The epigenetic regulators CBP and p300 facilitate leukemogenesis and represent therapeutic targets in acute myeloid leukemia.

Growing evidence links abnormal epigenetic control to the development of hematological malignancies. Accordingly, inhibition of epigenetic regulators is emerging as a promising therapeutic strategy. The acetylation status of lysine residues in histone tails is one of a number of epigenetic post-translational modifications that alter DNA-templated processes, such as transcription, to facilitate malignant transformation. Although histone deacetylases are already being clinically targeted, the role of histone lysine acetyltransferases (KAT) in malignancy is less well characterized. We chose to study this question in the context of acute myeloid leukemia (AML), where, using in vitro and in vivo genetic ablation and knockdown experiments in murine models, we demonstrate a role for the epigenetic regulators CBP and p300 in the induction and maintenance of AML. Furthermore, using selective small molecule inhibitors of their lysine acetyltransferase activity, we validate CBP/p300 as therapeutic targets in vitro across a wide range of human AML subtypes. We proceed to show that growth retardation occurs through the induction of transcriptional changes that induce apoptosis and cell-cycle arrest in leukemia cells and finally demonstrate the efficacy of the KAT inhibitors in decreasing clonogenic growth of primary AML patient samples. Taken together, these data suggest that CBP/p300 are promising therapeutic targets across multiple subtypes in AML.

Recurrent mutations, including NPM1c, activate a BRD4-dependent core transcriptional program in acute myeloid leukemia.

Recent evidence suggests that inhibition of bromodomain and extra-terminal (BET) epigenetic readers may have clinical utility against acute myeloid leukemia (AML). Here we validate this hypothesis, demonstrating the efficacy of the BET inhibitor I-BET151 across a variety of AML subtypes driven by disparate mutations. We demonstrate that a common 'core' transcriptional program, which is HOX gene independent, is downregulated in AML and underlies sensitivity to I-BET treatment. This program is enriched for genes that contain 'super-enhancers', recently described regulatory elements postulated to control key oncogenic driver genes. Moreover, our program can independently classify AML patients into distinct cytogenetic and molecular subgroups, suggesting that it contains biomarkers of sensitivity and response. We focus AML with mutations of the Nucleophosmin gene (NPM1) and show evidence to suggest that wild-type NPM1 has an inhibitory influence on BRD4 that is relieved upon NPM1c mutation and cytosplasmic dislocation. This leads to the upregulation of the core transcriptional program facilitating leukemia development. This program is abrogated by I-BET therapy and by nuclear restoration of NPM1. Finally, we demonstrate the efficacy of I-BET151 in a unique murine model and in primary patient samples of NPM1c AML. Taken together, our data support the use of BET inhibitors in clinical trials in AML.

Exercise intervention for a musculoskeletal disorder in an oral health student: a case report.

OBJECTIVES: to describe and evaluate the effect of an exercise self-management programme for an oral health student who was suffering from a musculoskeletal disorder which had developed in the clinical environment during the course of training. DESIGN: Single participant case report. SETTING: The University of Otago Faculty of Dentistry and School of Physiotherapy. METHODS: The participant was a 21-year-old female student, enrolled in the University of Otago Bachelor of Oral Health (BOH) course, who was suffering back and neck pain which had developed during her training. A series of chairside stretches, regular home-based stretches, aerobic conditioning and postural strengthening exercises was prescribed. Pain and function was assessed using standardised questionnaires at the commencement of the exercise programme, and again at 6 weeks and 13 weeks. RESULTS: The participant demonstrated an improvement in her outcome questionnaires exceeding the MCID scores after 13 weeks. CONCLUSIONS: This case report provides a basis for highlighting the importance of measuring and monitoring MSD in a student learning oral health skills; it supports the findings of previous studies showing that an exercise intervention has some benefit for MSD in oral health students.

The AAA+ ATPase RUVBL2 is a critical mediator of MLL-AF9 oncogenesis.

The most frequent chromosomal translocations in pediatric acute myeloid leukemia affect the 11q23 locus and give rise to mixed lineage leukemia (MLL) fusion genes, MLL-AF9 being the most prevalent. The MLL-AF9 fusion gene has been shown to induce leukemia in both mouse and human models. In this study, we demonstrate that leukemogenic activity of MLL-AF9 requires RUVBL2 (RuvB-like 2), an AAA+ ATPase family member that functions in a wide range of cellular processes, including chromatin remodeling and transcriptional regulation. Expression of RUVBL2 was dependent on MLL-AF9, as it increased upon immortalization of human cord blood-derived hematopoietic progenitor cells with the fusion gene and decreased following loss of fusion gene expression in conditionally immortalized mouse cells. Short hairpin RNA-mediated silencing experiments demonstrated that both the immortalized human cells and the MLL-AF9-expressing human leukemia cell line THP-1 required RUVBL2 expression for proliferation and survival. Furthermore, inhibition of RUVBL2 expression in THP-1 cells led to reduced telomerase activity and clonogenic potential. These data were confirmed with a dominant-negative Walker B-mutated RUVBL2 construct. Taken together, these data suggest the possibility of targeting RUVBL2 as a potential therapeutic strategy for MLL-AF9-associated leukemia.

MLL-AF9-mediated immortalization of human hematopoietic cells along different lineages changes during ontogeny.

The MLL-AF9 fusion gene is associated with aggressive leukemias of both the myeloid and lymphoid lineage in infants, whereas in adults, this translocation is mainly associated with acute myeloid leukemia. These observations suggest that differences exist between fetal and adult tissues in terms of the 'cell of origin' from which the leukemia develops. Here we show that depending on extrinsic cues, human neonatal CD34(+) cells are readily immortalized along either the myeloid or lymphoid lineage upon MLL-AF9 expression and give rise to mainly lymphoid leukemia in immunocompromised mice. In contrast, immortalization of adult bone marrow CD34(+) cells is more difficult to achieve and is myeloid-biased, even when MLL-AF9 is expressed in purified hematopoietic stem cells (HSCs). Transcriptome analysis identified enrichment of HSC but not progenitor gene signatures in MLL-AF9-expressing cells. Although not observed in adult cells, neonatal cells expressing MLL-AF9 were enriched for gene signatures associated with poor prognosis, resistance to chemotherapeutic agents and MYC signaling. These results indicate that neonatal cells are inherently more prone to MLL-AF9-mediated immortalization than adult cells and suggest that intrinsic properties of the cell of origin, in addition to extrinsic cues, dictate lineage of the immortalized cell.

Clinical working postures of bachelor of oral health students.

OBJECTIVE: To observe and describe the clinical working postures of final-year Bachelor of Oral Health (BOH) students. DESIGN: Pilot observational study. SETTING: The University of Otago Faculty of Dentistry and School of Physiotherapy. METHODS: Eight final-year BOH students voluntarily participated in this study, where postural data were collected using a digital video camera during a standard clinical treatment session. The postural data were analysed using 3D Match biomechanical software. RESULTS: Final-year BOH students who work in the seated position are exposed to neck flexion of greater than 35 degrees, together with trunk flexion greater than 20 degrees and bilateral elbow flexion greater than 90 degrees. CONCLUSIONS: The findings of this study agree with the findings of previous postural studies of dental professionals. Dental hygiene students, together with their clinical supervisors, need to be aware of the importance of good working posture early in their careers, and pay particular attention to the degree of neck flexion occurring for prolonged periods.

Recurring enteric peritonitis associated with non-perforating colon carcinoma.

Peritonitis of enteric origin may occur during treatment with peritoneal dialysis due to visceral perforation or injury or, in the absence of perforation, due to transmural migration of enteric bacteria across the bowel wall into the peritoneal cavity. To the best of our knowledge, peritonitis has not previously been reported associated with carcinomatous colon polyp in the absence of bowel wall perforation. We describe the case of a 31-year-old female who experienced recurring episodes of enteric peritonitis associated with a clinically occult adenocarcinoma of the colon, without having any other known risk factors for peritonitis. A 15 mm carcinomatous polyp was not visible on CT scan but was found at colonoscopy with polypectomy. She proceeded to transverse colectomy; the resected colon showed no evidence of bowel wall perforation. This case demonstrates that a non-perforating carcinomatous polyp of the colon may predispose to enteric peritonitis in the setting of peritoneal dialysis, and it emphasizes the importance of making an aggressive search for underlying pathology, in patients who present with recurring enteric peritonitis or unusual presentations of enteric peritonitis.

Exploring the idiotypes of insulin antibodies as markers for remission in Type 1 diabetes.

AIMS: Complete or partial remission can occur in newly diagnosed Type 1 diabetes patients. We created idiotype-specific reagents to explore the idiotypes of insulin antibodies (IA) in a patient in remission, and to compare with a patient who was not. METHODS: Phage display was used to create a library of phagotopes specific to insulin binding in four sera. Sera from a Type 1 diabetes subject deemed to have undergone remission were taken at diagnosis and again during remission. Sera from a non-remitter were taken at diagnosis and after 3 months on insulin. Phagotopes from the four sera were randomly selected and tested for insulin specificity in a radiobinding assay by using sera from remitters and non-remitters. RESULTS: IA-binding phagotope selected from serum during remission displaced insulin binding in all nine IA(+) remitters and all 10 IA(+) non-remitters. IA-binding phagotope selected from the non-remission patient (3 months after insulin therapy) displaced insulin binding in 8/9 IA(+) remitters and 8/10 IA(+) non-remitters. The consensus peptide sequences adduced from the phages were identical for both these phagotopes. Phagotopes derived from insulin autoantibody-positive individuals at diagnosis were unable to displace insulin binding in the IA(+) sera 3 months later, whether in remission or not. CONCLUSIONS: We have established the principle of using phage display in the investigation of insulin antibodies during remission in Type 1 diabetes. The immunological characteristics of IA 3 months after the introduction of insulin treatment were different from those at diagnosis of Type 1 diabetes (IAA). Using phage display technology, it was not possible to distinguish insulin antibodies according to remission status.

The use of phage display to distinguish insulin autoantibody (IAA) from insulin antibody (IA) idiotypes.

AIM/HYPOTHESIS: Radiobinding assays (RBA) are unable to differentiate insulin autoantibodies (IAA) from insulin antibodies (IA). We sought to establish whether random peptide phage display might generate reagents with which to distinguish IAA idiotopes from IA idiotopes. METHODS: Two insulin-binding sera were used to select phagotopes from a phage library. The first, designated IAS, came from an insulin-treated patient with the insulin autoimmune syndrome, and was known to contain both IA and a high titre of human insulin specific (B30 threonine dependent) IAA. The second, designated IDD, was taken from a newly-diagnosed IAA(+) Type 1 diabetic patient. Phage colonies selected by insulin-purified IgG extracts of IAS and IDD were selected at random for DNA sequencing, and tested for their reactivity with insulin antibodies and ability to distinguish disease-associated idiotopes. RESULTS: Seven phagotopes bound IAS and the phagotope designated IAS-9, corresponding to sequence KRSRLDV, gave the highest binding standard deviation (SD) score. Seven phagotopes bound IDD and the phagotope designated IDD-10, corresponding to sequence LGRGGSK, bound most strongly. IAS-9 was able to displace insulin binding in IAS and all of ten insulin-treated Type 2 diabetic patients, but not the IAA present in any of the eight patients with newly-diagnosed Type 1 diabetes. IDD-10, on the other hand, could displace insulin binding detected in the sera of eight patients with untreated Type 1 diabetes (IAA), but not in IAS or sera of the insulin-treated Type 2 diabetics. CONCLUSION: Phagotopes provide reagents which between them can distinguish positively as well as negatively diabetes-associated IAA from non-diabetes associated IAA and from IA.

Effects of rhizobium, arbuscular mycorrhizal fungi and anion content of simulated rain on subterranean clover.

An experiment was conducted to determine the extent to which rhizobia, mycorrhizal fungi, and anions in simulated rain affect plant growth response to acid deposition. Germinating subterranean clover seeds were planted in steam-pasteurized soil in pots and inoculated with Rhizobium leguminosarum, Glomus intraradices, Glomus etunicatum, R. leguminosarum + G. intraradices, R. leguminosarum + G. etunicatum, or no microbial symbionts. Beginning 3 weeks later, plants and the soil surface were exposed to simulated rain in a greenhouse on 3 days week(-1) for 12 weeks. Rain solutions were deionized water amended with background ions only (pH 5.0) or also adjusted to pH 3.0 with HNO3 only, H2SO4 only, or a 50/50 mixture of the two acids. Glomus intraradices colonized plant roots poorly, and G. intraradices-inoculated plants responded like nonmycorrhizal plants to rhizobia and rain treatments. Variation in plant biomass attributable to different rain formulations was strongest for G. etunicatum-inoculated plants, and the effect of rain formulation differed with respect to nodulation by rhizobia. The smallest plants at the end of the experiment were noninoculated plants exposed to rains (0.38 g mean dry weight total for 3 plants pot(-1)). Among nonnodulated plants infected by G. etunicatum, those exposed to HNO3 rain were largest, followed by plants exposed to HNO3 + H2SO4, pH 5.0, and H2SO4 rain, in that order. Among plants inoculated with both R. leguminosarum + G. etunicatum, however, the greatest biomass occurred with pH 5.0 rains, resulting in the largest plants in the study (1.00 g/3 plants). Treatment-related variation among root and shoot biomass data reflected those for whole-plant biomass. Based on quantification of biomass and N concentrations in shoot and root tissues, total N content of plants inoculated with G. etunicatum alone and exposed to the HNO3 + H2SO4 rains was approximately the same as plants inoculated with R. leguminosarum + G. entunicatum and exposed to pH 5 rains. Thus, the acid-mixture rains and rhizobia under no acid deposition provided approximately equal amounts of N in biomass. The significant interactions among rain formulation and the symbiotic status of the plants suggest that conclusions concerning the impact of acid deposition on plants in the environment cannot be considered reliable because most experiments on which such assessments are based have not tested confounding influences of microorganisms and precipitation characteristics.

Effects of ozone on three open-pollinated families of Pinus taeda L. grown in two substrates.

Seedlings from three open-pollinated loblolly pine (Pinus taeda L.) families grown in a mixture of commercial peat moss and grade 3 vermiculite (1:3 by volume) or a mixture of mineral soil and peat (1:1 by volume) were exposed to 0, 160 or 320 ppb ozone (O3) for 6h/day, 4 days/week for 8 weeks beginning 12 weeks after transplanting. Before exposures began, seedlings grown in the vermiculite-peat substrate were taller but smaller in diameter than those grown in the mineral soil-peat substrate. After 8 weeks of exposure, seedlings grown in the mineral soil-peat substrate were significantly larger in diameter and total biomass than those grown in the vermiculite-peat substrate. Primary needle and secondary needle injury increased with increasing O3 concentrations. Suppression of diameter growth, shoot weight and root weight was linear as O3 concentration increased. The effect of O3 on height or diameter growth or shoot biomass was not influenced by substrate type; but the suppression of root biomass due to O3 was dependent on substrate, with greater suppression in biomass occurring in the vermiculite-peat substrate. Foliar injury due to O3 was slightly greater in family 8-103, but growth suppression due to O3 was not significantly different among the families. Based on root biomass, response of seedlings to O3 was substrate-dependent.

De novo paracentric inversion in an X chromosome.

A 10 1/2 year old female with skeletal abnormalities was referred for genetic consultation because of learning disabilities and a suggestion of 'Turner-like' stigmata. Cytogenetic analysis revealed a paracentric inversion of an X(q13.1q26.1) chromosome.

De novo interstitial direct duplication of 15q: 46,XY,dir dup(15) (pter leads to q24::q14 leads to q21 X 1::q24 leads to qter).

A profoundly retarded, slightly dysmorphic male was re-examined cytogenetically by high resolution GTG banding and found to have a de novo interstitial direct duplication of 15q.