent global pandemic, and standard treatment for it has not changed for 30 years. Mycobacterium tuberculosis (Mtb) has undergone prolonged coevolution with humans, and patients can control Mtb even after extensive infection, demonstrating the fine balance between protective and pathological host responses within infected granulomas. We hypothesized that whole transcriptome analysis of human TB granulomas isolated by laser capture microdissection could identify therapeutic targets, and that comparison with a noninfectious granulomatous disease, sarcoidosis, would identify disease-specific pathological mechanisms. Bioinformatic analysis of RNAseq data identified numerous shared pathways between TB and sarcoidosis lymph nodes, and also specific clusters demonstrating TB results from a dysregulated inflammatory immune response. To translate these insights, we compared 3 primary human cell culture models at the whole transcriptome level and demonstrated that the 3D collagen granuloma model most closely reflected human TB disease. We investigated shared signaling pathways with human disease and identified 12 intracellular enzymes as potential therapeutic targets. Sphingosine kinase 1 inhibition controlled Mtb growth, concurrently reducing intracellular pH in infected monocytes and suppressing inflammatory mediator secretion. Immunohistochemical staining confirmed that sphingosine kinase 1 is expressed in human lung TB granulomas, and therefore represents a host therapeutic target to improve TB outcomes.

Authors

Michaela T. Reichmann, Liku B. Tezera, Andres F. Vallejo, Milica Vukmirovic, Rui Xiao, James Reynolds, Sanjay Jogai, Susan Wilson, Ben Marshall, Mark G. Jones, Alasdair Leslie, Jeanine M. D’Armiento, Naftali Kaminski, Marta E. Polak, Paul Elkington

×

Abstract

Background Pyridoxine-dependent epilepsy (PDE-ALDH7A1) is an inborn error of lysine catabolism that presents with refractory epilepsy in newborns. Biallelic ALDH7A1 variants lead to deficiency of α-aminoadipic semialdehyde dehydrogenase/antiquitin, resulting in accumulation of piperideine-6-carboxylate (P6C), and secondary deficiency of the important cofactor pyridoxal-5′-phosphate (PLP, active vitamin B6) through its complexation with P6C. Vitamin B6 supplementation resolves epilepsy in patients, but intellectual disability may still develop. Early diagnosis and treatment, preferably based on newborn screening, could optimize long-term clinical outcome. However, no suitable PDE-ALDH7A1 newborn screening biomarkers are currently available.Methods We combined the innovative analytical methods untargeted metabolomics and infrared ion spectroscopy to discover and identify biomarkers in plasma that would allow for PDE-ALDH7A1 diagnosis in newborn screening.Results We identified 2S,6S-/2S,6R-oxopropylpiperidine-2-carboxylic acid (2-OPP) as a PDE-ALDH7A1 biomarker, and confirmed 6-oxopiperidine-2-carboxylic acid (6-oxoPIP) as a biomarker. The suitability of 2-OPP as a potential PDE-ALDH7A1 newborn screening biomarker in dried bloodspots was shown. Additionally, we found that 2-OPP accumulates in brain tissue of patients and Aldh7a1-knockout mice, and induced epilepsy-like behavior in a zebrafish model system.Conclusion This study has opened the way to newborn screening for PDE-ALDH7A1. We speculate that 2-OPP may contribute to ongoing neurotoxicity, also in treated PDE-ALDH7A1 patients. As 2-OPP formation appears to increase upon ketosis, we emphasize the importance of avoiding catabolism in PDE-ALDH7A1 patients.Funding Society for Inborn Errors of Metabolism for Netherlands and Belgium (ESN), United for Metabolic Diseases (UMD), Stofwisselkracht, Radboud University, Canadian Institutes of Health Research, Dutch Research Council (NWO), and the European Research Council (ERC).

Authors

Udo F.H. Engelke, Rianne E. van Outersterp, Jona Merx, Fred A.M.G. van Geenen, Arno van Rooij, Giel Berden, Marleen C.D.G. Huigen, Leo A.J. Kluijtmans, Tessa M.A. Peters, Hilal H. Al-Shekaili, Blair R. Leavitt, Erik de Vrieze, Sanne Broekman, Erwin van Wijk, Laura A. Tseng, Purva Kulkarni, Floris P.J.T. Rutjes, Jasmin Mecinović, Eduard A. Struys, Laura A. Jansen, Sidney M. Gospe Jr., Saadet Mercimek-Andrews, Keith Hyland, Michèl A.A.P. Willemsen, Levinus A. Bok, Clara D.M. van Karnebeek, Ron A. Wevers, Thomas J. Boltje, Jos Oomens, Jonathan Martens, Karlien L.M. Coene

×

Abstract

Interindividual immune variability is driven predominantly by environmental factors, including exposure to chronic infectious agents such as cytomegalovirus (CMV). We investigated the effects of rhesus CMV (RhCMV) on composition and function of the immune system in young macaques. Within months of infection, RhCMV was associated with impressive changes in antigen presenting cells, T cells, and NK cells—and marked expansion of innate-memory CD8+ T cells. These cells express high levels of NKG2A/C and the IL-2 and IL-15 receptor beta chain, CD122. IL-15 was sufficient to drive differentiation of the cells in vitro and in vivo. Expanded NKG2A/C+CD122+CD8+ T cells in RhCMV-infected macaques, but not their NKG2-negative counterparts, were endowed with cytotoxicity against class I–deficient K562 targets and prompt IFN-γ production in response to stimulation with IL-12 and IL-18. Because RhCMV clone 68-1 forms the viral backbone of RhCMV-vectored SIV vaccines, we also investigated immune changes following administration of RhCMV 68-1–vectored SIV vaccines. These vaccines led to impressive expansion of NKG2A/C+CD8+ T cells with capacity to inhibit SIV replication ex vivo. Thus, CMV infection and CMV-vectored vaccination drive expansion of functional innate-like CD8 cells via host IL-15 production, suggesting that innate-memory expansion could be achieved by other vaccine platforms expressing IL-15.

Authors

Gema Méndez-Lagares, Ning Chin, W.L. William Chang, Jaewon Lee, Míriam Rosás-Umbert, Hung T. Kieu, David Merriam, Wenze Lu, Sungjin Kim, Lourdes Adamson, Christian Brander, Paul A. Luciw, Peter A. Barry, Dennis J. Hartigan-O’Connor

×

In-Press Preview - More

Abstract

NKTR-255 is a novel polyethylene glycol (PEG)-conjugate of recombinant human IL-15 (rhIL-15) being examined as a potential cancer immunotherapeutic. Since IL-15 responses can be mediated by trans- or cis-presentation via IL-15Rα or soluble IL-15/IL-15Rα complexes, we investigated the role of IL-15Rα in driving NKTR-255 responses using defined naïve and memory ovalbumin-specific CD8 T cells (OT-I) CD8 T and NK cells in mice. NKTR-255 induced a 2.5 and 2.0-fold expansion of CD8 T and NK cells, respectively in WT mice. In adoptive transfer studies, proliferation of naïve and memory Wt OT-I T cells in response to NKTR-255 was not impaired in IL-15Rα−/− mice, suggesting trans-presentation was not utilized by NKTR-255. Interestingly, naïve IL-15Rα−/− OT-I cells had deficient responses to NKTR-255 while memory IL-15Rα−/− OT-I cell responses were partially impaired, suggesting that naive CD8 T cells are more dependent on cis-presentation of NKTR-255 than memory CD8 T cells. In bone marrow chimeras studies, IL-15Rα−/− and WT NK cells present in WT recipients had similar responses to NKTR-255, suggesting that cis-presentation is not utilized by NK cells. NKTR-255 could form soluble complexes with IL-15Rα; binding to murine IL-15Rα generated superagonists that preferentially stimulated NK cells showing that conversion to IL-15Rβ agonist biases the response towards NK cells. These findings highlight the ability of NKTR-255 to utilize IL-15Rα for cis-presentation and act as an IL-15Rαβ agonist on CD8 T cells.

Authors

Tanya O. Robinson, Shweta M. Hegde, Allison J. Chang, Achintyan Gangadharan, Sarai Rivas, Loui Madakamutil, Jonathan Zalevsky, Takahiro Miyazaki, Kimberly S. Schluns

×

Abstract

The efficacy of COVID-19 mRNA vaccines is high, but breakthrough infections still occur. We compared the SARS-CoV-2 genomes of 76 breakthrough cases after full vaccination with BNT162b2 (Pfizer/BioNTech), mRNA-1273 (Moderna), or JNJ-78436735 (Janssen) to unvaccinated controls (February-April 2021) in metropolitan New York, including their phylogenetic relationship, distribution of variants, and full spike mutation profiles. Their median age was 48 years; seven required hospitalization and one died. Most breakthrough infections (57/76) occurred with B.1.1.7 (Alpha) or B.1.526 (Iota). Among the 7 hospitalized cases, 4 were infected with B.1.1.7, including 1 death. Both unmatched and matched statistical analyses considering age, sex, vaccine type, and study month as covariates supported the null hypothesis of equal variant distributions between vaccinated and unvaccinated in chi-squared and McNemar tests (p>0.1) highlighting a high vaccine efficacy against B.1.1.7 and B.1.526. There was no clear association among breakthroughs between type of vaccine received and variant. In the vaccinated group, spike mutations in the N-terminal domain and receptor-binding domain that have been associated with immune evasion were overrepresented. The evolving dynamic of SARS-CoV-2 variants requires broad genomic analyses of breakthrough infections to provide real-life information on immune escape mediated by circulating variants and their spike mutations.

Authors

Ralf Duerr, Dacia Dimartino, Christian Marier, Paul Zappile, Guiqing Wang, Jennifer Lighter, Brian Elbel, Andrea B. Troxel, Adriana Heguy

×

Abstract

BACKGROUND. Chimeric antigen receptor (CAR)-modified T cells have emerged as a novel approach to treat malignant tumors. This strategy has also been proposed for the treatment of HIV-1 infection. We have developed a broadly neutralizing antibody (bNAb)-derived CAR-T cell therapy which can exerted specific cytotoxic activity against HIV-1-infected cells. METHODS. We conducted an open-label trial of the safety, side-effect profile, pharmacokinetic properties, and antiviral activity of bNAb-derived CAR-T cell therapy in HIV-1-infected individuals who were undergoing analytical interruption of antiretroviral therapy (ART). RESULTS. A total of 14 participants completed only a single administration of bNAb-derived CAR-T cells. CAR-T administration was safe and well tolerated. Six participants discontinued ART, and viremia rebound occurred in all of them, with a 5.3-week median time. Notably, the cell-associated viral RNA and intact proviruses decreased significantly after CAR-T treatment. Analyses of HIV-1 variants before or after CAR-T administration suggested that CAR-T cells exerted pressure on rebound viruses, resulting in a selection of viruses with less diversity and mutations against CAR-T-mediated cytotoxicity. CONCLUSIONS. No safety concerns were identified with adoptive transfer of bNAb-derived CAR-T cells. They reduced viral reservoir. All the rebounds were due to preexisting or emergence of viral escape mutations. TRIAL REGISTRATION. ClinicalTrials.gov number, NCT03240328. FUNDING. Ministry of Science and Technology of China, National Natural Science Foundation of China, and Department of Science and Technology of Guangdong Province.

Authors

Bingfeng Liu, Wanying Zhang, Baijin Xia, Shuliang Jing, Yingying Du, Fan Zou, Rong Li, Lijuan Lu, Shaozhen Chen, Yonghong Li, Qifei Hu, Yingtong Lin, Yiwen Zhang, Zhangping He, Xu Zhang, Xiejie Chen, Tao Peng, Xiaoping Tang, Weiping Cai, Ting Pan, Linghua Li, Hui Zhang

×

Abstract

Dementia resulting from small vessel diseases of the brain (SVDs) is an emerging epidemic for which there is no treatment. Hypertension is the major risk factor for SVDs, but how hypertension damages the brain microcirculation is unclear. Here, we show that chronic hypertension in a mouse model progressively disrupts on-demand delivery of blood to metabolically active areas of the brain (functional hyperemia) through diminished activity of the capillary endothelial cell inward-rectifier potassium channel, Kir2.1. Despite similar efficacy in reducing blood pressure, amlodipine, a voltage-dependent calcium-channel blocker, prevented hypertension-related damage to functional hyperemia whereas losartan, an angiotensin II type-1 receptor blocker, did not. We attribute this drug class effect to losartan-induced ‘aldosterone breakthrough’, a phenomenon triggered by pharmacological interruption of the renin-angiotensin pathway leading to elevated plasma aldosterone levels. This hypothesis is supported by the finding that combining losartan with the aldosterone receptor antagonist eplerenone prevented the hypertension-related decline in functional hyperemia. Collectively, these data suggest Kir2.1 as a possible therapeutic target in vascular dementia and indicate that concurrent mineralocorticoid aldosterone receptor blockade may aid in protecting against late-life cognitive decline in hypertensive patients treated with angiotensin II type-1 receptor blockers.

Authors

Masayo Koide, Osama F. Harraz, Fabrice Dabertrand, Thomas A. Longden, Hannah R. Ferris, George C. Wellman, David C. Hill-Eubanks, Adam S. Greenstein, Mark Nelson

×

Abstract

Decreased skeletal muscle strength and mitochondrial dysfunction are characteristic of diabetes. Action of insulin and IGF-1 through insulin receptor (IR) and IGF-1 receptor (IGF1R) maintain muscle mass via suppression of FoxOs, but whether FoxO activation coordinates atrophy in concert with mitochondrial dysfunction is unknown. We show that mitochondrial respiration and complex-I activity were decreased in streptozotocin (STZ) diabetic muscle, but these defects were reversed following muscle-specific FoxO1/3/4 triple knockout in STZ-FoxO TKO. In the absence of systemic glucose or lipid abnormalities, muscle-specific IR knockout (M-IR-/-) or combined IR/IGF1R knockout (MIGIRKO) impaired mitochondrial respiration, decreased ATP production, and increased ROS. These mitochondrial abnormalities were not present in muscle-specific IR/IGF1R and FoxO1/3/4 quintuple knockout mice (M-QKO). Acute tamoxifen-inducible deletion of IR/IGF1R also decreased muscle pyruvate respiration, complex-I activity, and supercomplex assembly. Although autophagy was increased when IR/IGF1R were deleted in muscle, mitophagy was not increased. Mechanistically, RNA-seq revealed that complex-I core subunits were decreased in STZ-diabetic and MIGIRKO muscle, and these changes were not present with FoxO knockout in STZ-FoxO TKO and M-QKO. Thus, insulin-deficient diabetes or loss of insulin/IGF-1 action in muscle decreases complex-I driven mitochondrial respiration and supercomplex assembly, in part by FoxO-mediated repression of Complex-I subunit expression.

Authors

Gourav Bhardwaj, Christie M. Penniman, Jayashree Jena, Pablo A. Suarez Beltran, Collin Foster, Kennedy Poro, Taylor L. Junck, Antentor O. Hinton Jr., Rhonda Souvenir, Jordan D. Fuqua, Pablo E. Morales, Roberto Bravo-Sagua, William I. Sivitz, Vitor A. Lira, E. Dale Abel, Brian T. O'Neill

×

Advertisement

August 2021 JCI This Month

JCI This Month is a digest of the research, reviews, and other features published each month.

×

Review Series - More

Gut-Brain Axis

Series edited by Ted M. Dawson and Jean-Pierre Raufman

This collection of reviews focuses on the gut-brain axis, highlighting crosstalk between the gastrointestinal tract and the enteric and central nervous systems. While the enteric nervous system can exert independent control over the gut, multi-directional communication with the central nervous system, as well as intestinal epithelial, stromal, immune, and enteroendocrine cells can result in wide-ranging influences on health and disease. The gut microbiome and its metabolites add further complexity to this intricate interactive network. Reviews in this series take a critical approach to describing the role of gut-brain connections in conditions affecting both gut and brain, with the common goal of illuminating the importance of the central and enteric nervous system interface in disease pathogenesis and identifying nodes that offer therapeutic potential.

×