The different phonetic systems.

Typewritten sheets in cover. Thesis (M.A.)--Boston University Bibliography: p. 41-43

The Democratization and Development of Cell-free Protein Synthesis

Cell-free protein synthesis (CFPS) using crude lysates has developed into a robust platform technology over the last 60 years to express numerous types of recombinant proteins. The open-nature, elimination of reliance on cell viability, and focus of all energy towards production of the protein of interest represent substantial advantages of CFPS over in vivo protein expression methods. CFPS has provided new opportunities across a series of research fields that include metabolic engineering, therapeutic and vaccine development, education, biosensors, and many more. In recent years, optimizations of CFPS have even allowed the platform to reach the industrial level of protein production. Although there have been many advancements toward CFPS development, the democratization of the platform to a wide variety of educational, research, and industrial institutions has lacked due to an absence of resources for new users as well as a limited number of developments toward redesigning the tedious and time-consuming protocols to generate robust cell extract. To address these challenges to CFPS implementation, a comprehensive review spanning numerous cell lines with their respective applications, methodologies, and reaction formats were provided in addition to detailed protocols outlining the process of going from E. coli cells to a completed CFPS reaction. Together, these resources provide the scientific community with easily accessible resources for CFPS implementation. Moreover, the aforementioned protocols were redesigned from a four-day process into one that may be completed in under 24-hour’s time with very little researcher oversight. The resulting workflow maintained the robustness of prior methods but generated 400% more extract compared to traditional methods via a set-it-and-forget-it approach. To date, the works presented herein have garnered tremendous viewership from the CFPS research community with a substantial following among all three of the articles. Moving forward, I anticipate that these works will continue to bring new users into the CFPS field through the ease of access to these resources and through the advance of the simplistic and reproducible new workflow for preparation of robust E. coli cell extract

Some Notes on the Voges-Proskauer Reaction

The formation of acetyl-methyl-carbinol by members of the genus Aerobacter was detected in six-hour cultures. The procedure involved the heavy inoculation of young agar slant cultures into approximately three cc. Difeo M.R.-V.P. medium, incubation at 30° C. for six hours and using 0.3 per cent creatine in 40 per cent KOH as the test reagent. Seventy per cent of 202 cultures tested produced positive results within one hour after the addition of the test reagent and over ninety-nine per cent within four hours. The creatine KOH solution was satisfactorily effective for three weeks when kept at ordinary laboratory conditions and for six weeks when stored at ice-box temperatures

Influence of Bile and Brilliant Green on Rate of Growth of Colon Bacilli

Evaporated bile may inhibit or stimulate growth of colon bacilli depending on concentration of bile, reaction of the medium, and the strain of organism employed. With all samples of bile employed, a concentration of 2.0% was found to accelerate growth whereas 5.0% was inhibitory in acid media (about pH 6.0), and stimulating in alkaline media (pH 7.3-7.8)

Relative Costs of Home Made and Dehydrated Nutrient Agar

Batches of 1, 4, and 7 liters of nutrient agar were prepared from the ingredients according to Standard Methods, and the costs compared with those of similar batches made from commercial dehydrated nutrient agar. The costs of ingredients, labor, and loss due to filtration, were the only items considered

Some Notes on Trickling Filters in the Purification of Creamery Wastes

Creamery wastes differ markedly from ordinary sewage in that a considerable portion (about 50%) of its organic content is the carbohydrate lactose. The acid produced by the anaerobic decomposition of the milk sugar markedly inhibits, and may completely prevent proteolysis thus seriously interfering with bacterial action in the septic tanks

Quadriceps Muscle Layer Thickness in Kidney Transplant Recipients: A Potential Measure of Frailty and Sarcopenia

Introduction: Frailty and sarcopenia are related concepts that can impact outcomes after kidney transplantation. Measures of these two entities and new/emerging metrics of sarcopenia remain to be validated. Methods: In a prospective cohort study, kidney and kidney-pancreas transplant recipients were assessed at the time of transplant with the Physical Frailty Phenotype, bioimpedance analysis, quadriceps muscle layer thickness (QMLT), and CT. The impact on length of stay (LOS), prediction of frailty/sarcopenia, and relative concordance of metrics were analyzed. Results: Low QMLT, a putative surrogate of sarcopenia/frailty, was more frequently associated with longer LOS (\u3e14 days) after transplant. Additionally, QMLT was predictive of low muscle mass but insufficient at discriminating true sarcopenia, while CT of the abdominal muscles at the L3 level showed good discrimination for sarcopenia. Conclusions: Further exploration of QMLT and cut-offs for CT and functional metrics in the transplant population are required for future studies and risk stratification

Determination of biomembrane bending moduli in fully atomistic simulations.

The bilayer bending modulus (Kc) is one of the most important physical constants characterizing lipid membranes, but precisely measuring it is a challenge, both experimentally and computationally. Experimental measurements on chemically identical bilayers often differ depending upon the techniques employed, and robust simulation results have previously been limited to coarse-grained models (at varying levels of resolution). This Communication demonstrates the extraction of Kc from fully atomistic molecular dynamics simulations for three different single-component lipid bilayers (DPPC, DOPC, and DOPE). The results agree quantitatively with experiments that measure thermal shape fluctuations in giant unilamellar vesicles. Lipid tilt, twist, and compression moduli are also reported

Some Observations on the Germicidal Efficiencies of Alkalies

A technique is described for measuring the relative germicidal efficiencies of strong alakalies. For a given alkali, the germicidal efficiency increases with decreasing H +ion concentration, but the H +ion concentration is not suitable as an index of the germicidal powers of different alkalies. The addition of various salts to sodium hydroxide increased the germicidal effects of the alkali

Adapting Cell-Free Protein Synthesis as a Platform Technology for Education

Cell-free protein synthesis (CFPS) has emerged as an enabling biotechnology for research and biomanufacturing as it allows for the production of protein without the need for a living cell. Applications of CFPS include the construction of libraries for functional genomics and structural biology, the production of personalized medicine, and the expression of virus-like particles. The absence of a cell wall provides an open platform for direct manipulation of the reaction conditions and biological machinery. This project focuses on adapting the CFPS biotechnology to the classroom, making a hands-on bioengineering approach to learning protein synthesis accessible to students grades K-16 through developing an affordable ‘protein synthesis classroom kit’. To address the barrier of cost to entering the classroom, CFPS reaction was reformulated with the goal to reduce costs while optimizing protein yield. An inexpensive sugar was added to the reaction in varying concentrations for its potential to recycle inorganic phosphate and regenerate ATP. Phosphoenolpyruvate (PEP), an expensive energy source, was replaced with a lower concentration of 3-phosphoglyceric acid (3-PGA). We determined that adding the sugar within the range of 10-30mM did not have a significant effect on high-performing cell extracts grown in 2xYTPG for the PEP energy system, and had a slight boost to protein yield at a concentration of 10mM for cell extract grown in 2xYTP media. Although the 3-PGA system yielded less protein than the PEP system, the sugar combined with 3-PGA contributed greater percentage gains for cell extract grown in both media when compared to controls. Future work may include whether the sugar and 3-PGA worked in tandem or independently. Understanding gained from these experiments would contribute to optimizing protein yield and reduce the cost barrier for classroom use