Nucleotide depletion promotes cell fate transitions by inducing DNA replication stress.

Control of cellular identity requires coordination of developmental programs with environmental factors such as nutrient availability, suggesting that perturbing metabolism can alter cell state. Here, we find that nucleotide depletion and DNA replication stress drive differentiation in human and murine normal and transformed hematopoietic systems, including patient-derived acute myeloid leukemia (AML) xenografts. These cell state transitions begin during S phase and are independent of ATR/ATM checkpoint signaling, double-stranded DNA break formation, and changes in cell cycle length. In systems where differentiation is blocked by oncogenic transcription factor expression, replication stress activates primed regulatory loci and induces lineage-appropriate maturation genes despite the persistence of progenitor programs. Altering the baseline cell state by manipulating transcription factor expression causes replication stress to induce genes specific for alternative lineages. The ability of replication stress to selectively activate primed maturation programs across different contexts suggests a general mechanism by which changes in metabolism can promote lineage-appropriate cell state transitions.

Long Term Functional Outcome of Tibial Tuberosity Advancement vs. Tibial Plateau Leveling Osteotomy and Extracapsular Repair in a Heterogeneous Population of Dogs.

OBJECTIVE: To determine a long term function of tibial tuberosity advancement (TTA) for treatment of ruptured cranial cruciate ligament (CCL) in dogs, and to compare this to the long term function of previously reported tibial plateau leveling osteotomy (TPLO), extracapsular reconstruction (ECR), and a population of normal dogs. STUDY DESIGN: Prospective clinical trial. ANIMALS: Dogs with unilateral ruptured CCL treated with TTA (n = 14), TPLO (n = 15), and ECR (n = 23), and normal adult dogs (control, n = 80). MATERIALS AND METHODS: Force plate gait analysis was performed at 1 time point for the normal control group and preoperatively, and at 2 and 8 weeks and 6 and 12 months postoperatively for the treatment groups. Using serial force plates, symmetry indices (SI) were calculated between the operated and unoperated pelvic limbs for peak vertical force (PVF), contact time (CT), and vertical impulse (VI). Ground reaction forces (GRF) of the treatment and control group were compared using a general linear model. RESULTS: Walk SI for dogs with TTA were not significantly different from the control group at 12 months postoperatively. At the trot, neither TTA nor ECR achieved normal GRF. SI of the TPLO group were not different from the normal control group by 6-12 months postoperatively. CONCLUSION: At the walk, TTA achieves normal function by 12 months; however, at the trot TTA is indistinguishable from ECR. TPLO resulted in operated limb function that was similar to the control population by 6-12 months postoperatively at the walk and the trot.

Precision and accuracy of ground reaction force normalization in a heterogeneous population of dogs.

OBJECTIVE: To determine if currently used ground reaction force (GRF) normalization methods are accurate and precise enough to be used on a single-limb basis. STUDY DESIGN: Prospective clinical trial. ANIMALS: Clinically normal (n = 69) dogs and 40 dogs with unilateral ruptured cranial cruciate ligaments (CCL). METHODS: Pelvic limb GRFs of orthopedically normal dogs and those with unilateral ruptured CCL were collected. Normalization methods included none, body weight (BW), withers height (WH), WH and relative velocity (WH*F) and principal component 1 (PC1). Normalization methods were evaluated both by individual GRFs and additively. Binary logistic regression was performed for all normalization methods; sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) calculated. Stepwise backward logistic regression was used; significant values were retained in the final model. P < .05 was significant. RESULTS: Normalization of contact time (CT) by BW uniformly increased sensitivity, specificity, PPV, NPV, and accuracy. SI was the most accurate at both the walk and trot (accuracy 80-96%). Normalization by BW, WH, and WH*F all achieved similar results. When normalized GRFs were added, the accuracy increased only at the walk. CLINICAL SIGNIFICANCE: CT should be normalized to BW. SIs remain the gold standard, if SIs cannot be used, combining GRFs normalized to BW will result in high precision (80%) and high accuracy (89.5%) at the walk. At the trot, normalization by BW, WH and WH*F results in consistent results for the individual GRFs, though not all accuracies are >80%.

Long-term functional outcome of tibial plateau leveling osteotomy versus extracapsular repair in a heterogeneous population of dogs.

OBJECTIVE: To compare the long-term outcome of tibial plateau leveling osteotomy (TPLO) and extracapsular repair (ECR) for treatment of a ruptured cranial cruciate ligament (RCCL). STUDY DESIGN: Prospective clinical trial. ANIMALS: Normal adult dogs (control, n = 79); dogs with unilateral CCL disease (n = 38). METHODS: Dogs had TPLO (n = 15) or ECR (n = 23) for treatment of RCCL. Force plate gait analysis was performed for the control group at one time point and for treatment groups at serial points: preoperatively, 2 weeks, 8 weeks, 6 and 12 months postoperatively. Symmetry indices (SIs) were calculated between operated and unoperated pelvic limb for ground reaction forces (GRFs), including peak vertical force (PVF), contact time (CT), and vertical impulse (VI). GRFs of the treatment groups and control group were compared using a general linear model and Kaplan-Meier survival analysis. RESULTS: At 8 weeks, for PVF and VI, the TPLO group had more symmetric limb loading than the ECR group at the walk and trot. SIs of the TPLO group were not different from the control group by 6 months to 1 year postoperatively. SIs for the ECR group were less symmetrical than the control group at all time periods. Using survival analysis, median time to normal function was no different at the walk between groups, but was shorter for the TPLO group for VI and PVF. CONCLUSIONS: Dogs achieved normal limb loading faster after TPLO than ECR. TPLO resulted in operated limb function that was indistinguishable from the control population by 1 year postoperatively.