Selection and evaluation of single domain antibody against p19 subunit of IL-23 by phage display for potential use as an autoinflammatory therapeutic.

IL-23 is a double-subunit cytokine that plays an important role in shaping the immune response. IL-23 was found to be associated with several autoinflammatory diseases by generating sustained inflammatory loops that lead to tissue damage. Antibody neutralization of IL-23 was proven to be effective in ameliorating associated diseases. However, antibodies as large proteins have limited tissue penetration and tend to elicit anti-drug antibodies. Additionally, anti-IL-23 antibodies target only one subunit of IL-23 leaving the other one unneutralized. Here, we attempted to isolate a recycling single domain antibody by phage display. One of IL-23 subunits, p19, was expressed in E. coli fused to Gamillus protein to stabilize the α-helix-only p19. To remove Gamillus binders, two biopanning methods were investigated, first, preselection with Gamillus and second, challenge with IL-23 then on the subsequent round challenge with p19-Gam. The isolation of calcium-dependent and pH-dependent recycling binders was performed with EDTA and citrate buffers respectively. Both methods of panning failed to isolate high-affinity and specific p19 recycling binders, while from the second panning method, a high affinity and specific p19 standard binder, namely H11, was successfully isolated. H11 significantly inhibited the gene expression of IL-17 and IL-22 in IL-23-challenged PBMCs indicating H11 specificity and neutralizing ability for IL-23. The new binder due to its small size can overcome antibodies limitations, also, it can be further engineered in the future for antigen clearance such as fusing it to cell penetrating peptides, granting H11 the ability to clear excess IL-23 and enhancing its potential therapeutic effect.

Parents' perceptions of their children's mental health during COVID-19: Evidence from Canada.

OBJECTIVE: This study examined several factors affecting the perception of Canadian parents about their children's mental health during COVID-19. The contribution of this research included fresh evidence from examining the demographic and sociological factors influencing children's well-being during COVID-19 using the Canadian context. METHODS: We used a cross-sectional dataset from Statistics Canada titled Impacts of COVID-19 on Canadians - Parenting during the Pandemic (2020). We relied on an ordered logit model and computed the respective odds ratios. RESULTS: Our results showed that parents with a university degree and those working from home are less concerned about their children's mental health. Nonetheless, having a disabled child, belonging to a minority, having children aged six-to fourteen-years old, and having lost a job or experienced a drop in working hours increased parents' worry. Additionally, having worries about being connected with family and friends, being concerned about work-life balance, feeling lonely at home, and waiting for the reopening of childcare services all increased the likelihood of parents' anxiety about their child's mental health. When running the analysis by province, we saw that being an immigrant and belonging to a minority increased parents' worry only in Ontario and British Columbia. CONCLUSIONS: Policymakers are encouraged to foster working-from-home practices as working from home has been linked to less worry about child mental health, mainly among mothers, as indicated in our gender heterogeneity analysis. In addition, it is advised that families with a disabled child, and families belonging to a minority received additional support. Lastly, policymakers are advised to consider the social cost of preventive measures and incorporate this into any future preventative policymaking as the social impact variables were robust across all models.

Design, analysis, and characterization of a compact silicon photonic modulator with meandered phase shifters.

We present a C-band Mach-Zehnder modulator with meandered phase shifters and a compact footprint of 432 × 260 µm2 on the silicon-on-insulator platform. Electrode, p-n junction, and optical transit time are considered when performing the electro-optic bandwidth (EO BW) simulations. The simulation results prove that the dominant bandwidth limiting factor for this type of modulator is optical transit time. The insertion loss of the modulator without bias is 2.1 dB. The measured half-wave voltage (V π ) and 3-dB EO BW at -0.5 V bias are 6.4 V and 7.7 GHz, respectively. 53 Gbaud PAM-4 transmission over 2 km of standard single-mode fiber is achieved at a bit error rate (BER) below the 6.7% overhead hard-decision forward error correction BER threshold of 3.8×10 -3.

Generation of human scFv-IgG1Fc antibodies for detection of lymphatic filarial recombinant antigens, BmR1 and BmSXP.

Lymphatic filariasis is a neglected parasitic disease that affects millions in tropical and subtropical countries and is caused by Wuchereria and Brugia species. Specific and sensitive detection methods are essential in mapping infected areas where rapid tests are needed to cover underdeveloped and remote regions, which facilitates eliminating the disease as a public health problem. A few commercialized rapid tests based on antigen or antibody detection are available, but the former only detects infection by Wuchereria species and cross-reacts with nonlymphatic filaria, whereas antibody detection might provide positive results of previous infection. Here, we report the production of three different recombinant immunoglobulin gamma (IgG)1 antibodies based on scFvs previously generated via human antibody phage display technology, that is, anti-BmR1 clone 4, anti-BmXSP clone 5B, and anti-BmXSP clone 2H2. The scFv sequences were cloned into a pCMV-IgG1 vector, then transfected into a HEK293F cell line. The generated antibodies were found to be able to bind to their respective targets even at relatively low concentration. Conjugation of Fc to scFv induces binder stability and provides multiple labeling sites for probes and signaling molecules that can be used in rapid tests.

Interleukin 23 and autoimmune diseases: current and possible future therapies.

PURPOSE: IL-23 is a central proinflammatory cytokine with a wide range of influence over immune response. It is implicated in several autoimmune diseases due to the infinite inflammatory loops it can create through the positive feedbacks of both IL-17 and IL-22 arms. This made IL-23 a key target of autoimmune disorders therapy, which indeed was proven to inhibit inflammation and ameliorate diseases. Current autoimmune treatments targeting IL-23 are either by preventing IL-23 ligation to its receptor (IL-23R) via antibodies or inhibiting IL-23 signaling by signaling downstream mediators' inhibitors, with each approach having its own pros and cons. METHODS: Literature review was done to further understand the biology of IL-23 and current therapies. RESULTS: In this review, we discuss the biological features of IL-23 and its role in the pathogenesis of autoimmune diseases including psoriasis, rheumatoid arthritis and inflammatory bowel diseases. Advantages, limitations and side effects of each concept will be reviewed, suggesting several advanced IL-23-based bio-techniques to generate new and possible future therapies to overcome current treatments problems.

Training and experience effect on light-curing efficiency by dental practitioners.

BACKGROUND: Light-curing is a crucial step during the application of composite resin restorations. Composite's success depends on delivering enough light energy to the resin to achieve adequate polymerization. However, dentists are not recognizing the importance of proper light-curing technique. OBJECTIVES: To measure light energy delivered to simulated restorations by preclinical dental students and dentists in internship year. To evaluate the effect of experience and training on the clinician's ability to light-cure composite restorations. METHODS: A group of 50 preclinical dental students and a group of 50 internship dentists light-cured for 10 seconds, a simulated class III and class I restorations positioned in a patient simulator (MARC-Patient Simulator [BlueLight Analytics Inc., Canada]) that measured the irradiance and energy delivered by the curing light. Then participants received individualized training on optimizing their light-curing technique. They were retested after the training. Statistical analysis was done with two-way ANOVA and Tukey's test. RESULTS: Participants delivered an average of 60% more energy after the instructions, which is a significant improvement (P < 0.05). The number of participants that failed to deliver the minimum amount of energy (6 J/cm²) decreased significantly from 37.5% to 2.5%. There was a significant difference in the amount of energy delivered by the Preclinical and Internship groups (P < 0.05). CONCLUSION: Initially, many participants were not using the curing light properly. Light-curing technique improved with training and using a patient simulator. Experience can enhance the operator's ability to light-cure composite restorations. However, a training session can improve light-curing performance more than years of experience.