Neurofibromatosis 1 (NF1) mutation results in impaired function of human induced pluripotent stem cell-derived microglia

Neurofibromatosis type 1 (NF1) is an autosomal dominant condition caused by germline mutations in the NF1 gene. Children with NF1 are prone to the development of multiple nervous system abnormalities, including autism and brain tumors, which could reflect the effect of NF1 mutation on microglia function. Using heterozygous Nf1-mutant mice, we previously demonstrated that impaired purinergic signaling underlies deficits in microglia process extension and phagocytosis in situ. To determine whether these abnormalities are also observed in human microglia in the setting of NF1, we leveraged an engineered isogenic series of human induced pluripotent stem cells to generate human microglia-like (hiMGL) cells heterozygous for three different NF1 patient-derived NF1 gene mutations. While all NF1-mutant and isogenic control hiMGL cells expressed classical microglia markers and exhibited similar transcriptomes and cytokine/chemokine release profiles, only NF1-mutant hiMGL cells had defects in P2X receptor activation, phagocytosis and motility. Taken together, heterozygous NF1 mutation impairs a subset of human microglia functional properties, which could contribute to the neurological abnormalities seen in children with NF1

Terminal transferase expression in relapsed acute myeloid leukaemia

In three cases of acute myeloid leukaemia marked increases in expression of the nuclear enzyme, terminal deoxynucleotidyl transferase (TdT), were observed during disease relapse. The first case was heterogeneous at diagnosis, consisting of subpopulations of large TdT- myeloblasts and small TDT+ blasts; however, at relapse there was complete replacement by TdT+ lymphoblasts. The other cases at diagnosis were both typical acute myeloid leukaemia, TdT-; at relapse, one showed a mixture of TdT- myeloblasts and TdT+ lymphoblasts, while in the other, TdT was demonstrated on a subpopulation of myeloblasts. No chromosomal abnormalities were found. It is suggested that in the first two cases these phenomena may have been due to leukaemic involvement of a pluripotential stem cell, or that sub-clones with different properties may have coexisted. In the third case at relapse, TdT was expressed aberrantly by malignant myeloid cells

Standardized practices for RNA diagnostics using clinically accessible specimens reclassifies 75% of putative splicing variants

Purpose: Genetic variants causing aberrant premessenger RNA splicing are increasingly being recognized as causal variants in genetic disorders. In this study, we devise standardized practices for polymerase chain reaction (PCR)-based RNA diagnostics using clinically accessible specimens (blood, fibroblasts, urothelia, biopsy). Methods: A total of 74 families with diverse monogenic conditions (31% prenatal-congenital onset, 47% early childhood, and 22% teenage-adult onset) were triaged into PCR-based RNA testing, with comparative RNA sequencing for 19 cases. Results: Informative RNA assay data were obtained for 96% of cases, enabling variant reclassification for 75% variants that can be used for genetic counseling (71%), to inform clinical care (32%) and prenatal counseling (41%). Variant-associated mis-splicing was highly reproducible for 28 cases with samples from ≥2 affected individuals or heterozygotes and 10 cases with ≥2 biospecimens. PCR amplicons encompassing another segregated heterozygous variant was vital for clinical interpretation of 22 of 79 variants to phase RNA splicing events and discern complete from partial mis-splicing. Conclusion: RNA diagnostics enabled provision of a genetic diagnosis for 64% of recruited cases. PCR-based RNA diagnostics has capacity to analyze 81.3% of clinically significant genes, with long amplicons providing an advantage over RNA sequencing to phase RNA splicing events. The Australasian Consortium for RNA Diagnostics (SpliceACORD) provide clinically-endorsed, standardized protocols and recommendations for interpreting RNA assay data.Field, Ron Fleischer, Chiara Folland, Lucy Fox, Mary- Louise Freckmann, Clara Gaff, Melanie Galea, Roula Ghaoui, Himanshu Goel, Ilias Gornanitis, Thuong Ha, Bernadette Hanna, James Harraway, Rippei Hayashi, Ian Hayes, Alex Henderson, Luke Hesson, Erin Heyer, Michael Hildebrand, Michael Hipwell, Gladys Ho, Ari E. Horton, Cass Hoskins, Matthew F. Hunter, Matilda Jackson, Paul James, Kristi J. Jones, Justin Jong-Leong Wong, Sarah Josephi-Taylor, Himanshu Joshi, Karin Kassahn, Peter Kaub, Lucy Kevin, Edwin Kirk, Emma Krzesinski, Smitha Kumble, Sarah Kummerfeld, Nigel Laing, Chiyan Lau, Eric Lee, Sarah Leighton, Ben Lundie, Sebastian Lunke, Amali Mallawaarachchi, Chelsea Mayoh, Julie McGaughran, Ali- son McLean, Mary McPhillips, Cliff Meldrum, Edwina Middleton, Di Milnes, Kym Mina, David Mowat, Amy Nisselle, Emily Oates, Alicia Oshlack, Elizabeth E. Palmer, Gayathri Parasivam, Michael Parsons, Chirag Patel, Jason R. Pinner, Michael Quinn, John Rasko, Gina Ravenscroft, Anja Ravine, Krista Recsei, Matthew Regan, Jacqueline Rehn, Lisa G. Riley, Stephen Robertson, Anne Ronan, Tony Roscioli, Georgina Ryland, Simon Sadedin, Sarah A. San- daradura, Andreas Schreiber, Hamish Scott, Rodney Scott, Christopher Semsarian, Cas Simons, Emma Singer, Janine M. Smith, Renee Smyth, Amanda Spurdle, Zornitza Stark, Patricia Sullivan, Samantha Sundercombe, Tiong Y. Tan, Michel C. Tchan, Bryony A. Thompson, David Thorburn, John Toubia, Ronald Trent, Emma Tudini, Irina Voneague, Leigh Waddell, Logan Walker, Mathew Wallis, Nick War- nock, Robert Weatheritt, Deborah White, Susan M. White, Mark G. Williams, Meredith J. Wilson, Ingrid Winship, Lisa Worgan, Dale C. Wright, Kathy Wu, Alison Yeung, Andrew Ziolowski. Additional Information The online version of this article, https, doi.org, ., j. gim., ., ., contains supplementary material, which is available to authorized users. Authors Adam M. Bournazos, Lisa G. Riley, Shobhana Bommireddipalli, Lesley Ades, Lauren S. Akesson, Mohammad Al-Shinnag, Stephen I. Alexander, Alison D. Archibald, Shanti Balasubramaniam, Yemima Berman, Victoria Beshay, Kirsten Boggs, Jasmina Bojadzieva, Natasha J. Brown, Samantha J. Bryen, Michael F. Buckley, Belinda Chong, Mark R. Davis, Ruebena Dawes, Martin Delatycki, Liz Donaldson, Lilian Downie, Caitlin Edwards, Matthew Edwards, Amanda Engel, Lisa J. Ewans, Fathimath Faiz, Andrew Fennell, Michael Field, Mary-Louise Freckmann, Lyndon Gallacher, Russell Gear, Himanshu Goel, Shuxiang Goh, Linda Goodwin, Bernadette Hanna, James Harraway, Megan Higgins, Gladys Ho, Bruce K. Hopper, Ari E. Horton, Matthew F. Hunter, Aamira J. Huq, Sarah Josephi-Taylor, Himanshu Joshi, Edwin Kirk, Emma Krzesinski, Kishore R. Kumar, Frances Lemckert, Richard J. Leventer, Suzanna E. Lindsey-Temple, Sebastian Lunke, Alan Ma, Steven Macaskill, Amali Mallawaarachchi, Melanie Marty, Justine E. Marum, Hugh J. McCarthy, Manoj P. Menezes, Alison McLean, Di Milnes, Shekeeb Mohammad, David Mowat, Aram Niaz, Elizabeth E. Palmer, Chirag Patel, Shilpan G. Patel, Dean Phelan, Jason R. Pinner, Sulekha Rajagopalan, Matthew Regan, Jonathan Rodgers, Miriam Rodrigues, Richard H. Roxburgh, Rani Sachdev, Tony Roscioli, Ruvishani Samarasekera, Sarah A. Sandaradura, Elena Savva, Tim Schindler, Margit Shah, Ingrid B. Sinnerbrink, Janine M. Smith, Richard J. Smith, Amanda Springer, Zornitza Stark, Samuel P. Strom, Carolyn M. Sue, Kenneth Tan, Tiong Y. Tan, Esther Tantsis, Michel C. Tchan, Bryony A. Thompson, Alison H. Trainer, Karin van Spaendonck-Zwarts, Rebecca Walsh, Linda Warwick, Stephanie White, Susan M. White, Mark G. Williams, Meredith J. Wilson, Wui Kwan Wong, Dale C. Wright, Patrick Yap, Alison Yeung, Helen Young, Kristi J. Jones, Bruce Bennetts, Sandra T. Cooper, on behalf of the Australasian Consortium for RNA Diagnostic

Silagem de milho e grão de sorgo como suplementos para vacas de descarte terminadas em pastagem cultivada de estação fria Corn silage and sorghum grain supplementation to cull cows finished on cultivated winter pasture

Avaliou-se o efeito da suplementação com silagem de milho ou grão de sorgo no desempenho e características da carcaça e da carne de vacas de descarte, submetidas ao pastejo restrito em pastagem de aveia (Avena strigosa) + azevém (Lolium multiflorum). Foram utilizadas 30 vacas mestiças Charolês-Nelore, com idade média de oito anos, distribuídas em igual número e ao acaso em três tratamentos: silagem de milho (TSI) ou grão de sorgo moído (TSO) como suplemento, e não suplementação (TPH). A suplementação com silagem de milho proporcionou aos animais maior escore corporal ao final do experimento (4,35 pontos contra 4,15 do TPH e 4,22 pontos para o TSO). Não houve efeito do volumoso ou concentrado suplementar sobre o peso corporal, ganho de peso diário e ganho em escore corporal. Não houve efeito de tratamento (P>0,05) sobre pesos e rendimentos quentes e frios da carcaça, espessura de gordura subcutânea, conformação, espessura de coxão, área de Longissimus dorsi e percentagem de cortes comerciais. A suplementação resultou em maior percentagem de gordura na carcaça e influenciou a cor e o marmoreio da carne. A silagem de milho e o grão de sorgo servem como alternativa para a suplementação de vacas de descarte em pastagem de aveia e azevém, uma vez que melhora as características da carcaça, embora sem efeito sobre o desempenho quando comparado à pastagem exclusiva.<br>The effect of supplementation with corn silage or sorghum grain on the performance and characteristics of carcass and meat of cull cows, under temporary grazing on oats (Avena strigosa) + ryegrass (Lolium multiflorum) pasture was evaluated. Thirty crossbred Charolais-Nellore cull cows, with averaging eight-yearold,were randomly distributed in three treatments. The animals received corn silage (TSI) or sorghum grain (TSO) as supplement, while another lot did not receive supplementation (TPH). Animals supplemented with corn silage showed higher final body condition score (4.35 points versus 4.15 for TPH and 4.22 for TSO). No effect of supplementation on body weight, average daily weight gain; and body condition score gain were verified. Differences were not observed concerning carcasses weight, hot and cold yield of carcasses, fat thickness, conformation, cushion thickness, Longissimus dorsi area, and commercial cuts percentages. Supplemented animals showed better color and higher marbling of meat. Corn silage and sorghum grain are alternative for the supplementation of cull cows in oats and ryegrass pasture, since they showed better carcass characteristics, although no effect on the performance in relation to exclusive pasture was detected