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Purpose of Review: Currently, the increasing diversity of our society is poorly reflected in the urology workforce. In this review, we sought to address this disparity by highlighting key components involved in forming an academic urology department and training program that is focused on diversity, equity, and inclusion (DEI) as well as recruitment and retention of underrepresented in medicine (URiM) trainees and faculty. Recent Findings: We identified obstacles and provided approaches to enhance the ability of a department in creating a DEI-based curriculum and recruitment strategy with a key focus on understanding and addressing unconscious biases and microaggressions in the workplace. Summary: Substantive changes in the level of diversity within the urologic community can be made through the organization of a structured approach to increasing DEI. It starts with a commitment from each department to form achievable goals surrounding early mentorship of URiM students and trainees, an inclusive curriculum that is rooted in DEI, and targeted benchmarks for recruitment and retention of diverse staff. [ABSTRACT FROM AUTHOR]

Introduction: Bladder cancer care has been increasingly concentrated in high‐volume metropolitan medical centres (ie, "regionalisation" of care). We aimed to assess the potential role of geographic factors, including facility region and distance to treatment centre, as determinants of neoadjuvant chemotherapy (NAC) delivery in patients with non‐metastatic urothelial muscle‐invasive bladder cancer (MIBC) using nationally representative data from the United States. Methods: We queried the National Cancer Database to identify patients with cT2‐cT4a, N0M0 urothelial MIBC who underwent radical cystectomy (RC) from 2006 to 2015. Patients who received radiation therapy, single‐agent chemotherapy, adjuvant chemotherapy or systemic therapies other than multi‐agent chemotherapy were excluded. Multivariate logistic regression analysis was performed to identify independent predictors of receiving NAC. Results: A total of 5986 patients met the criteria for inclusion, of whom 1788 (29.9%) received NAC and 4108 received RC alone. Younger age, increased Charlson‐Deyo score, increased cT stage, increased annual income, increased distance from cancer treatment centre, treatment at an Academic Research Program or Integrated Network Cancer Program and a later year of diagnosis were independently predictive of NAC receipt. Older age, Medicare insurance and treatment in the East South Central or West South Central regions were independently associated with decreased odds of NAC receipt. Conclusions: Distance to treatment centre and United States geographic region were found to affect the likelihood of NAC receipt independently of other established predictors of success in this quality‐of‐care metric. Access to transportation and related resources merits consideration as additional pertinent social determinants of health in bladder cancer care. [ABSTRACT FROM AUTHOR]

Purpose: Pathologic upstaging in renal cell carcinoma (RCC) is common and confers a significant risk of poor surgical and survival outcomes. Preoperative predictors of upstaging are of great clinical relevance but empirical evidence specific to racial minorities remains scarce. Methods: National Cancer Database (NCDB) analysis of T3a‐specific upstaging among White, African‐American, Hispanic and Asian Pacific Islander (API) patients with AJCC cT1N0M0 RCC who underwent partial or radical nephrectomy between 2010 and 2015. Independent preoperative predictors of tumour upstaging were identified using multivariate logistic regression analyses. Results: A total of 81 002 patients met the criteria for inclusion (5.6% T3a‐specific upstaging). Increased age, increased Charlson‐Deyo comorbidity index, clinical stages cT1b and unspecified cT1, and increased Fuhrman nuclear grade were identified as independent risk factors for upstaging. Independent protective factors for upstaging were younger age, female sex, African‐American race and papillary, chromophobe, and unspecified RCC histologic subtypes. Significant risk factors and protective factors within individual racial subgroups were highly consistent with those observed in the overall study sample. All independent factors identified on race‐specific subgroup analyses were significant in the same direction relative to the overall study sample. Variables found to be non‐significant in the overall study sample remained non‐significant across all racial subgroup analyses. Conclusion: The present study of nationally representative data found no clinically significant differences in upstaging risk across individual racial subgroups relative to the overall study sample. Preoperative factors that can be used to predict pT3a‐specific tumour upstaging in CT1N0M0 RCC likely persist across different racial groups. [ABSTRACT FROM AUTHOR]

Purpose: Recent literature has separately identified multiple determinants of the use of neoadjuvant chemotherapy (NAC) and adherence to pelvic lymph node dissection (PLND) guidelines in the management of non-metastatic bladder cancer. However, such NAC/PLND analyses tend not to account for the other modality, despite the fact that NAC may impact the extent of dissectible lymph nodes. We aimed to determine the predictors of adequate PLND in patients with non-metastatic urothelial muscle-invasive bladder cancer (MIBC) undergoing radical cystectomy (RC) following receipt of NAC. Methods: We queried the National Cancer Database to identify patients from 2006–2015 with cT2-cT4a/N0M0 urothelial MIBC who underwent RC and were pre-treated with NAC. Multivariate logistic regression analysis was used to identify independent predictors of undergoing an adequate PLND (defined as > 8 nodes). Results: A total of 1518 patients met the criteria for inclusion (74.4% underwent adequate PLND). Adequate PLND was associated with treatment at an academic research facility (OR 2.762 [95% CI 2.119–3.599], p < 0.001). The likelihood of adequate PLND was significantly decreased in patients of older age (0.607 [0.441–0.835], p = 0.002 for age 70–79 years; 0.459 [0.245–0.860], p = 0.015 for age ≥ 80 years), a Charlson–Deyo score of 1 (0.722 [0.537–0.971], p = 0.031), and those who were uninsured (0.530 [0.292–0.964], p = 0.038). Conclusions: Established predictors of PLND may not necessarily be generalizable to all patients undergoing treatment for bladder cancer. The interplay between PLND and NAC merits further study, particularly in view of recent literature calling into question the survival benefit of PLND in patients pre-treated with NAC. [ABSTRACT FROM AUTHOR]

An optics-free method is proposed to generate x-ray radiation with spatially variant states of polarization via an afterburner extension to a free electron laser. Control of the polarization in the transverse plane is obtained through the overlap of different coherent transverse light distributions radiated from a bunched electron beam in two consecutive orthogonally polarised undulators. Different transverse profiles are obtained by emitting at a higher harmonic in one or both of the undulators. This method enables the generation of beams structured in their intensity, phase, and polarization—so-called Poincaré beams—at high powers with tunable wavelengths. Simulations are used to demonstrate the generation of two different classes of light with spatially inhomogeneous polarization—cylindrical vector beams and full Poincaré beams. [ABSTRACT FROM AUTHOR]

The article discusses the evolution in the implementation of stop and frisk policies in cities like New York, New York, Philadelphia, Pennsylvania, and Chicago, Illinois. Also cited are the case Terry v. Ohio in which the U.S. Supreme Court ruled that the action of the police in stopping and frisking two men due to suspicious behavior were warranted and affirmed a lower court's denial of suppression ruling.

Corynebacterium glutamicum(C. glutamicum) is a highly promising alternative prokaryotic host for recombinant protein expression, as it possesses several significant advantages overEscherichia coli (E. coli), the currently leading bacterial protein expression system. During the past decades, several experimental techniques and vector components for genetic manipulation ofC. glutamicumhave been developed and validated, including strong promoters for tightly regulating target gene expression, various types of plasmid vectors, protein secretion systems and methods of genetically modifying the host strain genome to improve protein production potential. This review critically discusses current progress in establishingC. glutamicumas a host for recombinant protein expression, and examines, in depth, some successful case studies of actual application of this expression system. The established “expression tool box” for developing novel constructs based onC. glutamicumas a host are also evaluated. Finally, the existing issues and solutions in process development withC. glutamicumas a host are specifically addressed. [ABSTRACT FROM PUBLISHER]

Promoter of alcohol oxidase I (PAOX1) is the most efficient promoter involved in the regulation of recombinant protein expression in Pichia pastoris (P. pastoris). PAOX1 is tightly repressed by the presence of glycerol in the culture medium; thus, glycerol must be exhausted before methanol can be taken up by P. pastoris and the expression of the heterologous protein can be induced. In this study, a candidate glycerol transporter (GT1, GeneID: 8197545) was identified, and its role was confirmed by further studies (e.g. bioinformatics analysis, heterologous complementation in Schizosaccharomyces pombe (S. pombe)). When GT1 is co-expressed with enhanced green fluorescent protein (EGFP), it localizes to the membrane and S. pombe carrying gt1 but not the wild-type strain can grow on medium containing glycerol as the sole carbon source. The present study is the first to report that AOX1 in the X-33⧠gt1 mutant can achieve constitutive expression in medium containing glycerol; thus, knocking down gt1 can eliminate the glycerol repression of PAOX1 in P. pastoris. These results suggest that the glycerol transporter may participate in the process of PAOX1 inhibition in glycerol medium. [ABSTRACT FROM AUTHOR]

Microalgae represents a promising biofuel feedstock as it can be cultivated on low-quality land and integrated with a variety of waste streams. Produced water is the largest waste stream generated in the oil and gas industry. The integration of the two systems is synergistic as algal cultivation requires large volumes of water and there is a need for remediation of produced water. Produced water contains inorganic and organic contaminants that could inhibit growth and lipid production of microalgae directly impacting the yield of the system. In this study, produced water from the Uintah Basin, Utah, oil and gas industry was substituted for water in the growth media for microalgae, Nannochloropsis salina. Experiments utilized produced water as the base for media preparation and evaluated microalgae productivity at different dilutions with deionized water, i.e., 0% (control), 25%, 50%, 75%, and 100% produced water, as well as the recycling of produced waterbased growth media. Results show that produced water severely inhib its the growth of microalgae, with a 102% decrease in the 100% produced water media. Recycling of growth media has the potential to positively impact productivity results at concentrations of produced water less than 25%. Discussion focuses on the importance of integration studies, the impact of reduced productivity with higher concentration of produced water, and the need for further experimental growth studies. [ABSTRACT FROM AUTHOR]

Contamination of food and feed by Aflatoxin B1 (AFB1) is a cause of serious economic and health problems. Different processes have been used to degrade AFB1. In this study, biological degradation of AFB1 was carried out using three Actinomycete species, Rhodococcus erythropolis ATCC 4277, Streptomyces lividans TK 24, and S. aureofaciens ATCC 10762, in liquid cultures. Biodegradation of AFB1 was optimised under a range of temperatures from 25 to 40 °C and pH values of 4.0 to 8.0. An initial concentration of 20 µg/mL of AFB1 was used in this study. The amount of AFB1 remaining was measured against time by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC), coupled with UV and mass spectrometry (LC-MS). All species were able to degrade the AFB1, and no significant difference was found between them. AFB1 remained in the liquid culture for R. erythropolis, S. lividans and S. aureofaciens were 0.81 µg/mL, 2.41 µg/mL and 2.78 µg/mL respectively, at the end of the first 24 h. Degradation occurred at all incubation temperatures and the pH with the optimal conditions for R. erythropolis was achieved at 30 °C and pH 6, whereas for S. lividans and S. aureofaciens the optimum conditions for degradation were 30 °C and pH 5. Analysis of the degradative route indicated that each microorganism has a different way of degrading AFB1. The metabolites produced by R. erythropolis were significantly different from the other two microorganisms. Products of degradation were identified through metabolomic studies by utilizing high-resolution mass spectral data. Mass spectrometric analysis indicated that the degradation of AFB1 was associated with the appearance of a range of lower molecular weight compounds. The pathway of degradation or chemical alteration of AFB1 was followed by means of high resolution Fourier transform mass spectrometry (HR-FTMS) analysis as well as through the MS2 fragmentation to unravel the degradative pathway for AFB1. AFB1 bio-degradation was coupled with the accumulation of intermediates of fatty acid metabolism and glycolysis. A plausible mechanism of degradation of AFB1 by Rhodococcus was hypothesized. [ABSTRACT FROM AUTHOR]

The American Committee to Keep Biafra Alive (ACKBA) was the largest and most influential organization in the United States that formed in response to the Nigerian civil war. While historians have pointed to the committee as an important source of activism that pushed the American government towards supporting more vigorous humanitarian relief, this is the first article to explore the development of the group from its inception and to look specifically at its claims of genocide. Not everyone at the time agreed that the Nigerian government was committing genocide against the people living in the secessionist state of Biafra, and that debate continues today. The ACKBA, appealing to genocide prevention and human rights, argued that the debate about the semantics of genocide got in the way of actually helping those that were suffering from famine as a result of the war. In the process, the committee offered a redefinition of genocide that wedded conceptions of Biafran identity to the Biafran state, which made the maintenance of ‘one Nigeria’, in the eyes of committee members, an act of genocide. In the end, this redefinition of genocide failed to bring more people in the United States towards supporting Biafran secession and might have, in the end, led to more confusion about genocide during the conflict. An analysis of the committee's activism highlights the often tenuous relationship between self-determination and genocide in the developing world and illustrates the growing limits of American political intervention in the global south. [ABSTRACT FROM PUBLISHER]

Background: Novel analytical tools, which shorten the long and costly development cycles of biopharmaceuticals are essential. Metabolic flux analysis (MFA) shows great promise in improving our understanding of the metabolism of cell factories in bioreactors, but currently only provides information post-process using conventional off-line methods. MFA combined with real time multianalyte process monitoring techniques provides a valuable platform technology allowing real time insights into metabolic responses of cell factories in bioreactors. This could have a major impact in the bioprocessing industry, ultimately improving product consistency, productivity and shortening development cycles. Results: This is the first investigation using Near Infrared Spectroscopy (NIRS) in situ combined with metabolic flux modelling which is both a significant challenge and considerable extension of these techniques. We investigated the feasibility of our approach using the industrial workhorse Pichia pastoris in a simplified model system. A parental P. pastoris strain (i.e. which does not synthesize recombinant protein) was used to allow definition of distinct metabolic states focusing solely upon the prediction of intracellular fluxes in central carbon metabolism. Extracellular fluxes were determined using off-line conventional reference methods and on-line NIR predictions (calculated by multivariate analysis using the partial least squares algorithm, PLS). The results showed that the PLS-NIRS models for biomass and glycerol were accurate: correlation coefficients, R2, above 0.90 and the root mean square error of prediction, RMSEP, of 1.17 and 2.90 g/L, respectively. The analytical quality of the NIR models was demonstrated by direct comparison with the standard error of the laboratory (SEL), which showed that performance of the NIR models was suitable for quantifying biomass and glycerol for calculating extracellular metabolite rates and used as independent inputs for the MFA (RMSEP lower than 1.5 × SEL). Furthermore, the results for the MFA from both datasets passed consistency tests performed for each steady state, showing that the precision of on-line NIRS is equivalent to that obtained by the off-line measurements. Conclusions: The findings of this study show for the first time the potential of NIRS as an input generating for MFA models, contributing to the optimization of cell factory metabolism in real-time. [ABSTRACT FROM AUTHOR]

The relationship between prior growth of food-spoilage yeast in high-sugar environments and their subsequent survival postpulsed UV (PUV) irradiation was investigated. Test yeast were separately grown to early stationary phase in YPD broth containing increasing concentrations of glucose (1-50% w/v) and were flashed with ≤40 pulses of broad-spectrum light at lamp discharge energy settings of 3.2, 7.2 and 12.8 J (equivalent to UV doses of 0.53, 1.09 and 3.36 μJ cm−2, respectively) and their inactivation measured. Findings showed that prior growth in high-sugar conditions (≥30% glucose w/v) enhanced the sensitivity of all nine representative strains of Zygosaccharomyces bailii, Z. rouxii and Saccharomyces cerevisiae yeast to PUV irradiation. Significant differences in inactivation amongst different yeast types also occurred depending on amount of UV dose applied, where the order of increasing sensitivity of osmotically stressed yeast to PUV irradiation was shown to be Z. rouxii, Z. bailii and > S. cerevisiae. For example, a 1.2-log order difference in CFU mL−1 reduction occurred between Z. bailii 11 486 and S. cerevisiae 834 when grown in 50% w/v sugar samples and treated with the uppermost test UV dosage of 3.36 μJ cm−2, where these two yeast strains were reduced by 3.8 and 5.0 log orders, respectively, after this PUV treatment regime compared to untreated controls. The higher the UV dose applied the greater the reduction in yeast numbers. For example, a 1.0-, 1.4- and 4.0-log order differences in CFU mL−1 numbers occurred for S. cerevisiae 834 grown in 15% w/v sugar samples and then treated with PUV dose of 0.53, 1.09 and 3.36 μJ cm−2, respectively. These findings support the development of PUV for the treatment of high-sugar foods that are prone to spoilage by osmotolerant yeast. [ABSTRACT FROM AUTHOR]

There is considerable interest in exploiting the novel physical and biological properties of microbial exopolysaccharides in industry and medicine. For economic and scientific reasons, large scale production under carefully monitored and controlled conditions is required. Producing exopolysaccharides in industrial fermenters poses several complex bioengineering and microbiological challenges relating primarily to the very high viscosities of such culture media, which are often exacerbated by the producing organism''s morphology. What these problems are, and the strategies for dealing with them are discussed critically in this review, using pullulan, curdlan, xanthan, and fungal ββ-glucans as examples of industrially produced microbial exopolysaccharides. The role of fermenter configuration in their production is also examined. [ABSTRACT FROM AUTHOR]

With intensities 108-1010 times greater than other laboratory sources, X-ray free-electron lasers are currently opening up new frontiers across many areas of science. In this Review we describe how these unconventional lasers work, discuss the range of new sources being developed worldwide, and consider how such X-ray sources may develop over the coming years. [ABSTRACT FROM AUTHOR]

Fungi are amongst the most industrially important microorganisms in current use within the biotechnology industry. Most such fungal cultures are highly aerobic in nature, a character that has been frequently referred to in both reactor design and fungal physiology. The most fundamentally significant outcome of the highly aerobic growth environment in fermenter vessels is the need for the fungal culture to effectively combat in the intracellular environment the negative consequences of high oxygen transfer rates. The use of oxygen as the respiratory substrate is frequently reported to lead to the development of oxidative stress, mainly due to oxygen-derived free radicals, which are collectively termed as reactive oxygen species (ROS). Recently, there has been extensive research on the occurrence, extent, and consequences of oxidative stress in microorganisms, and the underlying mechanisms through which cells prevent and repair the damage caused by ROS. In the present study, we critically review the current understanding of oxidative stress events in industrially relevant fungi. The review first describes the current state of knowledge of ROS concisely, and then the various antioxidant strategies employed by fungal cells to counteract the deleterious effects, together with their implications in fungal bioprocessing are also discussed. Finally, some recommendations for further research are made. [ABSTRACT FROM AUTHOR]

We tested the hypothesis that endogenously produced hydrogen sulfide (H2S) can potentially contribute to the adrenergic stress response in rainbow trout by initiating catecholamine secretion from chromaffin cells. During acute hypoxia (water P02 = 35 mmHg), plasma H2S levels were significantly elevated concurrently with a rise in circulating catecholamine concentrations. Tissues enriched with chromaffin cells (posterior cardinal vein and anterior kidney) produced H2S in vitro when incubated with L-cysteine. In both tissues, the production of H2S was eliminated by adding the cystathionine β-synthase inhibitor, aminooxyacetate. Cystathionine β-synthase and cystathionine γ-lyase were cloned and sequenced and the results of real-time PCR demonstrated that with the exception of white muscle, mRNA for both enzymes was broadly distributed within the tissues that were examined. Electrical field stimulation of an in situ saline-perfused posterior cardinal vein preparation caused the appearance of H2S and catecholamines in the outfiowing perfusate. Perfusion with the cholinergic receptor agonist carbachol (1 X 10-6 M) or depolarizing levels of KCI (1 X 10-2 M) caused secretion of catecholamines without altering H2S output, suggesting that neuronal excitation is required for H2S release. Addition of H2S (at concentrations exceeding 5 X 10-7 M) to the perfusion fluid resulted in a marked stimulation of catecholamine secretion that was not observed when Ca2+-free perfusate was used. These data, together with the finding that H2S-induced catecholamine secretion was unaltered by the nicotinic receptor blocker hexamethonium, suggest that H2S is able to directly elicit catecholamine secretion via membrane depolarization followed by Ca2+-mediated exocytosis. [ABSTRACT FROM AUTHOR]

Although host proteases are often considered to have a negative impact upon heterologous protein production by filamentous fungi, relatively little is known about the pattern of their appearance in recombinant fungal bioprocesses. In the present study, we investigated extracellular proteases from a filamentous fungus, Aspergillus niger B1-D, genetically modified to secrete hen egg white lysozyme (HEWL). Our findings indicate that extracellular protease activity is only detected after the carbon source is completely utilised in batch cultures. The proteases are predominantly acid proteases and have optimal temperature for activity at around 45°C. Their activity could be partially inhibited by protease inhibitors, indicating the existence of at least four kinds of proteases in these culture fluids, aspartic-, serine-, cysteine-, and metallo-proteases. Oxygen enrichment does not have any noticeable effects on extracellular protease activity except that the onset of protease activity appears earlier in oxygen enrichment runs. Oxygen enrichment stimulates HEWL production substantially, and we propose that it is related to fungal morphology. Thermal stress imposed by raising process temperature (from 25 to 30 and 35°C) in early exponential phase, led to appearance of protease activity in the medium following the heat shock. Continued cultivation at high temperatures significantly reduced HEWL production, which was associated with increased activity of the extracellular proteases in these cultures. [ABSTRACT FROM AUTHOR]

Inoculation of bioreactors with shake-flask cultures present the organism with an immediate shift from an environment with little O2 to one in which O2 is typically at 100% saturation. The inoculation of such shake-flasks cultures into bioreactors sparged with 1 vvm air or 1 vvm air/O2 mix i.e. 50% O2 enrichment is an oxidatively stressful event, as judged by immediate increases in the intracellular concentrations of superoxide anion radical (O) (from 4,600 to 11,600 RLU mg DCW−1 and 5,500 to 23,000 RLU mg DCW−1 respectively) and changes in the activities of the major antioxidant enzymes superoxide dismutase and catalase in all cultures. There are further effects on metabolic indices, particularly decreased nutrient consumption in oxygenated cultures (from 0.16 to 0.12 g starch g DCW h−1) and decreased protein production, indicating that inoculation of the bioreactor exerts a global burden on the cellular metabolic networks. [ABSTRACT FROM AUTHOR]

The use of at-line NIRS to monitor a high cell density fed-batch baker's yeast bioprocess was investigated. Quantification of the key analytes (biomass, ethanol and glucose) and the product quality indicator (percentage protein content) was studied. Biomass was quantitatively modelled using whole matrix samples (as was percentage protein content). The dominance of the whole matrix spectrum by biomass, and its associated light scattering effects, were overcome by use of filtrate samples and adapted (semi-synthetic) filtrate samples, which allowed successful ethanol and glucose modelling, respectively. Calibrations were rigorously challenged via external validation with large sample sets relative to the calibration sample size, ensuring model robustness and potential practical utility. The standard errors of calibration for biomass, glucose, ethanol and total intracellular protein were (g/l) 1.79, 0.19, 0.79 and 0.91, respectively, comparable to those of the primary assays. The calibration strategies necessary to generate quantitative models for this range of analytes in such a complex high cell density bioprocess fluid are discussed. Copyright © 2006 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]