Schulich students for emergency childcare: Results of a community initiative during COVID-19

The coronavirus disease (COVID-19) pandemic led to the abrupt suspension of many businesses and services in Ontario including schools and childcare centres. This resulted in many parents struggling to identify options for childcare, including frontline healthcare workers (HCWs). A volunteer initiative composed of medical and dental students was developed to provide emergency childcare support to HCWs in Southwestern Ontario. Experts in areas of public health, law, and bioethics were consulted to minimize the risks associated with participation and develop a clear policy to prioritize the health and safety of all participants. Social media was utilized to recruit volunteers and HCWs who were matched on a first-come, first-served basis. 59 medical and dental students were recruited to provide emergency support for 21 HCWs within the unique safety and ethical conditions posed by COVID-19. By reflecting on the strengths of this initiative and the challenges faced during its completion, key areas of improvement were identified that should be addressed in future initiatives of a similar nature

Antioxidants and NOX1/NOX4 inhibition blocks TGFβ1-induced CCN2 and α-SMA expression in dermal and gingival fibroblasts.

TGFbeta induces fibrogenic responses in fibroblasts. Reactive oxygen species (ROS)/nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) may contribute to fibrogenic responses. Here, we examine if the antioxidant N-acetylcysteine (NAC), the NOX inhibitor diphenyleneiodonium (DPI) and the selective NOX1/NOX4 inhibitor GKT-137831 impairs the ability of TGFbeta to induce profibrotic gene expression in human gingival (HGF) and dermal (HDF) fibroblasts. We also assess if GKT-137831 can block the persistent fibrotic phenotype of lesional scleroderma (SSc) fibroblasts. We use real-time polymerase chain reaction and Western blot analysis to evaluate whether NAC and DPI impair the ability of TGFbeta1 to induce expression of fibrogenic genes in fibroblasts. The effects of GKT-137831 on TGFbeta-induced protein expression and the persistent fibrotic phenotype of lesional scleroderma (SSc) fibroblasts were tested using Western blot and collagen gel contraction analyses. In HDF and HGF, TGFbeta1 induces CCN2, CCN1, endothelin-1 and alpha-smooth muscle actin (SMA) in a fashion sensitive to NAC. Induction of COL1A1 mRNA was unaffected. Similar results were seen with DPI. NAC and DPI impaired the ability of TGFbeta1 to induce protein expression of CCN2 and alpha-SMA in HDF and HGF. GKT-137831 impaired TGFbeta-induced CCN2 and alpha-SMA protein expression in HGF and HDF. In lesional SSc dermal fibroblasts, GKT-137831 reduced alpha-SMA and CCN2 protein overexpression and collagen gel contraction. These results are consistent with the hypothesis that antioxidants or NOX1/4 inhibition may be useful in blocking profibrotic effects of TGFbeta on dermal and gingival fibroblasts and warrant consideration for further development as potential antifibrotic agents

TGFβ1-induced CCN2 and αSMA protein expression in human dermal and gingival fibroblasts is reduced by N-acetylcysteine and diphenyleneiodonium.

<p>Human dermal fibroblasts (A) and human gingival fibroblasts (B) were serum-starved overnight. Cells were incubated with either N-acetylcysteine (10 mM) or diphenyleneiodonium (10 μM) for 45 min followed by treatment with or without TGFβ1 (4ng/ml) for 24 hours. Protein lysates were prepared and subjected to western blot analysis with the indicated antibodies. β-actin was used to normalize for protein loading. Representative western blots are shown (n = 3).</p

Inhibition of NOX4 reduces TGFβ1-induced CCN2 and αSMA protein expression in human dermal and gingival fibroblasts and the overexpression of CCN2 and α-SMA, as well as the contractility of lesional SSc dermal fibroblasts.

<p>Human dermal fibroblasts (A) and human gingival fibroblasts (B) were serum-starved overnight. Cells were incubated with GKT-137831 (30 μM) for 45 min followed by treatment with or without TGFβ1 (4ng/ml) for 24 hours. Protein lysates were prepared and subjected to western blot analysis with the indicated antibodies. β-actin was used to normalize for protein loading. Representative western blots are shown (n = 3). <b>(C)</b> Dermal fibroblasts cultured from healthy individuals (NF, normal fibroblasts) and those with scleroderma (systemic sclerosis, SSc) were serum-starved overnight. Cells were incubated with GKT-137831 (30μM) for 24 hours. Protein lysates were prepared and subjected to western blot analysis with the indicated antibodies. GAPDH was used to normalize for protein loading. Representative western blots containing lysates from three different patients are shown (n = 3). (D) NF and SSc fibroblasts were cultured within 3D-collagen lattices in the presence and absence of GKT-137831 (30 μM). After polymerization, the gels were mechanically detached from the wells, and the contraction of the gel was quantified. Results are expressed as a mean +/- SD (n = 3). One-Way ANOVA with post-hoc Tukey test was conducted. * = p<0.05 relative to SSc (-) GKT.</p

Diphenyleneiodonium reduces TGFβ1-induced mRNA expression of pro-fibrotic genes in human dermal and gingival fibroblasts.

<p>Human dermal fibroblasts (A) and human gingival fibroblasts (B) were serum-starved overnight. Cells were incubated with the broad-range NOX protein inhibitor, diphenyleneiodonium (DPI, 10 μM), for 45 min followed by treatment with or without TGFβ1 (4ng/ml). Total RNA was harvested either 6 or 24 (in the case of α-SMA, see text) hours later and subjected to RT-qPCR analysis for the indicated genes. Each sample was conducted in triplicate and gene expression was normalized to 18S (internal control). Results are expressed as a mean +/- SD (n = 4). One-Way ANOVA with post-hoc Tukey test was conducted. * = p<0.05 relative to control, Ŧ = p<0.05 relative to TGFβ1, Ø = p<0.05 relative to DPI+TGFβ1.</p

Students' participation in collaborative research should be recognised

Letter to the editor