Songbird organotypic culture as an in vitro model for interrogating sparse sequencing networks

Sparse sequences of neuronal activity are fundamental features of neural circuit computation; however, the underlying homeostatic mechanisms remain poorly understood. To approach these questions, we have developed a method for cellular-resolution imaging in organotypic cultures of the adult zebra finch brain, including portions of the intact song circuit. These in vitro networks can survive for weeks, and display mature neuron morphologies. Neurons within the organotypic slices exhibit a diversity of spontaneous and pharmacologically induced activity that can be easily monitored using the genetically encoded calcium indicator GCaMP6. In this study, we primarily focus on the classic song sequence generator HVC and the surrounding areas. We describe proof of concept experiments including physiological, optical, and pharmacological manipulation of these exposed networks. This method may allow the cellular rules underlying sparse, stereotyped neural sequencing to be examined with new degrees of experimental control

In-depth critical analysis of complications following robot-assisted radical cystectomy with intracorporeal urinary diversion

Background: Robot-assisted radical cystectomy with intracorporeal urinary diversion (iRARC) is an attractive option to open cystectomy, but the benefit in terms of improved outcomes is not established. Objective: To evaluate the early postoperative morbidity and mortality of patients undergoing iRARC and conduct a critical analysis of complications using standardised reporting criteria as stratified according to urinary diversion. Design, setting, and participants: A total of 134 patients underwent iRARC for bladder cancer at a single centre between June 2011 and July 2015. Intervention: Radical cystectomy with iRARC. Outcome measurements and statistical analysis: Patient demographics, pathologic data, and 90-d perioperative mortality and complications were recorded. Complications were reported according to the Clavien-Dindo (CD) classification and stratified according to urinary diversion type and either surgical or medical complications. The chi-square test and t test were used for categorical and continuous variables respectively. Multivariable logistic regression was performed on variables with significance in univariate analysis. Results and limitations: The 90-d all complication rate following ileal conduit and continent diversion was 68% and 82.4%, and major complications were 21.0% and 20.6% respectively. The 90-d mortality was 3% and 2.9% for ileal conduit and continent diversion patients, respectively. On multivariate analysis, the blood transfusion requirement was independently associated with major complications (p = 0.002) and all 30-d (p = 0.002) and 90-d (p = 0.012) major complications. Male patients were associated with 90-d major complications (p = 0.015). Critical analysis identified that surgical complications were responsible for 39.4% of all 90-d major complications. The incidence of surgical complications did not decline with increasing number of iRARC cases performed (p = 0.742, r = 0.31). Limitations of this study include its retrospective nature, limited sample size, and limited multivariate analysis due to the low number of major complications events. Conclusions: Although complications following iRARC are common, most are low grade. A critical analysis identified surgical complications as a cause of major complications. Addressing this issue could have a significant impact on lowering the morbidity associated with iRARC. Patient summary: We looked at the surgical outcomes in bladder cancer patients treated with minimally invasive robotic surgery. We found that surgical complications account for most major complications and previous surgical experience may be a confounding factor when interpreting results from a different centre even in a randomised trial setting

Leukemia-associated RhoGEF (LARG) is a novel RhoGEF in cytokinesis and required for the proper completion of abscission.

Proper completion of mitosis requires the concerted effort of multiple RhoGEFs. Here we show that leukemia-associated RhoGEF (LARG), a RhoA-specific RGS-RhoGEF, is required for abscission, the final stage of cytokinesis, in which the intercellular membrane is cleaved between daughter cells. LARG colocalizes with α-tubulin at the spindle poles before localizing to the central spindle. During cytokinesis, LARG is condensed in the midbody, where it colocalizes with RhoA. HeLa cells depleted of LARG display apoptosis during cytokinesis with unresolved intercellular bridges, and rescue experiments show that expression of small interfering RNA-resistant LARG prevents this apoptosis. Moreover, live cell imaging of LARG-depleted cells reveals greatly delayed fission kinetics in abscission in which a population of cells with persistent bridges undergoes apoptosis; however, the delayed fission kinetics is rescued by Aurora-B inhibition. The formation of a Flemming body and thinning of microtubules in the intercellular bridge of cells depleted of LARG is consistent with a defect in late cytokinesis, just before the abscission event. In contrast to studies of other RhoGEFs, particularly Ect2 and GEF-H1, LARG depletion does not result in cytokinetic furrow regression nor does it affect internal mitotic timing. These results show that LARG is a novel and temporally distinct RhoGEF required for completion of abscission

Heat pump integration for total site waste heat recovery

Total Site Heat Integration (TSHI) promotes energy recovery between processes to enhance overall energy efficiency of an industrial complex. Various industrial waste heat utilisation technologies have been studied to improve the energy efficiency of energy system. Vapour compression as an open loop heat pump system has good potential to be used to upgrade the waste heat to useful heat in Total Site systems. Vapour compression systems upgrade low grade waste heat by supplying a low quantity of high pressure steam (thermocompressor) or mechanical work (mechanical-compressor) to generate higher pressure steam, as is common with evaporation systems. The vapour compression system recovers the latent heat content of the industrial waste heat, which reduces cooling demand, decreasing the demand for high quality steam and reducing boiler load. This paper introduces an effective Total Site targeting methodology to integrate open cycle heat pump systems, i.e. vapour compression technologies, into an integrated industrial energy system for enhancing overall site energy efficiency. Industrial waste heat and high quality steam demand are able to be reduced simultaneously though this integration. The energy reduction and cost-benefit of thermo-compressor and mechanical-compressor installations are compared through a literature case study. The case study showed a deficit of heat at the MPS and a surplus of heat the LPS, which was identified as a candidate for compression according to the appropriate placement principle for heat pumps. For the case study, a four-stage mechanical vapour compression system and two-stage thermal vapour compression system resulted in an energy cost reductions of 343,859 USD/y and 168,829 USD/y

Modelling of Explosion Venting Fireballs

PresentationThe purpose of this study is to compare calculated sizes of explosion venting fireballs to correlations for a range of gas explosions in enclosures fitted with an explosion vent. Explosion vents are commonly used to protect process equipment containing flammable gases. When properly designed, an explosion vent can reduce the peak overpressure inside the enclosure so that the enclosure does not permanently deform or fail catastrophically. The fuel and combustion products that exit the vent, however, create an external fireball hazard. Deflectors can be used to reduce the extent of that external fireball, but their effectiveness has not been rigorously tested for vessels larger than 20 m3Recently, there have been questions about the accuracy of correlations to predict fireball dimensions from explosion vents and the effects of deflectors on those dimensions. The fireball size during venting is crucial in determining the thermal hazard area around the protected enclosure. The National Fire Protection Association standard NFPA 68 and International Standard EN14994 provide correlations for calculating the extent of the expected fireball during explosion venting scenarios. The formulas for gas-explosion fireball-sizes are empirical and based on data from a limited number of experiments. In this study, explosion-venting scenarios are modelled numerically using FLACS v10. FLACS is a Computational Fluid Dynamics (CFD) software widely used in the oil and gas industry to perform explosion consequence modelling (Gexcon 2015). The CFD model was used to evaluate the fireball extent and temperature. The model results provide the fireball shape and size for different scenarios including scenarios with deflectors. A number of parameters are varied, including: Enclosure size Type of fuel Presence of a deflagration deflector plate The study elucidates the relative importance of each of these parameters on the explosion fireball size. This can assist in designing future testing programs to examine fireball size and identifying parameters for use in improved fireball size correlations

Phosphorylation sites in BubR1 that regulate kinetochore attachment, tension, and mitotic exit

BubR1 kinase is essential for the mitotic checkpoint and also for kinetochores to establish microtubule attachments. In this study, we report that BubR1 is phosphorylated in mitosis on four residues that differ from sites recently reported to be phosphorylated by Plk1 (Elowe, S., S. Hummer, A. Uldschmid, X. Li, and E.A. Nigg. 2007. Genes Dev. 21:2205–2219; Matsumura, S., F. Toyoshima, and E. Nishida. 2007. J. Biol. Chem. 282:15217–15227). S670, the most conserved residue, is phosphorylated at kinetochores at the onset of mitosis and dephosphorylated before anaphase onset. Unlike the Plk1-dependent S676 phosphorylation, S670 phosphorylation is sensitive to microtubule attachments but not to kinetochore tension. Functionally, phosphorylation of S670 is essential for error correction and for kinetochores with end-on attachments to establish tension. Furthermore, in vitro data suggest that the phosphorylation status of BubR1 is important for checkpoint inhibition of the anaphase-promoting complex/cyclosome. Finally, RNA interference experiments show that Mps1 is a major but not the exclusive kinase that specifies BubR1 phosphorylation in vivo. The combined data suggest that BubR1 may be an effector of multiple kinases that are involved in discrete aspects of kinetochore attachments and checkpoint regulation

Associations of staple food consumption and types of cooking oil with waist circumference and body mass index in older Chinese men and women: a panel analysis

Background: The dietary landscape has changed rapidly in China in the past few decades. This research investigates the associations of older adults’ choices and consumption of staple foods and cooking oils with obesity related measurements. Methods: Panel data were extracted from the Chinese Longitudinal Health Longevity Survey from3253 older participants with 6506 observations. Ordinary least squares and ordered logistic regression models were estimated with the outcomes of obesity determined by waist circumference (WC) and body mass index (BMI), respectively. Results: Older men who consumed wheat had wider WCs (β=2.84 [95% confidence interval {CI} 1.55 to 4.13], p\u3c0.01) and higher BMIs (adjusted odds ratio 1.74 [95% CI 1.40 to 2.17], p\u3c0.01) than those who preferred rice. Female participants who used animal-based cooking oil had lower WCs and BMIs than their counterparts who consumed vegetable-based cooking oil. Increased consumption of staple foods was associated with increased rates of obesity in both sexes. Conclusion: Dieticians and nutritionists should design appropriate dietary plans to help reduce obesity and chronic diseases among older Chinese adults. Further clinical trials are needed to continue investigating this topic

Single-dose pharmacokinetic and toxicity analysis of pyrrole–imidazole polyamides in mice

Purpose: Pyrrole–imidazole (Py-Im) polyamides are programmable, sequence-specific DNA minor groove–binding ligands. Previous work in cell culture has shown that various polyamides can be used to modulate the transcriptional programs of oncogenic transcription factors. In this study, two hairpin polyamides with demonstrated activity against androgen receptor signaling in cell culture were administered to mice to characterize their pharmacokinetic properties. Methods: Py-Im polyamides were administered intravenously by tail vein injection. Plasma, urine, and fecal samples were collected over a 24-h period. Liver, kidney, and lung samples were collected postmortem. Concentrations of the administered polyamide in the plasma, excretion, and tissue samples were measured using LC/MS/MS. The biodistribution data were analyzed by both non-compartmental and compartmental pharmacokinetic models. Animal toxicity experiments were also performed by monitoring weight loss after a single subcutaneous (SC) injection of either polyamide. Results: The biodistribution profiles of both compounds exhibited rapid localization to the liver, kidneys, and lungs upon injection. Plasma distribution of the two compounds showed distinct differences in the rate of clearance, the volume of distribution, and the AUCs. These two compounds also have markedly different toxicities after SC injection in mice. Conclusions: The variations in pharmacokinetics and toxicity in vivo stem from a minor chemical modification that is also correlated with differing potency in cell culture. The results obtained in this study could provide a structural basis for further improvement of polyamide activity both in cell culture and in animal models