A TOG:αβ-tubulin complex structure reveals conformation-based mechanisms for a microtubule polymerase

Stu2p/XMAP215/Dis1 family proteins are evolutionarily conserved regulatory factors that use αβ-tubulin-interacting tumor overexpressed gene (TOG) domains to catalyze fast microtubule growth. Catalysis requires that these polymerases discriminate between unpolymerized and polymerized forms of αβ-tubulin, but the mechanism by which they do so has remained unclear. Here, we report the structure of the TOG1 domain from Stu2p bound to yeast αβ-tubulin. TOG1 binds αβ-tubulin in a way that excludes equivalent binding of a second TOG domain. Furthermore, TOG1 preferentially binds a curved conformation of αβ-tubulin that cannot be incorporated into microtubules, contacting α- and β-tubulin surfaces that do not participate in microtubule assembly. Conformation-selective interactions with αβ-tubulin explain how TOG-containing polymerases discriminate between unpolymerized and polymerized forms of αβ-tubulin and how they selectively recognize the growing end of the microtubule.     

Independent and parallel recruitment of preexisting mechanisms underlying C₄ photosynthesis

C? photosynthesis allows increased photosynthetic efficiency because carbon dioxide (CO?) is concentrated around the key enzyme RuBisCO. Leaves of C? plants exhibit modified biochemistry, cell biology, and leaf development, but despite this complexity, C? photosynthesis has evolved independently in at least 45 lineages of plants. We found that two independent lineages of C? plant, whose last common ancestor predates the divergence of monocotyledons and dicotyledons about 180 million years ago, show conserved mechanisms controlling the expression of genes important for release of CO(2) around RuBisCO in bundle sheath (BS) cells. Orthologous genes from monocotyledonous and dicotyledonous C? species also contained conserved regulatory elements that conferred BS specificity when placed into C? species. We conclude that these conserved functional genetic elements likely facilitated the repeated evolution of C? photosynthesis.     

Suppression of the acute inflammatory response of porcine alveolar- and liver macrophages

During infection and inflammation drug disposition and hepatic metabolism are markedly affected in mammals. Pro-inflammatory mediators play an important role in the suppression of (cytochrome-P450-mediated) drug metabolism. Inflammatory mediators like cytokines, nitric oxide (NO), reactive oxygen species (ROS) and eicosanoids are released by activated macrophages from various sources, including liver and lung. It was the aim of this study to investigate ways to suppress the activation of macrophages during the onset of the inflammatory cascade. Therefore porcine lung and liver macrophages were isolated, and incubated with lipopolysaccharide (LPS) to initiate an acute inflammatory response, represented by the release of high amounts of tumour necrosis factor-alpha (TNF-alpha) into the culture medium. Additionally the primary macrophages were coincubated with phosphodiesterase-IV-(PDE-IV)-inhibitors or beta-adrenoceptor agonists that in previous studies demonstrated strong suppressive effects on TNF-alpha release. Especially the beta-adrenoceptor agonists showed to be very potent TNF-alpha suppressants, which indicates that the beta-adrenoceptor might be an interesting target for suppression of activation of macrophages. This was strengthened by the observation that the beta-adrenoceptor expression was not altered during the onset of the inflammatory cascade.     

Ultrasound-guided transversus abdominis plane block in the dog: an anatomical evaluation

Objective To describe the ultrasound‐guided technique to the transversus abdominis plane (TAP) block in the dog and evaluate the spread of a local anesthetic/methylene blue solution. Study design Prospective experimental trial. Animals Ten adult Beagle cadavers weighing 11.1 ± 1.1 kg (mean ± SD). Methods Transversus abdominis plane (TAP) blocks were performed bilaterally by a single trained individual on unpreserved cadaver dogs using 10 mL of methylene blue/bupivacaine solution per site. Dissection of the abdominal wall was performed within 15–55 minutes of block to determine distribution of injectate and nerve involvement in the transversus abdominis fascial plane. Results The transversus abdominis fascial plane was adequately visualized via ultrasound and injected in twenty hemi‐abdominal walls. Segmental branches of T11, T12, T13, L1, L2, and L3 were adequately stained in 20%, 60%, 100%, 100%, 90%, and 30% of injections, respectively. Conclusions and clinical relevance This anatomical study suggests that the transversus abdominis plane (TAP) block would provide adequate regional anesthesia of the abdomen, potentially extending to the cranial and caudal limits of the abdomen. This supports the clinical potential of this block in veterinary medicine.     

Effects of modified protein diets in dogs with chronic renal failure

A maintenance diet containing 44.4% dry weight protein and 2 reduced protein diets containing 17.2% and 8.2% dry weight protein were studied in dogs with induced chronic renal failure to determine effects on their (1) clinical status, (2) renal function, (3) electrolyte and acid-base balance, (4) divalent ion balance, (5) nutritional status, (6) hematologic status, and (7) hemostasis. A cross-over design of feeding was utilized so that the effect of diet could be studied in each dog at a known state of renal dysfunction and so that reversibility of effects could be determined. The inulin clearance rate was higher when the 44.4% protein diet was fed than when the 8.2% or 17.2% protein diets were fed. However, consumption of lower protein diets was associated with greater physical activity and reduction in serum urea nitrogen concentrations, compared with consumption of the 44.4% protein maintenance diet. Varying degrees of hyperchloremic metabolic acidosis developed in dogs fed the 8.2% and 17.2% protein diets. Reduced total serum protein and albumin concentrations, and hypercholesterolemia developed in dogs fed the 8.2% protein diet. All effects were reversible after diets were changed. It was concluded that diet-induced alterations in renal function associated with moderate dietary protein restriction did not adversely effect clinical and biochemical status of dogs with renal failure. It was unclear whether the clinical and biochemical changes observed with more severe protein restriction resulted from diet-induced changes in renal function.