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Microglia reactivity entails a large-scale remodeling of cellular geometry, but the behavior of the microtubule cytoskeleton during these changes remains unexplored. Here we show that activated microglia provide an example of microtubule reorganization from a non-centrosomal array of parallel and stable microtubules to a radial array of more dynamic microtubules. While in the homeostatic state microglia nucleate microtubules at Golgi outposts, activating signaling induces recruitment of nucleating material nearby the centrosome, a process inhibited by microtubule stabilization. Our results demonstrate that a hallmark of microglia reactivity is a striking remodeling of the microtubule cytoskeleton and suggest that while pericentrosomal microtubule nucleation may serve as a distinct marker of microglia activation, inhibition of microtubule dynamics may provide a different strategy to reduce microglia reactivity in inflammatory disease.

Melanoma is a deadly form of cancer characterized by remarkable therapy resistance. Analyzing the transcriptome of MAPK inhibitor sensitive- and resistant-melanoma, we discovered that APAF-1 is negatively regulated by MITF in resistant tumors. This study identifies the MITF/APAF-1 axis as a molecular driver of MAPK inhibitor resistance. A drug-repositioning screen identified quinacrine and methylbenzethonium as potent activators of apoptosis in a context that mimics drug resistance mediated by APAF-1 inactivation. The compounds showed anti-tumor activity in in vitroand in vivomodels, linked to suppression of MITF function. Both drugs profoundly sensitize melanoma cells to MAPK inhibitors, regulating key signaling networks in melanoma, including the MITF/APAF-1 axis. Significant activity of the two compounds in inhibiting specific epigenetic modulators of MITF/APAF-1 expression, such as histone deacetylases, was observed. In summary, we demonstrate that targeting the MITF/APAF-1 axis may overcome resistance and could be exploited as a potential therapeutic approach to treat resistant melanoma.

Many proteins “bind” DNA through positively charged amino acids on their surfaces. However, to overcome significant energetic and topological obstacles, proteins that bend or package DNA might also modulate the stiffness that is generated by repulsions between phosphates within DNA. Much previous work describes how ions change the flexibility of DNA in solution, but when considering macromolecules such as chromatin in which the DNA contacts the nucleosome core each turn of the double helix, it may be more appropriate to assess the flexibility of DNA on charged surfaces. Mica coated with positively charged molecules is a convenient substrate upon which the flexibility of DNA may be directly measured with a scanning force microscope. In the experiments described below, the flexibility of DNA increased as much as fivefold depending on the concentration and type of polyamine used to coat mica. Using theory that relates charge neutralization to flexibility, we predict that phosphate repulsions were attenuated by ∼50% in the most flexible DNA observed. This simple method is an important tool for investigating the physiochemical causes and molecular biological effects of DNA flexibility, which affects DNA biochemistry ranging from chromatin stability to viral encapsulation.

Lumbosacral problems involving the cauda equina are commonly seen in companion animal practice. An understanding of the anatomic considerations and the corresponding clinical symptoms is important in diagnosing and treating these problems. Outcome depends a great deal on the nature and degree of involvement of the cauda equina.

The effect of feeding threonine-imbalanced and threonine-deficient purified diets (containing L-amino acids as the only source of dietary nitrogen) on food intake, weight gain and blood plasma amino acid pattern has been examined in growing kittens. The imbalance was created by adding 17.5% of amino acid mixture lacking threonine to a low amino acid basal diet containing 17.5% amino acid mixture including 0.4% threonine. A depression in food intake and weight gain occurred while feeding the imbalanced diet which was corrected by adding an additional 0.2% threonine to the imbalanced diet. There was no adaptation with time in the form of increased food intake or weight gain while feeding the imbalanced diet. Plasma threonine was consistently and similarly depressed (10 to 35% of normal) while feeding the basal, imbalanced and corrected diets and increased to normal when the standard diet with 1.4% threonine was fed. Plasma threonine and total amino acid concentrations of kittens fed the imbalanced diet did not differ from those observed in kittens fed the basal diet. Signs of neurologic dysfunction and/or lameness developed in 14 of the 17 kittens fed threonine-imbalanced or deficient diets, which resolved as dietary threonine was increased. J. Nutr. 110: 2444-2459, 1980.