bioenergetics-oxidative-burst-protocols-for-isolation-evaluation

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

Mitochondrial dysfunction is known to play a significant role in a number of pathological conditions such as atherosclerosis, diabetes, septic shock, and ...

University of Alabama at Birmingham

&#x82B3;&#x9999;&#x5316;&#x9176; (CYP19) &#x662F;&#x96CC;&#x6FC0;&#x7D20;&#x751F;&#x7269;&#x5408;&#x6210;&#x4E2D;&#x7684;&#x5173;&#x952E;&#x9176;&#x548C;&#x82B3;&#x9999;&#x5316;&#x9176;&#x6291;&#x5236;&#x5242; (AI) &#x4ECE;&#x4E8B;&#x5E7F;&#x6CDB;&#x7684;&#x5185;&#x5206;&#x6CCC;&#x6CBB;&#x7597;&#x7EDD;&#x7ECF;&#x540E;&#x5973;&#x6027;&#x4E73;&#x817A;&#x764C;&#x3002;&#x6211;&#x4EEC;&#x5047;&#x8BBE;&#x8FD9;&#x5355;&#x6838;&#x82F7;&#x9178;&#x591A;&#x6001;&#x6027; (SNPs) &#x5728; CYP19 &#x57FA;&#x56E0;&#x53EF;&#x80FD;&#x4F1A;&#x6539;&#x53D8; AI &#x4E2D;&#x8BBE;&#x7F6E;&#x7684;&#x65B0;&#x8F85;&#x52A9;&#x7597;&#x6CD5;&#x7684;&#x6709;&#x6548;&#x6027;&#x3002;&#x57FA;&#x56E0;&#x7EC4; DNA &#x6D4B;&#x5E8F;&#x4ECE; 52 &#x5987;&#x5973; pre-AI &#x548C; post-AI &#x6CBB;&#x7597;&#x5728;&#x6B64;&#x8BBE;&#x7F6E;&#x4E2D;&#x83B7;&#x5F97;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x57FA;&#x56E0;&#x7EC4; DNA &#x6765;&#x81EA; 82 &#x6837;&#x54C1;&#x7684;&#x4E73;&#x817A;&#x764C;&#x548C;&#x6B63;&#x5E38;&#x4E73;&#x817A;&#x7EC4;&#x7EC7; 19 &#x6837;&#x672C;&#x53D7;&#x5230;&#x4E86;&#x91CD;&#x6D4B;&#x5E8F;&#x670D;&#x52A1;&#x3002;&#x540C;&#x4E00;&#x4E2A;&#x75C5;&#x4EBA;&#x7684;&#x80BF;&#x7624;&#x548C;&#x80DA;&#x82BD;&#x7EBF; DNA &#x4E4B;&#x95F4;&#x89C2;&#x5BDF;&#x5230; CYP19 &#x5E8F;&#x5217;&#x4E2D;&#x7684;&#x6CA1;&#x6709;&#x5DEE;&#x5F02;&#x3002;&#x603B;&#x5171;&#x6709; 48 SNPs &#x786E;&#x5B9A;&#x4E86;&#x5305;&#x62EC; 4 &#x79CD;&#x65B0;&#x578B; SNPs &#x4E0E;&#x4EE5;&#x524D;&#x7684; resequencing &#x6570;&#x636E;&#x76F8;&#x6BD4;&#x3002;&#x57FA;&#x56E0;&#x578B;&#x548C;&#x8868;&#x578B;&#x7814;&#x7A76;&#x534F;&#x4F1A;&#xFF0C;&#x6211;&#x4EEC;&#x786E;&#x5B9A;&#x7684;&#x82B3;&#x9999;&#x5316;&#x9176;&#x6D3B;&#x6027;&#x3001; &#x96CC;&#x916E;&#x3001; &#x96CC;&#x4E8C;&#x9187;&#x548C;&#x60A3;&#x8005; pre-AI &#x548C; post-AI &#x6CBB;&#x7597;&#x80BF;&#x7624;&#x5927;&#x5C0F;&#x7684;&#x7EA7;&#x522B;&#x3002;&#x6211;&#x4EEC;&#x5B9A;&#x4E49;&#x4E24;&#x4E2A;&#x7D27;&#x5BC6;&#x76F8;&#x8FDE;&#x7684; Snp &#xFF08;rs6493497 &#x548C; rs7176005 &#x7684;&#x5916;&#x663E;&#x5B50; CYP19 1.1 5&#39; &#x4FA7;&#x7FFC;&#x5730;&#x533A;&#xFF09; AI &#x6CBB;&#x7597;&#x540E;&#x6709;&#x660E;&#x663E;&#x5173;&#x8054;&#x8F83;&#x5927;&#x7684;&#x6539;&#x53D8;&#xFF0C;&#x5728;&#x82B3;&#x9999;&#x5316;&#x9176;&#x6D3B;&#x6027;&#x3002;&#x5728; 200 &#x540D;&#x5987;&#x5973;&#x4E0E;&#x65E9;&#x671F;&#x4E73;&#x817A;&#x764C;&#x6CBB;&#x7597;&#x4F50;&#x5242;&#x90A3;&#x66F2;&#x7684;&#x968F;&#x8BBF;&#x7814;&#x7A76;&#xFF0C;&#x8FD9;&#x4E9B;&#x76F8;&#x540C;&#x7684;&#x4E24;&#x4E2A;&#x5355;&#x6838;&#x82F7;&#x9178;&#x591A;&#x6001;&#x6027;&#x4E5F;&#x4E0E;&#x60A3;&#x8005; pre-AI &#x548C; post-AI &#x7684;&#x5F85;&#x9047;&#x66F4;&#x9AD8;&#x8840;&#x6D46;&#x96CC;&#x4E8C;&#x9187;&#x76F8;&#x5173;&#x8054;&#x3002;&#x7535;&#x6CF3; shift &#x548C;&#x8BB0;&#x8005;&#x57FA;&#x56E0;&#x68C0;&#x6D4B;&#x8FD9;&#x4E9B;&#x4E24;&#x4E2A;&#x5355;&#x6838;&#x82F7;&#x9178;&#x591A;&#x6001;&#x6027;&#x5BF9;&#x8F6C;&#x5F55; CYP19 &#x8BC1;&#x5B9E;&#x53EF;&#x80FD;&#x529F;&#x80FD;&#x5F71;&#x54CD;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x663E;&#x793A;&#x800C;&#x4E24;&#x4E2A;&#x5E38;&#x89C1;&#x9057;&#x4F20;&#x591A;&#x6001;&#x6027;&#x7814;&#x7A76;&#x4E2D;&#x7684;&#x82B3;&#x9999;&#x5316;&#x9176;&#x57FA;&#x56E0; CYP19 &#x5F02;&#x82B3;&#x9999;&#x5316;&#x9176;&#x6291;&#x5236;&#x5242;&#x5BF9;&#x4E73;&#x817A;&#x764C;&#x60A3;&#x8005;&#x7684;&#x53CD;&#x5E94;&#x3002;

&#x82B3;&#x9999;&#x5316;&#x9176;&#x57FA;&#x56E0; CYP19 &#x529F;&#x80FD;&#x57FA;&#x56E0;&#x591A;&#x6001;&#x6027;&#x4E0D;&#x540C;&#x82B3;&#x9999;&#x5316;&#x9176;&#x6291;&#x5236;&#x5242;&#x7684;&#x65B0;&#x8F85;&#x52A9;&#x6CBB;&#x7597;&#x5BF9;&#x4E73;&#x817A;&#x764C;&#x60A3;&#x8005;&#x7684;&#x53CD;&#x5E94;&#x3002;

Aromatase (CYP19) is a critical enzyme in estrogen biosynthesis and aromatase inhibitors (AI) are employed widely for endocrine therapy in postmenopausal women with breast cancer. We hypothesized that single nucleotide polymorphisms (SNPs) in the CYP19 gene may alter the effectiveness of AI therapy in the neoadjuvant setting. Genomic DNA was obtained for sequencing from 52 women pre-AI and post-AI treatment in this setting. Additionally, genomic DNA obtained from 82 samples of breast cancer and 19 samples of normal breast tissue was subjected to resequencing. No differences in CYP19 sequence were observed between tumor and germ-line DNA in the same patient. A total of 48 SNPs were identified including 4 novel SNPs when compared with previous resequencing data. For genotype-phenotype association studies, we determined the levels of aromatase activity, estrone, estradiol, and tumor size in patients pre-AI and post-AI treatment. We defined two tightly linked SNPs (rs6493497 and rs7176005 in the 5'-flanking region of CYP19 exon 1.1) that were significantly associated with a greater change in aromatase activity after AI treatment. In a follow-up study of 200 women with early-stage breast cancer who were treated with adjuvant anastrozole, these same two SNPs were also associated with higher plasma estradiol levels in patients pre-AI and post-AI treatment. Electrophoretic mobility shift and reporter gene assays confirmed likely functional effects of these two SNPs on transcription of CYP19. Our findings indicate that two common genetic polymorphisms in the aromatase gene CYP19 vary the response of breast cancer patients to aromatase inhibitors.

Functional genetic polymorphisms in the aromatase gene CYP19 vary the response of breast cancer patients to neoadjuvant therapy with aromatase inhibitors.

Aromatase (CYP19) est une enzyme essentielle dans la biosynth&egrave;se des oestrog&egrave;nes et des inhibiteurs de l&#39;aromatase (IA) sont employ&eacute;s couramment pour l&#39;hormonoth&eacute;rapie chez les femmes m&eacute;nopaus&eacute;es atteintes d&#39;un cancer. Nous avons suppos&eacute; que simples de nucl&eacute;otide simple (SNP) dans le CYP19 g&egrave;ne peut-&ecirc;tre alt&eacute;rer l&#39;efficacit&eacute; de la th&eacute;rapie de AI dans le n&eacute;oadjuvante d&eacute;finissant. L&#39;ADN g&eacute;nomique a &eacute;t&eacute; obtenue pour le s&eacute;quen&ccedil;age d&#39;un traitement pr&eacute; et post-AI 52 femmes dans ce cadre. En outre, l&#39;ADN g&eacute;nomique provenant de 82 &eacute;chantillons de cancer du sein et 19 &eacute;chantillons de tissu mammaire normal a &eacute;t&eacute; soumis &agrave; la renum&eacute;rotation. Aucune diff&eacute;rence dans la s&eacute;quence CYP19 ont &eacute;t&eacute; observ&eacute;s entre la tumeur et la lign&eacute;e germinale ADN chez le m&ecirc;me patient. Un total de 48 SNP ont &eacute;t&eacute; identifi&eacute;s dont 4 SNP roman en comparaison avec les donn&eacute;es pr&eacute;c&eacute;dentes resequencing. Pour les &eacute;tudes de liaison g&eacute;notype-ph&eacute;notype, nous avons d&eacute;termin&eacute; les niveaux de l&#39;activit&eacute; d&#39;aromatase, oestrone, estradiol et taille de la tumeur dans le traitement de pr&eacute; et post-AI les patients. Nous avons d&eacute;fini deux SNP fortement li&eacute;s (rs6493497 et rs7176005 dans la r&eacute;gion flanquante 5&#39; de l&#39;exon CYP19 1.1) qui &eacute;tait significativement associ&eacute;s &agrave; un plus grand changement dans l&#39;activit&eacute; d&#39;aromatase apr&egrave;s traitement de l&#39;AI. Dans une &eacute;tude compl&eacute;mentaire de 200 femmes atteintes d&#39;un cancer du sein au stade pr&eacute;coce, qui ont &eacute;t&eacute; trait&eacute;s avec adjuvant anastrozole, ces m&ecirc;me deux SNP &eacute;tait aussi associ&eacute;es &agrave; des niveaux plus &eacute;lev&eacute;s d&#39;estradiol plasmatique dans le traitement de pr&eacute; et post-AI les patients. G&egrave;ne de Maj et reporter la mobilit&eacute; &eacute;lectrophor&eacute;tique essais confirm&eacute;s probables des effets fonctionnels de ces deux SNP sur la transcription de CYP19. Nos r&eacute;sultats indiquent que deux polymorphismes g&eacute;n&eacute;tiques communs dans le g&egrave;ne de l&#39;aromatase CYP19 varient la r&eacute;ponse des patients atteints de cancer du sein aux inhibiteurs de l&#39;aromatase.

Des polymorphismes g&eacute;n&eacute;tiques fonctionnelles du g&egrave;ne de l&#39;aromatase CYP19 varient la r&eacute;ponse des patients atteints de cancer du sein &agrave; th&eacute;rapie n&eacute;oadjuvante avec des inhibiteurs de l&#39;aromatase.

Aromatase (CYP19) ist eine kritische Enzym in &Ouml;strogen-Biosynthese und Aromatasehemmer (AI) werden weithin f&uuml;r endokrine Therapie bei postmenopausalen Frauen mit Brustkrebs eingesetzt. Wir vermutet, dass Einzel-Nukleotid-Polymorphismen (SNPs) in das CYP19 gen die Wirksamkeit der AI-Therapie in der neoadjuvanten Einstellung ver&auml;ndern kann. Genomischer DNA Sequenzierung von 52 Frauen Pre-AI und post-AI Behandlung in dieser Einstellung erfolgte. Dar&uuml;ber hinaus genomischer DNA aus 82 Proben von Brustkrebs und 19 Proben von normalen Brustgewebe Neusequenzierung unterworfen war. Keine Unterschiede in der Sequenz CYP19 wurden zwischen Tumor und Keimbahn DNA bei derselben Patientin beobachtet. Insgesamt 48 SNPs wurden identifiziert, darunter 4 Roman SNPs im Vergleich zu fr&uuml;heren resequencing Daten. F&uuml;r Genotyp-Ph&auml;notyp-Verein-Studien haben wir die Ebenen der Aromatase-Aktivit&auml;t, Estron, Estradiol und Tumor-Gr&ouml;&szlig;e in Pre-AI und post-AI Behandlung der Patienten festgestellt. Wir definiert zwei eng verkn&uuml;pfte SNPs (rs6493497 und rs7176005 in der 5&#39;-flankierenden Region CYP19 Exon 1.1), die nach der AI-Behandlung deutlich von einer gr&ouml;&szlig;eren Verschiebung im Aromatase-Aktivit&auml;t zugeordnet waren. In einer Follow-up Studie &uuml;ber 200 Frauen mit Fr&uuml;hstadium Brustkrebs Adjuvant Anastrozole behandelt wurden, waren diese gleichen zwei SNPs auch h&ouml;here &Ouml;stradiol-Plasmaspiegel bei Patienten Pre-AI und post-AI Behandlung zugeordnet. Elektrophoretische Mobilit&auml;t-Shift und Reporter-Gen-Proben best&auml;tigte m&ouml;glichen funktionelle Auswirkungen des diese zwei SNPs auf Transkription CYP19. Unsere Ergebnisse zeigen, dass zwei gemeinsame genetische Polymorphismen in der Aromatase-gen CYP19 die Reaktion von Brustkrebspatientinnen auf Aromatasehemmer variieren.

Funktionale genetische Polymorphismen in der Aromatase-gen CYP19 variieren die Reaktion von Brustkrebspatientinnen auf Neoadjuvante Therapie mit Aromatasehemmer.

Aromatasi (CYP19) sono un enzima fondamentale nella biosintesi degli estrogeni e inibitori dell&#39;aromatasi (AI) sono ampiamente impiegati per la terapia endocrina nelle donne in postmenopausa con cancro al seno. Abbiamo ipotizzato che polimorfismi a singolo nucleotide (SNPs) nel CYP19 gene pu&ograve; alterare l&#39;efficacia della terapia neoadiuvante impostazione AI. DNA genomico &egrave; stato ottenuto per il sequenziamento di trattamento pre-ai e post-AI 52 donne in questa impostazione. Inoltre, DNA genomic ottenuto da 82 campioni di tumore al seno e 19 campioni di tessuto mammario normale &egrave; stato sottoposto a risequenziazione. Non in sequenza CYP19 sono state osservate differenze tra DNA tumorale e germinale nel paziente stesso. Un totale di 48 SNPs sono stati identificati tra cui 4 SNPs romanzo rispetto ai precedenti dati resequencing. Per gli studi di associazione genotipo-fenotipo, abbiamo determinato i livelli di attivit&agrave; di aromatase, estrone, estradiolo e dimensioni del tumore in pazienti pre-ai e post-AI trattamento. Abbiamo definito due SNPs strettamente collegati (rs6493497 e rs7176005 in 5&#39;-regione di esone CYP19 1.1 di accompagnamento) che erano significativamente associati con un maggiore cambiamento nell&#39;attivit&agrave; di aromatase dopo trattamento AI. In uno studio di follow-up di 200 donne con cancro della mammella in fase iniziale, che sono state trattate con anastrozolo adiuvante, queste stesse due SNPs sono stati anche associati con livelli pi&ugrave; elevati di estradiolo al plasma nel trattamento di pazienti pre-ai e post-AI. Confermato probabili effetti funzionali di questi due SNPs su trascrizione di CYP19 analisi del gene di shift e reporter di mobilit&agrave; elettroforetica. I nostri risultati indicano che i due comuni polimorfismi genetici del gene dell&#39;aromatasi CYP19 variano la risposta di pazienti affetti da cancro al seno agli inibitori dell&#39;aromatasi.

Polimorfismi genetici funzionale del gene dell&#39;aromatasi CYP19 variano la risposta di pazienti affetti da cancro al seno alla terapia neoadiuvante con inibitori dell&#39;aromatasi.

&#x30A8;&#x30B9;&#x30C8;&#x30ED;&#x30B2;&#x30F3;&#x751F;&#x5408;&#x6210;&#x306B;&#x304A;&#x3051;&#x308B;&#x91CD;&#x8981;&#x306A;&#x9175;&#x7D20;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC; (CYP19) &#x3092;&#x3042;&#x308A;&#x3001;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x963B;&#x5BB3;&#x5264; &#xFF08;AI&#xFF09; &#x306F;&#x9589;&#x7D4C;&#x5F8C;&#x5973;&#x6027;&#x306E;&#x4E73;&#x304C;&#x3093;&#x306E;&#x5185;&#x5206;&#x6CCC;&#x7642;&#x6CD5;&#x306E;&#x5E83;&#x304F;&#x63A1;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x907A;&#x4F1D;&#x5B50; AI &#x8A2D;&#x5B9A;&#x30CD;&#x30AA;&#x30A2;&#x30B8;&#x30E5; &#x30D0;&#x30F3;&#x30C8;&#x7642;&#x6CD5;&#x306E;&#x6709;&#x52B9;&#x6027;&#x3092;&#x5909;&#x66F4;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C; CYP19 &#x3067;&#x3001;&#x5358;&#x4E00;&#x306E;&#x30CC;&#x30AF;&#x30EC;&#x30AA;&#x30C1;&#x30C9;&#x306E;&#x591A;&#x5F62; (SNPs) &#x3092;&#x4EEE;&#x5B9A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x30B2;&#x30CE;&#x30E0; DNA &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B7;&#x30F3;&#x30B0; 52 &#x5973;&#x6027;&#x306E;&#x30D7;&#x30EC; ai &#x3068; post-AI &#x306E;&#x6CBB;&#x7642;&#x3053;&#x306E;&#x8A2D;&#x5B9A;&#x304B;&#x3089;&#x5F97;&#x3089;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x307E;&#x305F;&#x3001;&#x4E73;&#x304C;&#x3093;&#x306E; 82 &#x306E;&#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x304B;&#x3089;&#x30B2;&#x30CE;&#x30E0; DNA &#x3092;&#x5F97;&#x3001;19 &#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x6B63;&#x5E38;&#x4E73;&#x623F;&#x7D44;&#x7E54;&#x306E;&#x30EA;&#x30B7;&#x30FC;&#x30AF;&#x30A8;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x3092;&#x53D7;&#x3051;&#x305F;&#x3002;&#x540C;&#x3058;&#x60A3;&#x8005;&#x306E;&#x816B;&#x760D;&#x304A;&#x3088;&#x3073;&#x751F;&#x6B96;&#x7CFB;&#x5217;&#x306E; DNA CYP19 &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x306E;&#x9055;&#x3044;&#x306F;&#x8A8D;&#x3081;&#x3089;&#x308C;&#x306A;&#x304B;&#x3063;&#x305F;&#x3002;48 SNPs &#x306E;&#x5408;&#x8A08;&#x306F;&#x3001;&#x524D;&#x306E;&#x9806;&#x5E8F;&#x5236;&#x5FA1;&#x30C7;&#x30FC;&#x30BF;&#x3068;&#x6BD4;&#x8F03;&#x3059;&#x308B;&#x3068; 4 &#x5C0F;&#x8AAC; SNPs &#x3092;&#x542B;&#x3080;&#x8B58;&#x5225;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x907A;&#x4F1D;&#x5B50;&#x578B;-&#x8868;&#x73FE;&#x578B;&#x5354;&#x4F1A;&#x306E;&#x7814;&#x7A76;&#x306F;&#x3001;&#x60A3;&#x8005;&#x306E;&#x30D7;&#x30EC; ai &#x3068; post-AI &#x306E;&#x6CBB;&#x7642;&#x306B;&#x304A;&#x3051;&#x308B;&#x816B;&#x760D;&#x5F84;&#x30FB; &#x30A8;&#x30B9;&#x30C8;&#x30E9;&#x30B8; &#x30AA;&#x30FC;&#x30EB;&#x3001;&#x30A8;&#x30B9;&#x30C8;&#x30ED;&#x30F3; &#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x6D3B;&#x6027;&#x306E;&#x30EC;&#x30D9;&#x30EB;&#x3092;&#x6C7A;&#x5B9A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306F; 2 &#x3064;&#x306E;&#x7DCA;&#x5BC6;&#x306B;&#x7D50;&#x3073;&#x3064;&#x3044;&#x305F; SNPs &#x3092;&#x5B9A;&#x7FA9; (rs6493497 &#x3068; rs7176005 &#x306E; 5&#39;-&#x96A3;&#x63A5;&#x9818;&#x57DF; CYP19 &#x30A8;&#x30AF;&#x30BD;&#x30F3; 1.1 &#x306E;) &#x306F;&#x3001;AI &#x6CBB;&#x7642;&#x5F8C;&#x95A2;&#x9023;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x6D3B;&#x6027;&#x306F;&#x5927;&#x304D;&#x304F;&#x5909;&#x5316;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x30A2;&#x30B8;&#x30E5;&#x30D0;&#x30F3;&#x30C8; &#x30A2;&#x30CA;&#x30B9;&#x30C8;&#x30ED;&#x30BE;&#x30FC;&#x30EB;&#x3068;&#x6271;&#x308F;&#x308C;&#x305F; 200 &#x4EBA;&#x306E;&#x5973;&#x6027;&#x65E9;&#x671F;&#x4E73;&#x304C;&#x3093;&#x60A3;&#x8005;&#x306E;&#x30D5;&#x30A9;&#x30ED;&#x30FC; &#x30A2;&#x30C3;&#x30D7;&#x7814;&#x7A76;&#x3067;&#x306F;, &#x3053;&#x308C;&#x3089;&#x306E;&#x540C;&#x3058; 2 &#x3064; SNPs &#x307E;&#x305F;&#x30D7;&#x30E9;&#x30BA;&#x30DE; &#x30A8;&#x30B9;&#x30C8;&#x30E9;&#x30B8; &#x30AA;&#x30FC;&#x30EB; &#x30EC;&#x30D9;&#x30EB;&#x306E;&#x9AD8;&#x3044;&#x60A3;&#x8005;&#x306E;&#x30D7;&#x30EC; ai &#x3068; post-AI &#x306E;&#x6CBB;&#x7642;&#x306B;&#x95A2;&#x9023;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x96FB;&#x6C17;&#x6CF3;&#x52D5;&#x306E;&#x79FB;&#x52D5;&#x6027;&#x30B7;&#x30D5;&#x30C8;&#x3068;&#x30EC;&#x30DD;&#x30FC;&#x30BF;&#x30FC;&#x907A;&#x4F1D;&#x5B50; CYP19 &#x306E;&#x8EE2;&#x5199;&#x306B;&#x95A2;&#x3059;&#x308B;&#x3053;&#x308C;&#x3089; 2 &#x3064;&#x306E; SNPs &#x306E;&#x78BA;&#x5B9A;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x9AD8;&#x3044;&#x6A5F;&#x80FD;&#x7684;&#x52B9;&#x679C;&#x8A66;&#x91D1;&#x3057;&#x307E;&#x3059;&#x3002;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x907A;&#x4F1D;&#x5B50; CYP19 &#x306E; 2 &#x3064;&#x306E;&#x5171;&#x901A;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x591A;&#x5F62;&#x4E73;&#x304C;&#x3093;&#x60A3;&#x8005;&#x306E;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x963B;&#x5BB3;&#x5264;&#x3078;&#x306E;&#x5FDC;&#x7B54;&#x3067;&#x7570;&#x306A;&#x308B;&#x3053;&#x3068;&#x304C;&#x793A;&#x5506;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;

&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x907A;&#x4F1D;&#x5B50; CYP19 &#x306E;&#x6A5F;&#x80FD;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x591A;&#x5F62;&#x4E73;&#x304C;&#x3093;&#x60A3;&#x8005;&#x306E;&#x5FDC;&#x7B54;&#x30CD;&#x30AA;&#x30A2;&#x30B8;&#x30E5; &#x30D0;&#x30F3;&#x30C8;&#x7642;&#x6CD5;&#x30A2;&#x30ED;&#x30DE;&#x30BF;&#x30FC;&#x30BC;&#x963B;&#x5BB3;&#x5264;&#x3068;&#x306B;&#x7570;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;

Neoplastic cells are genetically unstable. Strategies that target pathways affecting genome instability can be exploited to disrupt tumor cell growth, potentially with limited consequences to normal cells. Chromosomal instability (CIN) is one type of genome instability characterized by mitotic defects that increase the rate of chromosome mis-segregation. CIN is frequently caused by extra centrosomes that transiently disrupt normal bipolar spindle geometry needed for accurate chromosome segregation. Tumor cells survive with extra centrosomes because of biochemical pathways that cluster centrosomes and promote chromosome segregation on bipolar spindles. Recent work shows that targeted inhibition of these pathways prevents centrosome clustering and forces chromosomes to segregate to multiple daughter cells, an event triggering apoptosis that we refer to as anaphase catastrophe. Anaphase catastrophe specifically kills tumor cells with more than 2 centrosomes. This death program can occur after genetic or pharmacologic inhibition of cyclin dependent kinase 2 (Cdk2) and is augmented by combined treatment with a microtubule inhibitor. This proapoptotic effect occurs despite the presence of ras mutations in cancer cells. Anaphase catastrophe is a previously unrecognized mechanism that can be pharmacologically induced for apoptotic death of cancer cells and is, therefore, appealing to engage for cancer therapy and prevention.

Anaphase catastrophe is a target for cancer therapy.

&#x5176;&#x4ED6;&#x51E0;&#x4E2A;&#x9000;&#x884C;&#x6027;&#x75BE;&#x75C5;&#x5982;&#x5305;&#x6DB5;&#x4F53;&#x808C;&#x708E;&#x548C;&#x963F;&#x5C14;&#x8328;&#x6D77;&#x9ED8;&#x75C5;&#x7684;&#x7279;&#x5F81;&#x4E4B;&#x4E00;&#x662F;&#x6DC0;&#x7C89;&#x6837;&#x524D;&#x4F53;&#x86CB;&#x767D; (APP) &#x7684;&#x5F02;&#x5E38;&#x79EF;&#x805A;&#x548C;&#x5176;&#x6C34;&#x89E3;&#x6DC0;&#x7C89;&#x6837;&#x80BD;&#x3002;&#x66F4;&#x597D;&#x5730;&#x7406;&#x89E3;&#x4E0D;&#x5F53; APP &#x8868;&#x8FBE;&#x5BF9;&#x53D1;&#x5C55;&#x4E2D;&#x56FD;&#x5BB6;&#x7EC4;&#x7EC7;&#x75C5;&#x7406;&#x7684;&#x540E;&#x679C;&#xFF0C;&#x6211;&#x4EEC;&#x4E8E;&#x9AA8;&#x9ABC;&#x808C;&#xFF0C;&#x751F;&#x6210;&#x8F6C;&#x57FA;&#x56E0;&#x82CD;&#x8747;&#xFF0C;&#x8868;&#x8FBE;&#x91CE;&#x751F;&#x578B;&#x4EBA;&#x7C7B;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#xFF0C;&#x7136;&#x540E;&#x6267;&#x884C;&#x6210;&#x4EBA;&#x7684;&#x89E3;&#x5256;&#x3001; &#x751F;&#x7406;&#x3001; &#x884C;&#x4E3A;&#x5206;&#x6790;&#x3002;

&#x4EBA;&#x7C7B;&#x6DC0;&#x7C89;&#x6837;&#x524D;&#x4F53;&#x86CB;&#x767D;&#x5728;&#x679C;&#x8747;&#x7684;&#x9AA8;&#x9ABC;&#x808C;&#x4E2D;&#x8868;&#x8FBE;&#x5F0F;&#x7684;&#x7ED3;&#x679C;&#x5728;&#x5E74;&#x9F84;&#x548C;&#x6D3B;&#x52A8;&#x76F8;&#x5173;&#x808C;&#x8089;&#x8F6F;&#x5F31;&#x3002;

One of the hallmarks of Alzheimer's disease, and several other degenerative disorders such as Inclusion Body Myositis, is the abnormal accumulation of amyloid precursor protein (APP) and its proteolytic amyloid peptides. To better understand the pathological consequences of inappropriate APP expression on developing tissues, we generated transgenic flies that express wild-type human APP in the skeletal muscles, and then performed anatomical, electrophysiological, and behavioral analysis of the adults.

Expression of human amyloid precursor protein in the skeletal muscles of Drosophila results in age- and activity-dependent muscle weakness.

L&#x26;#39;une des caract&#xE9;ristiques de la maladie d&#x26;#39;Alzheimer, et de plusieurs autres maladies d&#xE9;g&#xE9;n&#xE9;ratives telles que la myosite &#xE0; inclusions, est l&#x26;#39;accumulation anormale de la prot&#xE9;ine pr&#xE9;curseur amylo&#xEF;de (APP) et de ses peptides amylo&#xEF;des prot&#xE9;olytiques. Afin de mieux comprendre les cons&#xE9;quences pathologiques de l&#x26;#39;expression inappropri&#xE9;e APP sur les tissus en voie de d&#xE9;veloppement, nous avons g&#xE9;n&#xE9;r&#xE9; des mouches transg&#xE9;niques qui expriment de type sauvage APP humain dans les muscles squelettiques, puis effectu&#xE9; anatomique, l&#x26;#39;analyse &#xE9;lectrophysiologique et comportementale des adultes.

L&#x26;#39;expression de la prot&#xE9;ine pr&#xE9;curseur amylo&#xEF;de humaine dans les muscles squelettiques de r&#xE9;sultats chez la drosophile dans l&#x26;#39;&#xE2;ge et d&#xE9;pendant de l&#x26;#39;activit&#xE9; Faiblesse musculaire

Eines der Kennzeichen der Alzheimer-Krankheit, und mehrere andere degenerativen Erkrankungen wie Einschlussk&ouml;rper-Myositis, ist die abnorme Ansammlung von Amyloid Vorl&auml;uferprotein (APP) und seine proteolytische Amyloid-Peptide. Um besser zu verstehen, die pathologischen Folgen der unangemessenen APP-Ausdruck auf die Entwicklung von Geweben, wir generiert transgenen fliegen, dass menschliche Ausdr&uuml;cken Wildtyp-APP in der Skelettmuskulatur und dann anatomische, Verhaltens- und elektrophysiologische Analyse der Erwachsenen durchgef&uuml;hrt.

Ausdruck der menschlichen Amyloid-Vorl&auml;uferprotein in der Skelettmuskulatur von Drosophila f&uuml;hrt Alter und leistungsabh&auml;ngige Muskelschw&auml;che.

Una delle caratteristiche della malattia di Alzheimer e diverse altre malattie degenerative come miosite, sono l&#39;accumulo anormale della proteina precursore dell&#39;amiloide (APP) e suoi peptidi amiloidi proteolitici. Per meglio comprendere le conseguenze patologiche di inappropriata espressione APP sullo sviluppo di tessuti, abbiamo generato mosche transgenici che esprimono selvaggio-tipo umano APP nei muscoli scheletrici e poi eseguite analisi anatomica, elettrofisiologico e comportamento degli adulti.

Espressione della proteina precursore dell&#39;amiloide umana nei muscoli scheletrici di Drosophila si traduce in debolezza muscolare et&agrave; e attivit&agrave; dipendente.

&#x30A2;&#x30EB;&#x30C4;&#x30CF;&#x30A4;&#x30DE;&#x30FC;&#x75C5;&#x306E;&#x7279;&#x5FB4;&#x306E;&#x4E00;&#x3064;&#x3067;&#x3042;&#x308A;&#x3001;&#x305D;&#x306E;&#x3088;&#x3046;&#x306A;&#x5C01;&#x5165;&#x4F53;&#x7B4B;&#x708E;&#x306E;&#x3088;&#x3046;&#x306A;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x4ED6;&#x306E;&#x5909;&#x6027;&#x75BE;&#x60A3;&#x306F;&#x3001;&#x30A2;&#x30DF;&#x30ED;&#x30A4;&#x30C9;&#x524D;&#x99C6;&#x4F53;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#xFF08;APP&#xFF09;&#x3068;&#x305D;&#x306E;&#x5206;&#x89E3;&#x30A2;&#x30DF;&#x30ED;&#x30A4;&#x30C9;&#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x306E;&#x7570;&#x5E38;&#x306A;&#x84C4;&#x7A4D;&#x3067;&#x3042;&#x308B;&#x3002;&#x512A;&#x308C;&#x305F;&#x958B;&#x767A;&#x7D44;&#x7E54;&#x3078;&#x306E;&#x4E0D;&#x9069;&#x5207;&#x306A;APP&#x767A;&#x73FE;&#x306E;&#x75C5;&#x7406;&#x5B66;&#x7684;&#x7D50;&#x679C;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3001;&#x6211;&#x3005;&#x306F;&#x9AA8;&#x683C;&#x7B4B;&#x3067;&#x306F;&#x3001;&#x91CE;&#x751F;&#x578B;&#x30D2;&#x30C8;APP&#x3092;&#x767A;&#x73FE;&#x3059;&#x308B;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30B8;&#x30A7;&#x30CB;&#x30C3;&#x30AF;&#x30CF;&#x30A8;&#x3092;&#x751F;&#x6210;&#x3057;&#x3001;&#x6210;&#x4EBA;&#x306E;&#x89E3;&#x5256;&#x5B66;&#x7684;&#x3001;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x3001;&#x884C;&#x52D5;&#x89E3;&#x6790;&#x3092;&#x884C;&#x3063;&#x305F;&#x3002;

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&#xD3EC;&#xD568; &#xC2E0;&#xCCB4; &#xADFC;&#xC721; &#xB4F1; &#xB2E4;&#xB978; &#xC5EC;&#xB7EC; &#xAC00;&#xC9C0; &#xD1F4;&#xD589; &#xC131; &#xC9C8;&#xD658; &#xBC0F; &#xC54C; &#xCE20; &#xD558;&#xC774; &#xBA38; &#xBCD1;&#xC758; &#xD2B9;&#xC9D5; &#xC911; &#xD558;&#xB098;&#xB294; &#xB179;&#xB9D0; &#xCCB4; &#xC804; &#xAD6C;&#xCCB4; &#xB2E8;&#xBC31;&#xC9C8; (&#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;)&#xC758; &#xBE44;&#xC815;&#xC0C1;&#xC801; &#xCD95;&#xC801;&#xACFC; &#xB2E8;&#xBC31;&#xC9C8; &#xB179;&#xB9D0; &#xCCB4; &#xD3A9; &#xD2F0; &#xB4DC;&#xC758;. &#xC870;&#xC9C1; &#xAC1C;&#xBC1C;&#xC5D0; &#xBD80;&#xC801; &#xC808; &#xD55C; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8; &#xC2DD;&#xC758; &#xBCD1; &#xC801;&#xC778; &#xACB0;&#xACFC; &#xB354; &#xC798; &#xC774;&#xD574; &#xD558;&#xAE30; &#xACE8;&#xACA9; &#xADFC;&#xC721;&#xC5D0; &#xC720;&#xC804;&#xC790; &#xBCC0;&#xD615; &#xD30C;&#xB9AC; &#xADF8; &#xC775;&#xC2A4;&#xD504;&#xB808;&#xC2A4; &#xC57C;&#xC0DD;-&#xD0C0;&#xC785; &#xC778;&#xAC04; &#xC560;&#xD50C; &#xB9AC; &#xCF00;&#xC774; &#xC158;&#xC744; &#xC0DD;&#xC131; &#xD558; &#xACE0; &#xC131;&#xC778;&#xC758; &#xD574;&#xBD80;&#xD559;, electrophysiological, &#xBC0F; &#xD589;&#xB3D9; &#xBD84;&#xC11D;&#xC744; &#xC218;&#xD589; &#xD569;&#xB2C8;&#xB2E4;.

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Una de las caracter&#xED;sticas de la enfermedad de Alzheimer y otros trastornos degenerativos como la miositis con cuerpos de inclusi&#xF3;n, es la acumulaci&#xF3;n anormal de la prote&#xED;na precursora amiloide (APP) y sus p&#xE9;ptidos amiloides proteol&#xED;ticas. Para entender mejor las consecuencias patol&#xF3;gicas de la expresi&#xF3;n de APP inadecuado de los tejidos en desarrollo, hemos generado moscas transg&#xE9;nicas que expresan de tipo salvaje APP humano en los m&#xFA;sculos esquel&#xE9;ticos, y luego realizaron an&#xE1;lisis anat&#xF3;micos, electrofisiol&#xF3;gicos y de comportamiento de los adultos.

La expresi&#xF3;n de la prote&#xED;na precursora de amiloide humana en los m&#xFA;sculos esquel&#xE9;ticos de los resultados de Drosophila en la edad y la actividad dependiente de la debilidad muscular

The antiangiogenic activity of rPAI-1(23), a truncated plasminogen activator inhibitor-1 (PAI-1) protein, induces vasa vasorum collapse and significantly reduces plaque area and plaque cholesterol in hypercholesterolemic low-density lipoprotein receptor-deficient/apolipoprotein B48-deficient mice.

Antiangiogenic activity of rPAI-1(23) promotes vasa vasorum regression in hypercholesterolemic mice through a plasmin-dependent mechanism.

Mesenchymal stem cells (MSCs) are an alluring therapeutic resource because of their plasticity, immunoregulatory capacity and ease of availability. Human BM-derived MSCs have limited proliferative capability, consequently, it is challenging to use in tissue engineering and regenerative medicine applications. Hence, placental MSCs of maternal origin, which is one of richest sources of MSCs were chosen to establish long-term culture from the cotyledons of full-term human placenta. Flow analysis established bonafied MSCs phenotypic characteristics, staining positively for CD29, CD73, CD90, CD105 and negatively for CD14, CD34, CD45 markers. Pluripotency of the cultured MSCs was assessed by in vitro differentiation towards not only intralineage cells like adipocytes, osteocytes, chondrocytes, and myotubules cells but also translineage differentiated towards pancreatic progenitor cells, neural cells, and retinal cells displaying plasticity. These cells did not significantly alter cell cycle or apoptosis pattern while maintaining the normal karyotype; they also have limited expression of MHC-II antigens and are Naive for stimulatory factors CD80 and CD 86. Further soft agar assays revealed that placental MSCs do not have the ability to form invasive colonies. Taking together all these characteristics into consideration, it indicates that placental MSCs could serve as good candidates for development and progress of stem-cell based therapeutics.

Long-term cultured human term placenta-derived mesenchymal stem cells of maternal origin displays plasticity.

Peripheral blood mononuclear cells and platelets have long been recognized as having the potential to act as sensitive markers for mitochondrial dysfunction in a broad range of pathological conditions. However, the bioenergetic function of these cells has not been examined from the same donors, yet this is important for the selection of cell types for translational studies. Here, we demonstrate the measurement of cellular bioenergetics in isolated human monocytes, lymphocytes, and platelets, including the oxidative burst from neutrophils and monocytes from individual donors. With the exception of neutrophils, all cell types tested exhibited oxygen consumption that could be ascribed to oxidative phosphorylation with each having a distinct bioenergetic profile and distribution of respiratory chain proteins. In marked contrast, neutrophils were essentially unresponsive to mitochondrial respiratory inhibitors indicating that they have a minimal requirement for oxidative phosphorylation. In monocytes and neutrophils, we demonstrate the stimulation of the oxidative burst using phorbol 12-myristate 13-acetate and its validation in normal human subjects. Taken together, these data suggest that selection of cell type from blood cells is critical for assessing bioenergetic dysfunction and redox biology in translational research.

Methods for defining distinct bioenergetic profiles in platelets, lymphocytes, monocytes, and neutrophils, and the oxidative burst from human blood.

Defective clearance of apoptotic cells is frequently associated with perpetuation of inflammatory conditions. Our results show a rapid activation of AMP-activated kinase (AMPK) in macrophages upon exposure to apoptotic cells or lysophosphatidylcholine, a specific phospholipid that is produced and released from dying cells. AMPK activation resulted from inhibition of mitochondrial oxygen consumption and ATP production and further depended on Ca(2+) mobilization and mitochondrial reactive oxygen species generation. Once activated, AMPK increased microtubule synthesis and chemokinesis and provided adaptation to energy demand during tracking and engulfment. Uptake of apoptotic cells was increased in lungs of mice that received lysophosphatidylcholine. Furthermore, inhibition of AMPK diminished clearance of apoptotic thymocytes in vitro and in dexamethasone-treated mice. Taken together, we conclude that the mitochondrial AMPK axis is a sensor and enhancer of tracking and removal of apoptotic cell, processes crucial to resolution of inflammatory conditions and a return to tissue homeostasis.

Mitochondria and AMP-activated protein kinase-dependent mechanism of efferocytosis.

The assessment of metabolic function in cells isolated from human blood for treatment and diagnosis of disease is a new and important area of translational research. It is now becoming clear that a broad range of pathologies which present clinically with symptoms predominantly in one organ, such as the brain or kidney, also modulate mitochondrial energetics in platelets and leukocytes allowing these cells to serve as "the canary in the coal mine" for bioenergetic dysfunction. This opens up the possibility that circulating platelets and leukocytes can sense metabolic stress in patients and serve as biomarkers of mitochondrial dysfunction in human pathologies such as diabetes, neurodegeneration and cardiovascular disease. In this overview we will describe how the utilization of glycolysis and oxidative phosphorylation differs in platelets and leukocytes and discuss how they can be used in patient populations. Since it is clear that the metabolic programs between leukocytes and platelets are fundamentally distinct the measurement of mitochondrial function in distinct cell populations is necessary for translational research.

Saranya Ravi

UAB Mitochondrial Medicine Laboratory,
Center for Free Radical Biology,
Department of Pathology,
UAB Mitochondrial Medicine Laboratory, Center for Free Radical Biology, Department of Pathology

University of Alabama at Birmingham

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

Saranya Ravi

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}UAB Mitochondrial Medicine Laboratory,Center for Free Radical Biology,Department of Pathology,UAB Mitochondrial Medicine Laboratory, Center for Free Radical Biology, Department of Pathology

University of Alabama at Birmingham

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

UAB Mitochondrial Medicine Laboratory,
Center for Free Radical Biology,
Department of Pathology,
UAB Mitochondrial Medicine Laboratory, Center for Free Radical Biology, Department of Pathology