St. Marianna University School of Medicine

AUTORES: Daniel J Klionsky1745,1749*, Kotb Abdelmohsen840, Akihisa Abe1237, Md Joynal Abedin1762, Hagai Abeliovich425,
\nAbraham Acevedo Arozena789, Hiroaki Adachi1800, Christopher M Adams1669, Peter D Adams57, Khosrow Adeli1981,
\nPeter J Adhihetty1625, Sharon G Adler700, Galila Agam67, Rajesh Agarwal1587, Manish K Aghi1537, Maria Agnello1826,
\nPatrizia Agostinis664, Patricia V Aguilar1960, Julio Aguirre-Ghiso784,786, Edoardo M Airoldi89,422, Slimane Ait-Si-Ali1376,
\nTakahiko Akematsu2010, Emmanuel T Akporiaye1097, Mohamed Al-Rubeai1394, Guillermo M Albaiceta1294,
\nChris Albanese363, Diego Albani561, Matthew L Albert517, Jesus Aldudo128, Hana Alg€ul1164, Mehrdad Alirezaei1198,
\nIraide Alloza642,888, Alexandru Almasan206, Maylin Almonte-Beceril524, Emad S Alnemri1212, Covadonga Alonso544,
\nNihal Altan-Bonnet848, Dario C Altieri1205, Silvia Alvarez1497, Lydia Alvarez-Erviti1395, Sandro Alves107,
\nGiuseppina Amadoro860, Atsuo Amano930, Consuelo Amantini1554, Santiago Ambrosio1458, Ivano Amelio756,
\nAmal O Amer918, Mohamed Amessou2089, Angelika Amon726, Zhenyi An1538, Frank A Anania291, Stig U Andersen6,
\nUsha P Andley2079, Catherine K Andreadi1690, Nathalie Andrieu-Abadie502, Alberto Anel2027, David K Ann58,
\nShailendra Anoopkumar-Dukie388, Manuela Antonioli832,858, Hiroshi Aoki1791, Nadezda Apostolova2007,
\nSaveria Aquila1500, Katia Aquilano1876, Koichi Araki292, Eli Arama2098, Agustin Aranda456, Jun Araya591,
\nAlexandre Arcaro1472, Esperanza Arias26, Hirokazu Arimoto1225, Aileen R Ariosa1749, Jane L Armstrong1930,
\nThierry Arnould1773, Ivica Arsov2120, Katsuhiko Asanuma675, Valerie Askanas1924, Eric Asselin1867, Ryuichiro Atarashi794,
\nSally S Atherton369, Julie D Atkin713, Laura D Attardi1131, Patrick Auberger1787, Georg Auburger379, Laure Aurelian1727,
\nRiccardo Autelli1992, Laura Avagliano1029,1755, Maria Laura Avantaggiati364, Limor Avrahami1166, Suresh Awale1986,
\nNeelam Azad404, Tiziana Bachetti568, Jonathan M Backer28, Dong-Hun Bae1933, Jae-sung Bae677, Ok-Nam Bae409,
\nSoo Han Bae2117, Eric H Baehrecke1729, Seung-Hoon Baek17, Stephen Baghdiguian1368,
\nAgnieszka Bagniewska-Zadworna2, Hua Bai90, Jie Bai667, Xue-Yuan Bai1133, Yannick Bailly884,
\nKithiganahalli Narayanaswamy Balaji473, Walter Balduini2002, Andrea Ballabio316, Rena Balzan1711, Rajkumar Banerjee239,
\nG abor B anhegyi1052, Haijun Bao2109, Benoit Barbeau1363, Maria D Barrachina2007, Esther Barreiro467, Bonnie Bartel997,
\nAlberto Bartolom e222, Diane C Bassham550, Maria Teresa Bassi1046, Robert C Bast Jr1273, Alakananda Basu1798,
\nMaria Teresa Batista1578, Henri Batoko1336, Maurizio Battino970, Kyle Bauckman2085, Bradley L Baumgarner1909,
\nK Ulrich Bayer1594, Rupert Beale1553, Jean-Fran¸cois Beaulieu1360, George R. Beck Jr48,294, Christoph Becker336,
\nJ David Beckham1595, Pierre-Andr e B edard749, Patrick J Bednarski301, Thomas J Begley1135, Christian Behl1419,
\nChristian Behrends757, Georg MN Behrens406, Kevin E Behrns1627, Eloy Bejarano26, Amine Belaid490,
\nFrancesca Belleudi1041, Giovanni B enard497, Guy Berchem706, Daniele Bergamaschi983, Matteo Bergami1401,
\nBen Berkhout1441, Laura Berliocchi714, Am elie Bernard1749, Monique Bernard1354, Francesca Bernassola1880,
\nAnne Bertolotti791, Amanda S Bess272, S ebastien Besteiro1351, Saverio Bettuzzi1828, Savita Bhalla913,
\nShalmoli Bhattacharyya973, Sujit K Bhutia838, Caroline Biagosch1159, Michele Wolfe Bianchi520,1378,1381,
\nMartine Biard-Piechaczyk210, Viktor Billes298, Claudia Bincoletto1314, Baris Bingol350, Sara W Bird1128, Marc Bitoun1112,
\nIvana Bjedov1258, Craig Blackstone843, Lionel Blanc1183, Guillermo A Blanco1496, Heidi Kiil Blomhoff1812,
\nEmilio Boada-Romero1297, Stefan B€ockler1464, Marianne Boes1423, Kathleen Boesze-Battaglia1835, Lawrence H Boise286,287,
\nAlessandra Bolino2063, Andrea Boman693, Paolo Bonaldo1823, Matteo Bordi897, J€urgen Bosch608, Luis M Botana1308,
\nJoelle Botti1375, German Bou1405, Marina Bouch e1038, Marion Bouchecareilh1331, Marie-Jos ee Boucher1901,
\nMichael E Boulton481, Sebastien G Bouret1926, Patricia Boya133, Micha€el Boyer-Guittaut1345, Peter V Bozhkov1141,
\nNathan Brady374, Vania MM Braga469, Claudio Brancolini1997, Gerhard H Braus353, Jos e M Bravo-San Pedro299,393,508,1374,
\nLisa A Brennan322, Emery H Bresnick2022, Patrick Brest490, Dave Bridges1939, Marie-Agn es Bringer124, Marisa Brini1822,
\nGlauber C Brito1311, Bertha Brodin631, Paul S Brookes1872, Eric J Brown352, Karen Brown1690, Hal E Broxmeyer480,
\nAlain Bruhat486,1339, Patricia Chakur Brum1893, John H Brumell446, Nicola Brunetti-Pierri315,1171,
\nRobert J Bryson-Richardson781, Shilpa Buch1777, Alastair M Buchan1819, Hikmet Budak1022, Dmitry V Bulavin118,505,1789,
\nScott J Bultman1792, Geert Bultynck665, Vladimir Bumbasirevic1470, Yan Burelle1356, Robert E Burke216,217,
\nMargit Burmeister1750, Peter B€utikofer1473, Laura Caberlotto1987, Ken Cadwell896, Monika Cahova112, Dongsheng Cai24,
\nJingjing Cai2099, Qian Cai1018, Sara Calatayud2007, Nadine Camougrand1343, Michelangelo Campanella1700,
\nGrant R Campbell1525, Matthew Campbell1249, Silvia Campello556,1876, Robin Candau1769, Isabella Caniggia1983,
\nLavinia Cantoni560, Lizhi Cao116, Allan B Caplan1656, Michele Caraglia1051, Claudio Cardinali1043, Sandra Morais Cardoso1579, Jennifer S Carew208, Laura A Carleton874, Cathleen R Carlin101, Silvia Carloni2002,
\nSven R Carlsson1267, Didac Carmona-Gutierrez1643, Leticia AM Carneiro312, Oliana Carnevali971, Serena Carra1318,
\nAlice Carrier120, Bernadette Carroll900, Caty Casas1324, Josefina Casas1116, Giuliana Cassinelli324, Perrine Castets1462,
\nSusana Castro-Obregon214, Gabriella Cavallini1841, Isabella Ceccherini568, Francesco Cecconi253,555,1884,
\nArthur I Cederbaum459, Valent ın Ce~na199,1281, Simone Cenci1323,2064, Claudia Cerella444, Davide Cervia1996,
\nSilvia Cetrullo1478, Hassan Chaachouay2028, Han-Jung Chae187, Andrei S Chagin634, Chee-Yin Chai626,628,
\nGopal Chakrabarti1502, Georgios Chamilos1601, Edmond YW Chan1142, Matthew TV Chan181, Dhyan Chandra1003,
\nPallavi Chandra548, Chih-Peng Chang818, Raymond Chuen-Chung Chang1653, Ta Yuan Chang345, John C Chatham1434,
\nSaurabh Chatterjee1910, Santosh Chauhan527, Yongsheng Che62, Michael E Cheetham1263, Rajkumar Cheluvappa1783,
\nChun-Jung Chen1153, Gang Chen598,1676, Guang-Chao Chen9, Guoqiang Chen1078, Hongzhuan Chen1077, Jeff W Chen1514,
\nJian-Kang Chen370,371, Min Chen249, Mingzhou Chen2104, Peiwen Chen1823, Qi Chen1674, Quan Chen172,
\nShang-Der Chen138, Si Chen325, Steve S-L Chen10, Wei Chen2125, Wei-Jung Chen829, Wen Qiang Chen979, Wenli Chen1113,
\nXiangmei Chen1133, Yau-Hung Chen1157, Ye-Guang Chen1250, Yin Chen1447, Yingyu Chen953,955, Yongshun Chen2135,
\nYu-Jen Chen712, Yue-Qin Chen1145, Yujie Chen1208, Zhen Chen339, Zhong Chen2123, Alan Cheng1702,
\nChristopher HK Cheng184, Hua Cheng1728, Heesun Cheong814, Sara Cherry1836, Jason Chesney1703,
\nChun Hei Antonio Cheung817, Eric Chevet1359, Hsiang Cheng Chi140, Sung-Gil Chi656, Fulvio Chiacchiera308,
\nHui-Ling Chiang958, Roberto Chiarelli1826, Mario Chiariello235,567,577, Marcello Chieppa835, Lih-Shen Chin290,
\nMario Chiong1285, Gigi NC Chiu878, Dong-Hyung Cho676, Ssang-Goo Cho650, William C Cho982, Yong-Yeon Cho105,
\nYoung-Seok Cho1064, Augustine MK Choi2095, Eui-Ju Choi656, Eun-Kyoung Choi387,400,685, Jayoung Choi1563,
\nMary E Choi2093, Seung-Il Choi2116, Tsui-Fen Chou412, Salem Chouaib395, Divaker Choubey1574, Vinay Choubey1936,
\nKuan-Chih Chow822, Kamal Chowdhury730, Charleen T Chu1856, Tsung-Hsien Chuang827, Taehoon Chun657,
\nHyewon Chung652, Taijoon Chung978, Yuen-Li Chung1194, Yong-Joon Chwae18, Valentina Cianfanelli254,
\nRoberto Ciarcia1775, Iwona A Ciechomska886, Maria Rosa Ciriolo1876, Mara Cirone1042, Sofie Claerhout1694,
\nMichael J Clague1698, Joan Cl aria1457, Peter GH Clarke1687, Robert Clarke361, Emilio Clementi1045,1398, C edric Cleyrat1781,
\nMiriam Cnop1366, Eliana M Coccia574, Tiziana Cocco1459, Patrice Codogno1375, J€orn Coers271, Ezra EW Cohen1533,
\nDavid Colecchia235,567,577, Luisa Coletto25, N uria S Coll123, Emma Colucci-Guyon516, Sergio Comincini1829,
\nMaria Condello578, Katherine L Cook2073, Graham H Coombs1929, Cynthia D Cooper2076, J Mark Cooper1395,
\nIsabelle Coppens601, Maria Tiziana Corasaniti1387, Marco Corazzari485,1884, Ramon Corbalan1566,
\nElisabeth Corcelle-Termeau251, Mario D Cordero1899, Cristina Corral-Ramos1289, Olga Corti507,1109, Andrea Cossarizza1767,
\nPaola Costelli1993, Safia Costes1518, Susan L Cotman721, Ana Coto-Montes946, Sandra Cottet566,1688, Eduardo Couve1301,
\nLori R Covey1015, L Ashley Cowart762, Jeffery S Cox1536, Fraser P Coxon1427, Carolyn B Coyne1846, Mark S Cragg1919,
\nRolf J Craven1679, Tiziana Crepaldi1995, Jose L Crespo1300, Alfredo Criollo1285, Valeria Crippa558, Maria Teresa Cruz1576,
\nAna Maria Cuervo26, Jose M Cuezva1277, Taixing Cui1907, Pedro R Cutillas987, Mark J Czaja27, Maria F Czyzyk-Krzeska1572,
\nRuben K Dagda2068, Uta Dahmen1404, Chunsun Dai800, Wenjie Dai1187, Yun Dai2059, Kevin N Dalby1940,
\nLuisa Dalla Valle1822, Guillaume Dalmasso1340, Marcello D’Amelio557, Markus Damme188, Arlette Darfeuille-Michaud1340,
\nCatherine Dargemont950, Victor M Darley-Usmar1433, Srinivasan Dasarathy205, Biplab Dasgupta202, Srikanta Dash1254,
\nCrispin R Dass242, Hazel Marie Davey8, Lester M Davids1560, David D avila227, Roger J Davis1731, Ted M Dawson604,
\nValina L Dawson606, Paula Daza1898, Jackie de Belleroche470, Paul de Figueiredo1180,1182,
\nRegina Celia Bressan Queiroz de Figueiredo135, Jos e de la Fuente1023, Luisa De Martino1775,
\nAntonella De Matteis1171, Guido RY De Meyer1443, Angelo De Milito631, Mauro De Santi2002,

autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Japanese mortality due to colorectal cancer is on the rise, surpassing 49,000 in 2015. Many new treatment methods have been developed during recent decades. The Japanese Society for Cancer of the Colon and Rectum Guidelines 2016 for the treatment of colorectal cancer (JSCCR Guidelines 2016) were prepared to show standard treatment strategies for colorectal cancer, to eliminate disparities among institutions in terms of treatment, to eliminate unnecessary treatment and insufficient treatment, and to deepen mutual understanding between health-care professionals and patients by making these Guidelines available to the general public. These Guidelines were prepared by consensus reached by the JSCCR Guideline Committee, based on a careful review of the evidence retrieved by literature searches, and in view of the medical health insurance system and actual clinical practice settings in Japan. Therefore, these Guidelines can be used as a tool for treating colorectal cancer in actual clinical practice settings. More specifically, they can be used as a guide to obtaining informed consent from patients and choosing the method of treatment for each patient. As a result of the discussions held by the Guideline Committee, controversial issues were selected as Clinical Questions, and recommendations were made. Each recommendation is accompanied by a classification of the evidence and a classification of recommendation categories based on the consensus reached by the Guideline Committee members. Here we present the English version of the JSCCR Guidelines 2016.

Abstract The number of deaths from colorectal cancer in Japan continues to increase. Colorectal cancer deaths exceeded 50,000 in 2016. In the 2019 edition, revision of all aspects of treatments was performed, with corrections and additions made based on knowledge acquired since the 2016 version (drug therapy) and the 2014 version (other treatments). The Japanese Society for Cancer of the Colon and Rectum guidelines 2019 for the treatment of colorectal cancer (JSCCR guidelines 2019) have been prepared to show standard treatment strategies for colorectal cancer, to eliminate disparities among institutions in terms of treatment, to eliminate unnecessary treatment and insufficient treatment and to deepen mutual understanding between healthcare professionals and patients by making these guidelines available to the general public. These guidelines have been prepared by consensuses reached by the JSCCR Guideline Committee, based on a careful review of the evidence retrieved by literature searches and in view of the medical health insurance system and actual clinical practice settings in Japan. Therefore, these guidelines can be used as a tool for treating colorectal cancer in actual clinical practice settings. More specifically, they can be used as a guide to obtaining informed consent from patients and choosing the method of treatment for each patient. Controversial issues were selected as clinical questions, and recommendations were made. Each recommendation is accompanied by a classification of the evidence and a classification of recommendation categories based on the consensus reached by the Guideline Committee members. Here, we present the English version of the JSCCR guidelines 2019.

BehÇet's disease is an inflammatory disorder of unknown cause, characterized by recurrent oral aphthous ulcers, genital ulcers, uveitis, and skin lesions.1,2 All these common manifestations are self-limiting except for the ocular attacks. Repeated attacks of uveitis can cause blindness.13 Behçet's disease is not a chronic, persistent inflammatory disease, but rather one consisting of recurrent attacks of acute inflammation. Involvement of the gastrointestinal tract, central nervous system, and large vessels is less frequent (Table 1), although it can be life-threatening.1,2,46 Susceptibility to Behçet's disease is strongly associated with the presence of the HLA-B51 allele.7,8 Environmental . . .

Takotsubo syndrome (TTS) is a poorly recognized heart disease that was initially regarded as a benign condition. Recently, it has been shown that TTS may be associated with severe clinical complications including death and that its prevalence is probably underestimated. Since current guidelines on TTS are lacking, it appears timely and important to provide an expert consensus statement on TTS. The clinical expert consensus document part I summarizes the current state of knowledge on clinical presentation and characteristics of TTS and agrees on controversies surrounding TTS such as nomenclature, different TTS types, role of coronary artery disease, and etiology. This consensus also proposes new diagnostic criteria based on current knowledge to improve diagnostic accuracy.

Takotsubo syndrome is an acute reversible heart failure syndrome that is increasingly recognized in modern cardiology practice. This Position Statement from the European Society of Cardiology Heart Failure Association provides a comprehensive review of the various clinical and pathophysiological facets of Takotsubo syndrome, including nomenclature, definition, and diagnosis, primary and secondary clinical subtypes, anatomical variants, triggers, epidemiology, pathophysiology, clinical presentation, complications, prognosis, clinical investigations, and treatment approaches. Novel structured approaches to diagnosis, risk stratification, and management are presented, with new algorithms to aid decision-making by practising clinicians. These also cover more complex areas (e.g. uncertain diagnosis and delayed presentation) and the management of complex cases with ongoing symptoms after recovery, recurrent episodes, or spontaneous presentation. The unmet needs and future directions for research in this syndrome are also discussed.

Abstract The genetic make-up of an individual contributes to the susceptibility and response to viral infection. Although environmental, clinical and social factors have a role in the chance of exposure to SARS-CoV-2 and the severity of COVID-19 1,2 , host genetics may also be important. Identifying host-specific genetic factors may reveal biological mechanisms of therapeutic relevance and clarify causal relationships of modifiable environmental risk factors for SARS-CoV-2 infection and outcomes. We formed a global network of researchers to investigate the role of human genetics in SARS-CoV-2 infection and COVID-19 severity. Here we describe the results of three genome-wide association meta-analyses that consist of up to 49,562 patients with COVID-19 from 46 studies across 19 countries. We report 13 genome-wide significant loci that are associated with SARS-CoV-2 infection or severe manifestations of COVID-19. Several of these loci correspond to previously documented associations to lung or autoimmune and inflammatory diseases 3–7 . They also represent potentially actionable mechanisms in response to infection. Mendelian randomization analyses support a causal role for smoking and body-mass index for severe COVID-19 although not for type II diabetes. The identification of novel host genetic factors associated with COVID-19 was made possible by the community of human genetics researchers coming together to prioritize the sharing of data, results, resources and analytical frameworks. This working model of international collaboration underscores what is possible for future genetic discoveries in emerging pandemics, or indeed for any complex human disease.

The Cognitive Abilities Screening Instrument (CASI) has a score range of 0 to 100 and provides quantitative assessment on attention, concentration, orientation, short-term memory, long-term memory, language abilities, visual construction, list-generating fluency, abstraction, and judgment. Scores of the Mini-Mental State Examination, the Modified Mini-Mental State Test, and the Hasegawa Dementia Screening Scale can also be estimated from subsets of the CASI items. Pilot testing conducted in Japan and in the United States has demonstrated its cross-cultural applicability and its usefulness in screening for dementia, in monitoring disease progression, and in providing profiles of cognitive impairment. Typical administration time is 15 to 20 minutes. Record form, manual, videotape of test administration, and quizzes to qualify potential users on the administration and scoring of the CASI are available upon request.

Primary ovarian insufficiency (POI) and polycystic ovarian syndrome are ovarian diseases causing infertility. Although there is no effective treatment for POI, therapies for polycystic ovarian syndrome include ovarian wedge resection or laser drilling to induce follicle growth. Underlying mechanisms for these disruptive procedures are unclear. Here, we explored the role of the conserved Hippo signaling pathway that serves to maintain optimal size across organs and species. We found that fragmentation of murine ovaries promoted actin polymerization and disrupted ovarian Hippo signaling, leading to increased expression of downstream growth factors, promotion of follicle growth, and the generation of mature oocytes. In addition to elucidating mechanisms underlying follicle growth elicited by ovarian damage, we further demonstrated additive follicle growth when ovarian fragmentation was combined with Akt stimulator treatments. We then extended results to treatment of infertility in POI patients via disruption of Hippo signaling by fragmenting ovaries followed by Akt stimulator treatment and autografting. We successfully promoted follicle growth, retrieved mature oocytes, and performed in vitro fertilization. Following embryo transfer, a healthy baby was delivered. The ovarian fragmentation-in vitro activation approach is not only valuable for treating infertility of POI patients but could also be useful for middle-aged infertile women, cancer patients undergoing sterilizing treatments, and other conditions of diminished ovarian reserve.

BACKGROUND: No treatment has surpassed platinum-based chemotherapy in improving overall survival in patients with previously untreated locally advanced or metastatic urothelial carcinoma. METHODS: We conducted a phase 3, global, open-label, randomized trial to compare the efficacy and safety of enfortumab vedotin and pembrolizumab with the efficacy and safety of platinum-based chemotherapy in patients with previously untreated locally advanced or metastatic urothelial carcinoma. Patients were randomly assigned in a 1:1 ratio to receive 3-week cycles of enfortumab vedotin (at a dose of 1.25 mg per kilogram of body weight intravenously on days 1 and 8) and pembrolizumab (at a dose of 200 mg intravenously on day 1) (enfortumab vedotin-pembrolizumab group) or gemcitabine and either cisplatin or carboplatin (determined on the basis of eligibility to receive cisplatin) (chemotherapy group). The primary end points were progression-free survival as assessed by blinded independent central review and overall survival. RESULTS: A total of 886 patients underwent randomization: 442 to the enfortumab vedotin-pembrolizumab group and 444 to the chemotherapy group. As of August 8, 2023, the median duration of follow-up for survival was 17.2 months. Progression-free survival was longer in the enfortumab vedotin-pembrolizumab group than in the chemotherapy group (median, 12.5 months vs. 6.3 months; hazard ratio for disease progression or death, 0.45; 95% confidence interval [CI], 0.38 to 0.54; P<0.001), as was overall survival (median, 31.5 months vs. 16.1 months; hazard ratio for death, 0.47; 95% CI, 0.38 to 0.58; P<0.001). The median number of cycles was 12 (range, 1 to 46) in the enfortumab vedotin-pembrolizumab group and 6 (range, 1 to 6) in the chemotherapy group. Treatment-related adverse events of grade 3 or higher occurred in 55.9% of the patients in the enfortumab vedotin-pembrolizumab group and in 69.5% of those in the chemotherapy group. CONCLUSIONS: Treatment with enfortumab vedotin and pembrolizumab resulted in significantly better outcomes than chemotherapy in patients with untreated locally advanced or metastatic urothelial carcinoma, with a safety profile consistent with that in previous reports. (Funded by Astellas Pharma US and others; EV-302 ClinicalTrials.gov number, NCT04223856.).

The clinical expert consensus statement on takotsubo syndrome (TTS) part II focuses on the diagnostic workup, outcome, and management. The recommendations are based on interpretation of the limited clinical trial data currently available and experience of international TTS experts. It summarizes the diagnostic approach, which may facilitate correct and timely diagnosis. Furthermore, the document covers areas where controversies still exist in risk stratification and management of TTS. Based on available data the document provides recommendations on optimal care of such patients for practising physicians.

Twenty-four edible plants were investigated for the presence of melatonin, heretofore considered to be a molecule found only in the animal kingdom. The amount of melatonin in different plants varied greatly with highest melatonin being present in plants of the rice family. Melatonin was identified by radioimmunoassay and verified by high performance liquid chromatography with fluorescence detection. Feeding a diet containing plant products rich in melatonin to chicks increased radioimmunoassayable levels of melatonin in their blood. Likewise, melatonin extracted from plants inhibited binding of [125I]iodomelatonin to rabbit brain. Thus, melatonin ingested in foodstuffs enters the blood and is capable of binding to melatonin binding sites in the brain of mammals.

OBJECTIVE: Interleukin-6 (IL-6) is a pleiotropic cytokine that regulates the immune response, inflammation, and hematopoiesis. Overproduction of IL-6 plays pathologic roles in rheumatoid arthritis (RA), and the blockade of IL-6 may be therapeutically effective for the disease. This study was undertaken to evaluate the safety and efficacy of a humanized anti-IL-6 receptor antibody, MRA, in patients with RA. METHODS: In a multicenter, double-blind, placebo-controlled trial, 164 patients with refractory RA were randomized to receive either MRA (4 mg/kg body weight or 8 mg/kg body weight) or placebo. MRA was administered intravenously every 4 weeks for a total of 3 months. The clinical responses were measured using the American College of Rheumatology (ACR) criteria. RESULTS: Treatment with MRA reduced disease activity in a dose-dependent manner. At 3 months, 78% of patients in the 8-mg group, 57% in the 4-mg group, and 11% in the placebo group achieved at least a 20% improvement in disease activity according to the ACR criteria (an ACR20 response) (P < 0.001 for 8-mg group versus placebo). Forty percent of patients in the 8-mg group and 1.9% in the placebo group achieved an ACR50 response (P < 0.001). The overall incidences of adverse events were 56%, 59%, and 51% in the placebo, 4-mg, and 8-mg groups, respectively, and the adverse events were not dose dependent. A blood cholesterol increase was observed in 44.0% of the patients. Liver function disorders and decreases in white blood cell counts were also observed, but these were mild and transient. There was no increase in antinuclear antibodies or anti-DNA antibodies. Anti-MRA antibodies were detected in 2 patients. CONCLUSION: Treatment with MRA was generally well tolerated and significantly reduced the disease activity of RA.

Although hormonal regulation of ovarian follicle development has been extensively investigated, most studies concentrate on the development of early antral follicles to the preovulatory stage, leading to the successful use of exogenous FSH for infertility treatment. Accumulating data indicate that preantral follicles are under stringent regulation by FSH and local intraovarian factors, thus providing the possibility to develop new therapeutic approaches. Granulosa cell-derived C-type natriuretic factor not only suppresses the final maturation of oocytes to undergo germinal vesicle breakdown before ovulation but also promotes preantral and antral follicle growth. In addition, several oocyte- and granulosa cell-derived factors stimulate preantral follicle growth by acting through wingless, receptor tyrosine kinase, receptor serine kinase, and other signaling pathways. In contrast, the ovarian Hippo signaling pathway constrains follicle growth and disruption of Hippo signaling promotes the secretion of downstream CCN growth factors capable of promoting follicle growth. Although the exact hormonal factors involved in primordial follicle activation has yet to be elucidated, the protein kinase B (AKT) and mammalian target of rapamycin signaling pathways are important for the activation of dormant primordial follicles. Hippo signaling disruption after ovarian fragmentation, combined with treating ovarian fragments with phosphatase and tensin homolog (PTEN) inhibitors and phosphoinositide-3-kinase stimulators to augment AKT signaling, promote the growth of preantral follicles in patients with primary ovarian insufficiency, leading to a new infertility intervention for such patients. Elucidation of intraovarian mechanisms underlying early folliculogenesis may allow the development of novel therapeutic strategies for patients diagnosed with primary ovarian insufficiency, polycystic ovary syndrome, and poor ovarian response to FSH stimulation, as well as for infertile women of advanced reproductive age.

BRCA1-BARD1 constitutes a heterodimeric RING finger complex associated through its N-terminal regions. Here we demonstrate that the BRCA1-BARD1 heterodimeric RING finger complex contains significant ubiquitin ligase activity that can be disrupted by a breast cancer-derived RING finger mutation in BRCA1. Whereas individually BRCA1 and BARD1 have very low ubiquitin ligase activities in vitro, BRCA1 combined with BARD1 exhibits dramatically higher activity. Bacterially purified RING finger domains comprising residues 1–304 of BRCA1 and residues 25–189 of BARD1 are capable of polymerizing ubiquitin. The steady-state level of transfected BRCA1 in vivo was increased by co-transfection of BARD1, and reciprocally that of transfected BARD1 was increased by BRCA1 in a dose-dependent manner. The breast cancer-derived BARD1-interaction-deficient mutant, BRCA1C61G, does not exhibit ubiquitin ligase activity in vitro. These results suggest that the BRCA1-BARD1 complex contains a ubiquitin ligase activity that is important in prevention of breast and ovarian cancer development. BRCA1-BARD1 constitutes a heterodimeric RING finger complex associated through its N-terminal regions. Here we demonstrate that the BRCA1-BARD1 heterodimeric RING finger complex contains significant ubiquitin ligase activity that can be disrupted by a breast cancer-derived RING finger mutation in BRCA1. Whereas individually BRCA1 and BARD1 have very low ubiquitin ligase activities in vitro, BRCA1 combined with BARD1 exhibits dramatically higher activity. Bacterially purified RING finger domains comprising residues 1–304 of BRCA1 and residues 25–189 of BARD1 are capable of polymerizing ubiquitin. The steady-state level of transfected BRCA1 in vivo was increased by co-transfection of BARD1, and reciprocally that of transfected BARD1 was increased by BRCA1 in a dose-dependent manner. The breast cancer-derived BARD1-interaction-deficient mutant, BRCA1C61G, does not exhibit ubiquitin ligase activity in vitro. These results suggest that the BRCA1-BARD1 complex contains a ubiquitin ligase activity that is important in prevention of breast and ovarian cancer development. ubiquitin ubiquitin-activating enzyme ubiquitin carrier protein SKP1-CUL1 (CDC53)- F-box protein complex anaphase-promoting complex polyacrylamide gel electrophoresis, GST, glutathioneS-transferase hemagglutinin dithiothreitol isopropyl-1-thio-β-d-galactopyranoside Germline mutations of BRCA1 predispose women to breast and ovarian cancers (1Zheng L. Li S. Boyer T.G. Lee W.H. Oncogene. 2000; 19: 6159-6175Crossref PubMed Scopus (136) Google Scholar). BRCA1 contains several domains that interact with a variety of molecules and is potentially responsible for multiple functions in DNA damage repair, transcription, and cell-cycle regulation (2Venkitaraman A.R. Science. 1999; 286: 1100-1102Crossref PubMed Scopus (48) Google Scholar, 3Haile D.T. Parvin J.D. J. Biol. Chem. 1999; 274: 2113-2117Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 4Xu X. Weaver Z. Linke S.P. Li C. Gotay J. Wang X.W. Harris C.C. Ried T. Deng C.X. Mol. Cell. 1999; 3: 389-395Abstract Full Text Full Text PDF PubMed Scopus (703) Google Scholar). BARD1 was identified in a yeast two-hybrid screen as a protein that interacts with BRCA1 (5Wu L.C. Wang Z.W. Tsan J.T. Spillman M.A. Phung A. Xu X.L. Yang M.C. Hwang L.Y. Bowcock A.M. Baer R. Nat. Genet. 1996; 14: 430-440Crossref PubMed Scopus (626) Google Scholar). Both BRCA1 and BARD1 proteins contain a RING finger (5Wu L.C. Wang Z.W. Tsan J.T. Spillman M.A. Phung A. Xu X.L. Yang M.C. Hwang L.Y. Bowcock A.M. Baer R. Nat. Genet. 1996; 14: 430-440Crossref PubMed Scopus (626) Google Scholar) and exist as homodimers or preferentially form stable heterodimers (6Meza J.E. Brzovic P.S. King M-C. Klevit R.E. J. Biol. Chem. 1999; 274: 5659-5665Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). The heterodimeric interaction is mediated by the flanking regions of the RING finger motif of the two molecules (6Meza J.E. Brzovic P.S. King M-C. Klevit R.E. J. Biol. Chem. 1999; 274: 5659-5665Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). Although a transcriptional function in the C terminus of BRCA1 has been recently reported (3Haile D.T. Parvin J.D. J. Biol. Chem. 1999; 274: 2113-2117Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar), the biochemical function of the heterodimeric RING finger constituted from the N termini of BRCA1 and BARD1 is not known. Previously, we and others identified a highly conserved small RING finger protein, ROC1 (also called Rbx1 and Hrt1), as an essential subunit of the SCF Ub1 ligase (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar, 8Tan P. Fuchs S.Y. Chen A. Wu K. Gomez C. Ronai Z. Pan Z.Q. Mol. Cell. 1999; 3: 527-533Abstract Full Text Full Text PDF PubMed Scopus (303) Google Scholar, 9Kamura T. Koepp D.M. Conrad M.N. Skowyra D. Moreland R.J. Iliopoulos O. Lane W.S. Kaelin Jr., W.G. Elledge S.J. Conaway R.C. Harper J.W. Conaway J.W. Science. 1999; 284: 657-661Crossref PubMed Scopus (670) Google Scholar, 10Seol J.H. Feldman R.M. Zachariae W. Shevchenko A. Correll C.C. Lyapina S. Chi Y. Galova M. Claypool J. Sandmeyer S. Nasmyth K. Deshaies R.J. Shevchenko A. Deshaies R.J. Genes Dev. 1999; 13: 1614-1626Crossref PubMed Scopus (360) Google Scholar). The Ub ligase (E3) catalyzes the formation of polyubiquitin chains onto substrate proteins via isopeptide bonds utilizing the Ubs that have been sequentially activated by enzymes E1 and E2. Polyubiquitinated substrates are then rapidly degraded by the 26 S proteasome (11Hershko A. Ciechanover A. Annu. Rev. Biochem. 1998; 67: 425-479Crossref PubMed Scopus (6959) Google Scholar). The SCF and the APC are the two major Ub ligase complexes that regulate ubiquitin-mediated proteolysis during G1/S and anaphase (12Peters J.M. Curr. Opin. Cell Biol. 1998; 10: 759-768Crossref PubMed Scopus (225) Google Scholar), and contain the small RING finger proteins ROC1 and APC11, respectively (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar, 8Tan P. Fuchs S.Y. Chen A. Wu K. Gomez C. Ronai Z. Pan Z.Q. Mol. Cell. 1999; 3: 527-533Abstract Full Text Full Text PDF PubMed Scopus (303) Google Scholar, 9Kamura T. Koepp D.M. Conrad M.N. Skowyra D. Moreland R.J. Iliopoulos O. Lane W.S. Kaelin Jr., W.G. Elledge S.J. Conaway R.C. Harper J.W. Conaway J.W. Science. 1999; 284: 657-661Crossref PubMed Scopus (670) Google Scholar, 10Seol J.H. Feldman R.M. Zachariae W. Shevchenko A. Correll C.C. Lyapina S. Chi Y. Galova M. Claypool J. Sandmeyer S. Nasmyth K. Deshaies R.J. Shevchenko A. Deshaies R.J. Genes Dev. 1999; 13: 1614-1626Crossref PubMed Scopus (360) Google Scholar). Point mutations in the RING finger domain of ROC1 completely disrupted the Ub ligase activity, suggesting an essential role of the domain for its activity (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar). APC11 also contains Ub ligase activity in vitro (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar). More recently, several large RING finger proteins, such as MDM2, c-Cbl, IAP, and AO7, with otherwise diverse structures and functions were linked to ubiquitination (13Honda R. Yasuda H. Oncogene. 2000; 19: 1473-1476Crossref PubMed Scopus (315) Google Scholar, 14Joazeiro C.A. Wing S.S. Huang H. Leverson J.D. Hunter T. Liu Y.C. Science. 1999; 286: 309-312Crossref PubMed Scopus (916) Google Scholar, 15Yang Y. Fang S. Jensen J.P. Weissman A.M. Ashwell J.D. Science. 2000; 288: 874-877Crossref PubMed Scopus (873) Google Scholar, 16Lorick K.L. Jensen J.P. Fang S. Ong A.M. Hatakeyama S. Weissman A.M. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 11364-11369Crossref PubMed Scopus (947) Google Scholar), suggesting a potentially broad and general function for RING fingers in activating Ub ligase activity. One of these RING proteins, BRCA1, has been closely scrutinized for Ub ligase activity. However, the ability of BRCA1 by itself to promote ubiquitin polymerization was found to be limited (16Lorick K.L. Jensen J.P. Fang S. Ong A.M. Hatakeyama S. Weissman A.M. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 11364-11369Crossref PubMed Scopus (947) Google Scholar). In this report, we have provided evidence demonstrating that the RING finger of BRCA1, in concert with BARD1, exhibits significant ubiquitin ligase activity. This activity can be disrupted by a breast cancer-derived RING finger mutation of BRCA1, suggesting a direct relationship between the ubiquitin ligase function of BRCA1 and breast cancer development. cDNAs for full-length human BARD1 and CstF-50 were amplified by polymerase chain reaction from a Hela cell cDNA library with pfu polymerase (Stratagene) and subcloned into the pcDNA3 vector inframe with appropriate tags. pGEX-BARD1-(1–111) was created by digesting the full-length BARD1 with HindIII, and subcloning into the pGEX-KG vector. pET-His6-BARD1-(14–189) and pET-His6-BARD1-(25–189) were created by digesting the full-length BARD1 with BamHI and PstI, andNcoI and PstI, respectively and subcloned into the pET-3E-His6 vector. Full-length BRCA1 cDNA was a generous gift from Dr. Wen-Hwa Lee. The N-terminal fragment (1) of BRCA1 was generated by digesting the full-length BRCA1 at itsKpnI restriction site and subcloning into either the pGEX vector or the pcDNA3 vector with the appropriate tag. pGEX-BRCA1-(1–342) was created by self-ligation ofBglII-digested pGEX-BRCA1-(1–772). pET-His6-BRCA1-(1–304) was created by digesting BRCA1-(1–772) with EcoRI and subcloning into the pET-3E-His6 vector. BRCA1 or BARD1 point mutations were introduced by site-directed mutagenesis (Stratagene). cDNA for each human E2/Ubc was amplified by polymerase chain reaction from a HeLa cell cDNA library and subcloned into the pET-3E-His6vector. cDNA for human E2F1 and cyclin B1 in the pcDNA3 expression vector were gifts from Dr. Yue Xiong. All the constructs used were verified by DNA sequencing. Cells (293T) were cultured in Dulbecco's modified Eagle's medium (Sigma) supplemented with 10#x0025; fetal bovine serum (Life Technologies, Inc.) and 1% antibiotic-antimycotic agent (Life Technologies, Inc.) in a 37 #x00B0;C incubator with 5#x0025; CO2. DNA was transfected using the standard calcium phosphate precipitation method. For each transfection, the total plasmid DNA was adjusted to 15 μg per 100-mm dish by adding the parental pcDNA3 vector. For immunoprecipitation, cells were harvested 36 h after transfection and lysed by incubating at 4 °C for 1 h with 0.6 ml per 100 mm dish of buffer A containing 15 mm Tris-HCl pH 7.5, 0.5m NaCl, 0.35#x0025; Nonidet P-40, 1 mmphenylmethylsulfonyl fluoride, 2 μg/ml aprotinin, 2 μg/ml leupeptin, 10 μg/ml trypsin inhibitor, and 150 μg/ml benzamidine. Lysed cells were then clarified by centrifugation at 100,000 ×g at 4 °C for 1 h. The supernatants (0.3 ml) were mixed with 3 μg of anti-Myc (9E10) or anti-HA (12CA5) antibody, and then the antibody-bound proteins were precipitated with protein A-agarose beads (7.5 μl). The proteins bound to the beads were used either for the Ub ligation assay or immunoblotting. For straight immunoblotting to analyze the steady-state levels of Myc-BRCA1 or HA-BARD1, the transfected cells were lysed and clarified as described above, and 50 μg of each sample was resolved by SDS-PAGE on a 7.5% gel, followed by immunoblotting. Rabbit E1 was purchased from Affiniti Research Products. His6-tagged Ub with a protein kinase C recognition site, His6-tagged E2 proteins, and His6-E2F1 were purified as previously described (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar, 17Ohta T. Xiong Y. Cancer Res. 2001; 61: 1347-1353PubMed Google Scholar). His6-BARD1-(14–189), His6-BARD1-(25–189), His6-BRCA1-(1–304), GST-BRCA1-(1–342) and GST-BARD1-(1–111) were produced in BL21/DE3 bacteria by induction with 0.4 mmIPTG for 12 h at 25 °C. Cells were lysed in buffer containing 50 mm Tris-HCl, pH 8.0, 0.5% NP-40, 1% Triton X-100, 50 mm NaCl, 1 mm DTT, 1 mm EDTA, 10% glycerol, and protease inhibitors, and the proteins were purified either with nickel beads (Qiagen) or glutathione-agarose beads (Sigma) according to the manufacturer's instructions. The procedure for the in vitroUb ligation assay was essentially the same as previously described (7Ohta T. Michel J.J. Schottelius A.J. Xiong Y. Mol. Cell. 1999; 3: 535-541Abstract Full Text Full Text PDF PubMed Scopus (402) Google Scholar,18Ohta T. Michel J.J. Xiong Y. Oncogene. 1999; 18: 6758-6766Crossref PubMed Scopus (44) Google Scholar). The BRCA1-BARD1 immunocomplexes immobilized on protein A-agarose beads were washed three times with buffer A and two times with buffer B containing 25 mm Tris-HCl, pH 7.5, 50 mm NaCl, 0.01% Nonidet P-40, 10% glycerol, and 1 mm EDTA, and added to a ubiquitin ligation reaction mixture (30 μl) that contained 50 mm Tris-HCl, pH 7.4, 5 mm MgCl2, 2 mm NaF, 10 nm okadaic acid, 2 mmATP, 0.6 mm DTT, 0.75 μg of 32P-Ub, 40 ng of E1, and 0.3 μg of E2 protein. After incubation for 30 min at 37 °C with shaking, the reactions were terminated by boiling in Laemmli SDS-loading buffer with 0.1 m DTT, and half of the sample was resolved by SDS-PAGE followed by autoradiography. For the in vitro Ub ligation assay using the purified RING finger domain of BRCA1 or BARD1, GST-fused proteins bound to glutathione-agarose beads, and eluted His6-tagged proteins were used instead of immunocomplexes. The activity of E2 was analyzed by the same procedure without E3. The discovery that the small RING finger proteins, ROC1 and ROC2, are ubiquitin ligases prompted us to determine whether BRCA1 and BARD1, two of the most important RING finger proteins implicated in breast cancer, also function as ubiquitin ligases. We first determined which E2/Ubc could be activated by BRCA1 or BARD1. Five representative mammalian E2s (UbcH1, UbcH2, Cdc34, UbcH5c and UbcH7) were purified from bacteria (Fig.1 A). Individual E2s were incubated with E1 and 32P-Ub in the presence of ATP to verify their Ub binding capacity. Four of the five E2s (UbcH2, Cdc34, UbcH5c, and UbcH7) were determined to be active enzymes by judging the ability to bind to 32P-Ub (Fig. 1 B, lanes 3–6, lower panel), and was dissociated by addition of DTT (lanes 3–6, upper panel). When the anti-Myc immunocomplex derived from Myc-BRCA1-(1–772) or Myc-BARD1-transfected 293T cells was added to the reaction, only UbcH5c among the five E2s was capable of promoting ubiquitin polymerization with the immunocomplex as determined by the appearance of a high molecular weight 32P smear (Fig. 1 C, lanes 4 and 9). Although a crystal structure study revealed the interaction between the RING finger of c-Cbl and UbcH7 (19Zheng N. Wang P. Jeffrey P.D. Pavletich N.P. Cell. 2000; 102: 533-539Abstract Full Text Full Text PDF PubMed Scopus (724) Google Scholar), UbcH7 was deficient in collaborating with the RING finger of BRCA1 or BARD1 to promote Ub polymerization (lanes 5 and 10). These results suggest that the BRCA1- and BARD1-associated protein complexes contain Ub ligase activities. We noted that the Ub ligase activities of the BRCA1 and BARD1 immunocomplexes, which were only detectable after long exposure of the film (Fig. 1 C), were very low and sought conditions for higher activity. Because it was reported that BRCA1 and BARD1 proteins preferentially formed stable heterodimers (6Meza J.E. Brzovic P.S. King M-C. Klevit R.E. J. Biol. Chem. 1999; 274: 5659-5665Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar), we next tested whether the BRCA1-BARD1 heterodimer complex formation affects Ub ligase activity. Surprisingly, when Myc-BRCA1-(1–772) was co-transfected with HA-BARD1, the anti-Myc immunocomplex exhibited significantly higher Ub ligase activity (Fig.2 A, lane 5) than that of either the Myc-BARD1 (lane 3) or Myc-BRCA1-(1–772) (lane 4) single transfection. is that the low Ub ligase activity with the immunocomplexes from single of BRCA1 or BARD1 be by with the RING finger protein. addition of Myc-BRCA1-(1–772) dramatically the Ub ligase activity of the immunocomplex (lane E2F1 and cyclin the proteins to interact with the N terminus of BRCA1 H. N. J. Oncogene. PubMed Scopus Google Scholar), as as a protein that to BARD1 Science. 1999; PubMed Scopus Google Scholar) not have a on Ub ligase activity of BRCA1 or BARD1 (Fig. 2 the that the with BRCA1-BARD1 is mediated by the structure of each protein by its and verified the between these proteins (lane verify the of the RING finger in activity and to the relationship between the BRCA1 function and the Ub ligase activity, we constructs containing BRCA1 with a mutation at P.S. J. King M.C. Klevit R.E. J. Biol. Chem. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). The mutation completely Ub ligase activity (Fig. 2 C, lane 3) when with the anti-Myc immunocomplex precipitated from cells with Myc-BRCA1-(1–772) and (lane and the activity of the immunocomplex to the level of the single transfection (lanes mutation in the RING finger also disrupted the activity to the same as (lanes 4 3 and the and the RING mutations to of and not the Ub ligase activity, a detectable of the activity was (Fig. 2 lanes 3 and the that the ligase activity in the immunocomplex is by proteins and also to determine the of the Ub ligase activity, the RING finger domain of BRCA1 and of BARD1 were purified from bacteria via or A). with the a mixture of the purified RING finger domains of BRCA1 and BARD1 exhibited significant Ub ligase activity (Fig. 3 B, lanes and either the BRCA1 (lanes 1 and 5) or BARD1 (lanes 2 and RING detectable Ub ligase activity. His6-E2F1 not the Ub ligase activity of BRCA1 (lane it interact with GST-BRCA1-(1–342) not RING finger domains comprising residues 1–304 of BRCA1 and residues 25–189 of BARD1 were capable of polymerizing ubiquitin (lane BRCA1C61G, the breast cancer-derived RING finger mutant, not with BARD1 to UbcH5c (lane these results that BARD1 and BRCA1 Ub ligase activity with the E2 The contained at and (Fig. the same as that in the assay (Fig. These are to be ubiquitin or ubiquitin bound to small such as The of the these not from the BRCA1-BARD1 complex when washed with buffer B not is a that these two molecules are ubiquitination substrates of each for We analyzed the steady-state levels of these proteins by Polyubiquitinated substrates are rapidly degraded by the 26 S proteasome the steady-state level of such substrates to However, the steady-state level of Myc-BRCA1-(1–772) was increased after co-transfection of in a A). the steady-state level of was increased when it was with Myc-BRCA1-(1–772) (Fig. These results that BRCA1 and BARD1 each the ability to the polyubiquitin suggesting that are not substrates by each for The RING finger motif was to be a DNA binding site when BRCA1 was first identified as a for breast and ovarian cancer, it was to the a DNA binding motif (1Zheng L. Li S. Boyer T.G. Lee W.H. Oncogene. 2000; 19: 6159-6175Crossref PubMed Scopus (136) Google Scholar). the of the domain in that BRCA1 is a and the RING finger in DNA However, binding between the RING finger of BRCA1 and DNA has not been the RING finger of BRCA1 has been to be important for a interaction with RING finger which was identified and the BARD1 (5Wu L.C. Wang Z.W. Tsan J.T. Spillman M.A. Phung A. Xu X.L. Yang M.C. Hwang L.Y. Bowcock A.M. Baer R. Nat. Genet. 1996; 14: 430-440Crossref PubMed Scopus (626) Google Scholar). A biochemical that the interaction between BRCA1 and BARD1 was mediated by regions the RING finger motif (6Meza J.E. Brzovic P.S. King M-C. Klevit R.E. J. Biol. Chem. 1999; 274: 5659-5665Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar), suggesting that the two RING fingers contained function the of the two was In the report, we evidence demonstrating that the RING heterodimer BRCA1-BARD1 functions as a Ub the BRCA1-BARD1 immunocomplex from BRCA1- and cells contains a significant ability to promote vitro, and of either of the two from the transfection this activity. the purified RING finger domains of BRCA1 and BARD1 exhibit high Ub ligase activity. of either of the two RING fingers this activity. in a breast cancer-derived RING finger mutation of BRCA1, the ability to promote Although it is that the major reaction in the study is BRCA1 and BARD1 are not substrates by each for each in A of RING finger ubiquitin ligases is R. Yasuda H. Oncogene. 2000; 19: 1473-1476Crossref PubMed Scopus (315) Google Scholar, 14Joazeiro C.A. Wing S.S. Huang H. Leverson J.D. Hunter T. Liu Y.C. Science. 1999; 286: 309-312Crossref PubMed Scopus (916) Google Scholar, 15Yang Y. Fang S. Jensen J.P. Weissman A.M. Ashwell J.D. Science. 2000; 288: 874-877Crossref PubMed Scopus (873) Google Scholar, 16Lorick K.L. Jensen J.P. Fang S. Ong A.M. Hatakeyama S. Weissman A.M. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 11364-11369Crossref PubMed Scopus (947) Google and T. Michel J.J. Xiong Y. Oncogene. 1999; 18: 6758-6766Crossref PubMed Scopus (44) Google Scholar). The of in the function of RING ubiquitin ligases to be BRCA1-BARD1 a of RING containing Ub ligase that a heterodimeric RING finger However, the of RING finger molecules in complexes of RING Ub ligases has not been to be determined whether either as or between two RING fingers a general for activating Ub ligase activity. this can be in immunocomplexes precipitated from cells transfected with and the S of the cell the steady-state levels of BRCA1 a and BARD1 with BRCA1 into Y. Xu X.L. Yang M.C. Bowcock A.M. Baer R. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). it is that an in vivo Ub ligase activity of BRCA1-BARD1 a in S the formation of the heterodimer is a in the of BRCA1 and in vivo Ub ligase activity to be the function of the BRCA1-BARD1 Ub the substrates for is proteins that are degraded or in S be substrates for the BRCA1-BARD1 Ub The high Ub ligase activity of BRCA1-BARD1 The that the two RING finger domains of BRCA1 and BARD1 are for biochemical function that the or the by mutations in the domains of two molecules be the mutations in in breast and ovarian cancers have been reported Tsan J.T. Y. Phung A. Spillman M.A. R. J.M. Baer R. Bowcock A.M. Mol. Genet. 1998; PubMed Scopus Google Scholar). BARD1 is potentially as important a as BRCA1 in of breast and ovarian each mutation of the two molecules and vitro Ub ligase activity, it be to the for breast and ovarian cancer in with point mutations in the RING finger domains of BRCA1 or BARD1. the that the BRCA1-BARD1 heterodimer complex as a Ub ligase a to into the of BRCA1 and BARD1 in We Dr. Yue Xiong and W. for and of the Dr. Lee for BRCA1 We also for

Colorectal cancer is a major cause of death in Japan, where it accounts for the largest number of deaths from malignant neoplasms among women and the third largest number among men. Many new methods of treatment have been developed during recent decades. The Japanese Society for Cancer of the Colon and Rectum Guidelines 2014 for treatment of colorectal cancer (JSCCR Guidelines 2014) have been prepared as standard treatment strategies for colorectal cancer, to eliminate treatment disparities among institutions, to eliminate unnecessary treatment and insufficient treatment, and to deepen mutual understanding among health-care professionals and patients by making these guidelines available to the general public. These guidelines have been prepared as a result of consensuses reached by the JSCCR Guideline Committee on the basis of careful review of evidence retrieved by literature searches and taking into consideration the medical health insurance system and actual clinical practice in Japan. They can, therefore, be used as a guide for treating colorectal cancer in clinical practice. More specifically, they can be used as a guide to obtaining informed consent from patients and choosing the method of treatment for each patient. As a result of the discussions of the Guideline Committee, controversial issues were selected as clinical questions, and recommendations were made. Each recommendation is accompanied by a classification of the evidence and a classification of recommendation categories, on the basis of consensus reached by Guideline Committee members. Here we present the English version of the JSCCR Guidelines 2014.

Longevity regulatory genes include the Forkhead transcription factor FOXO and the NAD-dependent histone deacetylase silent information regulator 2 (Sir2). Genetic studies demonstrate that Sir2 acts to extend lifespan in Caenorhabditis elegans upstream of DAF-16, a member of the FOXO family, in the insulin-like signaling pathway. However, the molecular mechanisms underlying the requirement of DAF-16 activity in Sir2-mediated longevity remain unknown. Here we show that reversible acetylation of Foxo1 (also known as FKHR), the mouse DAF-16 ortholog, modulates its transactivation function. cAMP-response element-binding protein (CREB)-binding protein binds and acetylates Foxo1 at the K242, K245, and K262 residues, the modification of which is involved in the attenuation of Foxo1 as a transcription factor. Conversely, Sir2 binds and deacetylates Foxo1 at residues acetylated by cAMP-response element-binding protein-binding protein. Sir2 is recruited to insulin response sequence-containing promoter and increases the expression of manganese superoxide dismutase and p27(kip1) in a deacetylase-activity-dependent manner. Our findings establish Foxo1 as a direct and functional target for Sir2 in mammalian systems.

Quality adjustment weights for quality-adjusted life years (QALYs) are available with the EQ-5D Instrument, which are based on a survey that quantified the preferences of the British public. However, the extent to which this British value set is applicable to other, especially non-European, countries is yet unclear. The objectives of this study are (a) to compare the valuations obtained in Japan and Britain, and (b) to explore a local Japanese value set. A diminished study design is employed, where 17 hypothetical EQ-5D health states are evaluated as opposed to 42 in the British study. The official Japanese version of the instrument and the Time Trade-Off method are used to interview 543 members of the public. The results are: firstly, the evaluations obtained in Japan and those from Britain differ by 0.24 on average on a [-1, +1] scale, and mean absolute error (MAE) in predicting the Japanese preferences with the British value set is 0.23. Secondly, comparable regressions suggest that the two peoples have systematically different preference structures (p<0.001 for 8 of 12 coefficients; F-test). Thirdly, using alternative models, the predictions are improved so that the local Japanese value set achieves MAE in the order of 0.01.

Despite major improvements in outcome over the past decades, acute myeloid leukemia (AML) remains a life-threatening malignancy in children, with current survival rates of ∼70%. State-of-the-art recommendations in adult AML have recently been published in this journal by Döhner et al. The primary goal of an international expert panel of the International BFM Study Group AML Committee was to set standards for the management, diagnosis, response assessment, and treatment in childhood AML. This paper aims to discuss differences between childhood and adult AML, and to highlight recommendations that are specific to children. The particular relevance of new diagnostic and prognostic molecular markers in pediatric AML is presented. The general management of pediatric AML, the management of specific pediatric AML cohorts (such as infants) or subtypes of the disease occurring in children (such as Down syndrome related AML), as well as new therapeutic approaches, and the role of supportive care are discussed.