Identification, Characterization, and Application of a Highly Sensitive Lactam Biosensor from Pseudomonas putida

Caprolactam is an important polymer precursor to nylon traditionally derived from petroleum and produced on a scale of 5 million tons per year. Current biological pathways for the production of caprolactam are inefficient with titers not exceeding 2 mg/L, necessitating novel pathways for its production. As development of novel metabolic routes often require thousands of designs and result in low product titers, a highly sensitive biosensor for the final product has the potential to rapidly speed up development times. Here we report a highly sensitive biosensor for valerolactam and caprolactam from Pseudomonas putida KT2440 which is >1000× more sensitive to an exogenous ligand than previously reported sensors. Manipulating the expression of the sensor oplR (PP_3516) substantially altered the sensing parameters, with various vectors showing Kd values ranging from 700 nM (79.1 μg/L) to 1.2 mM (135.6 mg/L). Our most sensitive construct was able to detect in vivo production of caprolactam above background at ∼6 μg/L. The high sensitivity and range of OplR is a powerful tool toward the development of novel routes to the biological synthesis of caprolactam

Best practices protocol for the evaluation of bulbar dysfunction: summary recommendations from the NEALS bulbar subcommittee symposium

Objective: The aim of this Symposium was to develop a consensus based, bulbar assessment protocol for implementation within NEALS clinics. Methods: A one-day symposium, held in April 2017, was organized into Speech and Swallowing sections to establish summary recommendations for the assessment of bulbar dysfunction within each group. Results: Summary recommendations included speech referrals and AAC evaluations at initial visit, CNS-BFS, maximum sustained phonation, and speaking rate. Dysarthria evaluation included the speech subsystem involvement of respiration, phonation, resonance, and articulation. Specific recommendations for swallowing were established for each of the following domains: dietary/oral intake, airway defense physiologic capacity, swallow safety screen, patient-reported swallow-related outcomes, oral sensorimotor exam, and pulmonary function. Practice parameters focused upon patient education and unresolved questions included the use of videofluoscopy, monitoring diet progression, and swallow safety screening. Conclusions: The working goal is to establish a clinical bulbar protocol, designed to be incorporated within ALS clinics and ultimately to formulate a best practice set of bulbar ALS guidelines, available for implementation throughout the international ALS community

Provisional best practices guidelines for the evaluation of bulbar dysfunction in amyotrophic lateral sclerosis

Introduction: Universally established comprehensive clinical bulbar scales objectively assessing disease progression in amyotrophic lateral sclerosis (ALS) are currently lacking. The goal of this working group project is to design a best practice set of provisional bulbar ALS guidelines, available for immediate implementation within all ALS clinics. Methods: ALS specialists across multiple related disciplines participated in a series of clinical bulbar symposia, intending to identify and summarize the currently accepted best practices for the assessment and management of bulbar dysfunction in ALS Results: Summary group recommendations for individual speech, Augmentative and Alternative Communication (AAC), and swallowing sections were achieved, focusing on the optimal proposed level of care within each domain. Discussion: We have identified specific clinical recommendations for each of the 3 domains of bulbar functioning, available for incorporation within all ALS clinics. Future directions will be to establish a formal set of bulbar guidelines through a methodological and evidence-based approach

Alimentação e ecomorfologia de duas espécies de piranhas (Characiformes: Characidae) do lago de Viana, estado do Maranhão, Brasil Feeding and ecomorphology of two species of piranhas (Characiformes: Characidae) from the Viana Lake, Maranhão state, Brazil

Serrasalmus aff. brandtii e Pygocentrus nattereri são duas espécies de piranhas comuns no Lago de Viana, um lago formado a partir das inundações do Rio Pindaré, tributário da margem esquerda do Rio Mearim. Uma amostra composta por 249 exemplares destas espécies foi estudada a fim de identificar a composição da dieta e as estratégias alimentares empregadas pelas duas. Os peixes foram coletados trimestralmente durante um ano, entre março de 1998 e março de 1999, usando redes de emalhar e tarrafas. A composição qualitativa da dieta foi analisada com uso do método de freqüência de ocorrência. As estratégias alimentares empregadas pelas espécies foram inferidas através do método gráfico de Costello (1990), modificado por Amundsen et al. (1996). Os resultados indicaram que peixes foi o item encontrado com maior freqüência nos estômagos de ambas as espécies, seguido de matéria vegetal, com maior participação na dieta de P. nattereri. Os recursos peixes e vegetais tenderam a apresentar relações inversas quando comparados com as diferentes classes de comprimento. A análise gráfica da dieta de P. nattereri e S. aff. brandtii sugere que a maioria dos indivíduos utiliza vários recursos simultaneamente. Uma análise multivariada de índices ecomorfológicos indicou que as espécies são segregadas com relação à habilidade natatória, posição ocupada na coluna d'água e tamanho relativo das presas.<br>Serrasalmus aff. brandtii and Pygocentrus nattereri are two species of piranhas, both common in the Viana Lake, which is formed by the flooding waters of the Pindaré River, a tributary of the left bank of the Mearim River. A sample composed of 249 specimens of these species was studied in order to identify diet composition and feeding strategies employed by the two species. The piranhas were captured thrimonthly, during one year from March 1998 to March 1999 using gill and fishing nets of different mesh sizes. The qualitative composition of the diet was analysed through the frequency of occurrence method. The feeding strategies employed by the piranhas were inferred using the graphic method of Costello (1990), modified by Amundsen et al. (1996). The results indicated that fish was the main food item in the stomach contents of the two species, followed by plant material, more in the P. nattereri. Fish and plant material showed different relationships in relation to piranhas's length classes. The graphic analysis of the feeding strategies employed by P. nattereri and S. aff. brandtii suggest a generalist habit, wide width niche with strong participation of the within-phenotype component, which indicated that the majority of individuals of these species use several resources simultaneously. A multivariate analysis of the ecomorphological index indicated that the species are discriminated by swimming ability, water column position and relative prey size