IN \u3ci\u3eVITRO\u3c/i\u3e ASSAY DEVELOPMENT AS AN ALTERNATIVE TO THE USE OF LABORATORY ANIMALS FOR MEASURING \u3ci\u3eCLOSTRIDIUM PERFRINGENS\u3c/i\u3e TYPE C TOXOID

Clostridium perfringens type C produces beta toxin, which is a primary virulence determinate that can cause necrotic enteritis in neonatal animals. Vaccines directed against C. perfringens toxins have shown to be efficacious in preventing disease. However, the production of a commercial vaccine requires not only an efficacious antigen, but also testing methods for quantifying antigens and potency on final product. This research was undertaken to fulfill a need for the reduction in small animal usage to produce and deliver vaccines. The current testing methods are performed in vivo according to the Code of Federal Regulations number 9 (9CFR) United States Department of Agriculture (USDA) and European Pharmacopeia (EP) monograph guidelines. With limited in vitro alternatives, the aim of this work was to develop an enzyme-linked immunosorbent assay (ELISA) to be able to measure C. perfringens type C beta toxoid, independent of laboratory animals. To accomplish this, investigational polyclonal and monoclonal antibody candidates were screened for their specificity to beta toxin and toxoid through western blot and ELISA. Additionally, through a neutralization assay, the capability to neutralize beta toxin was demonstrated. Through this process, one polyclonal antibody and one monoclonal antibody were selected and an in-process ELISA for quantifying beta toxoid was developed. By using the ELISA to quantify beta toxoid, the current practice of testing in animals could be reduced. Advisor: Greg A. Somervill

Diffusion Tensor Imaging for Assessment of Response to Neoadjuvant Chemotherapy in Patients With Breast Cancer.

In this study, the prognostic significance of tumor metrics derived from diffusion tensor imaging (DTI) was evaluated in patients with locally advanced breast cancer undergoing neoadjuvant therapy. DTI and contrast-enhanced magnetic resonance imaging were acquired at 1.5 T in 34 patients before treatment and after 3 cycles of taxane-based therapy (early treatment). Tumor fractional anisotropy (FA), principal eigenvalues (λ1, λ2, and λ3), and apparent diffusion coefficient (ADC) were estimated for tumor regions of interest drawn on DTI data. The association between DTI metrics and final tumor volume change was evaluated with Spearman rank correlation. DTI metrics were investigated as predictors of pathological complete response (pCR) by calculating the area under the receiver operating characteristic curve (AUC). Early changes in tumor FA and ADC significantly correlated with final tumor volume change post therapy (ρ = -0.38, P = .03 and ρ = -0.71, P < .001, respectively). Pretreatment tumor ADC was significantly lower in the pCR than in the non-pCR group (P = .04). At early treatment, patients with pCR had significantly higher percent changes of tumor λ1, λ2, λ3, and ADC than those without pCR. The AUCs for early percent changes in tumor FA and ADC were 0.60 and 0.83, respectively. The early percent changes in tumor eigenvalues and ADC were the strongest DTI-derived predictors of pCR. Although early percent change in tumor FA had a weak association with pCR, the significant correlation with final tumor volume change suggests that this metric changes with therapy and may merit further evaluation

Effectiveness of maxillary protraction using a hybrid hyrax-facemask combination: A controlled clinical study

OBJECTIVE: To evaluate the treatment effects of a hybrid hyrax-facemask (FM) combination in growing Class III patients. MATERIAL AND METHODS: A sample of 16 prepubertal patients (mean age, 9.5 ± 1.6 years) was investigated by means of pre- and posttreatment cephalograms. The treatment comprised rapid palatal expansion with a hybrid hyrax, a bone- and toothborne device. Simultaneously, maxillary protraction using an FM was performed. Mean treatment duration was 5.8 ± 1.6 months. The treatment group was compared with a matched control group of 16 untreated Class III subjects. Statistical comparisons were performed with the Mann-Whitney U-test. RESULTS: Significant improvement in skeletal sagittal values could be observed in the treatment group over controls: SNA: 2.4°, SNB: −1.7°, Co-Gn: −2.3 mm, Wits appraisal: 4.5 mm. Regarding vertical changes, maintenance of vertical growth was obtained as shown by a small nonsignificant increase of FMA and a small significant decrease of the Co-Go-Me angle. CONCLUSIONS: The hybrid hyrax-FM combination was found to be effective for orthopedic treatment in growing Class III patients in the short term. Favorable skeletal changes were observed both in the maxilla and in the mandible. No dentoalveolar compensations were found

The impact of sea-level rise on tidal characteristics around Australia

An established tidal model, validated for present-day conditions, is used to investigate the effect of large levels of sea-level rise (SLR) on tidal characteristics around Australasia. SLR is implemented through a uniform depth increase across the model domain, with a comparison between the implementation of coastal defences or allowing low-lying land to flood. The complex spatial response of the semi-diurnal M2 constituent does not appear to be linear with the imposed SLR. The most predominant features of this response are the generation of new amphidromic systems within the Gulf of Carpentaria and large-amplitude changes in the Arafura Sea, to the north of Australia, and within embayments along Australia's north-west coast. Dissipation from M2 notably decreases along north-west Australia but is enhanced around New Zealand and the island chains to the north. The diurnal constituent, K1, is found to decrease in amplitude in the Gulf of Carpentaria when flooding is allowed. Coastal flooding has a profound impact on the response of tidal amplitudes to SLR by creating local regions of increased tidal dissipation and altering the coastal topography. Our results also highlight the necessity for regional models to use correct open boundary conditions reflecting the global tidal changes in response to SLR.</p

Learning and attention increase visual response selectivity through distinct mechanisms

Selectivity of cortical neurons for sensory stimuli can increase across days as animals learn their behavioral relevance and across seconds when animals switch attention. While both phenomena occur in the same circuit, it is unknown whether they rely on similar mechanisms. We imaged primary visual cortex as mice learned a visual discrimination task and subsequently performed an attention switching task. Selectivity changes due to learning and attention were uncorrelated in individual neurons. Selectivity increases after learning mainly arose from selective suppression of responses to one of the stimuli but from selective enhancement and suppression during attention. Learning and attention differentially affected interactions between excitatory and PV, SOM, and VIP inhibitory cells. Circuit modeling revealed that cell class-specific top-down inputs best explained attentional modulation, while reorganization of local functional connectivity accounted for learning-related changes. Thus, distinct mechanisms underlie increased discriminability of relevant sensory stimuli across longer and shorter timescales

Comparison of base-line and chemical-induced transcriptomic responses in HepaRG and RPTEC/TERT1 cells using TempO-Seq

The utilisation of genome-wide transcriptomics has played a pivotal role in advancing the field of toxicology, allowing the mapping of transcriptional signatures to chemical exposures. These activities have uncovered several transcriptionally regulated pathways that can be utilised for assessing the perturbation impact of a chemical and also the identification of toxic mode of action. However, current transcriptomic platforms are not very amenable to high-throughput workflows due to, high cost, complexities in sample preparation and relatively complex bioinformatic analysis. Thus, transcriptomic investigations are usually limited in dose and time dimensions and are, therefore, not optimal for implementation in risk assessment workflows. In this study, we investigated a new cost-effective, transcriptomic assay, TempO-Seq, which alleviates the aforementioned limitations. This technique was evaluated in a 6-compound screen, utilising differentiated kidney (RPTEC/TERT1) and liver (HepaRG) cells and compared to non-transcriptomic label-free sensitive endpoints of chemical-induced disturbances, namely phase contrast morphology, xCELLigence and glycolysis. Non-proliferating cell monolayers were exposed to six sub-lethal concentrations of each compound for 24 h. The results show that utilising a 2839 gene panel, it is possible to discriminate basal tissue-specific signatures, generate dose-response relationships and to discriminate compound-specific and cell type-specific responses. This study also reiterates previous findings that chemical-induced transcriptomic alterations occur prior to cytotoxicity and that transcriptomics provides in depth mechanistic information of the effects of chemicals on cellular transcriptional responses. TempO-Seq is a robust transcriptomic platform that is well suited for in vitro toxicity experiments.Horizon 2020(H2020)68100

Ligand-induced type II interleukin-4 receptor dimers are sustained by rapid re-association within plasma membrane microcompartments

Biological and Soft Matter Physic