Lights, camera, action: Microbiology laboratory teaching in the spotlight

[EN] Broadening of access to higher education, leading to increasing class sizes, presents particular challenges in teaching specialised, laboratory-based subjects such as Microbiology. The Republic of Ireland has seen a 20% increase in undergraduate student numbers in the past ten years, with this trend set to continue in the near future. To complement traditional learning approaches, we have designed, produced and trialled a comprehensive suite of teaching videos that demonstrate common techniques taught in our Microbiology degree programme. The 42 videos, each of 4-9 minutes duration, were filmed in-house using a professional film maker. Videos were designed for viewing before linked laboratory sessions to increase student engagement, assist learners with little prior technical experience to process core concepts, and improve the quality of hands-on practical training in the laboratory. Student reaction to a pilot release was exceptionally positive, underlining the videos’ effectiveness for visual learners and the added value of the content due to its bespoke nature. The complete video collection will be amalgamated into our B.Sc. programme in 2019-20. The initiative is expected to enhance students’ experience in hands-on laboratory sessions, promote active learning by blending video into traditional teaching programmes, and support reflective study through their availability.The NUI Galway Student Project Fund is thanked for financial support.Lacey, K.; Wall, JG. (2019). Lights, camera, action: Microbiology laboratory teaching in the spotlight. En HEAD'19. 5th International Conference on Higher Education Advances. Editorial Universitat Politècnica de València. 939-946. https://doi.org/10.4995/HEAD19.2019.9429OCS93994

Recurrent lateral inhibitory spiking networks for speech enhancement

Automatic speech recognition accuracy is affected adversely by the presence of noise. In this paper we present a novel noise removal and speech enhancement technique based on spiking neural network processing of speech data. The spiking network has a recurrent lateral topology that is biologically inspired, specifically by the inhibitory cells of the cochlear nucleus. The network can be configured for different acoustic environments and it will be demonstrated how the connectivity results in enhancement of temporal correlation between similar frequency bands and removal of uncorrelated noise sources. Demonstration of the speech enhancement capability will be provided with data taken from the TIMIT database with different levels of additive Gaussian white noise. Future directions for further development of this novel approach to noise removal and signal processing will also be discussed

Poly(ethylene glycol)-Based Hyperbranched Polymer from RAFT and Its Application as a Silver-Sulfadiazine-Loaded Antibacterial Hydrogel in Wound Care

A multifunctional branched copolymer was synthesized by Reversible Addition−Fragmentation Chain Transfer polymerization (RAFT) of poly(ethylene glycol) diacrylate (PEGDA Mn = 575) and poly(ethylene glycol) methyl methacrylate (PEGMEMA Mn = 500) at a feed molar ratio of 50:50. Proton nuclear magnetic resonance spectros- copy (1HNMR) confirmed a hyperbranched molecular structure and a high degree of vinyl functionality. An in situ cross-linkable hydrogel system was generated via a "click" thiol−ene-type Michael addition reaction of vinyl functional groups from this PEGDA/PEGMEMA copolymer system in combination with thiol-modified hyaluronic acid. Furthermore, encapsulation of antimicrobial silver sulfadiazine (SSD) into the copolymer system was conducted to create an advanced antimicrobial wound care dressing. This hydrogel demon- strated a sustained antibacterial activity against the bacterial strains Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli in comparison to the direct topical application of SSD. In addition, in vitro toxicology evaluations demonstrated that this hydrogel?with low concentrations of encapsulated SSD-supported the survival of embedded human adipose derived stem cells (hADSCs) and inhibited growth of the aforementioned pathogens. Here we demonstrate that this hydrogel encapsulated with a low concentration (1.0% w/v) of SSD can be utilized as a carrier system for stem cells with the ability to inhibit growth of pathogens and without adverse effects on hADSCs

Self-assembly of a barnacle cement protein into intertwined amyloid fibres and determination of their adhesive and viscoelastic properties.

peer reviewedThe stalked barnacle Pollicipes pollicipes uses a multi-protein cement to adhere to highly varied substrates in marine environments. We investigated the morphology and adhesiveness of a component 19 kDa protein in barnacle cement gland- and seawater-like conditions, using transmission electron microscopy and state-of-the art scanning probe techniques. The protein formed amyloid fibres after 5 days in gland-like but not seawater conditions. After 7-11 days, the fibres self-assembled under gland-like conditions into large intertwined fibrils of up to 10 µm in length and 200 nm in height, with a distinctive twisting of fibrils evident after 11 days. Atomic force microscopy (AFM)-nanodynamic mechanical analysis of the protein in wet conditions determined E' (elasticity), E'' (viscosity) and tan δ values of 2.8 MPa, 1.2 MPa and 0.37, respectively, indicating that the protein is a soft and viscoelastic material, while the adhesiveness of the unassembled protein and assembled fibres, measured using peak force quantitative nanomechanical mapping, was comparable to that of the commercial adhesive Cell-Tak™. The study provides a comprehensive insight into the nanomechanical and viscoelastic properties of the barnacle cement protein and its self-assembled fibres under native-like conditions and may have application in the design of amyloid fibril-based biomaterials or bioadhesives

Insights into the mode of action of a putative zinc transporter CzrB in thermus thermophilus

peer-reviewedThis paper was obtained through PEER (Publishing and the Ecology of European Research) http://www.peerproject.euThe crystal structures of the cytoplasmic domain of the putative zinc transporter CzrB in the apoand zinc-bound forms reported herein are consistent with the protein functioning in vivo as a homodimer. NMR, X-ray scattering and size exclusion chromatography provide support for dimer formation. Full-length variants of CzrB in the apo and zinc-loaded states were generated by homology modelling with the Zn2+ / H+ antiporter YiiP. The model suggests a way in which zinc binding to the cytoplasmic fragment creates a docking site to which a metallochaperone can bind for delivery and transport of its zinc cargo. Since the cytoplasmic domain may exist in the cell as an independent, soluble protein a proposal is advanced that it functions as a metallochaperone and that it regulates the zinc-transporting activity of the full-length protein. The latter requires that zinc binding becomes uncoupled from the creation of a metallochaperone-docking site on CzrB

Intestinal-derived ILCs migrating in lymph increase IFNγ production in response to Salmonella Typhimurium infection

Innate lymphoid cells (ILCs) are enriched in mucosae and have been described as tissue-resident. Interestingly, ILCs are also present within lymph nodes (LNs), in the interfollicular regions, the destination for lymph-migratory cells. We have previously shown that LN ILCs are supplemented by peripheral tissue-derived ILCs. Using thoracic duct cannulations, we here enumerate the intestinal lymph ILCs that traffic from the intestine to the mesenteric LNs (MLNs). We provide, for the first time, a detailed characterisation of these lymph-migratory ILCs. We show that all ILC subsets migrate in lymph, and while global transcriptional analysis reveals a shared signature with tissue-resident ILCs, lymph ILCs express migration-associated genes including S1PRs, SELL (CD62L) and CCR7. Interestingly, we discovered that while Salmonella Typhimurium infections do not increase the numbers of migrating ILCs, infection changes their composition and cytokine profile. Infection increases the proportions of RORyt+ T-bet+ ILCs, levels of IFNγ, and IFNγ/GM-CSF co-expression. Infection-induced changes in migratory ILCs are reflected in colon-draining MLN ILCs, where RORyt+ T-bet+ ILCs accumulate and display corresponding increased cytokine expression. Thus, we reveal that ILCs respond rapidly to intestinal infection and can migrate to the MLN where they produce cytokines

CDK19 is disrupted in a female patient with bilateral congenital retinal folds, microcephaly and mild mental retardation

Microcephaly, mental retardation and congenital retinal folds along with other systemic features have previously been reported as a separate clinical entity. The sporadic nature of the syndrome and lack of clear inheritance patterns pointed to a genetic heterogeneity. Here, we report a genetic analysis of a female patient with microcephaly, congenital bilateral falciform retinal folds, nystagmus, and mental retardation. Karyotyping revealed a de novo pericentric inversion in chromosome 6 with breakpoints in 6p12.1 and 6q21. Fluorescence in situ hybridization analysis narrowed down the region around the breakpoints, and the breakpoint at 6q21 was found to disrupt the CDK19 gene. CDK19 was found to be expressed in a diverse range of tissues including fetal eye and fetal brain. Quantitative PCR of the CDK19 transcript from Epstein–Barr virus-transformed lymphoblastoid cell lines of the patient revealed ~50% reduction in the transcript (p = 0.02), suggesting haploinsufficiency of the gene. cdk8, the closest orthologue of human CDK19 in Drosophila has been shown to play a major role in eye development. Conditional knock-down of Drosophila cdk8 in multiple dendrite (md) neurons resulted in 35% reduced dendritic branching and altered morphology of the dendritic arbour, which appeared to be due in part to a loss of small higher order branches. In addition, Cdk8 mutant md neurons showed diminished dendritic fields revealing an important role of the CDK19 orthologue in the developing nervous system of Drosophila. This is the first time the CDK19 gene, a component of the mediator co-activator complex, has been linked to a human disease

Adaptive immunity and neutralizing antibodies against SARS-CoV-2 variants of concern following vaccination in patients with cancer: the CAPTURE study

Coronavirus disease 2019 (COVID-19) antiviral response in a pan-tumor immune monitoring (CAPTURE) (NCT03226886) is a prospective cohort study of COVID-19 immunity in patients with cancer. Here we evaluated 585 patients following administration of two doses of BNT162b2 or AZD1222 vaccines, administered 12 weeks apart. Seroconversion rates after two doses were 85% and 59% in patients with solid and hematological malignancies, respectively. A lower proportion of patients had detectable titers of neutralizing antibodies (NAbT) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOC) versus wild-type (WT) SARS-CoV-2. Patients with hematological malignancies were more likely to have undetectable NAbT and had lower median NAbT than those with solid cancers against both SARS-CoV-2 WT and VOC. By comparison with individuals without cancer, patients with hematological, but not solid, malignancies had reduced neutralizing antibody (NAb) responses. Seroconversion showed poor concordance with NAbT against VOC. Previous SARS-CoV-2 infection boosted the NAb response including against VOC, and anti-CD20 treatment was associated with undetectable NAbT. Vaccine-induced T cell responses were detected in 80% of patients and were comparable between vaccines or cancer types. Our results have implications for the management of patients with cancer during the ongoing COVID-19 pandemic