关键词： alternative hosts   automation   media blending   media development   Pichia pastoris  

摘要： Developing media to sustain cell growth and production is an essential and ongoing activity in bioprocess development. Modifications to media can often address host or product-specific challenges, such as low productivity or poor product quality. For other applications, systematic design of new media can facilitate the adoption of new industrially relevant alternative hosts. Despite manifold existing methods, common approaches for optimization often remain time and labor-intensive. We present here a novel approach to conventional media blending that leverages stable, simple, concentrated stock solutions to enable rapid improvement of measurable phenotypes of interest. We applied this modular methodology to generate high-performing media for two phenotypes of interest: biomass accumulation and heterologous protein production, using high-throughput, milliliter-scale batch fermentations of Pichia pastoris as a model system. In addition to these examples, we also created a flexible open-source package for modular blending automation on a low-cost liquid handling system to facilitate wide use of this method. Our modular blending method enables rapid, flexible media development, requiring minimal labor investment and prior knowledge of the host organism, and should enable developing improved media for other hosts and phenotypes of interest.

关键词： matrix algebra   independent measurements   Cauchy-Schwarz inequality   constrained Cramér-Rao bound   deterministic equality constraints   gradient methods   unbiased parameter estimation   Fisher information matrices   equality-constrained estimation   Gaussian noise   alternative derivations   gradient matrix   estimation error   estimation theory   zero-mean Gaussian noise   pseudomeasurements   parameter estimation   differentiable equality constraints  

摘要： This paper presents new alternative derivations of the well-known constrained Cramer-Rao bound for estimating parameters that satisfy a set of deterministic and differentiable equality constraints. Specifically, for unbiased parameter estimation, the equality constraints are treated as pseudo-measurements corrupted by independent zero-mean Gaussian noise with infinitely small covariance. In this way, the desired constrained Cramer-Rao bound can be established via utilising the property that the Fisher information matrices for independent measurements are additive. This paper also provides a new simple way for deriving the constrained Cramer-Rao bound when the parameter estimate has a specified bias through exploring that the estimation error lies in the null space of the gradient matrix of the constraints and invoking the Cauchy-Schwarz inequality.

关键词： Sustainable transportation   Life-cycle sustainability assessment   Electric vehicles   Spherical fuzzy sets   Analytic hierarchy process   Integrated combinative distance-based assessment (CODAS)  

摘要： Environmental, economic, and social comparisons of alternative fuel vehicles come with various conflicting objectives. For example, adopting electric vehicles might reduce greenhouse gas emissions but It can also cause more water consumption. To overcome such challenges when comparing alternatives, we developed an integrated sustainability assessment framework combining hybrid multi-regional input-output based life cycle sustainability assessment and multi-criteria decision-making methods including Spherical fuzzy sets. Fourteen different sustainability indicators, encompassing the economic, environmental, and social pillars of sustainability, are quantified for well-to-wheel (operation phase) impacts of internal combustion, hybrid, plug-in hybrid, and full battery electric vehicles. Qatar is investigated as a case study, where electricity generation is mainly from natural gas, but with high solar energy potential. The model favors battery electric vehicles charged through solar energy. The proposed framework is scalable and can be helpful towards developing effective national policies and strategies for sustainable transportation around the world.

关键词： Alternative fuel   Heavy-duty vehicle   Fleet   Organizational adoption   Innovation non-adoption decision   Technology acceptance  

摘要： Understanding heavy-duty vehicle (HDV) fleet operator behavior with respect to adoption of alternative fuel vehicles (AFVs) is critically important for accelerating diffusion of these technologies, and for achieving societal benefits through reduced emissions and improved public health. However, fleet operator perspectives have thus far received limited attention, leaving a key knowledge gap. This study aims to fill this gap by exploring HDV fleet operator decisions about alternative fuel adoption using both existing literature and new empirical data. To this end, we first develop an initial theoretical framework of AFV fleet adoption behavior in organizations based upon existing theories and literature. We then empirically improve the framework by investigating 20 organizations in California via in-depth qualitative interviews and project reports. A total of 29 adoption and 42 non-adoption cases were probed across various alternative fuel technologies, including natural gas, propane, electricity, hydrogen, biodiesel, and renewable diesel options. Content analysis of the qualitative data yielded 38 motivators or barriers related to AFV adoption, encompassing perceived technological characteristics, organization characteristics, and external environmental influences. The study results contribute theoretically and empirically to a better understanding of the demand-side aspects of AFV adoption by HDV fleet operators, particularly in California and in the other US states that follow California's environmental policies.

关键词： alternative splicing   SMRT   full-length transcriptome   RNA-Seq   plant virus   rice stripe virus   insect vector   small brown planthopper   JNK  

摘要： Alternative splicing (AS) is a frequent posttranscriptional regulatory event occurring in response to various endogenous and exogenous stimuli in most eukaryo-tic organisms. However, little is known about the effects of insect-transmitted viruses on AS events in insect vectors. The present study used third-generation sequencing technology and RNA sequencing (RNA-Seq) to evaluate the AS response in the small brown planthopper Laodelphax striatellus to rice stripe virus (RSV). The full-length tran-scriptome of L. striatellus was obtained using single-molecule real-time sequencing technology (SMRT). Posttranscriptional regulatory events, including AS, alternative polyadenylation, and fusion transcripts, were analyzed. A total of 28,175 nonredundant transcript isoforms included 24,950 transcripts assigned to 8,500 annotated genes of L. striatellus, and 5,000 of these genes (58.8%) had AS events. RNA-Seq of the gut sam-ples of insects infected by RSV for 8 d identified 3,458 differentially expressed tran-scripts (DETs);2,185 of these DETs were transcribed from 1,568 genes that had AS events, indicating that 31.4% of alternatively spliced genes responded to RSV infection of the gut. One of the c-Jun N-terminal kinase (JNK) genes, JNK2, experienced exon skipping, resulting in three transcript isoforms. These three isoforms differentially responded to RSV infection during development and in various organs. Injection of double-stranded RNAs targeting all or two isoforms indicated that three or at least two JNK2 isoforms facilitated RSV accumulation in planthoppers. These results implied that AS events could participate in the regulation of complex relationships between viruses and insect vectors. IMPORTANCE Alternative splicing (AS) is a regulatory mechanism that occurs after gene transcription. AS events can enrich protein diversity to promote the reactions of the organisms to various endogenous and exogenous stimulations. It is not known how insect vectors exploit AS event

关键词： Hemopatch   Laparoscopic partial nephrectomy   Veriset   Hemostasis   Complication  

摘要： Background Improvements in laparoscopic partial nephrectomy (LPN) in order to minimize perioperative warm ischemia time (WIT), complications, and consequently patient outcome are desirable. Veriset (TM) is a ready-to-use hemostatic patch of absorbable oxidized cellulose and hydrogel components that has earlier been implemented in vascular and hepatic surgery. We report our experience using this device in LPN. Methods Patients with a solitary malignant renal mass suspicious for renal cancer underwent LPN with either the use of Veriset (TM) hemostatic patch (n = 40) or conventional suture technique (n = 40). Patient characteristics, operation time and WIT, postoperative course and complications were recorded retrospectively. Tumor complexity was calculated according to the R.E.N.A.L. score. Outcome was determined according to the "trifecta" criteria (negative surgical margin, WIT < 25 min, no complications within 30 days). Results No significant differences with regard to clinical parameters and median R.E.N.A.L. score (6) were observed between both groups. Operation time (mean 127.1 min vs. 162. 8 min;p = 0.001) and WIT were both lower in the Veriset (TM) group (14.6 min vs. 20.6 min;p = 0.01). No differences in surgical margins (p = 0.602) and overall complication rates at 30 (p = 0.599) and 90 days (p = 0.611) postoperatively were noticed. The surgical outcome according to "trifecta" was achieved in 65% of patients using Veriset (TM) and in 57.5% of patients by suture closure, respectively. Conclusion The hemostatic Veriset (TM) patch can successfully be implemented in LPN. Handling and application appear favorable, thereby reducing operation time and WIT. The present results suggest that the device may represent an alternative to parenchyma suturing in LPN.

关键词： Life cycle assessment   Egg   Laying hen   Canada   Alternative hen housing  

摘要： The supply-managed Canadian egg industry produces over 789 million dozen eggs per year, the majority of which are produced in conventional cages (similar to 60%). Recently, the industry committed to a complete transition to alternative (i.e. enriched cage, single- and multi-tier free run, free range, and organic) production systems by 2036. This transition may have significant sustainability implications. Here, we present updated (i.e. based on 2019 data) LCA models and results representing the cradle to farm gate environmental impacts of Canadian egg production systems based on a much expanded data set compared to previous models. Generally, input and emission levels decreased across all housing systems, with few exceptions, compared to previously reported levels. Acidifying and GHG emissions decreased across most housing systems due to increases in feed- and pullet-use efficiency, while eutrophying emissions increased across all housing systems modeled due to differences in manure management systems. Feed inputs represented the greatest contributor to most impact categories (similar to 18% - 84%), followed by pullet production and manure management (similar to 10% - 37% and similar to 0.01% - 62%, respectively). Organic production systems had the lowest impacts in nine of the ten categories assessed, while free range systems generally performed the worst. Conventional cages generally had lower impacts than all non-organic systems. However, it can be expected that as farmers gain experience with alternative systems resource-use efficiency levels will increase and environmental impacts will decrease. Continued monitoring of environmental performance of Canadian egg production systems is therefore imperative to ensure net-positive outcomes during this housing system transition.

关键词： hepatitis B virus   nucleic acid testing   transfusion   blood safety  

摘要： Background -Multiplex viral nucleic acid testing (NAT) and a discriminatory testing algorithm have been used to detect viral infections in blood donors. Non-discriminated reactive (NDR) results may arise from low hepatitis B virus (HBV) DNA levels and are challenging for donor management by blood services. The aim of this study was to evaluate the performance and feasibility of alternative viral particle concentration methods to confirm and to characterise HBV infection status in NDR donors from Dalian, China, in order to improve routine donor management according to the potential residual risk estimate. Materials and methods -Individual donations were tested with ULTRIO Plus, and discriminated when reactive. Virions were concentrated from 12 and 6 mL plasma samples by ultracentrifugation (UC) and polyethylene glycol (PEG) precipitation, respectively. HBV DNA was detected with four nested polymerase chain reactions (95% limit of detection: 5-25 IU/mL). Amplified products were sequenced for definitive confirmation. Anti-HBc and anti-HBs were tested. Results -Of 77,556 donors, 79 (0.1%) were NAT NDR. After viral particle concentration by UC and PEG precipitation, HBV DNA was detected in 46 (58.2%) and 34 (43.0%) NDR donors, respectively, including 61.7% of samples that were repeatedly non-reactive with multiple NAT testing. Anti-HBc and anti-HBs (median titre: 37 mIU/mL) were detected in 87.3% and 46.8% of NDR donors, respectively. Sequencing confirmed HBV DNA in 65.8% of NDR donors, of whom 96.2% were occult HBV carriers with rare mutations in S and core proteins. Discussion -A HBV DNA confirmatory procedure with limited technical constraints was implemented successfully. The majority of NDR donors had occult HBV infections with extremely low viral DNA levels, which may constitute a potential residual threat for blood safety. Only a minority of anti-HBc+ NDR donors had anti-HBs levels high enough to consider their reinstatement as donors. The data support the permane

关键词： health care reform   prospective payment system   quality of health care   reimbursement   stroke  

摘要： Stroke contributes an estimated $28 billion to US health care costs annually, and alternative payment models aim to improve outcomes and lower spending over fee-for-service by aligning economic incentives with high value care. This systematic review evaluates historical and current evidence regarding the impacts of alternative payment models on stroke outcomes, spending, and utilization. Included studies evaluated alternative payment models in 4 categories: pay-for-performance (n=3), prospective payments (n=14), shared savings (n=5), and capitated payments (n=14). Pay-for-performance models were not consistently associated with improvements in clinical quality indicators of stroke prevention. Studies of prospective payments suggested that poststroke spending was shifted between care settings without consistent reductions in total spending. Shared savings programs, such as US Medicare accountable care organizations and bundled payments, were generally associated with null or decreased spending and service utilization and with no differences in clinical outcomes following stroke hospitalizations. Capitated payment models were associated with inconsistent effects on poststroke spending and utilization and some worsened clinical outcomes. Shared savings models that incentivize coordination of care across care settings show potential for lowering spending with no evidence for worsened clinical outcomes;however, few studies evaluated clinical or patient-reported outcomes, and the evidence, largely US-based, may not generalize to other settings.