What a long trip it's been: Trying to create a CSN-5 and GLH-1 mutant

Abstract only availableIt is known in Caenorhabditis elegans that non-membrane-bound cytoplasmic complexes called P granules are specific to the germline lineage. P granules contain four GLH proteins, germline RNA helicases, which are critical to the development of the germline lineage and the fertility of the nematode. The CSN-5, KGB-1, ZYX-1, and PAN-1 proteins all interact with the GLH proteins as found with a yeast two-hybrid screen, with CSN-5 and KGB-1 both critical for meiotic germline development. The Bennett lab has not been able to isolate a mutant csn-5, nor have they been able to create a knockout glh-1 mutant strain that is null. This summer we have been attempting to create both a new glh-1and a csn-5 mutant strain. In looking for the csn-5 and glh-1 deletion strains, I have been using a library technique that has been successfully used in the Bennett lab for several years. C. elegans wild type worms are grown to adults on plates and their eggs are harvested. Approximately 500,000-600,000 of these eggs are hatched and mutagenized by exposure to the chemical trimethylpsoralen and UV light. After 24 hours, 750,000-1 million progeny of the mutagenized worms are collected and distributed among 960 plates. Once adults, DNA is isolated from some of the worms on each plate and tested by PCR looking for a deletion band smaller than the wild type. However, this summer the PCR and gel electrophoresis have not quite gone as planned. Many problems arose with primers, solutions, and with the DNA. Much time has been spent trouble shooting. Recently gels have been working and three potential deletions were found and tested. Upon confirmation, it was discovered that none of the three are the deletions we were looking for.Life Sciences Undergraduate Research Opportunity Progra

Recognition-mediated hydrogel swelling controlled by interaction with a negative thermoresponsive LCST polymer

Most polymeric thermoresponsive hydrogels contract upon heating beyond the lower critical solution temperature (LCST) of the polymers used. Herein, we report a supramolecular hydrogel system that shows the opposite temperature dependence. When the non-thermosesponsive hydrogel NaphtGel, containing dialkoxynaphthalene guest molecules, becomes complexed with the tetra cationic macrocyclic host CBPQT4+, swelling occurred as a result of host–guest complex formation leading to charge repulsion between the host units, as well as an osmotic contribution of chloride counter-ions embedded in the network. The immersion of NaphtGel in a solution of poly(N-isopropylacrylamide) with tetrathiafulvalene (TTF) end groups complexed with CBPQT4+ induced positive thermoresponsive behaviour. The LCST-induced dethreading of the polymer-based pseudorotaxane upon heating led to transfer of the CBPQT4+ host and a concomitant swelling of NaphtGel. Subsequent cooling led to reformation of the TTF-based host–guest complexes in solution and contraction of the hydrogel

Stimulus-responsive delivery systems for enabling the oral delivery of protein therapeutics exhibiting high isoelectric point

textProtein therapeutics offer numerous advantages over small molecule drugs and are rapidly becoming one of the most prominent classes of therapeutics. Unfortunately, they are delivered almost exclusively by injection due to biological obstacles preventing high bioavailability via the oral route. In this work, numerous approaches to overcoming these barriers are explored. PH-Responsive poly(itaconic acid-co-N-vinylpyrrolidone) (P(IA-co-NVP)) hydrogels were synthesized, and the effects of monomer ratios, crosslinking density, microparticle size, protein size, and loading conditions were systematically evaluated using in vitro tests. P(IA-co-NVP) hydrogels demonstrated up to 69% greater equilibrium swelling at neutral conditions than previously-studied poly(methacrylic acid-co-N-vinylpyrrolidone) hydrogels and a 10-fold improvement in time-sensitive swelling experiments. Furthermore, P(IA-co-NVP) hydrogel microparticles demonstrated up to a 2.7-fold improvement in delivery of salmon calcitonin (sCT) compared to methacrylic acid-based systems, with a formulation comprised of a 1:2 ratio of itaconic acid to N-vinylpyrrolidone demonstrating the greatest delivery capability. Vast improvement in delivery capability was achieved using reduced ionic strength conditions during drug loading. Use of a 1.50 mM PBS buffer during loading yielded an 83-fold improvement in delivery of sCT compared to a standard 150 mM buffer. With this improvement, a daily dose of sCT could be provided using P(IA-co-NVP) microparticles in one standard-sized gel capsule. P(IA-co-NVP) was also tested with larger proteins urokinase and Rituxan. Crosslinking density provided a facile method for tuning hydrogels to accommodate a wide range of protein sizes. The effects of protein PEGylation were also explored. PEGylated sCT displayed lower release from P(IA-co-NVP) microparticles, but displayed increased apparent permeability across a Caco-2 monolayer by two orders of magnitude. Therefore, PEG-containing systems could yield high bioavailability of orally delivered proteins. Finally, a modified SELEX protocol for cellular selection of transcellular transport-initiating aptamers was developed and used to identify aptamer sequences showing enhanced intestinal perfusion. Over three selection cycles, the selected aptamer library showed significant increases in absorption, and from an initial library of 1.1 trillion sequences, 5-10 sequences were selected that demonstrated up to 10-fold amplification compared to the naïve library. These sequences could provide a means of overcoming the significant final barrier of intestinal absorption.Chemical Engineerin

Manufacturer changes lead to clinically important differences between two editions of the TNO stereotest

Purpose: Stereoacuity tests used in clinical practice should be repeatable and reproducible. However, it has been observed in a clinical setting that new editions of the TNO stereotest appear to give different values from those obtained using previous versions. The purpose of the present research was to investigate this observation. Methods: One hundred and twenty-one Dutch subjects, 88 (73%) females and 33 (27%) males, with an average age of 34.0 years (range 18–55) had their stereoacuity measured using two different versions of the TNO stereoacuity test (TNO 13 and TNO 15). The TNO was tested in a counterbalanced order so that consecutive subjects started with alternate editions to avoid bias. Results: There was a significant difference (p < 0.001) between the median value for stereoacuity measured with TNO 13 (30 s of arc) and TNO 15 (60 s of arc). The bias between the two test versions was -0.23 Log arcseconds (95% limits of the differences: 0.15 to -0.60 Log arcseconds). Conclusion: This study reveals that results obtained with two different editions of a commonplace stereoacuity test are not comparable. New versions come on the market at regular intervals and the assumption that they will give the same results as previous versions may not be valid. Besides the statistically significant difference between the TNO 13 and TNO 15, the Bland-Altman plot also showed a considerable bias and the 95% limits of the differences between the TNO 13 and TNO 15 are more than two steps on the Log arcsecond scale. This difference between two editions of the TNO stereotests is not clinically acceptable and therefore it is inappropriate to use the two versions of the test interchangeably. It is important in both research and clinical records to specify the edition of the TNO test used

Developing skills with pre-service teachers

Teachers need to equip students with visual, written and verbal communication skills

Examining the Interaction of Adverse Childhood Events, Dental Issues, Asthma and Autism

Dissertation supervised by Dr. Rebecca Kronk Purpose: Adverse childhood events (ACEs) have a profound and long-term effect on the health outcomes of many children and adults. However, few studies have focused on specific ACEs and health outcomes in children with autism spectrum disorder (ASD). The aim of the study was to investigate if ASD moderated the relationship between ACEs and specific health outcomes in children who had been the subject of a child welfare system report. Design and Methods: Using the second National Survey of Child and Adolescent Well-Being (NSCAW II), a secondary analysis was performed on a sample of 80 children with diagnosed autism and 5,698 children without autism. ACEs score criteria included physical abuse, sexual abuse, neglect, psychological aggression, domestic violence, parental substance abuse and mental health. Health outcome variables included dental issues and asthma. Child characteristics of age, gender, race and poverty level were also included in the analysis. Results: Chi-square analysis indicated a significantly higher percentage of children with ASD whose parents reported psychological abuse compared to children without autism ꭓ2 (28.03, df = 74, F = 11.08). Children with ASD have a noteworthy number of ACEs compared to children without autism (p = 0.00). Logistic regression found having an ACE increased the odds of having dental issues 2.80 times compared to not having an ACE (odds ratio [OR] = 2.8, standard error [SE] = 1.27, p \u3c 0.5). Conclusions: A diagnosis of ASD does not appear to moderate dental issues or asthma in this sample of children. Children with ASD are at risk of harmful and chronic health outcomes due to ACEs. Practice Implications: All healthcare providers should include ACE screening when assessing children with ASD. If completed early and regularly, preventative measures may be employed that help support families and may avoid entrance into the child welfare system

Cenobium 1982

Cenobium began publication in 1977 and continued through 1990. Produced by students, the yearbooks are a chronicle of the daily life and the special events experienced by veterinary students at the LSU School of Veterinary Medicine.https://digitalcommons.lsu.edu/cenobium/1005/thumbnail.jp

Oxygen persufflation as adjunct in liver preservation (OPAL): Study protocol for a randomized controlled trial

<p>Abstract</p> <p>Background</p> <p>Early graft dysfunction due to preservation/reperfusion injury represents a dramatic event after liver transplantation. Enhancement of donor organ criteria, in order to cope with the ever increasing donor shortage, further increases graft susceptibility to ischemic alterations.</p> <p>Major parts of post-preservation injury, however, occur at the time of warm reperfusion but not during ischemic storage; successful reperfusion of ischemic tissue in turn depends on an adequate redox and intracellular signal homeostasis. The latter has been shown experimentally to be favorably influenced by oxygen persufflation within short time spans. Thus viability of marginally preserved liver grafts could still be augmented by transient hypothermic reconditioning <b><it>even after </it></b>normal procurement and static cold storage. The present study is aimed to confirm the conceptual expectations, that hypothermic reconditioning by gaseous oxygen persufflation is a useful method to suppress injurious cellular activation cascades and to improve post-ischemic recovery of marginally preserved liver grafts.</p> <p>Methods/Design</p> <p>OPAL is a prospective single center randomized proof of concept study, including two parallel groups in a total of 116 liver transplant patients. The effect of an in hospital treatment of the isolated liver graft by 2 hours of oxygen persufflation immediately prior to transplantation will be assesses as compared to standard procedure (cold storage without further intervention). The primary endpoint is the peak transaminase serum level (AST) during the first three days after transplantation as a surrogate readout for parenchymal liver injury. Other outcomes comprise patient and graft survival, time of intensive care requirement, hepatic tissue perfusion 1h after revascularisation, early onset of graft dysfunction based on coagulation parameters, as well as the use of a refined scoring-system for initial graft function based on a multi-parameter (AST, ALT, Quick and bilirubin) score. Furthermore, the effect of OPAL on molecular pathways of autophagy and inflammatory cell activation will be evaluated. Final analysis will be based on all participants as randomized (intention to treat).</p> <p>Trial Registration</p> <p>Current Controlled Trials <a href="http://www.controlled-trials.com/ISRCTN00167887">ISRCTN00167887</a></p