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Sera from C3H mammary tumor-bearing mice contain cytotoxic antibodies for mouse mammary tumor virus (MMTV)-producing cells, based on 51Cr release in a complement-dependent serum cytotoxicity assay. The cytotoxic antibodies could be absorbed by purified C3H MMTV gp52 and C3H MMTV-infected cat cells (C3H [MMTV] CrFK) containing cell surface MMTV gp52. However, purified MMTV p27 and uninfected CrFK cat cells were negative. Absorption of the sera with GR (MMTV) CrFK cells also removed all of the cytotoxicity, whereas absorption with RIII (MMTV) CrFK cells was negative, even though all three infected cat cells contained equivalent amounts of gp52. The same C3H cytotoxic sera also neutralized the focus-forming capacity of a C3H MMTV pseudotype of Kirsten sarcoma virus containing MMTV gp52. In contrast, sera from mammary tumor-bearing GR and RIII mice did not neutralize the pseudotype. Furthermore, neutralization could be achieved only by anti-gp52 but not by anti-gp36, -p27, -p14, or -p10 C3H MMTV sera. The gp52’s of C3H, GR, and RIII MMTV could also be distinguished by using a type-specific competition radioimmunoassay employing 125I-gp52 of C3H MMTV and a hyperimmune rabbit anti-C3H MMTV serum. To demonstrate these differences directly, we studied the primary structure of gp52 on the surface of the C3H, GR, and RIII (MMTV) CrFK cells. Two-dimensional tryptic peptide maps of the cell surface lactoper-oxidase-catalyzed iodinated gp52’s revealed a greater number of peptides common to the gp52’s of C3H and GR MMTVs than to RIII MMTV gp52. These results demonstrate that gp52 is a major target antigen for both cytotoxic and neutralizing antibodies, that the cell surface and virion-associated gp52’s of C3H, GR, and RIII MMTV contain both group- and type-specific determinants, and that C3H and GR MMTV gp52’s are antigenically more related to each other than to RIII MMTV gp52. Furthermore, C3H mammary tumor-bearing mice develop type-specific antibodies capable of recognizing unique gp52 determinants and, therefore, are able to distinguish the gp52 of C3H MMTV from the gp52’s of GR and RIII MMTV.
Mice with Spontaneous Mammary Tumors Develop Type-Specific Neutralizing and Cytotoxic Antibodies Against the Mouse Mammary Tumor Virus Envelope Protein gp52
Gerald Schochetman, Larry O. Arthur, Cedric W. Long, and Richard J. Massey
The primary aim of this study was to develop an assessment of the fundamental, combined, and complex movement skills required to support childhood physical literacy. The secondary aim was to establish the feasibility, objectivity, and reliability evidence for the assessment.
An expert advisory group recommended a course format for the assessment that would require children to complete a series of dynamic movement skills. Criterion-referenced skill performance and completion time were the recommended forms of evaluation. Children, 8-12 years of age, self-reported their age and gender and then completed the study assessments while attending local schools or day camps. Face validity was previously established through a Delphi expert (n = 19, 21% female) review process. Convergent validity was evaluated by age and gender associations with assessment performance. Inter- and intra-rater (n = 53, 34% female) objectivity and test-retest (n = 60, 47% female) reliability were assessed through repeated test administration.
Median total score was 21 of 28 points (range 5-28). Median completion time was 17 s. Total scores were feasible for all 995 children who self-reported age and gender. Total score did not differ between inside and outside environments (95% confidence interval (CI) of difference: −0.7 to 0.6; p = 0.91) or with/without footwear (95%CI of difference: −2.5 to 1.9; p = 0.77). Older age (p < 0.001, η2 = 0.15) and male gender (p < 0.001, η2 = 0.02) were associated with a higher total score. Inter-rater objectivity evidence was excellent (intraclass correlation coefficient (ICC) = 0.99) for completion time and substantial for skill score (ICC = 0.69) for 104 attempts by 53 children (34% female). Intra-rater objectivity was moderate (ICC = 0.52) for skill score and excellent for completion time (ICC = 0.99). Reliability was excellent for completion time over a short (2-4 days; ICC = 0.84) or long (8-14 days; ICC = 0.82) interval. Skill score reliability was moderate (ICC = 0.46) over a short interval, and substantial (ICC = 0.74) over a long interval. Conclusion The Canadian Agility and Movement Skill Assessment is a feasible measure of selected fundamental, complex and combined movement skills, which are an important building block for childhood physical literacy. Moderate-to-excellent objectivity was demonstrated for children 8-12 years of age. Test-retest reliability has been established over an interval of at least 1 week. The time and skill scores can be accurately estimated by 1 trained examiner.
Agility course, Children, Dynamic motor skill, Locomotor skill, Object manipulation, Population assessment
Canadian Agility and Movement Skill Assessment (CAMSA): Validity, objectivity, and reliability evidence for children 8-12 years of age
Patricia E. Longmuir,a,b,* Charles Boyer,a Meghann Lloyd,c Michael M. Borghese,a,d Emily Knight,a Travis J. Saunders,a,e Elena Boiarskaia,f Weimo Zhu,f and Mark S. Tremblaya,b
The advent of comprehensive genomic profiling has markedly advanced the understanding of the biology of pediatric hematological malignancies, however, its application to clinical care is still unclear. We present our experience integrating genomic data into the clinical management of children with high-risk hematologic malignancies and blood disorders and describe the broad impact that genomic profiling has in multiple aspects of patient care.
The Precision in Pediatric Sequencing Program at Columbia University Medical Center instituted prospective clinical next-generation sequencing (NGS) for high-risk malignancies and blood disorders. Testing included cancer whole exome sequencing (WES) of matched tumor-normal samples or targeted sequencing of 467 cancer-associated genes, when sample adequacy was a concern, and tumor transcriptome (RNA-seq). A multidisciplinary molecular tumor board conducted interpretation of results and final tiered reports were transmitted to the electronic medical record according to patient preferences.
Sixty-nine samples from 56 patients with high-risk hematologic malignancies and blood disorders were sequenced. Patients carried diagnoses of myeloid malignancy (n = 25), lymphoid malignancy (n = 25), or histiocytic disorder (n = 6). Six patients had only constitutional WES, performed for a suspicion of an inherited predisposition for their disease. For the remaining 50 patients, tumor was sequenced with matched normal tissue when available. The mean number of somatic variants per sample was low across the different disease categories (2.85 variants/sample). Interestingly, a gene fusion was identified by RNA-seq in 58% of samples who had adequate RNA available for testing. Molecular profiling of tumor tissue led to clinically impactful findings in 90% of patients. Forty patients (80%) had at least one targetable gene variant or fusion identified in their tumor tissue; however, only seven received targeted therapy. Importantly, NGS findings contributed to the refinement of diagnosis and prognosis for 34% of patients. Known or likely pathogenic germline alterations were discovered in 24% of patients involving cancer predisposition genes in 12% of cases.
Incorporating whole exome and transcriptome profiling of tumor and normal tissue into clinical practice is feasible, and the value that comprehensive testing provides extends beyond the ability to target-specific mutations.
genomic, hematologic malignancies, pediatric leukemia, next-generation sequencing, targeted therapy
Precision Medicine in Children and Young Adults with Hematologic Malignancies and Blood Disorders: The Columbia University Experience
Lianna J. Marks,1,† Jennifer A. Oberg,2,† Danielle Pendrick,2 Anthony N. Sireci,3 Chana Glasser,4 Carrie Coval,5 Rebecca J. Zylber,2 Wendy K. Chung,5,6 Jiuhong Pang,3 Andrew T. Turk,3 Susan J. Hsiao,3 Mahesh M. Mansukhani,3,6 Julia L. Glade Bender,2,6 Andrew L. Kung,1 and Maria Luisa Sulis2,6,*