sj-docx-2-dhj-10.1177_20552076241247374 - Supplemental material for COVID-19 surveillance based on consumer wearable devices

Supplemental material, sj-docx-2-dhj-10.1177_20552076241247374 for COVID-19 surveillance based on consumer wearable devices by Chunbo Zhang, Aijun Sun, Jiping Liao, Chunbo Zhang, Kunyao Yu, Xiaoyu Ma and Guangfa Wang in DIGITAL HEALTH</p

Identification of the Biosynthetic Gene Cluster for the Anti-infective Desotamides and Production of a New Analogue in a Heterologous Host

The desotamides (DSAs) are potent antibacterial cyclohexapeptides produced by <i>Streptomyces scopuliridis</i> SCSIO ZJ46. We have identified the 39-kb <i>dsa</i> biosynthetic gene cluster by whole-genome scanning. Composed of 17 open reading frames, the cluster codes for four nonribosomal peptide synthetases and associated resistance, transport, regulatory, and precursor biosynthesis proteins. Heterologous expression of the <i>dsa</i> gene cluster in <i>S. coelicolor</i> M1152 afforded desotamides A and B and the new desotamide G. Cluster identification and its demonstrated amenability to heterologous expression provide the foundation for future mechanistic studies as well as the generation of new and potentially clinically significant DSA analogues

sj-docx-1-dhj-10.1177_20552076241247374 - Supplemental material for COVID-19 surveillance based on consumer wearable devices

Supplemental material, sj-docx-1-dhj-10.1177_20552076241247374 for COVID-19 surveillance based on consumer wearable devices by Chunbo Zhang, Aijun Sun, Jiping Liao, Chunbo Zhang, Kunyao Yu, Xiaoyu Ma and Guangfa Wang in DIGITAL HEALTH</p

Identification of the Grincamycin Gene Cluster Unveils Divergent Roles for GcnQ in Different Hosts, Tailoring the l‑Rhodinose Moiety

The gene cluster responsible for grincamycin (GCN, <b>1</b>) biosynthesis in <i>Streptomyces lusitanus</i> SCSIO LR32 was identified; heterologous expression of the GCN cluster in <i>S. coelicolor</i> M512 yielded P-1894B (<b>1b</b>) as a predominant product. The <i>ΔgcnQ</i> mutant accumulates intermediate <b>1a</b> and two shunt products <b>2a</b> and <b>3a</b> bearing l-rhodinose for l-cinerulose A substitutions. In vitro data demonstrated that GcnQ is capable of iteratively tailoring the two l-rhodinose moieties into l-aculose moieties, supporting divergent roles of GcnQ in different hosts

Cytotoxic and Antibacterial Marfuraquinocins from the Deep South China Sea-Derived Streptomyces niveus SCSIO 3406

Four new sesquiterpenoid naphthoquinones, marfuraquinocins A–D (<b>1</b>–<b>4</b>), and two new geranylated phenazines, phenaziterpenes A (<b>5</b>) and B (<b>6</b>), were isolated from the fermentation broth of Streptomyces niveus SCSIO 3406, which originated from a South China Sea sediment sample obtained from a depth of 3536 m. The structures of <b>1</b>–<b>6</b> were elucidated on the basis of extensive MS and one-dimensional and two-dimensional NMR spectroscopic analyses. In a panel of cytotoxicity and antibacterial assays, <b>1</b> and <b>3</b> were found to inhibit a NCI-H460 cancer cell line with IC₅₀ values of 3.7 and 4.4 μM, respectively. Compounds <b>1</b>, <b>3</b>, and <b>4</b> exhibited antibacterial activities against Staphylococcus aureus ATCC 29213 with equivalent MIC values of 8.0 μg/mL; compounds <b>3</b> and <b>4</b> each showed antibacterial activity against methicillin-resistant Staphylococcus epidermidis (MRSE) shhs-E1 with MIC values of 8.0 μg/mL

Schema for the mechanism of cardiac hypertrophy via AT1-R-dependent signaling pathway induced by AngII or mediated by mechanical stretch, respectively.

<p>Schema for the mechanism of cardiac hypertrophy via AT1-R-dependent signaling pathway induced by AngII or mediated by mechanical stretch, respectively.</p

Src/β-arrestin2 signal complex was required for mechanical stretch-mediated AT1-R signaling.

<p>(<b>A</b>) Western blotting analyses of time-dependent Src phosphorylation after treatment with β-arrestin1/2 siRNA or scrambled siRNA. (<b>B</b>) Coimmunoprecipitation analyses of Src and AT1-R in lysates of stretched cardiomyocytes in a time-dependent manner. (<b>C</b>) HA-tagged Src and FLAG-tagged β-arrestin2 (or dominant negative β-arrestin2-V54D) were transient transfected into HEK-293-AT1 cells, whole cell extracts were immunoprecipitated with anti-FLAG monoclonal antibody, and the proteins in the immunoprecipitates and in the total lysates were probed by Western blotting using an anti-HA antibody. (<b>D</b>) The intercellular location of Src kinase in GFP-tagged β-arrestin2 WT and dominant negative GFP-tagged β-arrestin2-V54D transfected HEK-293-AT1 cells was visualized by immunofluorescence after stretching cells for 10 min. (<b>E</b>) The effect of β-arrestin2 WT or β-arrestin2-V54D transfection on mechanical stretch-induced ERK1/2 phosphorylation. (<b>F</b>) The effect of SU6656 (5 mmol/L) on ERK1/2 phosphorylation in cardiomyocytes stimulated by mechanical stretch or AngII (10⁻⁷ mol/L). * <i>P</i><0.05 vs. both AngII and SU6656 treated groups; # <i>P</i><0.05 vs. stretched groups (n = 3 separated experiments).</p

The effects of different ARBs on the cardiac function and Src expression in AGT KO mice.

<p>AGT KO mice were induced by AngII or TAC for 2 weeks with or without the pretreatment of valsartan or candesartan. (<b>A</b>) Representative M-mode tracings of AGT KO hearts stimulated by AngII (10⁻⁵ mol/L, top line) or by TAC for 2 weeks (bottom line). (<b>B</b>) Representative recording of LVAWd, LVPWd and LVEF in AGT KO mice from each group. (<b>C</b>) The expression of Src in myocardium of AGT KO mice from each group was determined. Data were presented as mean ± s.e.m. from five to eight mice. * <i>P</i><0.05 vs. AngII-treated AGT KO mice; # <i>P</i><0.05 vs. TAC-treated AGT KO mice.</p

In vivo analyses of the cardiac function by echocardiography and hemodynamic measurements.

<p>Both AGT KO mice and the C57BL/6 WT littermates were pretreated with or without Src kinase inhibitor (SU6656), followed by TAC for 2 weeks. (<b>A</b>) Quantifications of LVAWd, LVPWd, LVEF and LVESP by representative M-mode tracing and hemodynamic recording from five mice. (<b>B</b>) Quantifications of cardiac immediate-early response genes in C57BL/6 mice and AGT KO mice with or without pretreatment of SU6656 (n = 5 separated experiments). * <i>P</i><0.05 vs. saline-treated TAC-operated AGT KO mice.</p

Mechanical stretch induced ERK1/2 signaling via activation of AT1-R, but not affected by G protein inhibition.

<p>(<b>A</b>) In vitro cultured cardiomyocytes were transfected with or without RGS4 plasmid, and then stimulated by stretch or AngII (10⁻⁷ mol/L) for 10 min, total proteins were collected and the expressions of phosphorylated ERK1/2 and total ERK1/2 were determined by Western blotting. (<b>B</b>) HA-tagged ERK2 was co-transfected with RGS4 plasmid into HEK-293 AT1 expressed cells for 24 h, ERK2 activity (indicated by MBP expression) was detected in cells induced by stretch or AngII, respectively. (<b>C</b>) Cardiomyocytes were pretreated with candesartan (10⁻⁶ mol/L) and then induced by stretch or AngII for 10 min, the expressions of phosphorylated ERK1/2 and total ERK1/2 were determined. * <i>P</i><0.05 vs. stretch-induced group (n = 3 separated experiments).</p