320 research outputs found
The Estimation of Parameters in Regression Functions Subject to Certain Restraints
We consider two types of problems in maximum likelihood estimation of parameters of linear functions subject to certain restraints. One is a family of lines with equal slopes or intercepts; the other is a pair of lines constrained to meet at a predetermined point. In the case of normally distributed errors with equal variances within each set, the solutions are identical with least squares solutions. In addition to linear functions, non-linear functions which are transformable to linearity may be treated under these methods
Exploring Counselor Educator Dispositions Related to Teaching
This study explores students’ preferences for counselor educator (CE) teaching dispositions. Forty-eight counselor education students completed a Q sort and answered post-sort qualitative questions. The study found four types of student preferences: a focus on experiential teaching, a focus on content and affect orientation, a focus on the educator-student relationship, and a focus on developing clinical skills. Also among the findings are a set of items that were unanimously unimportant to the participants: CE engagement in research and gatekeeping. These preferences are situated within the scholarship of teaching and learning and evidence-based practices in counselor education. Practical and research implications are shared
Supplemental chromium and revaccination effects on performance and health of newly weaned calves
Two trials were conducted to evaluate the effects of chromium (Cr) supplementation (4 mg/hd/day in a yeast form) or no supplementation, with or without revaccination with a modified live viral vaccine at 9 days postweaning o n performance, health, and ability to withstand an IBR challenge infection. In Trial 1, Cr supplementation had no effect on performance of newly weaned calves in a 28-day receiving study, but reduced the incidence of respiratory disease by 37%. Revaccinati on depressed dry matter intake and had no effect on animal health. In trial 2, blood plasma levels of cortisol and ACTH (stress hormones ) were measured at 6 and 26 days postweaning. Cortisol levels were unaffected by treatment or by time after weaning. Plasma ACTH conc e ntrations were lower at 26 vs 6 days postweaning, and were reduced at 26 days by revaccination. Despite some slight differences in rectal temperature, treatment did not appear to affect the animals\u27 ability to withstand a live IBR challenge. We concluded that supplemental Cr was beneficial in reducing the incidence of bovine respiratory disease, although mediation of stress hormones was not involved. Revaccination of newly weaned calves with a modified live viral vaccine showed no performance or health benefit
Clostridioides difficile binary toxin binding component (cdtb) increases virulence in a hamster model
Background
Clostridioides difficile is the leading cause of hospital-acquired gastrointestinal infection, in part due to the existence of binary toxin (CDT)-expressing hypervirulent strains. Although the effects of the CDT holotoxin on disease pathogenesis have been previously studied, we sought to investigate the role of the individual components of CDT during in vivo infection.
Methods
To determine the contribution of the separate components of CDT during infection, we developed strains of C difficile expressing either CDTa or CDTb individually. We then infected both mice and hamsters with these novel mutant strains and monitored them for development of severe illness.
Results
Although expression of CDTb without CDTa did not induce significant disease in a mouse model of C difficile infection, we found that complementation of a CDT-deficient C difficile strain with CDTb alone restored virulence in a hamster model of C difficile infection.
Conclusions
Overall, this study demonstrates that the binding component of C difficile binary toxin, CDTb, contributes to virulence in a hamster model of infection
Repeated cycles of Clostridium-directed enzyme prodrug therapy result in sustained antitumour effects in vivo
The unique properties of the tumour microenvironment can be exploited by using recombinant anaerobic clostridial spores as highly selective gene delivery vectors. Although several recombinant Clostridium species have been generated during the past decade, their efficacy has been limited. Our goal was to substantially improve the prospects of clostridia as a gene delivery vector. Therefore, we have assessed a series of nitroreductase (NTR) enzymes for their capacity to convert the innocuous CB1954 prodrug to its toxic derivative. Among the enzymes tested, one showed superior prodrug turnover characteristics. In addition, we established an efficient gene transfer procedure, based on conjugation, which allows for the first time genetic engineering of Clostridium strains with superior tumour colonisation properties with high success rates. This conjugation procedure was subsequently used to create a recombinant C. sporogenes overexpressing the isolated NTR enzyme. Finally, analogous to a clinical setting situation, we have tested the effect of multiple consecutive treatment cycles, with antibiotic bacterial clearance between cycles. Importantly, this regimen demonstrated that intravenously administered spores of NTR-recombinant C. sporogenes produced significant antitumour efficacy when combined with prodrug administration
Multi-timescale analysis of a metabolic network in synthetic biology: a kinetic model for 3-hydroxypropionic acid production via beta-alanine
A biosustainable production route for 3-hydroxypropionic acid (3HP), an important platform chemical, would allow 3HP to be produced without using fossil fuels. We are interested in investigating a potential biochemical route to 3HP from pyruvate through b -alanine and, in this paper, we develop and solve a mathematical model for the reaction kinetics of the metabolites involved in this pathway. We consider two limiting cases, one where the levels of pyruvate are never replenished, the other where the levels of pyruvate are continuously replenished and thus kept constant. We exploit the natural separation of both the time scales and the metabolite concentrations to make significant asymptotic progress in understanding the system without resorting to computationally expensive parameter sweeps. Using our asymptotic results, we are able to predict the most important reactions to maximize the production of 3HP in this system while reducing the maximum amount of the toxic intermediate compound malonic semialdehyde present at any one time, and thus we are able to recommend which enzymes experimentalists should focus on manipulating
Capturing the essence of folding and functions of biomolecules using Coarse-Grained Models
The distances over which biological molecules and their complexes can
function range from a few nanometres, in the case of folded structures, to
millimetres, for example during chromosome organization. Describing phenomena
that cover such diverse length, and also time scales, requires models that
capture the underlying physics for the particular length scale of interest.
Theoretical ideas, in particular, concepts from polymer physics, have guided
the development of coarse-grained models to study folding of DNA, RNA, and
proteins. More recently, such models and their variants have been applied to
the functions of biological nanomachines. Simulations using coarse-grained
models are now poised to address a wide range of problems in biology.Comment: 37 pages, 8 figure
Crowding Alone Cannot Account for Cosolute Effect on Amyloid Aggregation
Amyloid fiber formation is a specific form of protein aggregation, often resulting from the misfolding of native proteins. Aimed at modeling the crowded environment of the cell, recent experiments showed a reduction in fibrillation halftimes for amyloid-forming peptides in the presence of cosolutes that are preferentially excluded from proteins and peptides. The effect of excluded cosolutes has previously been attributed to the large volume excluded by such inert cellular solutes, sometimes termed “macromolecular crowding”. Here, we studied a model peptide that can fold to a stable monomeric β-hairpin conformation, but under certain solution conditions aggregates in the form of amyloid fibrils. Using Circular Dichroism spectroscopy (CD), we found that, in the presence of polyols and polyethylene glycols acting as excluded cosolutes, the monomeric β-hairpin conformation was stabilized with respect to the unfolded state. Stabilization free energy was linear with cosolute concentration, and grew with molecular volume, as would also be predicted by crowding models. After initiating the aggregation process with a pH jump, fibrillation in the presence and absence of cosolutes was followed by ThT fluorescence, transmission electron microscopy, and CD spectroscopy. Polyols (glycerol and sorbitol) increased the lag time for fibril formation and elevated the amount of aggregated peptide at equilibrium, in a cosolute size and concentration dependent manner. However, fibrillation rates remained almost unaffected by a wide range of molecular weights of soluble polyethylene glycols. Our results highlight the importance of other forces beyond the excluded volume interactions responsible for crowding that may contribute to the cosolute effects acting on amyloid formation
A Coarse-Grained Biophysical Model of E. coli and Its Application to Perturbation of the rRNA Operon Copy Number
We propose a biophysical model of Escherichia coli that predicts growth rate
and an effective cellular composition from an effective, coarse-grained
representation of its genome. We assume that E. coli is in a state of balanced
exponential steadystate growth, growing in a temporally and spatially constant
environment, rich in resources. We apply this model to a series of past
measurements, where the growth rate and rRNA-to-protein ratio have been
measured for seven E. coli strains with an rRNA operon copy number ranging from
one to seven (the wild-type copy number). These experiments show that growth
rate markedly decreases for strains with fewer than six copies. Using the
model, we were able to reproduce these measurements. We show that the model
that best fits these data suggests that the volume fraction of macromolecules
inside E. coli is not fixed when the rRNA operon copy number is varied.
Moreover, the model predicts that increasing the copy number beyond seven
results in a cytoplasm densely packed with ribosomes and proteins. Assuming
that under such overcrowded conditions prolonged diffusion times tend to weaken
binding affinities, the model predicts that growth rate will not increase
substantially beyond the wild-type growth rate, as indicated by other
experiments. Our model therefore suggests that changing the rRNA operon copy
number of wild-type E. coli cells growing in a constant rich environment does
not substantially increase their growth rate. Other observations regarding
strains with an altered rRNA operon copy number, such as nucleoid compaction
and the rRNA operon feedback response, appear to be qualitatively consistent
with this model. In addition, we discuss possible design principles suggested
by the model and propose further experiments to test its validity
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